Ophthalmic preparations for drug delivery and protection of the anterior segment of the eye

Abstract

The present application relates to a topical preparation comprising a compound-I or its free base and a second active agent selected from niacin, niacinamide and vitamin K and combinations thereof for treating ocular neovascularization. The present application also relates to a pharmaceutical composition comprising particles of the compound-I or its free base and a suspension preparation of a particle composition comprising the compound-I or its free base.

Description

[0001] This application is a divisional application of an invention patent application with an application date of September 17, 2015, application number 201580049902.7, and invention name “Ophthalmic preparation for drug delivery and protection of the anterior segment of the eye”. This application claims priority and benefits of USSN 62 / 051,794 filed on September 17, 2014, the contents of which are incorporated by reference in their entirety. Technical Field

[0002] Embodiments disclosed herein generally relate to the topical administration of a combination of a first active agent (e.g., a pharmaceutical compound or a salt thereof) and a second active agent to treat an eye disease or condition. The disclosed embodiments include an ophthalmic formulation comprising a first active agent or a salt thereof and a second active agent, wherein 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 of a subject while protecting the anterior segment of the eye of the subject. Background Art

[0003] Because of the ineffective delivery of active agents to the target site, the treatment of diseases or disorders of the posterior segment of the eye with topically applied active agents is still ineffective. Most topical drugs penetrate through the cornea. However, the cornea is not equally permeable to all topically applied active agents because the basic structure of the cornea determines the relative permeability of the active agent. Effectively, the largest barrier for active agents to penetrate is the corneal epithelium, which is rich in cell membranes and is therefore more easily penetrated by lipophilic agents. In contrast, since the corneal stroma is mainly composed of water, if the active agent is hydrophilic, it is easier to pass through. The endothelium represents a single layer, which is lipophilic in turn. Lipophilic or amphiphilic active agents, because they can appear to be charged or uncharged, have the best permeability to the cornea. Similar to the cornea, the conjunctival epithelium and the blood vessels within or below the conjunctival epithelium can be penetrated by the same type of lipophilic or biphasic agents. However, due to the nature of the lipophilic membranes in the conjunctiva and its inherent vascular system, most active agents generally do not penetrate the conjunctiva and enter the eye. Drugs that have limited permeability to the vascular tissue in the conjunctiva and subconjunctival region are excreted into the systemic circulation.

[0004] A major drawback of topical drops may be the need for high concentrations of active agent in the topical formulation in order to achieve meaningful therapeutic drug levels in the internal ocular tissues, due to the limited permeability of many topical drops through the corneal and conjunctival barriers. Depending on the active agent, the molecule itself or its high concentration, the topical formulation may be toxic to the anterior segment of the eye (including the conjunctiva, cornea and / or lens), causing various damage to the ocular surface, such as corneal epithelial defects and erosions.

[0005] The ocular side effects observed after treatment with anti-EGFR drug therapy (e.g., anti-EGFR cancer therapy) have inspired the important role of EGFR signaling pathways in maintaining and restoring the health of human corneal epithelium. Patients treated with anti-EGFR drug therapy may experience corneal changes such as epithelial degeneration and defects, ulcers, corneal epithelial thinning, erosion and / or corneal edema, keratitis, and perforation while undergoing therapy or even after stopping anti-EGFR therapy. The important role of EGFR signaling in the homeostasis and pathophysiology of corneal epithelium has been fully established. EGFR activation is necessary and sufficient for corneal epithelial migration, proliferation, and differentiation. In addition, EGFR is the main medium for wound healing during in vitro experiments with immortalized human corneal epithelial cells. Therefore, in addition to administering a preparation comprising an active agent (e.g., an active agent with toxic or anti-EGFR activity), treating ocular diseases or disorders (e.g., diseases or disorders of the posterior segment of the eye) will benefit from administering an agent that maintains EGFR activity.

[0006] The present application provides a novel formulation that avoids the problems encountered in the ocular delivery of existing topical therapeutic agents. The present application achieves a combined effect of reducing drug exposure to the cornea and anterior segment and protecting the cornea and anterior segment tissues while increasing the bioavailability of the posterior segment. By reducing corneal exposure, protecting corneal tissues and increasing the bioavailability of the posterior segment, the formulation of the present application improves ocular tolerance and increases the therapeutic index of the active agent. Summary of the invention

[0007] The present application relates to a pharmaceutical preparation in the form of a solution and / or suspension, which reduces exposure to a first active agent in the anterior segment of the eye (e.g., the ocular surface composed of the cornea and conjunctiva), and protects the eye (e.g., the anterior segment of the eye) by maintaining EGFR activity. The pharmaceutical preparation of the present application increases the bioavailability of the first active agent at the posterior segment of the eye (e.g., at the central choroid and / or central retina), and protects and / or improves the health of the anterior segment of the eye.

[0008] The present application provides a formulation comprising a first active agent and an optional second active agent in the form of a solution or suspension, which has excellent characteristics compared to a composition formed as a gel. The present application provides that the first active agent and / or the second active agent can be formulated together as a solution and / or a suspension. The increased level of the first active agent in the anterior segment of the eye limits the ocular tolerance of topical drops containing the first active agent, and can cause corneal epithelial defects and erosions. The presence of the second active agent can prevent damage that can be caused by exposure to the first active agent, treat any damage that can be caused by exposure to the first active agent, and / or improve the overall health of the ocular surface, particularly the corneal epithelium. The second active agent can be formulated together with the first active agent, or formulated as a separate formulation, which is administered in combination with a formulation comprising the first active agent.

[0009] The formulations of the present application reduce exposure of the first active agent at the anterior segment of the eye (such as the cornea or conjunctival surface), protect against and / or repair damage to the anterior segment of the eye (such as the cornea or conjunctival surface), and maintain sufficient concentrations of the first active agent required to bind to relevant receptors at the target tissue, and impart a therapeutic effect in the posterior segment of the eye (such as the choroid and retina).

[0010] The present application relates to formulations and methods that can be used to treat pathological conditions caused or exacerbated by: ocular angiogenesis, neovascularization and / or vascular leakage, for example, in 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) from any mechanism (e.g., high myopia, trauma, sickle cell disease; ocular histoplasmosis, angioid streaks, traumatic choroidal rupture, drusen of the optic nerve, or some retinal dystrophies); pathological retinal neovascularization and vascular leakage from any mechanism (e.g., sickle cell retinopathy, Eales disease, ocular ischemic syndrome, carotid cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic occlusive arteritis, birdshot retinochoroidopathy, retinal vasculitis, sarcoidosis, or toxoplasmosis); uveitis; retinal vein occlusion (central or branch); ocular trauma; surgically induced edema; surgically induced neovascularization; cystic 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 condition is caused or exacerbated by ocular angiogenesis and / or neovascularization.

[0011] The present application also relates to preparations and methods that can be used to prevent and / or treat corneal epithelial damage associated with diseases (including systemic diseases (e.g., cancer, diabetes, etc.) and eye diseases) or side effects from locally or systemically administered drugs (e.g., anti-EGFR agents or EGFR inhibitors). The preparation of the present application has at least one anti-angiogenic agent, anti-inflammatory agent or anti-vascular permeability agent for treating angiogenic eye disorders and an EGFR modulator (e.g., an agent that maintains EGFR activity or activates EGFR) for preventing and / or treating corneal epithelial damage.

[0012] According to an embodiment of the present application, the first active agent is an anti-angiogenic kinase inhibitor, and the second active agent is a kinase regulator (e.g., an activator). In one embodiment, the first active agent inhibits a kinase different from a kinase regulated (e.g., maintained or activated) by a second active agent. Examples of some kinase inhibitors that can be used to produce beneficial treatment results include inhibitors of receptor tyrosine kinases (e.g., but not limited to VEGFR, FGFR, Tie-2, and ephrin kinase receptors). Examples of kinase regulators that can be used to produce beneficial treatment results include activators of ErbB receptor tyrosine kinases (e.g., but not limited to, 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 regulator (e.g., an activator). In a further embodiment, the second active agent is nicotinic acid, nicotinamide, or vitamin K, or a combination thereof. In a further embodiment, the second active agent is nicotinic acid or nicotinamide. In another embodiment, the second active agent is vitamin K.

[0013] In some embodiments, the second active agent of the present application reduces or alleviates the inhibition of EGFR at the anterior segment of the eye caused by high concentrations of the first active agent of the present application (e.g., a transient high concentration of the first active agent after the first active agent is applied to the anterior segment of the eye).

[0014] Embodiments of the present application provide an ophthalmic preparation for treating ocular neovascularization, comprising a first active agent of formula I:

[0015] (I)

[0016] or a pharmaceutically acceptable salt thereof; a second active agent, wherein the second active agent is an EGFR modulator (e.g., an activator) such as niacin, niacinamide, or vitamin K, or a combination thereof; and a pharmaceutically acceptable excipient; the first active agent or a pharmaceutically acceptable salt thereof is present at about 0.02% to about 1.2% w / v such that the formulation forms a solution or suspension, and wherein:

[0017] X 1 is O or S;

[0018] R 1 Yes H, C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C(O)(C1-C 10 Alkyl), (CH2)t (C6-C 10 Aryl), (CH2) t (4-10 membered heterocyclic ring), C(O)(CH2) t (C6-C 10 Aryl) or C(O)(CH2) t (5-10 membered heterocyclic ring), wherein:

[0019] t is an integer from 0 to 5;

[0020] The alkyl group optionally includes 1 or 2 groups selected from O, S and N (R 6 ), provided that the two O atoms, the two S atoms, or the O and S atoms are not directly connected to each other;

[0021] Aryl and heterocyclic groups are optionally combined with C6-C 10 An aryl group, a C5-C8 saturated ring group or a 5-10 membered heterocyclic group is fused;

[0022] One or two carbon atoms in the above heterocyclic moiety are optionally substituted by an oxo (═O) moiety or an oxygen anion;

[0023] When t is an integer from 2 to 5, (CH2) t The moiety optionally includes a carbon-carbon double or triple bond; and

[0024] The above R 1 The group, except H, is optionally substituted by 1 to 3 R 4 group substitution;

[0025] R 2 It is H;

[0026] R 3 Yes (CH2) t (C6-C 10 aryl), wherein:

[0027] t is an integer from 0 to 5;

[0028] The aryl group is optionally 10 An aryl group, a C5-C8 saturated ring group or a 5-10 membered heterocyclic group is fused;

[0029] When t is an integer from 2 to 5, (CH2) t The moiety optionally includes a carbon-carbon double or triple bond; and

[0030] The above R 3 The group is optionally substituted by 1 to 5 R 4 group substitution;

[0031] Each R 4Independently selected from C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, OR 5 、C(O)R 5 、C(O)OR 5 NR 6 C(O)R 5 NR 6 C(O)OR 5 、OC(O)R 5 NR 6 S02R 5 、SO2NR 5 R 6 、C(O)NR 5 R 6 NR 5 R 6 、S(O) j R 7 (wherein j is an integer from 0 to 2), SO3H, NR 5 (CR 6 R 7 ) t OR 6 、(CH2) t (C6-C 10 Aryl), SO2(CH2) t (C6-C 10 Aryl), S(CH2) t (C6-C 10 Aryl), O(CH2) t (C6-C 10 Aryl), (CH2) t (5-10 membered heterocyclic ring) and (CR 6 R 7 ) m OR 6 ,in:

[0032] m is an integer from 1 to 5;

[0033] t is an integer from 0 to 5;

[0034] The alkyl group optionally includes 1 or 2 groups selected from O, S and N (R 6 ), provided that two O atoms, two S atoms, or O and S atoms are not directly connected to each other;

[0035] Aryl and heterocyclic groups are optionally combined with C6-C 10 An aryl group, a C5-C8 saturated ring group or a 5-10 membered heterocyclic group is fused;

[0036] One or two carbon atoms in the above heterocyclic moiety are optionally substituted by an oxo (═O) moiety or an oxygen anion; and

[0037] The above R 4 The alkyl, aryl and heterocyclic moieties of the group are optionally substituted by 1-3 groups independently selected from halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, NR 6 S02R 5 、SO2NR 5 R 6 、C(O)R 5 、C(O)OR 5 、OC(O)R 5 NR 6 C(O)R 5 、C(O)NR 5 R 6 NR 5 R 6 , (CR 6 R 7 ) m OR 6 (wherein m is an integer from 1 to 5), OR 5 , and R 5 is substituted with a substituent listed in the definition of ; and

[0038] R 5 , R 6 and R 7 Each is independently H or C1-C6 alkyl.

[0039] In one embodiment, R 3 Yes (CH2) t (C6-C 10 aryl), wherein t is an integer from 1 to 3, and R 3 Optionally 1-4 R 4 Group substitution.

[0040] In a further embodiment, R 3 is benzyl, which is optionally substituted with 1 to 4 substituents independently selected from halogen and C1-C4 alkyl. In a further embodiment, R 3 is benzyl substituted with 1 to 4 substituents independently selected from methyl, fluoro, chloro and bromo.

[0041] In one embodiment, R 1 Yes (CH2) t (5-10 membered heterocycle), wherein t is an integer from 0 to 5, which is optionally substituted with 1 or 2 substituents independently selected from C1-C4 alkyl, hydroxy and hydroxymethyl.

[0042] The present application provides R in Formula I 1 The heterocyclic portion of the group is selected from morpholino, pyrrolidinyl, imidazolyl, piperazinyl, piperidinyl and 2,5-diaza-bicyclo[2.2.1]hept-2-yl, R 1 The t variable range of the group is 2 to 5, and R 1 The group is optionally substituted with one or more hydroxy groups.

[0043] For example, R in Formula I of the present application 1 The heterocyclic portion of the group is pyrrolidine.

[0044] In a further embodiment of the present application, the first active agent is:

[0045] .

[0046] In a further embodiment of the present application, the first active agent is the hydrochloride salt of the compound of formula II, i.e., compound-I:

[0047] (Compound-I).

[0048] In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In a further embodiment, the second active agent is niacin, niacinamide, or vitamin K, or a combination thereof. In a further embodiment, the second active agent is vitamin K.

[0049] Embodiments of the present application provide formulations comprising 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, such as Compound-I. In some embodiments, for topical administration, the concentration of Compound-I or its free base (Formula II) in the formulation is 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%, 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. In some embodiments, for topical administration, the concentration of Compound-I or its free base (Formula II) in the formulation 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 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 1.1%, About 0.6%, about 0.4% - about 0.6%, 0.1% - about 0.5%, about 0.2% - about 0.5%, about 0.3% - about 0.5%, about 0.4% - about 0.5%, 0.1% - about 0.4%, about 0.2% - about 0.4%, about 0.3% - about 0.4% w / v. In some embodiments, the formulation includes 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).

[0050] In some embodiments, the present application provides a solution of a first active agent (eg, Compound-I) and a second active agent (eg, niacin, niacinamide, or vitamin K, or a combination thereof) comprising one or more solubilizing agents.

[0051] 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.

[0052] In some embodiments, the solubilizer in the formulation can be a cyclodextrin, such as 2-hydroxypropyl-β-cyclodextrin, methyl-β-cyclodextrin, random methylated-β-cyclodextrin, ethylated-β-cyclodextrin, triacetyl-β-cyclodextrin, peracetylated-β-cyclodextrin, carboxymethyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, 2-hydroxy-3-(trimethylammonio)propyl-β-cyclodextrin, glucosyl-β-cyclodextrin, maltosyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, branched-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, random methylated-γ-cyclodextrin, trimethyl-γ-cyclodextrin, or a combination thereof.

[0053] In one embodiment, the solubilizer in the formulation is 2-hydroxypropyl-β-cyclodextrin or β-cyclodextrin sulfobutyl ether.

[0054] In one embodiment, the formulation may further comprise one or more of benzalkonium chloride (BAK), sodium chloride, and a pH adjuster.

[0055] 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 the first active agent, or a pharmaceutically acceptable salt thereof, and a buffer, such as 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 tromethamine, and optionally further comprises about 0.005% w / v benzalkonium chloride (BAK).

[0056] In some embodiments, the second active agent is an EGFR modulator (e.g., an activator). For example, the second active agent includes, but is not limited to, nicotinic acid (or nicotinic acid / vitamin B3), nicotinamide, and vitamin K or a combination thereof. "Vitamin K" as defined herein includes one or more members of the naturally occurring or synthetic vitamin K family and prodrugs thereof. The vitamin K family consists of vitamin K1 (also known as phylloquinone, phytomenadione, or phytonadione), vitamin K2, and any vitamin K2 homologue. Vitamin K2 homologues are known as menaquinones, characterized by the number of isoprenoid residues in their side chains. Synthetic vitamin K includes, but is not limited to, vitamin K3 (i.e., menaquinone), vitamin K4, and 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 homolog. 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).

[0057] In some embodiments, for topical administration, the concentration of the second active agent in the formulation is about 0.00001% to about 5.0% w / v. In some embodiments, for topical administration, the concentration of the second active agent in the formulation 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. In some embodiments, for topical administration, the concentration of the second active agent in the formulation is about 0.00001% - about 0.0001%, 0.000012% - about 0.0001%, 0.000014% - about 0.0001%, 0.000016% - about 0.0001%, 0.000018% - about 0.0001%, 0.00002% - about 0.0001%, 0.00003% - about 0.0001%, 0.00004% - about 0.0001%, 0.00005% - about 0.0001%, 0.00006% - about 0.0001%, 0.00007% - about 0.0001%, 0.00008% - about 0.0001%, 0.00009% - About 0.0001%, 0.000016% - about 0.00009%, 0.000018% - about 0.00009%, 0.00002% - about 0.00009%, 0.00003% - about 0.00009%, 0.00004% - about 0.00009%, 0.00005% - about 0.00009%, 0.00006% - about 0.00009%, 0.00007% - about 0.00009%, or 0.00008% - about 0.00009% w / v. In some embodiments, the formulation comprises 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 or a pharmaceutically acceptable salt thereof.

[0058] In some embodiments, the concentration of the second active agent in the formulation 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.

[0059] The present application provides formulations having a pH value of about 4.5 to about 7.5 at or below about 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 about 40° C.

[0060] In another embodiment, the present application provides a formulation comprising a first active agent (e.g., Formula I or Compound-I or its free base (Formula II)) and a second active agent (e.g., niacin, niacinamide or vitamin K or a combination thereof) for the preparation of a medicament for entering the posterior segment of the eye and / or for treating and / or improving a posterior segment disease or vascular disease or inflammatory disease of the eye as described herein, such as 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) from any mechanism (e.g., high myopia, trauma, sickle cell disease; ocular histoplasmosis, angioid streaks, traumatic choroidal rupture, drusen of the optic nerve and some retinal dystrophies); pathological retinal neovascularization from any mechanism (e.g., sickle cell retinopathy, Eales disease, ocular ischemic syndrome, carotid cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic occlusive arteritis; birdshot retinochoroidopathy, retinal vasculitis, sarcoidosis and toxoplasmosis); uveitis; retinal vein occlusion (central or branch); ocular trauma; surgically induced edema; surgically induced neovascularization; cystic macular edema; ocular ischemia; retinopathy of prematurity; Coat's disease; sickle cell retinopathy and / or neovascular glaucoma. In one embodiment, the eye disease is AMD. In one embodiment, the eye disease is caused or exacerbated by ocular angiogenesis and / or neovascularization.

[0061] In another embodiment, the present application relates to a preparation for preparing a drug suitable for entering the posterior segment of the eye and / or treating and / or improving the posterior segment disease of the eye or vascular lesions or inflammatory diseases. In one embodiment, the preparation 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 a further embodiment, the first active agent is a compound of formula II or a pharmaceutically acceptable salt thereof. In a further embodiment, the first active agent is compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In a further embodiment, the second active agent is selected from nicotinic acid, nicotinamide and vitamin K, and combinations thereof. The preparation may further comprise one or more pharmaceutically acceptable excipients.

[0062] In another embodiment, the application relates to a preparation for the preparation of a drug for treating and / or improving the symptoms of an ocular disease or condition (e.g., a posterior segment of the eye or a vascular lesion or an inflammatory disease). In one embodiment, the preparation 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 a further embodiment, the first active agent is a compound of formula II or a pharmaceutically acceptable salt thereof. In a further embodiment, the first active agent is compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In a further embodiment, the second active agent is selected from nicotinic acid, nicotinamide and vitamin K, and combinations thereof. The preparation may further comprise one or more pharmaceutically acceptable excipients.

[0063] In yet another embodiment, the application relates to a preparation for entering the posterior segment of the eye and / or for treating and / or improving the posterior segment disease of the eye. In one embodiment, the preparation 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 a further embodiment, the first active agent is a compound of formula II or a pharmaceutically acceptable salt thereof. In a further embodiment, the first active agent is compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In one embodiment, the second active agent is selected from nicotinic acid, nicotinamide and vitamin K, and combinations thereof. The preparation may further comprise one or more pharmaceutically acceptable excipients.

[0064] In yet another embodiment, the application relates to a preparation for the treatment and / or improvement of ocular diseases or conditions (e.g., posterior segment vasculopathy or inflammatory diseases of the eye) in a method for the symptoms. In one embodiment, the preparation 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 a further embodiment, the first active agent is a compound of formula II or a pharmaceutically acceptable salt thereof. In a further embodiment, the first active agent is compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In a further embodiment, the second active agent is selected from nicotinic acid, nicotinamide and vitamin K, and combinations thereof. The preparation may further comprise one or more pharmaceutically acceptable excipients.

[0065] In another embodiment, the present application relates to a combination therapy for accessing the posterior segment of the eye and / or for treating and / or improving posterior segment diseases 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 a further embodiment, the first active agent is a compound of formula II or a pharmaceutically acceptable salt thereof. In a further embodiment, the first active agent is compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In a further 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 before administering the second active agent. In another embodiment, the first active agent is administered after administering the second active agent.

[0066] In another embodiment, the present application relates to a combination therapy for treating and / or improving symptoms of an ocular disease or condition (e.g., a posterior segment vasculopathy or inflammatory disease 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 a further embodiment, the first active agent is a compound of formula II or a pharmaceutically acceptable salt thereof. In a further embodiment, the first active agent is compound-I. In one embodiment, the second active agent is an EGFR modulator (e.g., an activator). In a further 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 before administering the second active agent. In another embodiment, the first active agent is administered after administering the second active agent.

[0067] In some embodiments, the exposure time of the first active agent (e.g., Compound-1) and the second active agent is 1 to 90 days or longer 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-1) and the second active agent is longer 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 the formulation comprising the first active agent (e.g., Compound-1) and the second active agent to the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) or longer than 90 days. For example, the dosage regimen involves applying the preparation once a day, twice a day, three times a day or four times a day for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months) or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months). For example, the dosage regimen involves applying the preparation once, twice, three times or four times every other day (i.e., on the 1st, 3rd, 5th, 7th day, etc.) for up to 90 days. For example, the dosage regimen involves applying once on the 1st day, once or twice on the 2nd to 90th day. For example, the dosage regimen involves applying once, twice, three times or four times on the 1st day, and then continuing once a day for 2-90 days. For example, the dosage regimen involves applying once, twice, three times, four times on the 1st day, and then continuing once, twice, three times or four times every other day (i.e., on the 1st, 3rd, 5th, 7th day, etc.) for up to 90 days. For example, a dosage regimen involves applying once or twice a day for 1, 2, 3, 4 or 5 consecutive days. For twice or three times a day dosing regimen, the subject receives a topical ocular dose of the first active agent (e.g., compound-1) and the second active agent preparation on the 1st and 4th day, separated by about 4, 6 or 8 hours. In another embodiment, the subject receives a topical ocular dose of the first active agent (e.g., compound-1) and the second active agent preparation for four consecutive days, separated by about 4, 6 or 8 hours. In some embodiments, the subject receives one or two topical ocular doses of the first active agent (e.g., compound-1) and the second active agent preparation every day for 5 consecutive days. In other embodiments, the subject receives one or two topical ocular doses of the first active agent (e.g., compound-1) and the second active agent preparation for 5-90 consecutive days. In some embodiments, the subject receives one or two topical ocular doses of the first active agent (e.g., compound-1) and the second active agent preparation for at least 25 consecutive days. In one embodiment, the subject receives one or two topical ocular doses of at least 90 consecutive days or more.

[0068] For example, a formulation comprising about 1 mg / mL BID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 1 mg / mL QD of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 1 mg / mL TID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 1 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 2 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 3 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 3 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, a formulation comprising about 3 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 3 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 4 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 4 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 4 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 4 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL TID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL QID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 6 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 6 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 6 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 6 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL BID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL QD of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, a formulation comprising about 8 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 9 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 9 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 9 mg / mL TID is applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 9 mg / mL QID is applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 10 mg / mL QD is applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months). The dosage regimen of 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months) can be any regimen related to the continuous or alternate days described in the above paragraph. In some embodiments, the preparation of the application is applied by QD or BID. In some embodiments, when administered at a low dose (e.g., 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, or 5 mg / mL), the formulation of the present application is administered QD, BID, TID, or QID, and when administered at a high dose (e.g., 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, or 10 mg / mL), the formulation of the present application is administered QD or BID.

[0069] In some embodiments, a formulation of a first active agent (e.g., Formula II or Compound-I) and a second active agent is administered to one or both eyes of the subject. For example, a formulation of the present application comprising about 0.2% - about 1.0% (w / v) of a compound of Formula II or about 0.1% -1.2% (w / v) of a compound-I and a second active agent is administered once a day (QD), twice a day (BID), three times a day (TID), or four times a day (QID) to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a compound of Formula II or Compound-I is compounded with a complexing agent such as cyclodextrin (e.g., hydroxypropyl-β-cyclodextrin (HP-β-CD, KLEPTOSE® HPB) (%)) at a ratio of about 1:8, wherein about 2% - 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, e.g., 10 mM phosphate buffer. The desired weight-gram molecular osmotic concentration of the formulation may be about 200-about 300 mOsm, which is achieved by adding a salt (e.g., sodium chloride) in an amount sufficient to achieve the weight-gram molecular osmotic concentration. The pH of the formulation may be about 6.0.

[0070] The present application relates to a pharmaceutical composition comprising particles of an active agent of the present application (eg, a first active agent (eg, 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.

[0071] 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 as described herein (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. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 This is a series of immunoblots of EGFR phosphorylation in immortalized corneal epithelial cells (hTCEpi cells) treated with various concentrations of the first active agent of the present application or control (AG1478, EGFR kinase inhibitor) and EGF (top panel: immunoblot of total EGFR, middle panel: immunoblot showing phosphorylation of tyrosine 1068 of EGFR, bottom panel: immunoblot showing phosphorylation of tyrosine 1045 of EGFR).

[0073] Figure 2is a series of immunoblots of EGFR phosphorylation in immortalized corneal epithelial cells (hTCEpi cells) treated with various concentrations of the first active agent of the present application, alone or after treatment with vitamin K3 (50 μM), niacin (10 μM) or niacinamide (10 μM) (first row: immunoblot of total EGFR, second row: immunoblot showing phosphorylation of tyrosine 1068 of EGFR, third row: immunoblot showing phosphorylation of tyrosine 1045 of EGFR).

[0074] Figure 3A is a series of micrographs showing the migration / proliferation of immortalized corneal epithelial cells (hTCEpi cells) treated with the indicated concentration of the first active agent of the present application or a control (AG1478, an EGFR kinase inhibitor) for 30 minutes, followed by treatment with the first active agent or control together with the indicated concentration of EGF or VEGF for 16 hours. Figure 3B It is quantitative Figure 3A A series of bar graphs showing the migration / proliferation of hTCEpi cells in FIG.

[0075] Figure 4A is a series of micrographs 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 various concentrations of the first active agent of the present application and subsequently treated with vitamin K3, the first active agent and EGF together. Figure 4B It is quantitative Figure 4A A series of bar graphs showing the migration / proliferation of hTCEpi cells in FIG.

[0076] Figure 5A is a series of images of epithelial wounds at initial wounding (0 hours) and after wounding of corneas treated with vehicle, the indicated concentrations of the compound of Formula I or II, or AG1478. Figure 5B and 5C is a quantitative analysis of corneal injury 16 hours after wounding of corneas treated with vehicle, a compound of Formula I or II at a specified concentration, or AG1478. Figure 5B ) or 24 hours ( Figure 5C ) at . Figure 5D is a graph showing the time course wound healing of corneas treated with vehicle, the indicated concentrations of compounds of Formula I or II, or AG1478.

[0077] Fig. 6A is a series of images of epithelial wounds at initial wounding (0 hours) and after wounding of corneas treated with vehicle, menadione, a compound of Formula I or II, or a combination of menadione and a compound of Formula I or II. Figure 6Bis a series of bar graphs quantifying wound healing at 16 hours or 24 hours post-wounding of corneas treated with vehicle, menadione, a compound of Formula I or II, or a combination of menadione and 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 a combination of menadione and a compound of Formula I or II.

[0078] Fig. 7A is a series of immunoblots showing total VEGFR2, phosphorylated VEGFR2, and α-tubulin (as a loading control) in human retinal endothelial cells treated with the indicated concentrations of menadione for 4 hours, followed by treatment with various concentrations of compounds of Formula I or II (1 nM, 10 nM, 100 nM, or 1 μM) for 30 minutes, and then treated with 10 ng / ml VEGF. Figure 7B is a graphical representation quantifying the growth of human retinal endothelial cells following treatment with the indicated concentrations of menadione for 4 hours, followed by treatment with various concentrations of a compound of Formula I or II (0.1 nM, 1 nM, 10 nM, or 100 nM) for 30 minutes, and then treated overnight with 10 ng / ml VEGF.

[0079] Figure 8 is a series of immunoblots showing total EGFR2, phosphorylated EGFR, and α-tubulin (as a loading control) in hTCEpi cells treated with the indicated concentrations of menadione for 4 h and subsequently incubated with 8.0 nM EGF for the indicated time periods.

[0080] Fig. 9A Shown is the particle size distribution of particles comprising 5% compound of Formula II and 1% Pluronic F-127 produced using small milling media at high roller speeds for long periods of time. Fig. 9B Shown is the particle size distribution of particles comprising 5% compound of Formula II and 1% Pluronic F-127 produced using large milling media at a slow roller speed for a short period of time. Fig. 9C Shown is the particle size distribution of particles comprising 3% Compound-I, 0.6% Tris HCl, and 2% glycerol produced without milling. Fig.9D Shown is the particle size distribution of particles comprising 0.4% of the compound of formula II, 10 μM menadione, 0.08% Pluronic F-127 and 2.5% glycerol after storage at 40°C for 7 days.

[0081] Fig.10 Shown is the particle size distribution of particles containing 5% menadione and 1% HPMC.

[0082] Figure 11 shows the particle size distribution of particles comprising 5% of the compound of Formula II and 1% Pluronic F-127 produced using large milling media at a low roller speed for a short period of time. DETAILED DESCRIPTION

[0083] The materials, compounds, compositions, articles and methods described herein can be more easily understood by reference to the detailed description of specific aspects of the subject matter disclosed below and the embodiments included therein. Before disclosing and describing the materials, compounds, compositions, articles, devices and methods of the present invention, it should be understood that the following aspects are not limited to specific methods or specific reagents and can therefore vary. It should also be understood that the terms used herein are only for the purpose of describing specific aspects and are not intended to be limiting.

[0084] In addition, throughout this specification, reference is made to various publications. The disclosures of these publications are incorporated herein by reference in their entireties in order to more fully describe the state of the art to which the disclosures pertain. For materials contained in references discussed in sentences relying on references, the disclosed references are also individually and specifically incorporated herein by reference.

[0085] The present application provides a composition or formulation containing a first active agent and / or a second active agent for treating an ocular condition caused by endothelial cell proliferation, enhanced vascular permeability, inflammation, angiogenesis or neovascularization, and a second active agent for preventing and / or treating damage to the anterior portion of the eye caused by the first active agent, systemic diseases and / or ocular diseases.

[0086] The application also relates to a combination of a first active agent (e.g., a compound of formula I or II) and a second active agent (e.g., nicotinic acid, nicotinamide, vitamin K, or a combination thereof). 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 before the second active agent or a pharmaceutically acceptable salt thereof is administered. In another embodiment, a compound of formula I or II or a pharmaceutically acceptable salt thereof is administered after the second active agent or a pharmaceutically acceptable salt thereof is administered.

[0087] The preparation of the present application can be used to prevent or inhibit the formation of new blood vessels and vascular leakage associated with eye disorders, while preventing or inhibiting corneal diseases. In some cases, the preparation of the present application causes the regression of new blood vessels. In short, within the context of the present application, the first active agent should be understood as any molecule that is synthetic or naturally present, and it acts to inhibit blood vessel growth, reduce vascular permeability and / or reduce inflammation.

[0088] The preparation of the present application can also be used to prevent and / or treat corneal epithelial damage caused by systemic diseases (e.g., cancer, diabetes, etc.), eye diseases, or side effects from drugs (e.g., anti-EGFR agents, or compounds of formula I or II with anti-EGFR activity) administered topically or systemically. In short, within the context of the present application, the second active agent should be understood as any molecule that is synthetic or naturally present, which acts to protect from and / or repair corneal edema, ulcers, or any other corneal abnormalities. Specifically, the present application provides a preparation comprising a first active agent and a second active agent, each of which is a therapeutically effective amount.

[0089] General Definition

[0090] In this specification and the claims that follow, reference is made to various terms which shall be defined to have the following meanings. Unless otherwise stated, all percentages, ratios and proportions herein are by weight. Unless otherwise stated, all temperatures are in degrees Celsius (° C.).

[0091] "Pharmaceutically acceptable" means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to a subject with the relevant active compound without causing clinically unacceptable biological effects or interacting in a deleterious manner with any other components of the pharmaceutical composition or formulation containing it.

[0092] Unless expressly stated to the contrary, weight percentages of components are based on the total weight of the formulation or composition in which the component is contained.

[0093] As used herein, "effective amount" means "an amount of one or more disclosed compounds that is effective at dosages and for periods of time necessary to achieve a desired or therapeutic result". The effective amount may vary according to factors known in the art such as the disease state, age, sex, and weight of the person or animal being treated. Although specific dosage regimens may be described in the examples herein, it will be understood by those skilled in the art that the dosage regimen may be altered to provide the optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. In addition, the formulations of the present disclosure may be administered at a frequency necessary to achieve a therapeutic amount.

[0094] The effective amount or effective dose in humans can be determined from the effective amount or effective dose in animals (e.g., experimental animals). For example, the effective dose in humans can be calculated based on the conversion rate shown in the following table.

[0095] Animal doses were converted to human equivalent doses (HED) by using the body surface area exponent of 0.67.

[0096]

[0097] "Excipient" is used herein to include any other compound that may be included in or combined with one or more disclosed inhibitors, which is not a therapeutic or biologically active compound. Therefore, an excipient should be pharmaceutically or biologically acceptable or relevant (e.g., an excipient should generally be nontoxic to a subject). An "excipient" includes a single such compound, and is also intended to include a variety of excipients. For purposes of this application, the terms "excipient" and "carrier" are used interchangeably throughout the specification of this application, and the terms are defined herein as "ingredients in practice for preparing safe and effective pharmaceutical compositions."

[0098] As used herein, "subject" means an individual. Thus, "subjects" may include domesticated animals (e.g., cats, dogs, etc.), livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratory animals (e.g., mice, rabbits, rats, guinea pigs, etc.), and birds. "Subjects" may also include primates or humans.

[0099] "Reduce" or other forms of the word, such as "reducing" or "reduction" means a reduction in an event or characteristic (e.g., vascular leakage or tissue swelling). It should be understood that this is usually related to some standard or expected value, in other words, it is relative, but does not always need to refer to a standard or relative value.

[0100] The term "treat" or other forms of the word such as "treated" or "treatment" is used herein to refer to the administration of a compound or formulation of the present application to improve a disease or condition in a host and / or reduce, inhibit or eliminate specific features or events associated with the condition (e.g., vascular leakage or corneal ulcer).

[0101] Insofar as the methods of the present application relate to preventing a disease, it should be understood that the term "prevention" does not require that the disease state be completely prevented. Instead, as used herein, the term prevention refers to the ability of a technician to identify a population susceptible to a disease so that the administration of the compounds of the present application can be performed before the onset of the disease. The term does not mean to completely avoid a disease state.

[0102] The term "ameliorating" or other forms of the word such as "ameliorate" is used herein to refer to the administration of the therapeutic agent of the present application to alleviate one or more symptoms of a disease or disorder in a host and / or to reduce, inhibit or eliminate specific symptoms associated with the disease or disorder before and / or after administration of the therapeutic agent.

[0103] The first active agent compound disclosed affects vascular leakage or pathological neovascularization by inhibiting receptor tyrosine kinases. The second active agent compound disclosed affects corneal epithelial damage by modulating (eg, activating) receptor tyrosine kinases.

[0104] Throughout the description and claims of this specification, the word "comprise" and other forms of the word, such as "comprising" and "comprises", mean including but not limited to, and are not intended to exclude, for example, other additives or components.

[0105] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0106] "Optional" or "optionally" means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0107] The term "about" refers to any minimal 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 the preparation of the formulation and in the treatment of a disease or condition. For example, but not limited to, if the concentration varies between 0.005% and 5.0% (e.g., ±0.0005%), the concentration of the therapeutic agent will be effective. The term "about" with respect to the concentration range of the therapeutic agent / active agent (e.g., a first active agent or a second active agent) of the present application also refers to any change in the amount or range, which will be an effective amount or range.

[0108] Ranges can be expressed herein as from "about" a particular value, and / or to "about" another particular value. When expressing this range, another aspect includes from a particular value and / or to another particular value. Similarly, when a value is expressed as an approximation by using the antecedent "about", it is understood that the particular value forms another aspect. It should also be understood that the endpoints of each range are significant relative to another endpoint and independently of another endpoint. It should also be understood that there are multiple values ​​disclosed herein, and 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, "about 10" is also disclosed. It should also be understood that when a value is disclosed, "less than or equal to" the value, "greater than or equal to" the value, and the possible range between the values ​​are also disclosed, as appropriately understood by the technician. For example, if the value "10" is disclosed, "less than or equal to 10" and "greater than or equal to 10" are also disclosed. It should also be understood that throughout the application, data is provided in a variety of different formats, and the data represents the range of any combination of endpoints and starting points and data points. For example, if a specific data point "10" and a specific 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, and between 10 and 15 are considered disclosed. It is also understood that every unit between two specific units is also disclosed. For example, if 10 and 15 are disclosed, 11, 12, 13, and 14 are also disclosed.

[0109] Unless otherwise stated, the term "halogen" as used herein includes fluorine, chlorine, bromine or iodine. Preferred halogen groups are fluorine, chlorine and bromine.

[0110] Unless otherwise indicated, the term "alkyl" as used herein includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. 1-6 Alkyl is intended to include C1, C2, C3, C4, C5 and C6 alkyl. Examples of alkyl include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl and n-hexyl. In certain embodiments, straight or branched alkyl has 6 or less carbon atoms (e.g., for straight chain, C1-C6, for branched chain, C3-C6) in its main chain, and in another embodiment, straight or branched alkyl has four or less carbon atoms. Similarly, cycloalkyl has 3-8 carbon atoms in its ring structure, and in other embodiments, cycloalkyl has 5 or 6 carbons in the ring structure.

[0111] The alkyl group can be substituted by replacing hydrogen on one or more carbon atoms of the hydrocarbon backbone. Such substituents may include, for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, phosphate, phosphonate, phosphinate, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and urea), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclic, alkylaryl or aromatic or heteroaromatic moiety. The cycloalkyl groups can 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 group substituted with an aryl group, for example, phenylmethyl (benzyl).

[0112] As used herein, the term "alkenyl", unless otherwise indicated, includes unsaturated aliphatic groups of similar length and possibly replacing the above-mentioned alkyl, but containing at least one double bond. For example, the term "alkenyl" includes straight chain alkenyl (e.g., vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched alkenyl, cycloalkenyl (e.g., alicyclic) groups (e.g., cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl substituted cycloalkenyl, and cycloalkyl or cycloalkenyl substituted alkenyl. In certain embodiments, straight or branched alkenyl has six or less carbon atoms in its main chain (e.g., for straight chain, C2-C6, for side chain, C3-C6). Similarly, cycloalkenyl can have 3-8 carbon atoms in its ring structure, and in some embodiments, cycloalkenyl has 5 or 6 carbons in the ring structure. The term "C2-C6" includes alkenyl groups containing 2 to 6 carbon atoms. The term "C3-C6" includes alkenyl groups containing 3 to 6 carbon atoms. Alkenyl groups can be substituted by replacing hydrogen on one or more hydrocarbon backbone carbon atoms. Such substituents may include, for example, alkynyl, halogen, hydroxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, 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, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[0113] As used herein, the term "alkynyl", unless otherwise indicated, includes unsaturated aliphatic groups that are similar in length and may replace the above-mentioned alkyl, but contain at least one triple bond. For example, "alkynyl" includes straight chain alkynyl (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl), branched chain alkynyl and cycloalkyl or cycloalkenyl substituted alkynyl. In certain embodiments, straight chain or branched chain alkynyl has six or less carbon atoms (e.g., for straight chain, C2-C6, for branched chain, C3-C6) in its main chain. The term "C2-C6" includes alkynyl containing 2 to 6 carbon atoms. The term "C3-C6" includes alkynyl containing 3 to 6 carbon atoms. Alkynyl can be substituted by replacing the hydrogen on one or more hydrocarbon main chain carbon atoms. Such substituents may include, for example, halogen, hydroxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, 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, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[0114] Unless otherwise indicated, the term "alkoxy", as used herein, includes O-alkyl groups, wherein "alkyl" is as defined above.

[0115] Unless otherwise indicated, the term "aryl", as used herein, includes 5- and 6-membered "non-conjugated", or monocyclic, aromatic groups, as well as "conjugated", or polycyclic systems having at least one aromatic ring. Examples of aryl groups include benzene, phenyl, and the like. In addition, the term "aryl" includes polycyclic aromatic groups, for example, tricyclic, bicyclic, such as naphthalene.

[0116] Aryl groups having heteroatoms in the ring structure may be referred to as "aromatic heterocycles," "aryl heterocycles," "heterocycles," "heteroaryls," or "heteroaromatics."

[0117] The aryl or heteroaryl groups may be substituted at one or more ring positions with such substituents as described above, for example, halogen, hydroxy, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonate, thiocarboxylate ... The aryl radicals may be substituted or substituted with alkyl or heteroaryl radicals, phosphonate, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl or aromatic or heteroaromatic moieties. The aryl radical may also be fused or bridged with an aliphatic ring or a heterocyclic ring that is not aromatic to form a polycyclic system (e.g., tetralin, methylenedioxyphenyl).

[0118] Unless otherwise indicated, the term "4-10 membered heterocyclic group" as used herein includes aromatic and non-aromatic heterocyclic groups containing one or more heteroatoms each selected from O, S and N, wherein each heterocyclic group has 4-10 atoms in its ring system. Non-aromatic heterocyclic groups include groups having only 4 atoms in their ring system, but aromatic heterocyclic groups must have at least 5 atoms in their ring system. An example of a 4-membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5-membered heterocyclic group is thiazolyl, and an example of a 10-membered heterocyclic group is quinolinyl.

[0119] Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothiophenyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl and quinolizinyl.

[0120] Examples of aromatic heterocyclic groups are pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl and furopyridinyl.

[0121] The above groups as derived from the compounds listed above may be C-attached or N-attached where possible. For example, a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). The "4-10 membered heterocycle" portion may be substituted.

[0122] Unless otherwise indicated, the phrase "pharmaceutically acceptable salt" as used herein includes salts of acidic or basic groups that may be present in the compound of the first active agent (e.g., Formula I or II) or the second active agent. The compounds of Formula I or II and the second active agent that are basic in nature are capable of forming a 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 the second active agent are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate [i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid)] salts.

[0123] Those compounds of the first active agent (e.g., Formula I or II) and the second active agent that are acidic in nature are capable of forming basic salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts, particularly sodium and potassium salts.

[0124] In some embodiments, the salt is an acid addition salt, such as an HCl salt.

[0125] Certain compounds of formula I or II and certain second active agents may have asymmetric centers and therefore exist in different enantiomeric forms. The present 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. The present application relates to the use of all such geometric isomers and tautomers and mixtures thereof.

[0126] The present application also includes isotopically labeled compounds and pharmaceutically acceptable salts thereof, which are identical to those described in Formula I or II, but in fact one or more atoms are replaced by atoms 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 the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 35 S. 18 F and 36 Cl. Compounds of formula I or II, their conjugates, and pharmaceutically acceptable salts of the compounds or conjugates containing the above-mentioned isotopes and / or other isotopes of other atoms are within the scope of the present application. Certain isotopically labeled compounds of the present application (e.g., compounds of formula I or II), for example, incorporating radioactive isotopes such as 3 H and 14 C, can be used in drug and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes are particularly preferred because they are easy to prepare and detect. In addition, heavier isotopes such as deuterium (i.e. 2 H) substitution may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased half-life in vivo or reduced dosage requirements, and may therefore be preferred in some cases. Isotopically labeled compounds of formula I or II of the present application and their esters or lipid conjugates can generally be prepared by carrying out the methods disclosed in the following schemes and / or examples and preparations, by substituting readily available isotopically labeled reagents for non-isotopically labeled reagents.

[0127] The application also covers the pharmaceutical preparations containing the derivatives of formula I or II compounds or their pharmaceutically acceptable salts and the second active agent or their pharmaceutically acceptable salts. Formula I or II compounds or their pharmaceutically acceptable salts and the second active agent or their pharmaceutically acceptable salts with free amino groups or acylamino groups can be converted into conjugated derivatives, wherein the polypeptide chain of amino acid residues or two or more (for example, two, three or four) amino acid residues is covalently connected to the free amino groups of formula I or II compounds or their pharmaceutically acceptable salts or the second active agent or their pharmaceutically acceptable salts by amide or ester bonds. Amino acid residues include but are not limited to 20 kinds of naturally occurring amino acids generally represented by three-letter symbols, and also include 4-hydroxyproline, hydroxylysine, desmosine (demosine), isodesmosine, 3-methylhistidine, norvaline, β-alanine, γ-aminobutyric acid, citrulline homocysteine, homoserine, ornithine and methionine sulfone.

[0128] Additional types of derivatives are also contemplated.Amides and ester moieties can be incorporated into groups, including but not limited to ethers, amines and carboxylic acid functional groups. Free hydroxyls can be derivatized using groups (including but not limited to hemisuccinates, phosphates, dimethylaminoacetates and phosphoryloxymethoxycarbonyl), as described in D. Fleisher, et al., Advanced Drug Delivery Reviews (1996) 19, 115. Also included are carbamate conjugates of hydroxyls and amino groups, as are carbonate conjugates and sulfates of hydroxyls. Hydroxyl derivatization is also contemplated to be (acyloxy) methyl and (acyloxy) ethyl ethers, wherein acyl groups can be alkyl esters, which are optionally substituted by groups (including but not limited to ethers, amines and carboxylic acid functional groups), or wherein acyl groups are amino acid esters as described above. This type of derivative is described in RP Robinson et al., J. Medicinal Chemistry (1996) 39, 10.

[0129] The term "kinase" refers to any enzyme that catalyzes the addition of a phosphate group to a protein residue; for example, serine and threonine kinases catalyze the addition of phosphate groups to serine and threonine residues.

[0130] The terms "VEGFR kinase" and "VEGFR" refer to any vascular endothelial growth factor receptor.

[0131] The terms "VEGF signaling" and "VEGF cascade" refer to the upstream and downstream components of the VEGF signaling cascade.

[0132] 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).

[0133] The terms "EGF signaling" and "EGF cascade" refer to the upstream and downstream components of the EGF signaling cascade.

[0134] 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 deleterious to the recipient thereof.

[0135] The term "administration of a compound" or "administering a compound" refers to the act of providing a compound or pharmaceutical formulation of the present application to a subject in need of treatment.

[0136] The term "vasostasis" refers to the maintenance of homeostatic vascular function, resulting in normal physiological function.

[0137] The term "vasostatic agent" refers to an agent that seeks to address conditions in which vasostasis is impaired by preventing the loss of vasostasis or restoring or maintaining vasostasis.

[0138] In this application, "composition" and "formulation" are used interchangeably and refer to the conventional understanding of compositions or formulations as known in the art.

[0139] The present application relates to ophthalmic preparations. In some embodiments, the ophthalmic preparations of the present application are gel preparations or semi-gel preparations or both.

[0140] A "gel" according to the present application is a semisolid dosage form of the present application, which contains suspended particles. A semisolid is not pourable; it does not flow or fit into its container at room temperature. A semisolid does not flow under low shear stresses 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 uniformly distributed throughout a liquid.

[0141] In some embodiments, "gel" is a semi-solid system composed of a suspension of small inorganic particles or a suspension of organic molecules interpenetrating liquids. "Gel" is classified as a single-phase or two-phase system. "Gel" is also composed of the following: the state of matter between the intermediate phase or liquid and the solid, which represents a partially ordered structure, which is the state of the active agent in the "gel drops" of the present embodiment. The two-phase gel is composed of a network of small discrete particles. In a two-phase system, if the particle size of the suspended material is large, the gel is sometimes referred to as a paste (e.g., bentonite paste). Both gel and paste are thixotropic, forming a semi-solid after standing and becoming a liquid after stirring. Semi-solid preparations should be shaken before application to ensure uniformity, and should be labeled as such (see suspension). Single-phase gel is composed of organic macromolecules uniformly distributed in the liquid, in a manner such that there is no obvious boundary between the dispersed macromolecules and the liquid. Single-phase gel can also be composed of organic low molecular weight (LMW) molecules, wherein the component responsible for gelling is the actual active ingredient. These so-called "LMW hydrogels" are different from traditional gelling agents for water, such as high molecular weight synthetic polymers, polysaccharides and proteins. High molecular weight gelling agents are highly ordered and unidirectional due to hydrogen bonding, while the forces that govern LMW hydrogels are mainly non-directional van der Waals (hydrophobic) interactions. In practice, LMW hydrogels are observed as highly anisotropic (usually fibrous) structures that propagate throughout the liquid, producing physically branched or entangled networks. Therefore, the gel can be disordered to slightly ordered, showing some birefringent, liquid crystal characteristics. The gel is applied topically, or, after vibration, in the form of a hydrogel as an eye drop.

[0142] A semisolid "gel" according to the present application is a semisolid according to the USP definition and the references cited therein. The apparent viscosity of a semisolid formulation increases with concentration. The clinical dosage strengths of this formulation range from a low strength of ≤1 mg / mL (0.1%) to a high strength of ≤6 mg / mL (0.6%). The low strength dose is the least viscous and falls into the category of a "solution", while the higher strengths are more viscous and meet the definition of a gel.

[0143] A "jelly" according to the present application is a type of gel which is a semisolid system consisting of a suspension of small inorganic particles or large organic molecules interpenetrated by a liquid, wherein the structural bonding matrix contains a large portion of the liquid, usually water.

[0144] A "solution" according to the present application is a clear, uniform liquid dosage form containing one or more chemical substances dissolved in a solvent or a mixture of mutually miscible solvents. A solution is a liquid preparation containing one or more dissolved chemical substances in a suitable solvent or a mixture of mutually miscible solvents. Because the molecules of the drug substance in the solution are uniformly dispersed, using a solution as a dosage form generally ensures uniform dosage after administration and good accuracy when the solution is diluted or otherwise mixed.

[0145] A "liquid" according to the present application is a dosage form consisting of a pure chemical in its liquid state. A liquid is pourable; it flows at room temperature and fits into its container. A liquid exhibits Newtonian or pseudoplastic flow behavior.

[0146] A "suspension" according to the present application is a liquid dosage form containing solid particles dispersed in a liquid carrier.

[0147] The compound of the first active agent (for example, Formula I or II) and the second active agent is formulated into a therapeutic preparation as a natural or salt form. Pharmaceutically acceptable non-toxic salts include base addition salts derived from inorganic bases (such as, for example, sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide or ferric hydroxide) and organic bases (such as isopropylamine, trimethylamine, 2-ethylamino-ethanol, histidine, procaine, etc.) (formed with free carboxyl or other anionic groups). Such salts are formed as acid addition salts with any free cationic group, and are usually formed with inorganic acids (such as, for example, hydrochloric acid, sulfuric acid or phosphoric acid) or organic acids (such as acetic acid, citric acid, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, tartaric acid, mandelic acid, etc.). The salt of the present application includes an amine salt formed by protonating an amino group with an inorganic acid (such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, etc.). The salt of the present application also includes an amine salt formed by protonating an amino group with a suitable organic acid (such as p-toluenesulfonic acid, acetic acid, etc.). Additional excipients contemplated for use in the practice of this application are those available to those of ordinary skill in the art, for example, those found in U.S. Pharmacopoeia Volume XXII and National Formulary Volume XVII, US Pharmacopoeia Convention, Inc., Rockville, Md. (1989) (the relevant contents of which are incorporated herein by reference). In addition, polymorphs of the present compounds are included in this application.

[0148] Embodiments of the present application provide an ophthalmic composition or formulation for treating ocular neovascularization, comprising a first active agent of Formula I:

[0149]

[0150] or a pharmaceutically acceptable salt thereof; a second active agent or a pharmaceutically acceptable salt thereof, wherein the second active agent is niacin, niacinamide, or vitamin K, or a combination thereof; and a pharmaceutically acceptable excipient; the first active agent or the pharmaceutically acceptable salt is present at about 0.02% to about 1.2% w / v, wherein:

[0151] X 1 is O or S;

[0152] R 1 Yes H, C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C(O)(C1-C 10 Alkyl), (CH2) t (C6-C 10 Aryl), (CH2) t (4-10 membered heterocyclic ring), C(O)(CH2) t (C6-C 10 Aryl) or C(O)(CH2) t (5-10 membered heterocyclic ring), wherein:

[0153] t is an integer from 0 to 5;

[0154] The alkyl group optionally includes 1 or 2 groups selected from O, S and N (R 6 ), provided that two O atoms, two S atoms, or O and S atoms are not directly connected to each other;

[0155] Aryl and heterocyclic groups are optionally combined with C6-C 10 An aryl group, a C5-C8 saturated ring group or a 5-10 membered heterocyclic group is fused;

[0156] One or two carbon atoms in the above heterocyclic moiety are optionally substituted by an oxo (═O) moiety or an oxygen anion;

[0157] When t is an integer from 2 to 5, (CH2) t The moiety optionally includes a carbon-carbon double or triple bond; and

[0158] The above R 1 The group, except H, is optionally substituted by 1 to 3 R 4 group substitution;

[0159] R 2 It is H;

[0160] R 3 Yes (CH2) t (C6-C 10 aryl), wherein:

[0161] t is an integer from 0 to 5;

[0162] The aryl group is optionally 10 An aryl group, a C5-C8 saturated ring group or a 5-10 membered heterocyclic group is fused;

[0163] When t is an integer from 2 to 5, (CH2) t The moiety optionally includes a carbon-carbon double or triple bond; and

[0164] Each R 4 Independently selected from C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, OR 5 、C(O)R 5 、C(O)OR 5 NR 6 C(O)R 5 NR 6 C(O)OR 5 、OC(O)R 5 NR 6 S02R 5 、SO2NR 5 R 6 、C(O)NR 5 R 6 NR 5 R 6 、S(O) j R 7 (wherein j is an integer from 0 to 2), SO3H, NR 5 (CR 6 R 7 ) t OR 6 、(CH2) t (C6-C 10 Aryl), SO2(CH2) t (C6-C 10 Aryl), S(CH2) t (C6-C 10 Aryl), O(CH2) t (C6-C 10 Aryl), (CH2) t (5-10 membered heterocyclic ring) and (CR 6 R 7 ) m OR 6 ,in:

[0165] m is an integer from 1 to 5;

[0166] t is an integer from 0 to 5;

[0167] The alkyl group optionally includes 1 or 2 groups selected from O, S and N (R 6 ), provided that two O atoms, two S atoms, or O and S atoms are not directly connected to each other;

[0168] Aryl and heterocyclic groups are optionally combined with C6-C 10 An aryl group, a C5-C8 saturated ring group or a 5-10 membered heterocyclic group is fused;

[0169] One or two carbon atoms in the above heterocyclic moiety are optionally substituted by an oxo (═O) moiety or an oxygen anion; and

[0170] The above R 4 The alkyl, aryl and heterocyclic moieties of the group are optionally substituted by 1-3 groups independently selected from halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, NR 6 S02R 5 、SO2NR 5 R 6 、C(O)R 5 、C(O)OR 5 、OC(O)R 5 NR 6 C(O)R 5 、C(O)NR 5 R 6 NR 5 R 6 , (CR 6 R 7 ) m OR 6 (wherein m is an integer from 1 to 5), OR 5 , and R 5 is substituted with a substituent listed in the definition of ; and

[0171] R 5 , R 6 and R 7 Each is independently H or C1-C6 alkyl.

[0172] In one embodiment, R 3 Yes (CH2) t (C6-C 10 aryl), wherein t is an integer from 1 to 3, and R 3 Optionally 1-4 R 4 Group substitution.

[0173] In a further embodiment, R 3is benzyl, which is optionally substituted with 1 to 4 substituents independently selected from halogen and C1-C4 alkyl. In a further embodiment, R 3 is benzyl substituted with 1 to 4 substituents independently selected from methyl, fluoro, chloro and bromo.

[0174] In one embodiment, R 1 Yes (CH2) t (5-10 membered heterocycle), wherein t is an integer from 0 to 5, which is optionally substituted with 1 or 2 substituents independently selected from C1-C4 alkyl, hydroxy and hydroxymethyl.

[0175] The present application provides R in Formula I 1 The heterocyclic portion of the group is selected from morpholino, pyrrolidinyl, imidazolyl, piperazinyl, piperidinyl and 2,5-diaza-bicyclo[2.2.1]hept-2-yl, R 1 The t variable range of the group is 2 to 5, and R 1 The group is optionally substituted with one or more hydroxy groups.

[0176] For example, R in Formula I of the present application 1 The heterocyclic portion of the group is pyrrolidine.

[0177] In a further embodiment of the present application, the first active agent is:

[0178] .

[0179] The compound of the present application is 3-[(4-bromo-2,6-difluorophenyl)methoxy]-5-[[[[4-(1-pyrrolidinyl)butyl]amino]carbonyl]amino]-4-isothiazolecarboxamide hydrochloride, molecular formula: C 20 H 24 BrF2N5O3S·HCl, molecular weight: 568.86 g / mol, and has the characteristics that the molecule does not contain an asymmetric center and is not chiral. The compound of the present application is represented by Compound-I:

[0180]

[0181] (Compound-I).

[0182] Compound-I of the present application is an inhibitor of the tyrosine kinase activity of VEGFR-2, which blocks the autophosphorylation of the receptor stimulated by VEGF and endothelial cell proliferation. It is selective (>500x) relative to the concentration required for inhibiting epidermal growth factor receptor (EGFR) and insulin receptor (IR) tyrosine kinase. Compound-I is described in U.S. Patent 6,235,764. In some embodiments, the compound of Formula I or II is a VEGFR-2 inhibitor.

[0183] The second active agent of the present application is an EGFR modulator (eg, activator) of the tyrosine kinase activity of EGFR.

[0184] 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.

[0185] In one embodiment, the compound of Formula I or II or its pharmaceutically acceptable salt and the second active agent or its pharmaceutically acceptable salt are administered simultaneously. Alternatively, the compound of Formula I or II or its pharmaceutically acceptable salt is administered before the second active agent or its pharmaceutically acceptable salt is administered. In another embodiment, the compound of Formula I or II or its pharmaceutically acceptable salt is administered after the second active agent or its pharmaceutically acceptable salt is administered.

[0186] General Features

[0187] The compound-I of the present application has the characteristics as shown in Table 1. The embodiment provides three preparations of the compound-I or its free base - the compound of formula II.

[0188] Table 1A: General properties of Compound-1 API

[0189]

[0190] The composition of Compound-I formulations is listed in Table 1B. The formulation materials are listed in Table 1C.

[0191] Table 1B: Compound-I formulations: gel drop, suspension, and solution.

[0192]

[0193] Table 1C: Preparation Materials

[0194]

[0195] The composition of the Compound-I formulation and the second active agent are listed in Table ID. The formulation materials are listed in Table IE.

[0196] Table ID: Compound-I formulations: gel drop, suspension, and solution.

[0197]

[0198] Table 1E: Preparation Materials

[0199]

[0200] Ophthalmic solution

[0201] The present application provides a formulation of a first active agent (e.g., Compound-I and / or its free base (Formula II compound)) and / or a second active agent formed as a solution having a viscosity similar to that of water. The solution includes a pharmaceutically acceptable agent / excipient, such as but not limited to cyclodextrin. The solution thus formed is a clear and colorless solution suitable for topical application to the eye.

[0202] The solutions of the present application reduce anterior segment exposure of the first active agent; thus they allow for increased concentration of the first active agent (eg, compound of Formula I or II) in solution, and increased frequency of delivery, thereby facilitating the maintenance of high concentrations of the first active agent in the posterior segment of the eye.

[0203] The solutions of the present application contain 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, such as Compound-I. In some embodiments, for topical administration, the concentration of Compound-I or its free base (Formula II) in the solution is 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%, 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. In some embodiments, for topical administration, the concentration of Compound-I or its free base (Formula II) in the formulation 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 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 1.0%, About 0.6%, about 0.4% - about 0.6%, 0.1% - about 0.5%, about 0.2% - about 0.5%, about 0.3% - about 0.5%, about 0.4% - about 0.5%, 0.1% - about 0.4%, about 0.2% - about 0.4%, about 0.3% - about 0.4% w / v. In some embodiments, the solution includes 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).

[0204] The solution of the present application may also include about 0.00001% to about 5.0% w / v of a second active agent or a pharmaceutically acceptable salt thereof, such as niacin, niacinamide, or vitamin K, or a combination thereof. The solution of the present application may also include 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 a second active agent or a pharmaceutically acceptable salt thereof, such as niacin, niacinamide, or vitamin K, or a combination thereof. In some embodiments, for topical administration, the concentration of the second active agent in the solution is about 0.00001% - about 0.0001%, 0.000012% - about 0.0001%, 0.000014% - about 0.0001%, 0.000016% - about 0.0001%, 0.000018% - about 0.0001%, 0.00002% - about 0.0001%, 0.00003% - about 0.0001%, 0.00004% - about 0.0001%, 0.00005% - about 0.0001%, 0.00006% - about 0.0001%, 0.00007% - about 0.0001%, 0.00008% - about 0.0001%, 0.00009% - about 0.0001%, 0.000016% - about 0.00009%, 0.000018% - about 0.00009%, 0.00002% - about 0.00009%, 0.00003% - about 0.00009%, 0.00004% - about 0.00009%, 0.00005% - about 0.00009%, 0.00006% - about 0.00009%, 0.00007% - about 0.00009%, 0.00008% - about 0.00009% w / v. In some embodiments, the solution comprises 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.

[0205] The solution of the present application may also include 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 acceptable salt thereof, such as niacin, niacinamide or vitamin K or a combination thereof. In some embodiments, the concentration of the second active agent is about 1 μM.

[0206] In some embodiments, the formulation comprises cyclodextrins for improving the solubility of the first active agent (e.g., Compound-1). Cyclodextrins, oligosaccharides consisting of 6 to 8 glucose units connected by one or four bonds, increase the solubility of active agents with poor or low solubility in water or aqueous solutions (e.g., in PBS buffer). Cyclodextrins form hydrophilic complexes with hydrophobic active agents.

[0207] One or more cyclodextrins can be used in the solution of the application. Non-limiting examples of cyclodextrins used in the preparation of the application are, for example: 2-hydroxypropyl-β-cyclodextrin, methyl-β-cyclodextrin, random methylation-β-cyclodextrin, ethylation-β-cyclodextrin, triacetyl-β-cyclodextrin, peracetylation-β-cyclodextrin, carboxymethyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, 2-hydroxy-3-(trimethylammonium)propyl-β-cyclodextrin, glucosyl-β-cyclodextrin, maltosyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, branched-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, random methylation-γ-cyclodextrin, trimethyl-γ-cyclodextrin or a combination thereof.

[0208] In some embodiments, the solution of the compound of formula II or compound-I containing cyclodextrin is a transparent and colorless solution and has a viscosity similar to that of water. In some embodiments, the application provides a solution comprising compound-I, one or more cyclodextrins and a second active agent for topical administration, and it is topically applied to the eye.

[0209] The ophthalmic solution of the present application comprises cyclodextrin and a pharmaceutical excipient selected at a concentration equal to or lower than the optimal concentration of the ophthalmic solution. The excipient of the present application is, for example, benzalkonium chloride (BAK) and NaCl. In some embodiments, the ophthalmic solution comprises about 0.001-about 0.005% w / v benzalkonium chloride (BAK). The amount of BAK varies according to the needs of the application.

[0210] The ophthalmic solution comprises, for example, but not limited to, about 0.005% - 5.0% compound-1 or its free base, about 2-about 25% cyclodextrin, such as, but not limited to, hydroxypropyl-β-cyclodextrin (HPβCD) or methyl cyclodextrin (KLEPTOSE® HPB), and / or sulfobutyl ether-β-cyclodextrin (CAPTISOL®), about 0.1 to about 0.7% salt, such as, but not limited to, NaCl), and / or about 0.005% antimicrobial agent, such as, but not limited to, benzalkonium chloride (BAK). The formulation comprises compound-1 or its free base and cyclodextrin in a ratio of 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, or 1:10 to 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% - 1% Tris. In some embodiments, the ophthalmic solution comprises about 0.05% - 0.5% Tris. In some embodiments, the ophthalmic solution comprises about 0.05% - 0.2% Tris. In some embodiments, the ophthalmic solution comprises about 0.1% - 0.15% Tris. In some embodiments, the ophthalmic solution comprises about 1% Tris. In some embodiments, the ophthalmic solution further comprises about 0.005% -5.0% of a second active agent.

[0211] The ophthalmic solution of the present embodiment includes, for example but not limited to: about 0.3% to about 5.0% Compound-I (about 3 mg / mL to about 50.0 mg / mL), about 0.05% sodium dihydrogen phosphate monohydrate, about 2% glycerol; about 0.4% Compound-I, about 7% HPβCD, about 0.7% NaCl, about 0.005% BAK; about 0.4% Compound-I, about 4% HPβCD, about 0.7% NaCl, about 0.005% BAK; about 0.4% Compound-I, about 7% HPβCD, about 1% tromethamine, about 0.4% NaCl, about 0.005% BAK; and about 0.6% Compound-I, about 7% HPβCD, about 0.7% NaCl, about 0.005% BAK. For a concentration of about 0.005% to about 5.0% Compound-I, the cyclodextrin is present in a corresponding molar ratio. In some embodiments, the ophthalmic solution further comprises about 0.005% - 5.0% of a second active agent.

[0212] Additional ophthalmic solutions include, for example, but are not limited to: about 0.4% compound of Formula II (free base), about 7.15% HPβCD, about 0.7% NaCl; about 0.1% compound of Formula II (free base), about 1.79% HPβCD, about 0.85% NaCl; about 0.2% compound of Formula II (free base), about 3.57% HPβCD, about 0.8% NaCl; about 0.6% compound of Formula II (free base), about 10.72% HPβCD, about 0.6% NaCl; about 0.4% compound of Formula II (free base), about 8.41% HPβCD, about 0.65% NaCl; about 0.4% compound-I, about 10.51 HPβCD, about 0.65% NaCl; about 0.4% compound of Formula II (free base), about 10.51% HPβCD, about 0.15% NaCl, about 1.0% tromethamine (Tris); and / or about 0.1% compound of formula II (free base), about 2.63% HPβCD, about 0.8% NaCl; about 0.6% compound-I (as free base), about 15.77% HPβCD, about 0.37% NaCl. For about 0.005% to about 5.0% concentration of formula II, cyclodextrin is present in a corresponding molar ratio. In some embodiments, the ophthalmic solution also contains about 0.005% - 5.0% of a second active agent.

[0213] In some embodiments, the ophthalmic solution includes about 1.0% to about 25% cyclodextrin. For example, but not limited to, the Compound-I formulation includes about 2.0% to about 3.0% HPβCD, about 3.0% to about 5.0% HPβCD, about 5.0% to about 10% HPβCD, or about 10% to about 25% HPβCD. In some embodiments, the ophthalmic solution further comprises about 0.005% to 5.0% of a second active agent.

[0214] In additional embodiments, the ophthalmic solutions are formulated, for example, but not limited to: 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.

[0215] In some embodiments, ophthalmic solutions comprising cyclodextrins are clear and colorless, and are extremely viscous, moderately viscous, or have a viscosity similar to that of water.

[0216] In some embodiments, the ophthalmic solutions of the present application have a pH of about 4.5 to about 7.5 at or below about 40°C.

[0217] In some embodiments, the ophthalmic solutions of the present application have a pH of about 5.0 to about 7.0 at or below about 40°C.

[0218] For example, the ophthalmic solution of the present application has a pH value of about 6.0 at or below about 40°C.

[0219] The ophthalmic solution of the present application may contain various additives that are usually incorporated, such as buffers (e.g., phosphate buffers, borate buffers, citrate buffers, tartrate buffers, acetate buffers, amino acids, sodium acetate, sodium citrate, etc.), tonicity agents (e.g., sugars such as sorbitol, glucose and mannitol, polyols such as glycerol, concentrated glycerol, PEG and propylene glycol, salts such as sodium chloride, etc.), preservatives or antibacterial agents (e.g., benzalkonium chloride, Benzatkonium chloride, parabens such as methylparaben or ethylparaben, benzyl alcohol, phenylethyl paraben, etc.), alcohol, sorbic acid or its salt, thimerosal, chlorobutanol, etc.), solubilizing aids or particle stabilizers (e.g., water-soluble polymers such as polyvinyl pyrrolidone, surfactants such as tyloxapol, polysorbates, poloxamers, etc.), pH adjusters (e.g., hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide, potassium hydroxide, ammonium hydroxide, etc.), thickeners (e.g., HEC, hydroxypropyl cellulose, methyl cellulose, HPMC, carboxymethyl cellulose and salts thereof), chelating agents (e.g., sodium edetate, sodium citrate, concentrated sodium phosphate, etc.) and second active agent stabilizers (e.g., EDTA, propyl gallate and combinations thereof).

[0220] The ophthalmic solution of the present application comprises cyclodextrin and may further comprise additional excipients, such as, but not limited to, about 0.5% to about 3% surfactants and emulsifiers, such as, but not limited to, polysorbate 80 or its equivalent excipients; about 0.05% to about 0.4% non-ionic liquid polymers of the alkylaryl polyether alcohol type, such as, but not limited to, tyloxapol; and / or about 0.05% to about 0.6% hydrophilic non-ionic surfactants, such as, but not limited to, poloxamers, such as poloxamer 407.

[0221] In some embodiments, the ophthalmic solution comprises about 0.01 - about 0.5%, about 0.02 - about 0.5%, about 0.04 - about 0.5%, about 0.06 - about 0.5%, about 0.08 - about 0.5%, about 0.08 - about 0.4%, about 0.08 - about 0.3%, about 0.08 - about 0.2%, about 0.08 - about 0.18%, about 0.08 - about 0.16%, about 0.08 - about 0.14%, or about 0.08 - 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.

[0222] In some embodiments, the ophthalmic solution comprises about 0.001 - about 0.5%, about 0.002 - about 0.5%, about 0.005 - about 0.5%, about 0.01 - about 0.5%, about 0.02 - about 0.5%, about 0.03 - about 0.5%, about 0.04 - about 0.5%, about 0.01 - about 0.4%, about 0.01 - about 0.3%, about 0.01 - about 0.2%, about 0.01 - about 0.1%, about 0.01 - about 0.08%, about 0.01 - 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.

[0223] Concentrations in various ocular tissues—delivered as an ophthalmic solution

[0224] The ophthalmic solution containing cyclodextrin improves the bioavailability of the first active agent of the present application in the posterior segment of the eye. Without being bound by theory, in one embodiment, the formulation containing cyclodextrin forms a clear and colorless solution, which reduces the corneal exposure of the active agent, for example, the exposure of Compound-1, by about 5-15 times compared to the corneal exposure of the equimolar gel drop formulation.

[0225] Without being bound by theory, in one embodiment, an ophthalmic solution comprising cyclodextrin increases the therapeutic index of Compound-1 during topical ocular administration. After administration, the hydrophilic complex of cyclodextrin-Compound-1 is pharmacologically inert at the cornea. Without being bound by theory, in some embodiments, the cyclodextrin-Compound-1 complex increases the corneal tolerability of Compound-1. Without being bound by theory, in some embodiments, the spontaneous dissociation of cyclodextrin from Compound-1 at the peripheral vasculature increases bioavailability at target tissues, such as the choroid or retina.

[0226] Unlike other formulations of Compound-I that contribute to corneal toxicity in some embodiments, cyclodextrin-based ophthalmic solutions containing similar concentrations of Compound-I reduce corneal exposure, thereby increasing the therapeutic index and corresponding benefits to patients. In one embodiment, the use of a cyclodextrin-based solution of Compound-I provides a reduction in corneal exposure by about 10 times compared to gel drops of equimolar concentration. In some embodiments, a cyclodextrin-based solution of Compound-I reduces corneal exposure of Compound-I by 5 times, 20 times, 30 times, 40 times, or 50 times. In one embodiment, topical ocular administration of about 0.005% to about 5.0% Compound-I as a cyclodextrin-based solution for 1-90 days or 3-9 months has no adverse or toxic effects at the cornea, choroid, and / or retina. In yet another embodiment, topical ocular administration of about 0.6% to about 5.0% Compound-I as a cyclodextrin-based solution for 1-90 days has no adverse or toxic effects at the cornea, choroid, and / or retina.

[0227] The reduction in corneal exposure is associated with increased bioavailability and therapeutic index of the active agent at the posterior segment of the eye (e.g., at the retina or choroid). For example, when about 0.1% to about 5.0% of a cyclodextrin-containing Compound-I formulation is topically applied to the eye for at least 30 days or more than 60 days, no toxic effects on the cornea or other parts of the eye attributable to the first active agent or a suitable carrier are observed.

[0228] In one embodiment, when the formulation comprises about 0.4% (about 4 mg / mL) of Compound-I or its free base and cyclodextrin, when topically applied to the eye, the central choroidal concentration is about 0.2 μM to about 0.9 μM, the central retinal concentration of the active agent is about 0.02 μM to about 0.4 μM, the aqueous humor concentration of the active agent is about 0.003 μM to about 0.009 μM, and the corneal concentration of the active agent is 6 μM to 40 μM. The cyclodextrin used in the formulation is, for example, but not limited to, KLEPTOSE® HPB or CAPTISOL®.

[0229] In some embodiments, a cyclodextrin-based solution of Compound-1 or its free base increases the bioavailability of the active agent at the central choroid and central retina while reducing the concentration at the cornea. In some embodiments, topical delivery of Compound-1 or its free base formulated in the presence of cyclodextrin reduces the corneal concentration by about 5-fold to about 15-fold compared to the corneal concentration of an equimolar gel drop.

[0230] Without being bound by theory, in some embodiments, the combined effect of reducing corneal drug exposure to avoid poor ocular tolerability while increasing posterior segment bioavailability increases the therapeutic index and corresponding benefit to the patient.

[0231] In some embodiments, the exposure time of compound-I and the second active agent is 1 to 90 days or longer 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 preparation comprising compound-I and a second active agent, wherein the subject is administered a second active agent as a separate preparation or as part of a compound-I preparation for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves administering the preparation once a day, twice a day, three times a day, or four times a day for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves administering a compound of formula I or II and a second active agent once, twice, three times, or four times every other day (i.e., on the 1st, 3rd, 5th, 7th day, etc.) for up to 90 days. For example, the dosage regimen involves administering once on the 1st day and once or twice on the 2nd day-90th day. For example, the dosage regimen involves applying once, twice, three times or four times on day 1, followed by once a day for 2-90 days. For example, the dosage regimen involves applying once, twice, three times, four times on day 1, followed by once, twice, three times or four times every other day (i.e., on days 1, 3, 5, 7, etc.) for up to 90 days. For example, one 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 a day dosage regimen, the subject receives a topical ocular dose of the compound-I preparation and the second active agent on day 1 and day 4, separated by about 4, 6 or 8 hours. In another embodiment, the subject receives a topical ocular dose of the compound-I preparation and the second active agent for four consecutive days, separated by about 4, 6 or 8 hours. In some embodiments, the subject receives one or two doses of a topical ocular dose of the compound-I preparation and the second active agent every day for 5 consecutive days. In yet other embodiments, the subject receives one or two doses of a topical ocular dose of the compound-I preparation and the second active agent for 5-90 consecutive days. In some embodiments, the subject receives one or two topical ocular doses of a Compound-1 formulation and a second active agent for at least 25 consecutive days. In one embodiment, the subject receives one or two topical ocular doses for at least 90 consecutive days or longer. The second active agent can be administered separately or as part of a Compound-1 formulation. When administered separately, the second active agent or a pharmaceutical salt thereof can be administered alone or as a formulation.

[0232] For example, a formulation comprising about 1 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 1 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 1 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 1 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 2 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 3 mg / mL BID of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 3 mg / mL QD of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, a formulation comprising about 3 mg / mL TID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 3 mg / mL QID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 4 mg / mL BID of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and / or the second active agent comprising about 4 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 4 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 4 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL BID of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL QD of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL TID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 5 mg / mL QID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, a formulation comprising about 6 mg / mL BID of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 6 mg / mL QD of a first active agent (e.g., Compound-1) and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 6 mg / mL TID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 6 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 7 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 7 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, a formulation comprising about 8 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 9 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 9 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 9 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 9 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 10 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). A dosage regimen of 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) can be any regimen involving continuous or alternate days described in the above paragraphs. In some embodiments, when administered at a low dose (e.g., 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, or 5 mg / mL), the formulation of the present application is administered QD, BID, TID, or QID, and when administered at a high dose (e.g., 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, or 10 mg / mL), the formulation of the present application is administered QD or BID.

[0233] In one embodiment, about 1 mg / mL BID of the first active agent (e.g., Compound-I) formulation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 1 mg / mL QD of the first active agent (e.g., Compound-I) formulation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 2 mg / mL BID of the first active agent (e.g., Compound-I) formulation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 2 mg / mL QD of the first active agent (e.g., Compound-I) formulation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 3 mg / mL BID of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 3 mg / mL QD of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 4 mg / mL BID of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 4 mg / mL QD of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 5 mg / mL BID of the first active agent (e.g., Compound-1) formulation and the second active agent are administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 5 mg / mL QD of the first active agent (e.g., Compound-1) formulation and the second active agent are administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, about 6 mg / mL BID of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 6 mg / mL QD of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 7 mg / mL BID of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 7 mg / mL QD of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 8 mg / mL BID of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 8 mg / mL QD of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 9 mg / mL BID of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 9 mg / mL QD of the first active agent (e.g., Compound-I) preparation and the second active agent are applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 10 mg / mL BID of the first active agent (e.g., Compound-1) formulation and the second active agent are administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, about 10 mg / mL QD of the first active agent (e.g., Compound-1) formulation and the second active agent are administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). The dosage regimen of 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) can be any regimen involving continuous or alternate days described in the above paragraphs.

[0234] The present application provides cyclodextrin-based solutions containing hydroxypropyl-β-cyclodextrin (HP-β-CD, KLEPTOSE® HPB) or CAPTISOL® that are well tolerated when topically administered for 30-90 days or 4-6 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% HP-β-CD or CAPTISOL® is well tolerated by the subject.

[0235] In some embodiments, the preparation of Formula II or Compound-I and the second active agent is applied to one or both eyes of the object. For example, the preparation of the compound-I preparation containing about 0.2%-about 1.0% (w / v) of Formula II or about 0.1%-1.2% (w / v) of the present application and the second active agent is applied once a day (QD), twice a day (BID), three times a day (BID) or four times a day (QID) to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months). In some embodiments, Formula II or Compound-I preparation and the second active agent are applied to one or both eyes of the object. For example, about 0.2% to about 1.0% (w / v) of a compound of Formula II or about 0.1%-1.2% (w / v) of a Compound-I formulation and a second active agent are administered to one or both eyes of a subject once a day (QD), twice a day (BID), three times a day (BID), or four times a day (QID) for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).

[0236] In some embodiments, the compound of formula II or compound-I is compounded with a complexing agent such as cyclodextrin (e.g., Kleptose® HPB (%)) at a ratio of about 1:8, wherein about 2% - 13% (w / v) cyclodextrin (e.g., Kleptose® HPB (%) is added to the formulation. The formulation may also include about 0.1% - about 0.2% of a buffer, for example, 10mM phosphate buffer. The desired weight-gram molecular osmotic concentration of the formulation is about 200-about 300 mOsm, which is achieved by adding a salt (e.g., sodium chloride) in an amount sufficient to achieve the weight-gram molecular osmotic concentration. The pH of the formulation is about 6.0 at about 40°C or below about 40°C. The dosage regimen of 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) can be any regimen involving continuous or alternate days described in the above paragraphs.

[0237] Ophthalmic suspension

[0238] The present application provides a suspension of the active agent comprising a first active agent (e.g., compound-I) and a pharmaceutically acceptable excipient. The present application also provides a suspension comprising a first active agent (e.g., compound-I), a second active agent and a pharmaceutically acceptable excipient and a first active agent and a second active agent. For example, the compound-I suspension and the second active agent suspension may include but are not limited to buffer, acid and alkali, such as but not limited to HCl and NaOH. In one embodiment, the suspension of compound-I or its free base may include a buffer, such as but not limited to tromethamine (Tris). In another embodiment, the suspension of compound-I or its free base and the second active agent may include a buffer, such as but not limited to tromethamine (Tris). The suspension based on tromethamine of formula II compound or compound-I and the second active agent can be used for topical application to the eye.

[0239] The suspension of the present application contains 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, such as Compound-I. In some embodiments, for topical administration, the concentration of Compound-I or its free base (Formula II) in the suspension is 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%, about 1.0- about 2.0%, about 2.0- about 3.0%, about 3.0- about 4.0%, or about 4.0- about 5.0% w / v. In some embodiments, for topical administration, the concentration of Compound-I or its free base (Formula II) in the formulation 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 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 1.0%, About 0.6%, about 0.4% - about 0.6%, 0.1% - about 0.5%, about 0.2% - about 0.5%, about 0.3% - about 0.5%, about 0.4% - about 0.5%, 0.1% - about 0.4%, about 0.2% - about 0.4%, about 0.3% - about 0.4% w / v. In some embodiments, the suspension includes 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).

[0240] The suspension of the present application may also contain about 0.00001% to about 5.0% w / v of a second active agent or a pharmaceutically acceptable salt thereof, such as niacin, niacinamide, or vitamin K, or a combination thereof. The suspension of the present application may also contain 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 a second active agent or a pharmaceutically acceptable salt thereof, such as niacin, niacinamide, or vitamin K, or a combination thereof. In some embodiments, for topical administration, the concentration of the second active agent in the suspension is about 0.00001% - about 0.0001%, 0.000012% - about 0.0001%, 0.000014% - about 0.0001%, 0.000016% - about 0.0001%, 0.000018% - about 0.0001%, 0.00002% - about 0.0001%, 0.00003% - about 0.0001%, 0.00004% - about 0.0001%, 0.00005% - about 0.0001%, 0.00006% - about 0.0001%, 0.00007% - about 0.0001%, 0.00008% - About 0.0001%, 0.00009% - about 0.0001%, 0.000016% - about 0.00009%, 0.000018% - about 0.00009%, 0.00002% - about 0.00009%, 0.00003% - about 0.00009%, 0.00004% - about 0.00009%, 0.00005% - about 0.00009%, 0.00006% - about 0.00009%, 0.00007% - about 0.00009%, or 0.00008% - about 0.00009% w / v. In some embodiments, the suspension comprises 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.

[0241] The suspension of the present application may also contain 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 acceptable salt thereof, such as niacin, niacinamide or vitamin K or a combination thereof. In some embodiments, the concentration of the second active agent is about 1 μM.

[0242] The ophthalmic solution of the present application may contain various additives that are usually incorporated, such as buffers (e.g., phosphate buffers, borate buffers, citrate buffers, tartrate buffers, acetate buffers, amino acids, sodium acetate, sodium citrate, etc.), tonicity agents (e.g., sugars such as sorbitol, glucose and mannitol, polyols such as glycerol, concentrated glycerol, PEG and propylene glycol, salts such as sodium chloride, etc.), preservatives or antibacterial agents (e.g., benzalkonium chloride, Benzatkonium chloride, parabens such as methylparaben or ethylparaben, benzyl alcohol, phenethyl alcohol, sorbic acid, etc.). or its salts, thimerosal, chlorobutanol, etc.), solubilizing aids or particle stabilizers (e.g., cyclodextrins and their derivatives, water-soluble polymers such as polyvinyl pyrrolidone, surfactants such as tyloxapol, polysorbates, poloxamers, etc.), pH adjusters (e.g., hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide, potassium hydroxide, ammonium hydroxide, etc.), thickeners (e.g., HEC, hydroxypropyl cellulose, methyl cellulose, HPMC, carboxymethyl cellulose and their salts), chelating agents (e.g., sodium edetate, sodium citrate, concentrated sodium phosphate, etc.) and second active agent stabilizers (e.g., EDTA, propyl gallate and combinations thereof).

[0243] The ophthalmic suspension of the present application comprises pharmaceutical excipients selected at a concentration equal to or lower than the optimal concentration of the ophthalmic solution. The excipients of the present application include, for example but not limited to, sodium phosphate monohydrate, glycerol and benzalkonium chloride (BAK).

[0244] In some embodiments, the ophthalmic suspension comprises about 0.01 - about 0.5%, about 0.02 - about 0.5%, about 0.04 - about 0.5%, about 0.06 - about 0.5%, about 0.08 - about 0.5%, about 0.08 - about 0.4%, about 0.08 - about 0.3%, about 0.08 - about 0.2%, about 0.08 - about 0.18%, about 0.08 - about 0.16%, about 0.08 - about 0.14%, or about 0.08 - 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.

[0245] In some embodiments, the ophthalmic suspension comprises about 0.001 - about 0.5%, about 0.002 - about 0.5%, about 0.005 - about 0.5%, about 0.01 - about 0.5%, about 0.02 - about 0.5%, about 0.03 - about 0.5%, about 0.04 - about 0.5%, about 0.01 - about 0.4%, about 0.01 - about 0.3%, about 0.01 - about 0.2%, about 0.01 - about 0.1%, about 0.01 - about 0.08%, about 0.01 - 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.

[0246] 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, such as Compound-I, and a second active agent, and may also comprise tromethamine (i.e., Tris). In some embodiments, the ophthalmic suspension comprises about 0.05% - 1% Tris. In some embodiments, the ophthalmic suspension comprises about 0.2% - 1.0% Tris. In some embodiments, the ophthalmic suspension comprises about 0.4% - 0.8% Tris. Suspensions based on tromethamine of Compound-I or its free base and / or a second active agent may comprise additional buffers and excipients, such as, but not limited to, phosphate buffer. The suspension may further comprise one or more surfactants and emulsifiers, such as but not limited to polysorbate 80 or its equivalent excipients; one or more non-ionic liquid polymers of the alkylaryl polyether alcohol type, such as but not limited to tyloxapol; and / or one or more hydrophilic non-ionic surfactants, such as but not limited to poloxamers, such as poloxamer 407.

[0247] The present application provides a suspension of the agent of the present application formulated in the presence of an excipient such as, but not limited to, povidone, polysorbate 80 (PS80), polyethylene glycol (PEG) 400, tyloxapol, poloxamer, glycerol, and BAK / Tris buffer.

[0248] In one embodiment, the suspension of Compound-I or its free base comprises about 0.1-0.5% phosphate buffer. In one embodiment, the suspension of Compound-I or its free base and the second active agent comprises about 0.1-0.5% phosphate buffer. In some embodiments, the pH of the tromethamine-based suspension is between pH 4-7, for example pH 6.0. In some embodiments, the suspension prepared in Tris further comprises 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.

[0249] In some such embodiments, the suspension of the present application further comprises about 0.01-about 1%, or about 1 to about 2.0% w / v glycerol. In a specific embodiment, the suspension comprises about 2% w / v glycerol.

[0250] In some embodiments, the suspension of the present application also comprises about 0.001- about 0.005% w / v benzalkonium chloride (BAK). The amount of BAK can be varied according to any observed side effects. BAK can damage cells on the surface of the eye, and therefore, the amount in the formulation can be varied to achieve the optimal level of eye penetration of Compound-1 without compromising the integrity of the eye cell layer and increasing toxicity.

[0251] In some embodiments, the suspension optionally includes a buffer. When used, the buffer can be, for example, sodium monophosphate alkaline, phosphoric acid and Tris buffer. The compound-I concentration in the suspension is about 0.005% - about 5.0% w / v. The suspension prepared in the absence of additional buffer further includes about 0.005% BAK and about 2% glycerol (pH 6.0). In another embodiment, the suspension prepared in the absence of additional buffer includes about 1% polysorbate 80, about 0.1% tyloxapol, about 0.2% poloxamer 407, about 0.005% BAK, about 2.0% glycerol (pH 6.0).

[0252] In a suspension 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, about 2.0% glycerol (pH 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.

[0253] The suspension prepared in tromethamine (Tris) alone comprises about 1% polysorbate 80, about 0.1% tyloxapol, about 0.2% poloxamer 407, about 0.6% Tris, about 0.005% BAK, and about 2.0% glycerol (pH 6.0). In another embodiment, the 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, about 0.005% BAK (pH 6.0). In these suspensions, 1N HCl and / or 1N NaOH are used for titration to the appropriate pH.

[0254] The suspension of the present application 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, such as 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 dihydrogen phosphate monohydrate and / or about 0.3% to about 1.0% Tris. Alternatively, the suspension of the present application 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, such as 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% - about 0.05%, about 0.05 - about 0.09%, or about 0.09 - about 0.2%. w / v sodium phosphate monobasic monohydrate and / or about 0.3% to about 1.0% Tris.

[0255] In a specific embodiment, the suspension comprises about 0.14% or about 0.2% w / v Tris-buffer. In additional embodiments, the suspension is prepared in 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% glycerol and about 0.005% 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% of the second active agent, about 5% Cremophor RH40, about 2.0% glycerol and about 0.005% BAK.

[0256] In some embodiments, the suspension of the present application has a pH of about 4.0 to about 7.5 at or below about 40°C.

[0257] In some embodiments, the suspension of the present application has a pH of about 5.0 to about 7.0 at or below about 40°C.

[0258] For example, the suspension of the present application has a pH value of about 6.0 at or below about 40°C.

[0259] In some embodiments, the first active agent may be formulated as a solution according to embodiments described herein, and the second active agent may be formulated as a suspension according to embodiments described herein.

[0260] In other embodiments, the first active agent may be formulated as a suspension according to embodiments described herein, and the second active agent may be formulated as a solution according to embodiments described herein.

[0261] Concentrations in various ocular tissues—delivered as an ophthalmic suspension

[0262] In some embodiments, a suspension of Compound-1 or its free base provides similar concentrations of the first active agent at the central choroid and central retina compared to the concentration of the first active agent delivered in gel drop form (discussed below).

[0263] In some embodiments, a Tris-based suspension of Compound-1 or its free base, with or without a second active agent, increases the bioavailability of the first active agent at the central choroid and central retina, while reducing the concentration at the cornea and preventing and / or treating corneal damage and / or disease (e.g., corneal edema, ulcers, abnormalities, etc.). In some embodiments, topical delivery of Compound-1 or its free base formulated in Tris-base reduces the corneal concentration of Compound-1 by about 5-10 times, 10-20 times, 20-30 times, 30-40 times, or about 50-100 times compared to the corneal concentration of an equimolar Compound-1 or its free base delivered as a gel drop.

[0264] The combined effect of reducing corneal drug exposure to avoid poor ocular tolerance while maintaining or increasing posterior segment bioavailability to increase inhibition of receptor tyrosine kinases (RTKs), such as VEGFR, significantly increases the therapeutic index and corresponding benefit to the patient. Additional prevention and / or treatment of damage to the anterior surface of the eye with a second active agent such as an EGFR modulator (e.g., an activator) further improves the therapeutic index and corresponding benefit to the patient.

[0265] About 0.005% to about 5.0% w / v suspension of Compound-I of the present application is well tolerated in the eye when applied once or twice daily for 30-90 days or 4-6 months.

[0266] Gel drops

[0267] 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 dihydrogen phosphate monohydrate to provide the required buffering capacity, and about 0.005% to about 2.0% compound-I and / or about 0.005% to about 5% of a second active agent in a free-flowing filterable formulation without the need for surfactant additives.

[0268] The gel drop formulation of the present application contains about 0.005% to about 2.0% w / v of a first active agent of Formula I or II or a pharmaceutically acceptable salt thereof, such as Compound-I. For topical administration, the concentration of Compound-I or its free base (Formula II) in the gel drop is 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. 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 / v of Compound-I or its free base (Formula II).

[0269] The gel drop formulation of the present application may also include about 0.00001% to about 5.0% w / v of a second active agent or a pharmaceutically acceptable salt thereof, such as niacin, niacinamide, or vitamin K, or a combination thereof. The gel drop formulation of the present application may also include 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 a second active agent or a pharmaceutically acceptable salt thereof, such as niacin, niacinamide, or vitamin K, or a combination thereof. For topical administration, the concentration of the second active agent in the gel drop is about 0.00001% - about 0.0001%, 0.000012% - about 0.0001%, 0.000014% - about 0.0001%, 0.000016% - about 0.0001%, 0.000018% - about 0.0001%, 0.00002% - about 0.0001%, 0.00003% - about 0.0001%, 0.00004% - about 0.0001%, 0.00005% - about 0.0001%, 0.00006% - about 0.0001%, 0.00007% - about 0.0001%, 0.00008% - about 0.0001%, 0.00009% - About 0.0001%, 0.000016% - about 0.00009%, 0.000018% - about 0.00009%, 0.00002% - about 0.00009%, 0.00003% - about 0.00009%, 0.00004% - about 0.00009%, 0.00005% - about 0.00009%, 0.00006% - about 0.00009%, 0.00007% - about 0.00009%, or 0.00008% - about 0.00009% w / v. In some embodiments, the gel drops comprise 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.

[0270] The gel drop formulation of the present application may also include 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 acceptable salt thereof, such as niacin, niacinamide or vitamin K or a combination thereof. In some embodiments, the concentration of the second active agent is about 1 μM.

[0271] In some embodiments, the gel drop ophthalmic composition of the application includes glycerol as a tonicity agent. Some embodiments of the application provide an ophthalmic composition comprising mannitol. The amount of any change in the solubility of Compound-I and the glycerol or mannitol content of the level of about 2.0-about 2.5% glycerol provide an osmotic concentration of about 225-about 300 mOsm / kg, depending on the phosphate concentration. In additional embodiments, glycerol is about 2%, and phosphate is about 0.05% of the gel drop ophthalmic composition. The concentration of glycerol and phosphate of the application is that the tonicity level of the ophthalmic composition is about 240mOsm / kg.

[0272] The gel drop ophthalmic composition of the present application may also include benzalkonium chloride (BAK). In some embodiments, the BAK content is about 0.005%, which is sufficient to protect the ophthalmic composition from microbial contamination. In some embodiments of the present application, BAK is not required for the use of the ophthalmic composition in a sterile, disposable product.

[0273] In some embodiments, the gel drop ophthalmic formulation of Compound-I comprises: about 0.005% to about 2.0% Compound-I or its free base, about 0.05% sodium phosphate, about 2% glycerol as a tonicity adjuster, about 0.005% BAK as a preservative, water (purified, i.e., distilled or deionized) as a vehicle, and sodium hydroxide to adjust the pH to 6.0. In one embodiment, no other excipients are added.

[0274] The gel drops of the present application include about 0.005% - about 2.0% of an active agent of Formula I or II or a pharmaceutically acceptable salt thereof, such as Compound-I, and about 0.01% - about 0.05%, about 0.05- about 0.09% or about 0.09- about 0.2% w / v sodium dihydrogen phosphate monohydrate. In a specific embodiment, the gel drops include about 0.05%, about 0.05-0.2% or about 0.2% w / v sodium dihydrogen phosphate monohydrate buffer. Other equivalent buffer systems well known in the art are also used in the gel drops of the present application. In one embodiment, Compound-I or its free base is formulated as about 0.4% to about 2.0% of the first active agent, about 5% Cremophor RH40, about 2.0% glycerol and about 0.005% BAK.

[0275] In one embodiment, the gel drops of Compound-I comprise about 0.3% to about 2.0% (3-20 mg / mL) Compound-I, about 0.05% to about 0.2% sodium phosphate and about 2% glycerol. The pH of the composition is between pH 5.0-7.0.

[0276] The gel drop ophthalmic formulation of compound-1 and a second active agent includes: about 0.005% to about 2.0% compound-1 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%) of the second active agent, about 0.05% sodium phosphate, about 2% glycerol as a tonicity adjuster, about 0.005% BAK as a preservative, water (purified, i.e., distilled or deionized) as a vehicle, and sodium hydroxide to adjust the pH to 6.0. In one embodiment, no other excipients are added.

[0277] The gel drops of the present application include about 0.005% - about 2.0% of an active agent of Formula I or II or a pharmaceutically acceptable salt thereof, such as Compound-I, about 0.00001% - about 5% (e.g., about 0.00001% to about 0.0002% or about 0.00001% to about 0.0001%) of a second active agent, and about 0.01% - about 0.05%, about 0.05- about 0.09% or about 0.09- about 0.2% w / v sodium dihydrogen phosphate monohydrate. In a specific embodiment, the gel drops include about 0.05%, about 0.05-0.2% or about 0.2% w / v sodium dihydrogen phosphate monohydrate buffer. Other equivalent buffer systems well known in the art are also used in the gel drops of the present application. In one embodiment, Compound-I or its free base is formulated at 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.

[0278] In one embodiment, the gel drops of Compound-I comprise about 0.3% to about 2.0% (3-20 mg / mL) Compound-I, about 0.005% to about 5.0% of a second active agent, about 0.05% to about 0.2% sodium phosphate, and about 2% glycerol. The pH of the composition is between pH 5.0-7.0.

[0279] The present application provides gel drops of the agents of the present application (e.g., a first active agent and a second active agent) formulated in the presence of an excipient, such as, but not limited to, examples povidone, polysorbate 80 (PS80), polyethylene glycol (PEG) 400, tyloxapol, poloxamer, glycerol, and BAK / phosphate buffer.

[0280] Eye drops

[0281] Disclosed herein is a formulation comprising a first active agent, e.g., a compound of formula I or II, and / or a second active agent, e.g., nicotinic acid, nicotinamide, or vitamin K, or a combination thereof, in a form of pharmaceutically acceptable, convenient, safe, and effective drug delivery to a patient for a few minutes as an eye drop. Standard eye drops used in therapy according to U.S. federal regulations are sterile, have a pH of about 6.0-7.4, and contain a preservative if used more than once, but have a limited shelf life after opening, typically one month. If the eye drops are packaged in a sterile, disposable unit dose dispenser, the preservative can be omitted.

[0282] A method of eye drop preparation includes the purest form (e.g., greater than 99% purity) of a disclosed compound of formula I or II and / or the purest form (e.g., greater than 99% purity) of a second active agent, and the compound and / or the second active agent are mixed with a buffer and a tonicity regulator for adjusting physiological pH and molar osmotic pressure concentration. Examples of buffers for maintaining or adjusting pH include, but are not limited to, acetate buffer, citrate buffer, phosphate buffer, and borate buffer. Examples of tonicity regulators are sodium chloride, mannitol, and glycerol. In some embodiments, other pharmaceutically acceptable ingredients are also added.

[0283] The formulated solution is then divided equally into a plurality of discrete, sterile disposable boxes each suitable for unit dose administration, or a single medicine box for unit dose administration. This single disposable box is a specific volume dispenser such as a cone or cylinder, which has a container having a side wall that can be squeezed in the radial direction of the longitudinal axis so as to dispense the container contents at one end of the container.

[0284] The application provides eye drops solution / suspension packaged in multiple doses or single doses, for example as a plastic bottle with an eye dropper. In multiple doses, a preservative is needed to prevent microbial contamination after opening the container. Suitable preservatives include, but are not limited to, benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, phenylethyl alcohol, disodium edetate, sorbic acid, polyquaternium-1 or other agents known to those skilled in the art, all of which are intended to be used in the application. This type of preservative is usually used at a level of 0.001 to about 1.0% weight / volume.

[0285] Without wishing to be bound by theory, the formulation of the present application in an eye drop provides pulsatile access to the drug. The route by which Compound-1 gains access to the posterior segment is not by direct diffusion through the cornea, followed by diffusion through the aqueous humor, vitreous humor, retina, and ultimately the choroid. Instead, the Compound-1 compound achieves significant bioavailability in the posterior segment following topical instillation using a circumferential approach around the sphere, rather than through the sphere.

[0286] Under certain clinical conditions, eye drop solutions / suspensions may be formulated with other agents to reduce the irritation of other ingredients and promote clinical response. Such agents include, but are not limited to, vasoconstrictors such as phenylephrine, oxymetazoline, naphtholine, or tetrahydrozoline; mast cell stabilizers such as olopatadine; antihistamines such as azelastine; antibiotics such as tetracycline; steroidal anti-inflammatory drugs such as betamethasone; nonsteroidal anti-inflammatory drugs such as diclofenac; immunomodulators such as imiquimod or interferon; and antiviral agents such as valacyclovir, cidofovir, and trifluridine. The dosage for the above purposes varies, but is an amount that effectively suppresses discomfort, itching, irritation, or pain in the eye. When the composition is administered topically, a "pharmaceutically effective amount" of the compound of Formula I or II can generally be in a concentration range of 0.05 mg / mL to about 10 mg / mL, and a "pharmaceutically effective amount" of the second agent can generally be in a concentration range of 0.05 mg / mL to about 10 mg / mL, wherein 1 to 4 drops of the composition are administered 1 to 4 times per day as a unit dose. The most common method of ocular drug delivery is to instill the drops into the cornea (i.e., "eye drops").

[0287] The key requirement is that the formulation is sterile and produced in a sterile environment. An ideal disclosed compound for an ophthalmic solution / suspension should be soluble and / or miscible in an aqueous medium at normal eye pH and tonicity. In addition, the disclosed compound should be stable, non-toxic, long-acting, and sufficiently effective to resist dilution of the drug concentration by blinking and tearing.

[0288] Dosage form

[0289] The preparation of the present application can be suitable for ophthalmic applications. In one embodiment, the preparation is a solution. The solution of the present application can 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 about 6.0 and can be stored at + 5 ° C. The drug product can be provided in a container closure system consisting of a translucent ophthalmic dispenser bottle with a dropper tip and a cap.

[0290] In some embodiments, the clinical concentration of the 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, about 0.2-about 1.0 mg / mL, about 0.3-about 1.0 mg / mL, about 0.4-about 1.0 mg / mL, about 0.5-about 1.0 mg / mL, about 0.6-about 1.0 mg / mL, about 0.7-about 1.0 mg / mL, about 0.8 – about 1.0 mg / mL, about 0.9-about 1.0 mg / mL, about 1.0-about 2.0 mg / mL, about 2.0-about 3.0 mg / mL, about 3.0-about 4.0 mg / mL, about 4.0-about 5.0 mg / mL, about 5.0-about 6.0 mg / mL, about 5.0-about 10.0 mg / mL, about 10-about 20 mg / mL, about 20-about 30 mg / mL, about 30 to about 40 mg / mL, or about 40 to about 50 mg / mL.

[0291] In other embodiments, the clinical concentration of the Compound-1 and the second active agent ophthalmic solution or suspension is independently equal to or less than about 0.1 mg / mL, equal to or less than about 0.2 mg / mL, about 0.2-about 1.0 mg / mL, about 0.3-about 1.0 mg / mL, about 0.4-about 1.0 mg / mL, about 0.5-about 1.0 mg / mL, about 0.6-about 1.0 mg / mL, about 0.7-about 1.0 mg / mL, about 0.8 – about 1.0 mg / mL, about 0.9-about 1.0 mg / mL, about 1.0-about 2.0 mg / mL, about 2.0-about 3.0 mg / mL, about 3.0-about 4.0 mg / mL, about 4.0-about 5.0 mg / mL, about 5.0-about 6.0 mg / mL, about 5.0-about 10.0 mg / mL, about 10-about 20 mg / mL, about 20-about 30 mg / mL, about 30 to about 40 mg / mL, or about 40 to about 50 mg / mL.

[0292] In one embodiment of the present application, the strength of Formula I or II compound is about 0.005% - about 5.0% (about 0.5- about 50 mg / mL). After ocular administration 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 preparation of the present application is realized for pathological choroid and retinal neovascularization desired pharmacological activity (or concentration). The application provides that after topical ocular administration with optimal dose (e.g., about 0.005% to about 5.0%), pharmacological activity concentration is achieved and maintained in central choroid target tissue. In one embodiment, the first active agent (Formula II or Compound-I) is formulated to a concentration of about 0.005-about 5.0% w / v, the second active agent (niacin, niacinamide or vitamin K or a combination thereof) is formulated to a concentration of about 0.005-about 5.0% w / v, and the combination is administered once or twice per day to each eye for more than 60 consecutive days. The plasma concentrations observed after topical administration are substantially below the levels expected to produce systemic toxicity.

[0293] Table 2: In vitro summary of the pharmacodynamic properties of Compound-I

[0294] In vitro assay <![CDATA[IC 50 = nM (ng / mL)]]> Inhibition of recombinant VEGFR-2 tyrosine kinase using exogenous substrates 10.55 (6) Inhibition of recombinant FGFR-2 tyrosine kinase using exogenous substrates 8.79 (5) Inhibition of recombinant PDGFR tyrosine kinase using exogenous substrates 2636.67 (1500) Inhibition of recombinant EGFR tyrosine kinase using exogenous substrates 5853.40 (3330) Inhibition of recombinant IR tyrosine kinase using exogenous substrates 10283.00 (5850) Inhibition of VEGF-stimulated VEGFR-2 autophosphorylation in intact cells 5.27 (3) Inhibition of VEGF-stimulated mitogenesis in HUVECs 14.06 (8)

[0295] In some embodiments, Compound-I exhibits potent inhibition of the tyrosine kinase activity of several pro-angiogenic growth factor receptors, wherein IC 50 Less than about 100 nM (see Table 3). Compound-I also blocks high affinity VEGF receptors, such as VEGFR-1 / Flt-1, but with lower potency (where IC 50 is about 122nM (69.41 ng / mL).

[0296] Table 3: In vitro inhibition of tyrosine kinases using a 10-point titration curve (257 nM-5000 nM) of Compound-I

[0297] Kinase <![CDATA[IC of Compound-I 50 = nM (ng / mL)]]> AURKB (Aurora B) 207 (117.76) FGFR-1 8.50 (4.84) FGFR-2 3.08 (1.75) FGFR-3 33.9 (19.29) FGFR4 500 (284.45) FLT1 (VEGFR-1) 122 (69.41) FLT3 419 (238.37) FLT4 (VEGFR-3) 54.2 (30.83) FYN 161 (91.59) KDR (VEGFR-2) 1.27 (0.72) PDGFRA (PDGFRα) 3120 (1774.97) PDGFRB (PDGFRβ) 1860 (1058.16) TEK (Tie2) 10.1 (5.75) RET 11.1 (6.31)

[0298] Although VEGFR inhibition appears to be essential for reducing vascular permeability and further preventing neovascularization, simultaneous inhibition of VEGF signaling and inhibition of other growth factor signaling pathways (e.g., PDGF and angiopoietin / Tie2) may be associated with unique therapeutic outcomes. The therapeutic outcome of more extensive inhibition of signaling pathways may contribute to the regression of newly established pathological blood vessels in the posterior segment of the eye.

[0299] In some embodiments, about 300nM (about 170.67 ng / mL) of Compound-I inhibits VEGFR-2 kinase function (see Table 4). A similar set of pro-angiogenic growth factor receptors (including FGFRs1-3, Tie-2, and EphB-4) was also observed to block substantially. An unexpected finding is that about 300nM concentrations of Compound-I inhibit VEGFR-2 kinase function, which falls within the typical range found in the central choroid and retina after five days of topical ocular delivery.

[0300] Table 4: In vitro inhibition of tyrosine kinases by 300 nM (170.67 ng / mL) of Compound-I.

[0301] Kinase Average % inhibition at 300 nM Compound-I EPHB-4 87 FGFR-1 96 FGFR-2 103 FGFR-3 (K650E variant) 104 FLT4 (VEGFR-3) 86 KDR (VEGFR-2) 104 RET 98 RET (Y791F mutation) 97 TEK (Tie2) 96

[0302] Table 5: In vitro inhibition of tyrosine kinases by 1 µM (568.9 ng / mL) of Compound-I.

[0303] Kinase Average % inhibition at 1 μM Compound-I ABL1 92 ABL1 E255K 90 ABL1 G250E 89 ABL1 T315I 101 ABL1 Y253F 93 ACVR1B (ALK4) 98 AURKB (Aurora B) 82 BRAF V599E 85 EPHA-1 81 EPHA-8 85 EPHB-1 83 EPHB-4 80 FGFR-1 98 FGFR-2 99 FGFR-3 96 FGFR-3 K650E 100 FGR 91 FLT-1 (VEGFR-1) 86 FLT-4 (VEGFR-3) 95 KDR (VEGFR-2) 98 LCK 97 LYN A 81 LYN B 91 MAP4K4 (HGK) 100 MAP4K5 (KHS1) 94 MAPK14 (p38α) 86 MINK1 100 PDGFRA T674I 86 PTK6 (Brk) 88 RET 98 RET Y791F 94 SNF1LK2 82 SRC 91 SRC N1 83 TEK (Tie2) 99 YES1 98

[0304] Overview of APIs and Drug Products

[0305] Drug product: Compound-I and / or a second active agent ophthalmic preparation for clinical studies is manufactured with a dosage strength of 0.05% - 1.0% of Compound-I and about 0.00001% - about 5% (e.g., about 0.00001% - about 0.0002%, or about 0.00001% - about 0.0001%) of the second active agent. In some embodiments, the dose of Compound-I in the preparation is 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.8% or 1.0%. In some embodiments, the dose of the second active agent in the preparation 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%. In a clinical setting, Compound-I and / or a second active agent ophthalmic preparation (solution or suspension) is used for daily, single use, topical application to the eye. In addition to the first and second active ingredients, in some embodiments, the drug product may also contain about 0.005% BAK as a preservative, purified water as a vehicle, and adjusted to pH 6.0 with sodium hydroxide.

[0306] Sodium phosphate based gel drops

[0307] The ophthalmic effect of Compound-1 in sodium phosphate-based formulations (listed in Table 6) is derived from the self-gelling properties of the API in a buffer such as sodium phosphate. The self-forming thixotropic gel of Compound-1 that spontaneously forms 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 supersaturated state, insoluble particles of Compound-1 are also observed within the gel.

[0308] The current state of the art predicts that applying a gel with increased viscosity to the surface of the eye will increase corneal residence time. Increasing corneal residence time is thus beneficial to the absorption of ophthalmic drugs. As a result, the intraocular drug concentration of the viscous gel will increase compared to non-viscous formulations such as aqueous solutions. One method of increasing viscosity is to use various viscosity-increasing excipients, such as carboxymethyl cellulose, which actually achieves increased intraocular absorption of different raw materials after topical ocular administration. The application provides a thixotropic gel of compound-1 and / or a second active agent formed in the absence of any viscosity-increasing excipient. For example, when compound-1 or compound-1 and a second active agent are dissolved in a simple buffer such as sodium phosphate, a thixotropic gel is formed. The thixotropic gel formed in the absence of any viscosity-increasing excipient is formulated as a gel drop.

[0309] The present application provides dose-dependent and dose-frequency-dependent delivery of Compound-I to posterior segment tissues.

[0310] The gel drop formulations of the present application (listed in Table 6) differ from each other in several aspects, such as first active agent concentration, sodium phosphate concentration, presence or absence of tonicity (glycerol) or preservatives (benzalkonium chloride / BAK), solubilizing surfactants (polysorbate 80, tyloxapol and / or poloxamer), and pH.

[0311] Tromethamine-based suspensions

[0312] The present application provides a suspension of compound-1 and / or a second active agent in a tromethamine-based formulation. In some embodiments, the suspension of compound-1 and / or a second active agent in a tromethamine-based formulation has an insoluble form of the first active raw material drug equal to or greater than 95%. This feature is distinguished from the soluble or semi-soluble state of compound-1 in gel drops (when the concentration of the first active agent increases, the gel drops (gel) are not completely soluble) or cyclodextrin-based formulations. The tromethamine-based formulation of compound-1 shows turbidity that increases with increasing concentration of the first active agent. Topical drops of compound-1 and / or a second active agent suspension (which is a combination of soluble and insoluble first active agent components) are beneficial in terms of safety / tolerance and efficacy when applied to the eye.

[0313] The present application provides compound-I in a tromethamine-based suspension to detect cellular ICs of various pro-angiogenic RTKs. 50 See, e.g., Table 7.

[0314] The corneal safety and tolerability of topical compound-1 is a direct result of the amount of the soluble (relative to insoluble) first active agent applied to the corneal surface and the resulting corneal tissue concentration. In some embodiments, subjects receiving topical ocular administration of a tromethamine-based suspension are able to tolerate a higher level of the first active agent concentration in the formulation compared to an equimolar formulation of sodium phosphate-based gel drops. The corneal safety and tolerability of topical compound-1 is also a result of administering 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 damage or disease caused by inhibition of EGFR.

[0315]

[0316]

[0317]

[0318] The present application provides the ocular bioavailability of compound-I in the posterior segment after applying a tromethamine-based suspension. The ocular bioavailability of compound-I in the posterior segment is proportional to the total amount of drug compound-I applied (insoluble plus soluble, see Table 7). Although insoluble drug particles are not easily utilized by anterior segment tissues; the inherent and unique physicochemical properties of compound-I allow insoluble and soluble components to enter posterior segment tissues, such as choroid and retina. Therefore, a tromethamine-based suspension is used to achieve a drug concentration that is even higher than that achieved by a gel drop formulation containing an equal amount of the first active agent. Therefore, a tromethamine-based suspension provides: a) improved corneal tolerance and b) maintaining or increasing the bioavailability of the posterior segment, particularly the choroid, the main target tissue for treating neovascularization (wet) AMD. In addition, a second active agent (e.g., niacin, niacinamide or vitamin K or a combination thereof) prevents and / or treats corneal damage potentially associated with the administration of Compound-I, thereby increasing the therapeutic index of Compound-I, wherein the second active agent is a modulator (e.g., activator) of EGFR that prevents or treats corneal damage or disease caused by inhibition of EGFR.

[0319] Cyclodextrin-based solutions

[0320] Cyclodextrins, which are cyclic oligosaccharides composed of six to eight glucose units (α-, β- and γ-CD) joined by one to four bonds, are well known for their ability to act as solubilizers for relatively poorly soluble drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol. , 36: 30-42 (2008).

[0321] In some embodiments, the proposed clinical formulation, 2-hydroxypropyl-β-cyclodextrin (HP-β-CD, also known as KLEPTOSE® HPB) at a molar ratio of equal to or greater than 1:6 or sulfobutyl ether-β-cyclodextrin (SBE-β-CD, also known as CAPTISOL®) at a ratio of equal to or greater than 1:2 in the free base of Compound-1 or the second active agent ophthalmic solution provides a solubility that meets the clinical dosage strength of 0.1-1.2% Compound-1.

[0322] In some embodiments, the cyclodextrin-based solution of compound-1 or its free base and / or the second active agent not only has improved solubility of the first active agent into a homogeneous solution, but, after topical ocular administration, also has a novel and previously unobserved characteristic of a significantly increased therapeutic index of the first active agent at the posterior segment of the eye. The solution of compound-1 and / or the second active agent of the present application reduces the anterior segment exposure of compound-1, thereby increasing the concentration of the first active agent in the solution and increasing the frequency of its delivery to maintain a high posterior segment concentration. Both of these beneficial features are related to the known properties of cyclodextrins 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 damage or disease caused by systemic disease (e.g., cancer, diabetes), eye disease, or inhibition of EGFR by 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., a compound of Formula I or II).

[0323] When formulated with Compound-1 or its free base and / or a second active agent, cyclodextrin can form a clear, colorless solution that exhibits a water-like viscosity. After topical ocular administration, the Compound-1 / cyclodextrin complex has an appearance as pharmacologically inactive and metabolically inert. The Compound-1 / cyclodextrin complex imparts corneal tolerance until the cyclodextrin spontaneously dissociates from the first active agent, thereby achieving high concentrations of Compound-1 at its intended site of action in the posterior segment of the eye (e.g., choroid and retina).

[0324] In some embodiments, compared with a gel drop formulation of similar drug concentration, a cyclodextrin-based solution of compound-I reduces the corneal exposure of compound-I. Compared with the administration of an equimolar formulation of gel drops, the use of a cyclodextrin-based solution of compound-I provides about a 10-fold reduction in corneal concentration. In some embodiments, after 20-30 days of topical ocular administration of about 0.2-2.0%, for example 0.6% compound-I as a cyclodextrin-based solution, no adverse results are attributed to the test article or vehicle. The present application provides a higher concentration of compound-I in the posterior segment target tissue, such as at the central choroid and central retina when a cyclodextrin-based solution of compound-I is topically applied. In some embodiments, reducing corneal drug exposure to avoid poor eye tolerance and increasing the bioavailability of the posterior segment to increase the combined effect of RTK inhibition significantly increases the therapeutic index and the corresponding benefits to the treated patients. In other embodiments, the combined effect of reducing corneal drug exposure via a second active agent to avoid poor ocular tolerance and prevent or treat corneal damage or disease, while increasing posterior segment bioavailability to increase RTK inhibition can significantly increase the therapeutic index and corresponding benefit to treated patients.

[0325] The present application provides a suspension-based formulation (see Example 3) and a cyclodextrin formulation of Compound-1 that extend the therapeutic window due to significantly reduced exposure (reduced by about 10-100 times or 1-2 log). Reduced exposure improves corneal safety / tolerance, which allows higher concentrations or frequency of dosing of Compound-1 to be topically applied. Higher concentrations enable Compound-1 to reach higher target tissue concentrations at the back of the eye, which can improve the therapeutic efficacy of Compound-1.

[0326] In some embodiments, topical ocular administration of ophthalmic gel drops is associated with high corneal tissue exposure (≥100uM) and corresponding adverse observations in the anterior segment, such as discomfort, corneal and conjunctival inflammation, corneal epithelial erosion and / or thinning and degeneration. In contrast, repeated topical ocular administration of compound-I ophthalmic solution produces corneal exposure of 1 / 5 to 1 / 10 of the ophthalmic gel drops of equimolar doses, and there are no adverse clinical or histopathological results. Compared with equimolar doses of ophthalmic gel drops, topical ocular administration of compound-I ophthalmic solution also achieves the same or higher target therapeutic exposure in the central choroid. In summary, reducing corneal exposure and correspondingly improving ocular tolerance, while maintaining or promoting drug delivery to the target tissue of the posterior segment, together with the combination of improved physicochemical stability, provides greater benefits to the subject than ophthalmic gel drops formulations.

[0327] 1- to 5-day PK results with topical ocular Compound-I in a cyclodextrin-based solution

[0328] The present application provides ocular pharmacokinetics of various formulations and dosage regimens of Compound-I after topical ocular administration. Three dosage strengths of nine (9) different topical ocular formulations of Compound-I were used for administration once a day (qd) or twice a day (bid) for 1, 2, 3, 4, or 5 consecutive days. Subjects each received approximately 30 μL bilateral topical ocular doses of one of the three (3) Compound-I formulations or the vehicle formulation using a positive displacement pipette.

[0329] 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 (qd) for one (1) or four (4) days. On Days 1 to 4, Groups 3, 7, 9, 12, 14, and 16 received bid dosing at 7:00 AM and 3:00 PM, approximately 8 hours apart, for four (4) days. Some subjects received bid dosing of vehicle-only formulations for five (5) consecutive days.

[0330] In some embodiments, eye sampling is performed at about 0.5, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 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 after the dose relative to the day 1 dose. Aqueous humor, corneal, central and peripheral retinal, and central and peripheral choroidal samples are collected to monitor the effect of the treatment. Aqueous humor, corneal, central retinal, and central choroidal samples are assayed.

[0331] Tables 8A-C list the 1 to 5 day PK results with topical ocular Compound-I in a cyclodextrin based solution.

[0332] Table 8A: Ophthalmic Formulations

[0333]

[0334] Table 8B lists the mean Compound-I concentrations in aqueous humor, retina, choroid, and cornea (LLOQ: lower limit of quantitation; LLOQ is the lowest analyte concentration that can be quantified with acceptable precision and accuracy).

[0335] Table 8B

[0336]

[0337] AH LLOQ = 0.000903 μM

[0338] Central retina LLOQ = 0.0181 μM

[0339] Peripheral retina LLOQ = 0.00873 μM (Groups 1 - 8); LLOQ = 0.00898 μM (Groups 12 - 16)

[0340] Central choroid LLOQ = 0.175 μM

[0341] Peripheral choroid LLOQ = 0.0349 μM (Groups 1 - 8); LLOQ = 0.0359 μM (Groups 12 - 16)

[0342] Cornea LLOQ = 0.0181 μM (Groups 1 - 5); LLOQ = 0.0453 μM (Groups 6 - 8, 10 - 13, 15, 16A17); LLOQ = 0.0873 μM (Groups 4, 9, 16B)

[0343] N / A = Not applicable; samples not measured according to the study protocol.

[0344] *Based on the average of n = 1.

[0345] Table 8C presents an overview of the 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 the statistical calculations. When all values at a given time point were <LLOQ, <LLOQ was reported as the mean.

[0346] Table 8C

[0347]

[0348] AH LLOQ = 0.000903 μM

[0349] Central retina LLOQ = 0.0181 μM

[0350] Peripheral retina LLOQ = 0.00873 μM (Groups 1 - 8); LLOQ = 0.00898 μM (Groups 12 - 16)

[0351] Central choroid LLOQ = 0.175 μM

[0352] Peripheral choroid LLOQ = 0.0349 μM (Groups 1 - 8); LLOQ = 0.0359 μM (Groups 12 - 16)

[0353] Corneal LLOQ = 0.0181 μM (Groups 1 - 5); LLOQ = 0.0453 μM (Groups 6 - 8, 10 - 13, 15, 16A17); LLOQ = 0.0873 μM (Groups 4, 9, 16B)

[0354] N / A = Not applicable; samples not measured according to the study protocol.

[0355] *Based on the mean of n = 1.

[0356] Five-day PK results with topical ocular Compound-I in a cyclodextrin-based solution

[0357] This application provides the ocular pharmacokinetics of various dosing regimens of a topical ophthalmic solution of Compound - I containing hydroxypropyl - β - cyclodextrin ("HDβCD") after ocular dose administration. Different topical ophthalmic solutions of Compound - I were administered once daily (q.d.) or twice daily (b.i.d.) for 4 or 5 consecutive days. Each subject received a 30 μL bilateral topical ophthalmic dose of one of four Compound - I dose strengths.

[0358] All doses were administered within ±1 hour of the scheduled dose time, except for some subjects who received on Day 1. For subjects, ocular sampling was performed 1 hour after the administration of Compound - I, after the first daily dose on Day 5, except for some subjects, in which ocular sampling was performed 24 hours after the first daily dose on Day 4.

[0359] Aqueous humor, corneal, central and peripheral retina, and central and peripheral choroid samples were collected. Corneal, central retina, and central choroid samples were assayed; aqueous humor, peripheral retina, and peripheral choroid samples were not assayed.

[0360] Table 9 (A - B) presents the 5 - day PK results for topical ocular Compound - I in cyclodextrin - based solutions.

[0361] Table 9A: Ophthalmic Formulations

[0362]

[0363]

[0364] Table 9B presents an overview of the mean ocular tissue concentrations of Compound - I in the central retina, central choroid, and cornea. Any values <LLOQ were excluded from the statistical calculations. When all values at a given time point were <LLOQ, <LLOQ was reported as the mean.

[0365] Table 9B: Mean Compound - I Concentrations in the Retina, Choroid, and Cornea

[0366]

[0367] Concentration of Compound-I in various intraocular fluids and tissues (in μM)

[0368] In some embodiments, after topical ocular administration of a solution of about 0.4% (about 4 mg / mL) of Compound-1 and cyclodextrin, the concentration of the first active agent in various tissues and fluids of the eye is measured. The average concentration of Compound-1 is measured in the central choroid, central retina, aqueous humor, and cornea. Compound-1 is in a solution (0.4% or 4 mg / mL) containing 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 Tables 10A-B.

[0369] In some embodiments, after topical ocular administration of a solution of about 0.4% (about 4 mg / mL) Compound-I and cyclodextrin, the central choroidal concentration of Compound-I is about 0.2 μM to about 0.8 μM. The central retinal concentration of Compound-I is about 0.05 μM to about 0.15 μM. In some embodiments, after topical ocular administration of a solution of about 0.4% (about 4 mg / mL) Compound-I and cyclodextrin, the aqueous humor concentration of Compound-I is 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 a solution of Compound-I that is topically applied to the eye.

[0370] In some embodiments, the average Compound-I ocular tissue concentration following twice daily topical administration of a 0.3% Compound-I ophthalmic gel drop formulation with and without benzalkonium chloride is highest in the cornea, with about 200 μM to about 350 μM in the cornea, about 2.0 μM to about 5.0 μM in the peripheral choroid, about 0.2 μM to about 0.7 μM in the central choroid, about 0.05 μM to about 0.5 μM in the peripheral retina, and about 0.01 μM to about 0.05 μM in the aqueous humor.

[0371] In some embodiments, the Tris-based suspension formulation of Compound-I is well tolerated, without any corneal findings, and only a few occasional episodes of mild conjunctivitis. In some embodiments, for twice daily topical administration of a Tris-based suspension of 0.3% Compound-I with or without benzalkonium chloride, the average Compound-I ocular tissue concentration evaluated at 1 hour ± 15 minutes after the first daily topical ocular dose on day 30 is highest in the cornea, for example, about 2.00 μM to about 4.0 μM. The peripheral choroidal concentration from the same dose is about 0.7 μM to about 1.5 μM; the central choroidal concentration is about 0.3 μM to about 0.4 μM; the peripheral retinal concentration is about 0.08 μM to about 0.09 μM; the central retinal concentration is about 0.04 μM to about 0.07 μM; and the aqueous humor concentration is about 0.001 μM to about 0.002 μM.

[0372] The present application provides cyclodextrin-based solutions of Compound-I (e.g., solutions containing hydroxypropyl-β-cyclodextrin (HP-β-CD, KLEPTOSE® HPB)) that are well tolerated when about 0.1% Compound-I (in a solution with about 2.0% to about 2.5% HP-β-CD) is topically applied twice a day in a subject for up to 30 days, about 0.2% Compound-I (in a solution with about 4.0% to about 4.5% HP-β-CD) is topically applied twice a day, about 0.4% Compound-I (in a solution with about 8.0% to about 8.5% HP-β-CD) is topically applied once or twice a day, and about 0.6% Compound-I (in a solution with up to about 14% HP-β-CD) is topically applied once or twice a day. Furthermore, in additional embodiments, about 0.4% w / v Compound-1 in Kleptose® HPB, Kleptose® HP, or Captisol® cyclodextrin-based solutions were well tolerated when administered twice daily for up to 24 days.

[0373] The present application provides dose-limited corneal toxicity observed with compound-I ophthalmic gel drop formulations. In some embodiments, ophthalmic gel drops make the corneal concentration of compound-I about five times to about fifteen times that of a cyclodextrin-based solution, and the corneal concentration of compound-I is about fifty times to about one hundred times that of a Tris-based suspension. The Tris-based suspension and cyclodextrin-based solution of the compound-I of the present application are well tolerated, with no evidence of obvious ocular toxicity. In some embodiments, about 0.005% to about 5.0% w / v of a cyclodextrin-based solution of compound-I or a Tris-based suspension of at least 30 days of daily administration once or twice is well tolerated in the subject. The present application provides the highest central choroidal concentration of compound-I using a cyclodextrin-based solution compared to an equimolar dose of gel and / or a Tris-based formulation.

[0374] Table 10A: Average concentrations of Compound-I in various intraocular fluids and tissues (in μM)

[0375]

[0376] Table 10B: Study Design

[0377]

[0378] Table 11 shows the corneal and central choroidal concentrations of Compound-I formulations.

[0379] Table 11:

[0380]

[0381] Phase I Protocol for a Dose Escalation Study in Patients with Neovascular AMD

[0382] The application provides a Phase I study involving 12 weeks, open label, dose escalation, multicenter trials to assess safety, tolerability and pharmacokinetics after topical ocular administration of compound-I in patients with neovascular age-related macular degeneration (AMD). Up to a total of 60 patients are treated 1 to 2 times a day with topical ocular administration of compound-I ophthalmic solution for three months, wherein three dose-escalating monotherapy groups and one adjuvant therapy group are planned, which use Lucentis® plus a single intravitreal injection of the maximum tolerated monotherapy dose (15 patients per treatment group). Patients who meet the pre-specified vision and CNV damage criteria confirmed by an independent reading center are allowed to stop topical ocular administration and receive standard of care treatment at the same time.

[0383] The present application provides 3 dosage strengths ranging from 0.1% to 1.0% (w / v) (as Compound-I) ophthalmic solution for clinical studies. The strengths are about 0.1%, about 0.3%, about 0.6%, and about 1.0% (w / v) Compound-I HCl.

[0384] Preparation

[0385] Non-limiting examples of formulations of the present application are summarized in Table 12.

[0386] Table 12: Overview of product compositions tested in the product screening study

[0387]

[0388] a Molar ratio of Compound-I:cyclodextrin.

[0389] b KLEPTOSE® HPB

[0390] c CAPTISOL®.

[0391] EGFR tyrosine phosphorylation assay in cells to determine the EGFR activity of compounds of Formula I or II

[0392] An EGFR tyrosine assay in corneal epithelial cells was run to determine whether higher concentrations of EGF could overcome inhibition of EGFR kinase activity. Cells were serum starved and then pretreated with various concentrations of a compound of Formula I or II (e.g., Compound-I or a control) and subsequently treated with EGF. Cells were then harvested and immunoblotted to determine phosphorylated EGFR and total EGFR concentrations, which were used to determine receptor activity (IC 50 ).

[0393] EGFR tyrosine phosphorylation assay in cells to determine the combination of a compound of Formula I or II and a second active agent EGFR activity

[0394] The EGFR tyrosine assay in corneal epithelial cells was run to determine whether vitamin K or niacin / nicotinamide can overcome the inhibition of EGFR and whether EGFR activity is increased. Cells were serum starved and then pretreated with various concentrations of a compound of Formula I or II (e.g., Compound-I) and saturating concentrations of vitamin K or niacin / nicotinamide, followed by treatment with EGF. Cells were harvested and immunoblotted to determine phosphorylated EGFR (tyrosine 1068 and tyrosine 1045) and total EGFR concentrations, which were used to determine receptor activity (IC 50 ).

[0395] Determination of the effects of various concentrations of compounds of formula I or II and EGF on cell migration / proliferation in cells (in vitro Wound healing)

[0396] The assay of cell migration / proliferation in corneal epithelial cells was run to determine cell migration / proliferation in the presence of various concentrations of Formula I or II compounds and EGF. The cells were plated with silicone plugs. The cells were then serum starved and pretreated with various concentrations of Formula I or II compounds or controls. The silicone plugs were then removed to create a cell-free area and the cells were treated with EGF. Cell migration was quantified from micrographs.

[0397] Assaying Cell Migration / Proliferation in Cells Treated with a Compound of Formula I or II and / or a Second Active Agent

[0398] An assay for cell migration / proliferation in corneal epithelial cells was run to determine cell migration / proliferation in the presence of various concentrations of a compound of Formula I or II, vitamin K, or nicotinic acid / nicotinamide. Cells were plated with silicone plugs. Cells were then serum starved and pretreated with various concentrations of a compound of Formula I or II or a control. The plugs were removed and cells were treated with 1) EGF, 2) EGF and vitamin K, or 3) EGF and nicotinic acid / nicotinamide. Cell migration was quantified from micrographs.

[0399] Determination of the in vivo effects of compounds of formula I or II on basal and EGF-mediated corneal wound healing

[0400] The effect of the rate of the basis and ligand stimulation of corneal wound healing by the compound of formula I or II was determined in mice. The cornea of ​​C57 / Bl mice was injured, then pretreated with the compound of formula I or II, and then EGF was added. The wound size was monitored and quantified by fluorescein staining and fluorescence photography.

[0401] Determination of the in vivo effects of a compound of Formula I or II and a second active agent on basal and EGF-mediated corneal wound healing Certainly

[0402] The effects of a compound of Formula I or II and / or a second active agent on corneal wound healing were determined in mice. The corneas of C57 / Bl mice were wounded and then pretreated with a compound of Formula I or II, followed by the addition of EGF, vitamin K or nicotinic acid / nicotinamide. Wound size and closure were monitored.

[0403] Treatment dose

[0404] The preparations of the present application are effective in treating (i.e., lesions are stabilized or regressed) or preventing choroidal and retinal neovascularization (NV) in the eyes of mammalian subjects. Compound-I of the present application inhibits receptor tyrosine kinases at a specific dose, and the second active agent inhibits ErbB receptor tyrosine kinases at a specific dose. In some embodiments, the compound-I preparation inhibits receptor tyrosine kinases at a specific dose, including VEGFR, FGFR, Tie2 and EphB-4. The preparations of the present application have a synergistic effect on the inhibition of several RTKs at a specific dose, and are effective in treating or regressing NV in the posterior segment of the eye. In some embodiments, the second active agent directly or indirectly regulates (e.g., activates) ErB receptor tyrosine kinases, including EGFR, HER2, HER3 and Erb4. The preparations of the present application can have a synergistic effect on the activation of several ErB receptor tyrosine kinases at a specific dose, and are effective in preventing or treating corneal damage or disease.

[0405] In one embodiment, the present application provides a method for treating (i.e., lesions are stable or subsided) or preventing choroidal and retinal neovascularization (NV) in the eye by administering 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 to a subject in need, wherein the 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 before the second active agent or a pharmaceutically acceptable salt thereof is administered. In another embodiment, a compound of formula I or II or a pharmaceutically acceptable salt thereof is administered after the second active agent or a pharmaceutically acceptable salt thereof is administered. In another embodiment, the present application provides a method for treating (i.e., lesions are stable or subsided) or preventing choroidal and retinal neovascularization (NV) in the eye by administering a therapeutically effective amount of a compound of formula I or II or a pharmaceutically acceptable salt thereof to a subject in need, and then administering a therapeutically effective dose of a preparation described herein.

[0406] In another embodiment, the present application provides a method for treating (i.e., lesions are stable or subsided) or preventing choroidal and retinal neovascularization (NV) in the eye and preventing or treating corneal damage or disease by administering 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 to a subject in need, wherein the 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 before the second active agent or a pharmaceutically acceptable salt thereof is administered. In another embodiment, a compound of formula I or II or a pharmaceutically acceptable salt thereof is administered after the second active agent or a pharmaceutically acceptable salt thereof is administered. In another embodiment, the present application provides a method for treating (i.e., lesions are stable or subsided) or preventing choroidal and retinal neovascularization (NV) in the eye and preventing or treating corneal damage or disease by administering a therapeutically effective amount of a compound of formula I or II or a pharmaceutically acceptable salt thereof to a subject in need, and then administering a therapeutically effective dose of a preparation described herein.

[0407] In a further embodiment, the formulation of the present application is effective in treating NV and treating and / or preventing corneal damage or disease caused by systemic diseases, eye diseases or administration of compounds of formula I or II when about 0.005% to about 5.0% (about 0.05-about 50 mg / mL) of compound-I and a second active agent are delivered topically to the eye once, twice, three times and four times a day. The formulation of compound-I or its free base (Formula II) and a second active agent for treating or eliminating NV and treating and / or eliminating corneal damage 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. When delivered to a subject exposed to atmospheric oxygen to induce oxygen-induced retinopathy (OIR) or NV, the solution or suspension, for example, can effectively reduce the average area of ​​preretinal NV per retina with little or no corneal damage or disease. Compound-I formulations and / or suspensions prevent or treat NV by inhibiting several receptor tyrosine kinases (RTKs), including VEGFR-2. Second active agents prevent or treat corneal damage or disease by modulating (eg, activating) EGFR.

[0408] Any of the disclosed diseases or conditions described herein can be treated or prevented by achieving a target tissue concentration of about 200 nM to about 2 μM of a disclosed compound or pharmaceutically acceptable salt, formulation and / or suspension thereof. One embodiment of the present application relates to a method for treating pathological angiogenesis in the posterior segment of the eye, achieving a target tissue concentration of about 200 nM to about 2 μM of a disclosed compound or pharmaceutically acceptable salt and / or formulation thereof. Another iteration of this embodiment relates to achieving a target tissue concentration of one or more disclosed compounds or pharmaceutically acceptable salts and / or formulations thereof of about 300 nM to about 2 μM.

[0409] In one embodiment of the present application, about 0.2-about 1.0% (about 2-about 10 mg / mL) of compound-I formulated as a solution or suspension can effectively inhibit VEGFR-2 kinase function and provide a substantial blockade of a group of pro-angiogenic growth factor receptors (including FGFRs1-3, Tie-2 and EphB-4) after administration. The concentration of 2-10 mg / mL of compound-I in the formulation provides effective pharmacologically effective concentrations of drugs to the central choroid and retina after 1-5 days of local ocular delivery.

[0410] In some embodiments, the exposure time of compound-I is 1 to 90 days or longer 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 preparation comprising compound-I to a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves administering the preparation once a day, twice a day, three times a day, or four times a day for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). For example, the dosage regimen involves administering the preparation once, twice, three times, or four times every other day (i.e., on the 1st, 3rd, 5th, 7th day, etc.) for up to 90 days. For example, the dosage regimen involves administering once on the 1st day, once or twice on the 2nd-90th day. For example, the dosage regimen involves administering once, twice, three times, or four times on the 1st day, followed by once a day for 2-90 days. For example, the dosage regimen involves applying once, twice, three times, four times on day 1, followed by applying once, twice, three times, or four times every other day (i.e., on days 1, 3, 5, 7, etc.) for up to 90 days. For example, one 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 a day dosage regimen, the subject receives a topical ocular dose of the compound-I preparation on day 1 and day 4, separated by about 4, 6, or 8 hours. In another embodiment, the subject receives a topical ocular dose of the compound-I preparation for four consecutive days, separated by about 4, 6, or 8 hours. In some embodiments, the subject receives one or two doses of a topical ocular dose of the compound-I preparation every day for 5 consecutive days. In yet other embodiments, the subject receives one or two doses of a topical ocular dose of the compound-I preparation for 5-90 consecutive days. In some embodiments, the subject receives one or two doses of a topical ocular dose of the compound-I preparation for at least 25 consecutive days. In one embodiment, the subject receives one or two topical ocular doses for at least 90 consecutive days or more.

[0411] In some embodiments, the application provides a preparation of a compound-I or its free base and / or a second active agent topically applied to the anterior segment of the eye of the object to treat AMD, pathological CNV and / or pathological NV. For example, the preparation is applied to the eye of the object 1, 2, 3 or 4 times a day. In a specific embodiment, the preparation is applied to the eye of the object 2 or 3 times a day. For example, the preparation is applied to one or both eyes of the object. For example, the preparation of the first active agent comprising about 1 mg / ml of the present disclosure is applied twice a day (BID) to one or both eyes of the object. 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 of the first active agent is administered to one or both eyes of a subject.

[0412] For example, a preparation comprising about 1 mg / mL BID of compound-1 and / or a second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a preparation comprising about 1 mg / mL QD of compound-1 and / or a second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a preparation comprising about 1 mg / mL TID of compound-1 and / or a second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a preparation comprising about 1 mg / mL QID of compound-1 and / or a second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 2 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 2 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 2 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 2 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the formulation of the compound-1 and / or the second active agent comprising about 3 mg / mL BID is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the formulation of the compound-1 and / or the second active agent comprising about 3 mg / mL QD is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the formulation of the compound-1 and / or the second active agent comprising about 3 mg / mL TID is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 3 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 4 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 4 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 4 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 4 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 5 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 5 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 5 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the formulation of Compound-1 and / or a second active agent comprising about 5 mg / mL QID is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the formulation of Compound-1 and / or a second active agent comprising about 6 mg / mL BID is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the formulation of Compound-1 and / or a second active agent comprising about 6 mg / mL QD is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 6 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the compound-1 and / or the second active agent comprising about 6 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., compound-1) and the second active agent comprising about 7 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., compound-1) and the second active agent comprising about 7 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 7 mg / mL TID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 7 mg / mL QID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL BID of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL QD of the first active agent (e.g., Compound-1) and the second active agent is administered to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL TID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, a formulation comprising about 8 mg / mL QID of a first active agent (e.g., Compound-1) and a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 9 mg / mL BID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 9 mg / mL QD is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 9 mg / mL TID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-1) and the second active agent comprising about 9 mg / mL QID is applied to one or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In one embodiment, the preparation of the first active agent (e.g., Compound-I) and the second active agent comprising about 10 mg / mL QD is applied to one eye or both eyes of the object for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months). The dosage regimen of 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months or 12 months) can be any scheme related to the continuous or alternate days described in the above paragraph. In some embodiments, when applied with low doses (e.g., 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL or 5 mg / mL), QD, BID, TID or QID apply the preparation of the application, and when applied with high doses (e.g., 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL or 10 mg / mL), QD or BID apply the preparation of the application.

[0413] In other embodiments, 1 mg / mL BID of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 1 mg / mL QD of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 2 mg / mL BID of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 2 mg / mL QD of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 3 mg / mL BID of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 3 mg / mL QD of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 4 mg / mL BID of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 4 mg / mL QD of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 5 mg / mL BID of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 5 mg / mL QD of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 6 mg / mL BID of the Compound-I formulation and / or the second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months).In some embodiments, 6 mg / mL QD of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 7 mg / mL BID of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 7 mg / mL QD of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 8 mg / mL BID of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 8 mg / mL QD of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 9 mg / mL BID of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 9 mg / mL QD of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, 10 mg / mL BID of Compound-I formulation and / or a second active agent is administered to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a 10 mg / mL QD Compound-I formulation and / or a second active agent is administered to one or both eyes of a subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). The dosage regimen of 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) can be any regimen involving continuous or alternate days as described in the above paragraphs.

[0414] The application provides formulations as shown in Table 13 for administration to one or both eyes of a subject.

[0415] Table 13

[0416]

[0417] *QS = quantity sufficient to achieve osmolality.

[0418] In some embodiments, a formulation of Formula II or Compound-I is administered to one or both eyes of the subject. For example, a formulation of the present disclosure comprising about 0.2% - about 1.0% (w / v) of a compound of Formula II or about 0.1% - 1.2% (w / v) of Compound-I is administered once a day (QD) or twice a day (BID) to one or both eyes of the subject for 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months). In some embodiments, a compound of Formula II or Compound-I is compounded with a complexing agent such as cyclodextrin (e.g., Kleptose® HPB (%)) at a ratio of about 1:8, wherein about 2% - 13% (w / v) of cyclodextrin (e.g., Kleptose® HPB (%) is added to the formulation. The formulation also comprises about 0.1% - about 0.2% buffer, e.g., 10 mM phosphate buffer. The desired weight-gram molecular osmotic concentration of the preparation is about 200-about 300 mOsm, which is achieved by adding a salt (e.g., sodium chloride) in an amount sufficient to achieve the weight-gram molecular osmotic concentration. The pH of the preparation is about 6.0 at about 40°C or below about 40°C. 1 to 90 days or longer than 90 days (e.g., 4 months, 6 months, 8 months, or 12 months) of dosage regimen can be any regimen involving continuous or alternate days described in the above paragraphs. In some embodiments, the compound of formula II or the compound-I preparation also includes a second active agent. In a specific embodiment, the compound of formula II or the compound-I preparation is administered together with a second active agent.

[0419] The second active agent (e.g., niacin, niacinamide or vitamin K or a combination thereof) directly or indirectly modulates (e.g., activates) ErbB receptor tyrosine kinases (RTKs) at a specific dose, including EGFR, HER2, HER3 and HER4. The formulations of the present application simultaneously have a synergistic effect on the modulation (e.g., activation) of one or more ErbB RTKs at a specific dose, and are effective in preventing or treating corneal damage or diseases (e.g., corneal ulcers, corneal epithelial defects, keratitis, etc.) in the anterior segment of the eye.

[0420] The methods of the present application are combined with standard of care including, but not limited to, laser therapy and treatment with injectable anti-neovascular agents.

[0421] Granular compositions and preparations containing granular compositions

[0422] 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 of 100 nm to 100 μm. In some embodiments, the particles have an average diameter of 20 μm to 90 μm. In some embodiments, the particles have an average diameter of 20 μm to 80 μm. In some embodiments, the particles have an average diameter of 20 μm to 70 μm. In some embodiments, the particles have an average diameter of 30 μm to 70 μm. In some embodiments, the particles have an average diameter of 30 μm to 60 μm. In some embodiments, the particles have an average diameter of 30 μm to 50 μm. In some embodiments, the particles have an average diameter of 30 μm to 40 μm. In some embodiments, the particles have an average diameter of 50 μm to 70 μm. In some embodiments, the particles have an average diameter of 50 μm to 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 of 100 nm to 8 μm. In some embodiments, the particles have an average diameter of 100 nm to 200 nm. In some embodiments, the particles have an average diameter of up to 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 of 1 μm to 5 μm. In some embodiments, the particles have an average diameter of 2 μm to 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.

[0423] 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.

[0424] 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., niacin, niacinamide, vitamin K, or a combination thereof). 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).

[0425] In some embodiments, the first active agent (e.g., Formula II or Compound-I) and the second active agent (e.g., niacin, niacinamide, vitamin K, or a combination thereof) are roll-milled together 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 roll-milled separately, and then the particles comprising the first active agent and the particles comprising the second active agent are mixed or roll-milled together. In some embodiments, the first active agent or the second active agent is roll-milled first, and then the particles are added to the other active agent for further roll-milling.

[0426] In some embodiments, the pharmaceutical composition of the particles of the active agent (e.g., the first active agent or the second active agent) of the present application also includes one or more excipients. The excipient can be selected from any suitable excipient known in the art, for example, for the preparation of ophthalmic preparations. In some embodiments, the excipient is selected from polysorbate (Tween) 80, poloxamer (Pluronic) F-127, hydroxypropyl methylcellulose (hydroxypropyl methylcellulose or HPMC), polyvidone (PVP K-29 / 32 or K-30) and tyloxapol and combinations thereof. In some embodiments, the excipient is selected from HPMC, Tween 80, Pluronic F-127 and tyloxapol and combinations thereof.

[0427] In some embodiments, the particles comprising the first active agent (eg, Formula II or Compound-I) and the particles comprising the second active agent (eg, niacin, niacinamide, vitamin K, or a combination thereof) comprise the same excipients.

[0428] In some embodiments, the pharmaceutical composition comprising particles of an active agent (eg, a first active agent or a second active agent) of the present application further comprises a surfactant, such as benzalkonium chloride (BAC).

[0429] In some embodiments, the pharmaceutical composition of the particles of the active agent (e.g., the first active agent or the second active agent) of the present application also includes an excipient (e.g., for enhancing the bioavailability of the active agent). In some embodiments, the excipient is hydroxyethylcellulose (HEC). In some embodiments, 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%.

[0430] In some embodiments, the particles of the present application comprise Formula II (ie, the free base of Compound-I). In other embodiments, the particles of the present application comprise Compound-I.

[0431] In some embodiments, the particles of the present application are prepared by roller milling. Factors that may affect particle size include, but are not limited to, the use of a free base or salt of an active agent (e.g., Compound I versus Formula II), the addition of excipients, the speed of the rollers during milling, the size of the milling media, and the duration of milling.

[0432] In some embodiments, the particles of the present application are prepared without roller milling.

[0433] In some embodiments, the particles of the active agent (e.g., the first active agent (e.g., Formula II or Compound-I) and / or the second active agent) of the present application are sterilized. In some embodiments, the sterilization is performed with gamma radiation. In some embodiments, the particles of the active agent (e.g., the first active agent (e.g., Formula II or Compound-I) and / or the second active agent) of the present application are stable after sterilization (e.g., maintaining substantially no impurities or degradation products caused by sterilization).

[0434] 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 as described herein (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.

[0435] 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 the 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 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.

[0436] In some embodiments, the suspension formulation of particles comprising 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 thereof). 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.

[0437] In some embodiments, the suspension formulation of particles comprising the first active agent (e.g., Formula II or Compound-I) further comprises an excipient selected from polysorbate (Tween) 80, poloxamer (Pluronic) F-127, hydroxypropyl methylcellulose (hydroxypropyl methylcellulose or HPMC), povidone (PVP K-29 / 32 or K-30) and tyloxapol and combinations thereof. 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 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.

[0438] In some embodiments, the suspension formulation of particles comprising the first active agent (e.g., Formula II or Compound-I) further comprises a buffer. In some embodiments, the buffer is selected from phosphate buffer, borate buffer, citrate buffer, tartrate buffer, acetate buffer, amino acids, sodium acetate, sodium citrate, Tris buffer, etc. In some embodiments, the buffer is Tris. In some embodiments, the buffer is present at a concentration of about 0.1% - about 2%, about 0.2% - about 1.8%, about 0.3% - about 1.6%, about 0.4% - about 1.4%, about 0.4% - about 1.2%, about 0.4% - about 1%, about 0.4% - about 0.8%, about 0.4% - about 0.7% or about 0.5% - about 0.7%. In some embodiments, the suspension formulation comprises about 0.6% Tris.

[0439] In some embodiments, the suspension formulation of particles comprising the first active agent (e.g., Formula II or Compound-I) also includes a weight-gram osmotic concentration regulator. In some embodiments, the weight-gram osmotic concentration regulator is glycerol. In some embodiments, the weight-gram osmotic concentration regulator is present in an amount of about 1% - about 10%, about 2% - about 10%, about 3% - about 10%, about 4% - about 10%, about 5% - about 10%, about 2% - about 9%, about 2% - about 8%, about 2% - about 7%, about 2% - about 6%, about 2% - about 5%, about 2% - about 4% or about 2% - about 3%. In some embodiments, the suspension formulation includes about 2% or about 2.5% glycerol.

[0440] In some embodiments, the suspension formulation comprising particles of the first active agent (e.g., Formula II or Compound-I) has a pH of less than 7.5. In some embodiments, the pH is about 6.0 to about 7.0. In some embodiments, the pH is about 6.0 or about 7.0.

[0441] In some embodiments, the suspension formulation of particles comprising the first active agent (e.g., Formula II or Compound-I) further comprises hydroxyethylcellulose (HEC). In some embodiments, 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%.

[0442] One of the first active agent, the second active agent, and the one or more excipients described herein can be present in combination with the remainder of the first active agent, the second active agent, and the one or more excipients described herein at any concentration or level described herein.

[0443] In some embodiments, the formulation of the present application comprises about 0.1% to about 1.2% (or any range between as described herein) of the first active agent, about 0.4% to about 0.8% (or any range between as described herein) of Tris, about 2% to about 4% (or any range between as described herein) of glycerol, about 0.1% to about 0.3% (or any range between as described herein) of HEC and about 0.04% to about 0.09% (or any range between as described herein) of HPMC. In some embodiments, the first active agent is Formula II or Compound-I. In some embodiments, the formulation of the present application comprises about 0.4% of Formula II or Compound-I, about 0.6% of Tris, about 2% of glycerol, about 0.2% of HEC and about 0.08% of HPMC. In some embodiments, the formulation comprises particles of the first active agent, wherein the particles have an average diameter of 30 μm to 60 μm. In some embodiments, the formulation comprises particles of a first active agent, wherein the particles have an average diameter of 50 μm to 60 μm. In some embodiments, the formulation comprises particles of a 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.

[0444] In some embodiments, the formulations of the present application include about 0.1% to about 1.2% (or any range therebetween as described herein) of a first active agent, about 0.00001% to about 0.0001% (or any range therebetween as described herein) of a second active agent, about 0.4% to about 0.8% (or any range therebetween as described herein) of Tris, about 2% to about 4% (or any range therebetween as described herein) of glycerol, about 0.1% to about 0.3% (or any range therebetween as described herein) of HEC, and about 0.04% to about 0.09% (or any range therebetween as described herein) of HPMC. 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 about 0.4% of Formula II or Compound-I, about 0.000086% of Vitamin K3, about 0.6% of Tris, about 2% of glycerol, about 0.2% of HEC, and about 0.08% of HPMC. In some embodiments, the formulation comprises particles of a first active agent, wherein the particles have an average diameter of 30 μm to 60 μm. In some embodiments, the formulation comprises particles of a first active agent, wherein the particles have an average diameter of 50 μm to 60 μm. In some embodiments, the formulation comprises particles of a first active agent, wherein the particles have an average diameter of about 50 μm or 60 μm. In some embodiments, the particles further comprise a 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.

[0445] Indications and treatments

[0446] Methods for treating diseases or conditions of the eye are disclosed. The disclosed methods involve treating, preventing or controlling ocular neovascularization (NV) or treating diseases or conditions associated with the pathogenesis of NV by administering to a subject one or more disclosed compounds (e.g., a first active agent (e.g., a compound of Formula I or II) and optionally a second active agent) and formulations thereof.

[0447] One aspect of the disclosed methods relates to treating or preventing NV by administering to a subject an effective amount of one or more disclosed compounds of Formula I or II or pharmaceutically acceptable salts and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) and / or formulations thereof. One embodiment of this aspect relates to a method for treating NV by administering to a subject a) an effective amount of one or more disclosed compounds of Formula I or II or pharmaceutically acceptable salts and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) and / or formulations thereof and optionally b) one or more carriers or compatible excipients.

[0448] The disclosed methods involve preventing or controlling pathological ocular neovascularization (NV) or treating diseases or conditions associated with the onset of NV by administering to a subject one or more disclosed compounds of Formula I or II and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof) and formulations thereof.

[0449] The present embodiment provides the use of a formulation of Compound-I or its free base (Formula II) and an optional second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) for the preparation of a medicament for treating a subject suffering from a posterior segment disease, vascular disease or inflammatory disease 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) from any mechanism (e.g., high myopia, trauma, sickle cell disease; ocular histoplasmosis, angioid streaks, traumatic choroidal rupture, drusen of the optic nerve and some retinal dystrophies); from any mechanism (e.g., sickle cell retinal neovascularization); from any mechanism (e.g., In one embodiment, the eye disease is AMD. In one embodiment, the eye disease is caused by or exacerbated by ocular angiogenesis and / or neovascularization.

[0450] In one aspect of the application, the formulation is used to treat age-related macular degeneration (AMD) (including neovascular (wet / exudative) AMD, dry AMD and geographic atrophy). The solution or suspension is used to treat neovascular (exudative or wet) AMD. In another embodiment, the solution or suspension is used to treat dry AMD. In yet another embodiment, the solution or suspension is used to treat geographic atrophy.

[0451] The formulations of the present application prevent, delay or treat the onset of pathological choroidal neovascularization (CNV) in a subject arising from any mechanism (e.g., high myopia, trauma, sickle cell disease; ocular histoplasmosis, angioid streaks, traumatic choroidal rupture, drusen of the optic nerve, and some retinal dystrophies).

[0452] 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.

[0453] One aspect of the method relates to treating or preventing ocular neovascularization by administering to a subject an effective amount of one or more disclosed compounds of Formula I or II or a pharmaceutically acceptable salt thereof and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof). One embodiment of this aspect relates to a method for treating ocular edema and neovascularization by administering to a subject a) an effective amount of one or more disclosed compounds of Formula I or II or a pharmaceutically acceptable salt thereof and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) and / or a formulation thereof and optionally b) a composition of one or more carriers or compatible excipients.

[0454] The disclosed methods also relate to preventing or controlling ocular edema or treating a disease or condition associated with the onset of ocular edema by administering to a subject one or more disclosed compounds of Formula I or II and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof).

[0455] One aspect of the method relates to treating or preventing ocular edema by administering to a subject an effective amount of one or more disclosed compounds of Formula I or II or a pharmaceutically acceptable salt thereof and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof). One embodiment of this aspect relates to a method for treating ocular edema by administering to a subject a) an effective amount of one or more disclosed compounds of Formula I or II or a pharmaceutically acceptable salt thereof and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) and / or a formulation thereof and optionally b) a composition of one or more carriers or compatible excipients.

[0456] Another disclosed method involves preventing or controlling retinal edema or retinal neovascularization or treating a disease or condition associated with the onset of retinal edema or retinal neovascularization by administering to a subject one or more disclosed compounds of formula I or II and optionally a second active agent (e.g., vitamin K, niacin or nicotinamide or a combination thereof). One aspect of the method involves treating or preventing retinal edema or retinal neovascularization by administering to a subject an effective amount of one or more disclosed compounds of formula I or II or a pharmaceutically acceptable salt thereof and optionally a second active agent (e.g., vitamin K, niacin or nicotinamide or a combination thereof). An embodiment of this aspect relates to a method for treating retinal edema or retinal neovascularization by administering to a subject a) an effective amount of one or more disclosed compounds of formula I or II or a pharmaceutically acceptable salt thereof and optionally a second active agent (e.g., vitamin K, niacin or nicotinamide or a combination thereof) and / or a formulation thereof and optionally b) a composition of one or more carriers or compatible excipients.

[0457] Another embodiment of this aspect is directed to a method for delaying or preventing the progression of non-proliferative retinopathy to proliferative retinopathy by administering to a subject a) an effective amount of one or more disclosed compounds of Formula I or II, or a pharmaceutically acceptable salt, and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof) and / or a formulation thereof, and optionally b) a composition of one or more carriers or compatible excipients.

[0458] One aspect of the disclosed method relates to diseases that are a direct or indirect result of diabetes, especially diabetic macular edema and diabetic retinopathy. The ocular vascular system of diabetes becomes unstable over time, leading to conditions such as non-proliferative retinopathy, macular edema, and proliferative retinopathy. When fluid leaks into the center of the macula (the part of the eye where sharp, straight vision occurs), accumulations of fluid and associated proteins begin to deposit above or below the macula. This causes swelling, which causes the central vision of the subject to gradually become distorted. The condition is called "macular edema". Another condition that may occur is non-proliferative retinopathy, in which vascular changes outside the macular area of ​​the eye, such as microaneurysms, can be observed. During proliferative DR, pathological new blood vessels grow in the retina and from the retina upward into the vitreous, where these abnormal blood vessels can change the retinal morphology in the macula, and / or bleed into the vitreous and obscure the visual axis.

[0459] Another disclosed method involves treating, preventing or managing diabetic retinopathy, or treating a disease or condition associated with the onset of diabetic retinopathy, by administering to a subject one or more disclosed compounds of Formula I or II and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof).

[0460] 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 disclosed compounds of Formula I or II, or a pharmaceutically acceptable salt thereof, and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof). One embodiment of this aspect relates to a method for treating diabetic retinopathy by administering to a subject a) an effective amount of one or more disclosed compounds of Formula I or II, or a pharmaceutically acceptable salt thereof, and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof) and / or a formulation thereof and optionally b) a composition of one or more carriers or compatible excipients.

[0461] Diabetic proliferative retinopathy is characterized by neovascularization. The new blood vessels are fragile and prone to bleeding. The result is scarring of the retina and occlusion or complete blockage of the light path through the eye due to the abnormal formation of new blood vessels. Typically, subjects with diabetic macular edema have diabetic retinopathy in the non-proliferative phase; however, it is not uncommon for subjects to begin to show macular edema only when the proliferative phase develops.

[0462] Yet another disclosed method involves preventing or controlling diabetic macular edema or treating a disease or condition associated with the onset of diabetic macular edema by administering to a subject one or more disclosed compounds of Formula I or II and optionally a second active agent (e.g., vitamin K, niacin or niacinamide, or a combination thereof).

[0463] One aspect of the method relates to treating or preventing diabetic macular edema by administering to a subject an effective amount of one or more disclosed compounds of Formula I or II or a pharmaceutically acceptable salt and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) or a formulation thereof. One embodiment of this aspect relates to a method for treating diabetic macular edema by administering to a subject a) an effective amount of one or more disclosed compounds of Formula I or II or a pharmaceutically acceptable salt and optionally a second active agent (e.g., vitamin K, niacin or niacinamide or a combination thereof) and / or a formulation thereof and b) a composition of one or more carriers or compatible excipients.

[0464] Another aspect of the disclosed method relates to treating or preventing NV and treating and / or preventing corneal damage or disease by administering to a subject an effective amount of one or more disclosed compounds of formula I or II or pharmaceutically acceptable salts, a second active agent and / or formulations thereof. One embodiment of this aspect relates to a method for treating NV and treating and / or preventing corneal damage or disease by administering to a subject a) an effective amount of one or more disclosed compounds of formula I or II or pharmaceutically acceptable salts and / or formulations thereof, b) a second active agent and optionally c) a composition of one or more carriers or compatible excipients. In one embodiment, the corneal damage or disease is caused by a compound of formula I or II.

[0465] Reagent test kit

[0466] Also disclosed is a kit for the disclosed compounds and compositions for drug delivery to people, mammals or cells. The kit may include a composition comprising one or more formula I or II compounds of one or more packaged unit doses, and a second activating agent to be delivered to people, mammals or cells of one or more packaged unit doses. Unit dose ampoules or multidose containers (wherein the formula I or II compounds or the second activating agent to be delivered are packaged before use) may include sealed containers, which encapsulate an activating agent or a pharmaceutically acceptable salt or its preparation suitable for its pharmaceutically effective dose or the amount of multiple effective doses. The compound may be packaged as a sterile preparation, and the sealed container is designed to keep the sterility of the preparation until use.

[0467] The kit of the present application has a disposable eye drop dispenser bottle for delivering ophthalmic preparations. In an alternative embodiment, the kit of the present application has a multiple eye drop dispenser bottle. The multi-dose dispenser bottle has an appropriate amount of anti-infective agent and / or preservative, such as but not limited to 0.005% BAK. The eye dispenser of the present application has a top and a cap. The container of the present application has a translucent LDPE eye dispenser bottle, which has an LDPE dropper and a HDPE cap. As required and / or as used in the art, the container can be other types and forms.

[0468] The following examples illustrate, but are not limiting of, the methods and compositions of the present application. Other suitable modifications and adaptations of the various conditions and parameters normally encountered in therapy and apparent to those skilled in the art are within the spirit and scope of the embodiments.

[0469] General approach

[0470] Roller Grinding

[0471] The horizontal roller mill is composed of a plurality of motor-driven rollers contained in a metal housing. A single container placed between the rollers will rotate at a rpm determined by the speed of the rollers and the diameter of the container. The drug slurry consisting of API and optional stabilizer, water and / or grinding media is added to the container and then placed between the rollers. The medium used is a bead of various sizes (e.g., 800 microns to 3000 microns), and can be made of yttrium zirconium oxide. After grinding, the dispersion is separated from the medium by transferring the contents to a centrifuge tube insert equipped with a screen. After centrifugation (e.g., centrifugation for about 5 minutes with about 300 x G), the dispersion is collected below the screen (which retains the culture medium).

[0472] Optical Microscopy (OM)

[0473] The morphology and size distribution of the particle composition can be assessed by optical microscopy, and micrographs of the particles can be taken, for example, using an Olympus BX51 system equipped with an oil immersion 100x objective (1000x magnification). A calibration bar (from 1 um to 100 μm) can be set as a comparator on each micrograph.

[0474] Particle size distribution (PSD)

[0475] The particle size distribution can be analyzed, for example, using a Horiba LA-950 V2 using laser diffraction light scattering. General assumptions are made in terms of setting conditions and refractive index values. The distribution is based on volume. The sample density is adjusted to the general range of the percent transmission on the blue LED light source. A small sample chamber (filled with water) can be used instead of a flow-through chamber to minimize the sample amount.

[0476] calculate

[0477] Use the following equation to determine the volume of diluent (50:50 methanol:water) required to prepare choroid, retina, and cornea samples at specific tissue concentrations.

[0478] volume 稀释剂 =(Mass 组织 / concentration 组织 )- Volume 组织 Equation 1

[0479] Where: Concentration 组织 = Expected tissue concentration (mg / mL)

[0480] quality 组织 = Mass of tissue (mg)

[0481] volume 稀释剂 = Volume of diluent (50:50 methanol:water) (mL)

[0482] volume 组织 = volume of tissue (mL), assuming a density of 1.0 g / mL.

[0483] To calculate the concentration in ng / g (ng drug / g tissue), the following equation was used:

[0484] concentration ng / g = Concentration ng / mL x (volume 总 / quality 组织 ) Equation 2

[0485] Where: Concentration ng / g = Calculated concentration (ng drug / g tissue)

[0486] concentration ng / mL = Calculated concentration (ng drug / mL homogenate)

[0487] volume 总 = Total volume of tissue homogenate (mL)

[0488] quality 组织 = mass of tissue (g).

[0489] To then calculate the concentration of drug in tissue* in μM (μmol drug / volume of tissue), the following equation was used, assuming a tissue density of 1 g / mL:

[0490] concentration µM = Concentration ng / g / MW Equation 3

[0491] Where: Concentration µM = Calculated concentration (µmol drug / volume of tissue)

[0492] concentration ng / g = Calculated concentration (ng drug / g tissue)

[0493] MW = molecular weight (g / mole)

[0494] *Tissue = choroid, retina, or cornea.

[0495] To calculate the concentration of drug in the fluid** in μM (μmol drug / volume of fluid), the following equation was used:

[0496] concentration µM = Concentration ng / mL / MW Equation 4

[0497] Where: Concentration µM= Calculated concentration (µmol of drug / volume of fluid)

[0498] Concentration ng / mL = Calculated concentration (ng of drug / mL of fluid)

[0499] MW = Molecular weight (g / mole)

[0500] ** Fluid = Plasma.

[0501] Note: For the calculation of the reported statistical values (mean, standard deviation, and percent coefficient of variation), any sample concentration <LLOQ within the body of this report is discarded or excluded.

[0502] Draize eye irritation scoring system for cornea, iris and conjunctiva

[0503]

[0504] Draize et al. (1944) Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes

[0505] If the cornea, iris, or conjunctiva is normal, a score of 0 is assigned for each parameter. Example

[0506] Compound-I is a potent and selective small molecule inhibitor of VEGFR-2 and other pro-angiogenic RTKs such as fibroblast growth factor receptors (FGFR-1-3), Tie-2, and ephrin receptor B4 (EPHB-4). After systemic administration in murine corneal and rat growth plate models, Compound-I was shown to inhibit phosphorylation of specific RTKs, endothelial cell proliferation, and pathological angiogenesis, and to inhibit the growth of human tumor xenografts in athymic mice. With regard to potential ophthalmic indications, the examples of this application described below demonstrate that topical ocular delivery of Compound-I provides significant inhibition of pathological retinal and choroidal neovascularization in clinically relevant rodent models. An overview of these data is presented below.

[0507] The following studies were conducted to measure the effects of the disclosed compounds on vascular leakage and neovascularization in retinal tissue.

[0508] Example 1

[0509] Main pharmacodynamics

[0510] In vitro efficacy pharmacology of compound ICompound-I potently inhibits the tyrosine kinase activity of vascular endothelial growth factor receptor-2 (VEGF-2) and a select subset of other pro-angiogenic RTKs during various in vitro assays. Specifically, Compound-I compounds blocked VEGF-stimulated VEGFR-2 phosphorylation in whole cells and proliferation of cultured endothelial cells. Compound-I was expressed at 50% inhibitory concentrations (IC ) of 10.55 nM (6 ng / mL) and 8.79 nM (5 ng / mL), respectively. 50 ) to inhibit the recombinant tyrosine kinase activity of VEGFR-2 and FGFR-2; and IC 50 = 5.27 nM (3 ng / mL) inhibits autophosphorylation of VEGFR-2 in intact cells. This inhibition is selective relative to many other tyrosine kinases, e.g., VEGFR-2IC 50 They are approximately 1 / 500 and 1 / 1000 of the epidermal growth factor receptor (EGFR) and insulin receptor (IR) tyrosine kinases, respectively (see Table 2).

[0511] Compound-I exhibited potent inhibition of the tyrosine kinase activity of several pro-angiogenic growth factor receptors using a 10-point titration curve ranging from 257 to 5000 nM (146-2845 ng / mL), as measured by IC 50 <100 nM (56.89 ng / mL) (see Table 3). The IC 50 As follows: recombinant KDR (human isoform of VEGFR-2) = 1.27nM (0.72 ng / mL), Tie-2 = 10.10nM (5.75 ng / mL), and FGFR1-3 = 8.50nM (4.84 ng / mL), 3.08nM (1.75 ng / mL) and 33.9nM (19.29 ng / mL), respectively. This compound also blocks the other high affinity VEGF receptor VEGFR-1 / Flt-1, but with lower potency: IC 50 = 122nM (69.41 ng / mL).

[0512] Although VEGFR inhibition appears to be essential for reducing vascular permeability and further preventing neovascularization, simultaneous inhibition of VEGF signaling and inhibition of other growth factor signaling pathways (e.g., PDGF and angiopoietin / Tie2) may be associated with unique therapeutic outcomes. The therapeutic outcome of more extensive inhibition of signaling pathways may contribute to the regression of newly established pathological blood vessels in the posterior segment of the eye.

[0513] Compound-1 at 300 nM (170.67 ng / mL) completely inhibited VEGFR-2 kinase function (see Table 4) and provided a substantial blockade of a similar set of angiogenic growth factor receptors (including FGFRs-1-3, Tie-2, and EphB-4). An unexpected finding was that a 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 after five days of topical ocular delivery in rabbits and dogs.

[0514] Overview of APIs and Drug Products

[0515] Drug substance: The active pharmaceutical ingredient (API), Formula II hydrochloride (Compound-I, CP-547,632-01), is a small molecule in a single polymorph. The API raw material is consistently manufactured with a purity of more than 99.7%. Any impurity ≥ 0.15% in the drug substance is appropriately qualified in toxicology studies, and the current specification for new unknown individual impurities is set at NMT 0.2%. The final drug substance and drug product are analyzed using standard methods.

[0516] Drug product: Compound-I ophthalmic preparations for clinical studies are manufactured in dosage strengths of 0.05%-1.0% (as Compound-I). The strengths used for GLP batches are 0% (placebo), 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.8% and 1.0% Compound-I. In clinical trials, Compound-I ophthalmic preparations (solutions or suspensions) are used for daily, single-use, topical administration to the eye. In addition to the active ingredient, the drug product may also contain 0.005% BAK as a preservative, purified water as a vehicle, and sodium hydroxide to adjust the pH to pH 6.0.

[0517] Example 2

[0518] Sodium phosphate based gel drops

[0519] The ophthalmic effect of compound-1 in sodium phosphate-based formulations (listed in Table 6) is derived from the self-gelling properties of the API in a buffer such as sodium phosphate. The self-forming thixotropic gel of compound-1 that spontaneously forms from a clear solution is formed by increasing the concentration of the API in the sodium phosphate. This gel initially appears clear, and then shows an increase in consistency / viscosity at higher API concentrations, and gradually becomes more opaque, i.e., turbid. Once the API concentration in the phosphate buffer reaches a supersaturated state, insoluble particles of compound-1 are also observed in the gel.

[0520] The gel with increased viscosity is applied to the surface of the eye to increase the corneal residence time. Increasing the corneal residence time is conducive to the absorption of ophthalmic drugs. As a result, compared with non-viscous preparations such as aqueous solutions, the intraocular drug concentration of viscous gel increases. A method of increasing viscosity is to use various viscosity-increasing excipients, for example, carboxymethyl cellulose, which actually increases the intraocular absorption of different raw materials after topical ocular administration. However, in this study, in the absence of any viscosity-increasing excipients, a thixotropic gel of compound-1 is unexpectedly formed. For example, when compound-1 is dissolved in a simple buffer such as sodium phosphate, a thixotropic gel is formed. In the present embodiment, the thixotropic gel formed in the absence of any viscosity-increasing excipient is formulated as a gel drop.

[0521] Gel drops of compound-1 were applied to the eyes of Dutch striped rabbits. Gel drops of compound-1 were applied to Dutch striped rabbits for 4 or 5 consecutive days, with three doses per day. The concentration of compound-1 at the target tissue was measured 1 hour after the last dose applied. The delivery of compound-1 to the posterior segment tissue was dose-dependent and dose-frequency-dependent.

[0522] The gel drop formulations (listed in Table 6) differed in several aspects, such as API concentration, sodium phosphate concentration, presence or absence of tonicity (glycerol) or preservatives (benzalkonium chloride / BAK), solubilizing surfactants (polysorbate 80, tyloxapol and / or poloxamer), and pH.

[0523] Example 3

[0524] Tromethamine-based suspensions

[0525] Compound-1 (about 1 mg / mL to about 10 mg / mL) forms a suspension in a tromethamine-based formulation. The suspension of compound-1 in a tromethamine-based formulation has an active drug substance in an insoluble form of >95%. This feature is distinguished from the soluble or semi-soluble state of compound-1 in gel drops (when the concentration of the active agent increases, the gel drops (gel) are not completely soluble) or cyclodextrin-based formulations. The tromethamine-based formulation of compound-1 shows an increased turbidity with an increased concentration of the active agent. It is expected that topical drops of compound-1 suspensions (which are a combination of soluble and insoluble active agent components) will provide unique benefits with respect to safety / tolerance and efficacy.

[0526] A tromethamine-based suspension of Compound-I was administered to Dutch striped rabbits for 4 or 5 consecutive days, with dosing three times per day. Ocular tissue and plasma concentrations of Compound-I were measured 1 hour after the last administered dose. Compound-I in a tromethamine-based suspension was compared with its cellular IC for various pro-angiogenic RTKs. 50See Table 7.

[0527] The corneal safety and tolerability of topical compound-1 is a direct result of the amount of soluble (relative to insoluble) active agent applied to the corneal surface and the resulting corneal tissue concentration. Compared with the equimolar formulation of sodium phosphate-based gel drops, animals receiving topical ocular administration of tromethamine-based suspensions are able to tolerate higher levels of active agent concentrations in the formulation. The results obtained from both Dutch striped rabbits and beagles show that when the corneal concentration of compound-1 exceeds 100 μM, ocular side effects such as discomfort and inflammation and, in some cases, corneal thinning are more consistently observed.

[0528] The administration of tromethamine-based suspension has an unexpected effect on the ocular bioavailability of compound-I in the posterior segment. It is observed that the ocular bioavailability of compound-I in the posterior segment is proportional to the total amount of the drug administered (insoluble plus soluble, see Table 7). Although insoluble drug particles are not easily utilized by the anterior segment tissue; the inherent and unique physicochemical properties of compound-I allow insoluble and soluble components to enter the posterior segment tissue, such as choroid and retina. Therefore, a suspension based on tromethamine is achieved that is higher than the drug concentration achieved by the gel drop formulation containing an equal amount of active agent. Therefore, a suspension based on tromethamine provides: a) improved corneal tolerance and b) increase in the posterior segment, particularly the bioavailability of the choroid (the main target tissue for the treatment of neovascularization (wet) AMD).

[0529] Example 4

[0530] Cyclodextrin-based solutions

[0531] Cyclodextrins, which are cyclic oligosaccharides composed of six to eight glucose units (α-, β- and γ-CD) joined by one to four bonds, are well known for their ability to act as solubilizers for relatively poorly soluble drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol. , 36: 30-42 (2008).

[0532] Clinical formulations of ophthalmic solutions of Compound-I or its free base in 2-hydroxypropyl-β-cyclodextrin (HP-β-CD, KLEPTOSE® HPB) at a molar ratio equal to or greater than 1:6 or in sulfobutyl ether-β-cyclodextrin (SBE-β-CD, CAPTISOL®) at a ratio equal to or greater than 1:2 provide solubility at clinical dosage strengths of 0.1-1.0% Compound-I.

[0533] The cyclodextrin-based solution of Compound-I or the free base not only improves the solubility of the active agent to a homogeneous solution, but also features a novel and previously unobserved significantly increased therapeutic index of the active agent at the posterior segment of the eye after topical ocular administration. The solution reduces anterior segment exposure, thereby providing an increased concentration of the active agent in the solution and an increased delivery frequency, which maintains a high posterior segment concentration. Both of these beneficial features are related to the known properties of cyclodextrins to form hydrophilic complexes with hydrophobic drugs. See Stella & He, Cyclodextrins, Toxicol. Pathol. , 36:30-42 (2008). When formulated with Compound-1 or its free base, cyclodextrin forms a clear, colorless solution and exhibits a watery viscosity. After topical ocular administration, the Compound-1 / cyclodextrin complex has the appearance of being pharmacologically inactive and metabolically inert. The Compound-1 / cyclodextrin complex imparts corneal tolerance until the cyclodextrin spontaneously dissociates from the active agent, thereby achieving high concentrations of Compound-1 at its intended site of action in the posterior segment of the eye (e.g., choroid and retina).

[0534] During the topical ocular administration study lasting 1 to 30 days in Dutch striped rabbits, at similar drug concentrations, the cyclodextrin-based solution of compound-I exhibited significantly reduced corneal exposure than the gel drop formulation (see Example 2). Compared with the administration of an equimolar formulation of gel drops, the use of a cyclodextrin-based solution of compound-I provides about a 10-fold reduction in corneal concentration. Therefore, after 30 days of topical ocular administration of 0.6% compound-I as a cyclodextrin-based solution, no adverse results were attributed to the test article or vehicle. Compound-I based cyclodextrin solutions also achieve equal or significantly higher drug concentrations in the posterior segment target tissues, such as at the central choroid and central retina. Reducing corneal drug exposure to avoid poor eye tolerance, while increasing the posterior segment bioavailability to increase the combined effect of RTK inhibition significantly increases the therapeutic index and the corresponding benefits experienced by patients.

[0535] For both suspension-based formulations (see Example 3) and cyclodextrin formulations, the therapeutic window is expanded due to significantly reduced exposure (reduced by 10-100 times or 1-2 log). Reduced exposure improves corneal safety / tolerance, which allows compound-1 to be topically administered to be administered at higher concentrations or frequencies. Higher concentrations enable the drug to reach higher target tissue concentrations at the back of the eye, which improves the therapeutic efficacy of compound-1.

[0536] This study shows that in rabbits and dogs, topical ocular administration of ophthalmic gel drops is associated with high corneal tissue exposure (≥100uM) and corresponding adverse observations in the anterior segment such as discomfort, corneal and conjunctival inflammation, corneal epithelial erosion and / or thinning and degeneration. In contrast, repeated topical ocular administration of compound-I ophthalmic solution produces corneal exposure of about 1 / 5 to 1 / 10 of that of an equimolar dose of ophthalmic gel drops, and there are no adverse clinical or histopathological results. In addition, topical ocular administration of compound-I ophthalmic solution achieves the same or higher target therapeutic exposure in the central choroid compared to an equimolar dose of ophthalmic gel drops. In summary, reducing corneal exposure and correspondingly improving ocular tolerance, while maintaining or promoting drug delivery to the posterior segment target tissue, together with a combination of improved physicochemical stability, will provide patients with greater benefits than ophthalmic gel drops formulations.

[0537] Example 5

[0538] 1- to 5-day PK results with topical ocular Compound-I in a cyclodextrin-based solution

[0539] Ocular pharmacokinetics were studied after topical ocular dosing of various formulations and dosage regimens of Compound-I in Dutch striped rabbits. Nine (9) different topical ocular formulations with three doses of Compound-I were administered once a day (qd) or twice a day (bid) for 1, 4, or 5 consecutive days. The study design (see Tables 10A-B) consisted of seventy-two (72) rabbits, each receiving 30 μL bilateral topical ocular doses of one of three (3) Compound-I formulations or vehicle formulation using a positive displacement pipette.

[0540] 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 (qd) for one (1) or four (4) days. On Days 1 to 4, Groups 3, 7, 9, 12, 14, and 16 received bid dosing at 7:00 AM and 3:00 PM, approximately 8 hours apart, for four (4) days. Animals in Groups 18 and 19 received bid dosing of vehicle-only formulations for five (5) consecutive days.

[0541] For Group 1, ocular sampling was performed at 0.5, 1, 2, 4, 8 or 24 hours after dosing relative to Day 1. For Groups 2 and 6, ocular sampling was performed at 1, 8 and 24 hours after dosing relative to the morning of Day 5. For Groups 3, 7, 8, 11 and 13, ocular sampling was performed at 1 and 24 hours after dosing relative to the morning of Day 5. For Groups 4, 9, 10, 12, 14, 15, 16 and 17, ocular sampling was performed at 1 hour after dosing relative to the morning of Day 5. For Group 5, ocular sampling was performed at 0.5, 1, 2, 4, 8 and 24 hours after dosing relative to Day 1. Animals in Groups 18 and 19 were followed for five (5) days of clinical observation only.

[0542] Aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid samples were collected. Aqueous humor, cornea, central retina, and central choroid samples were then assayed. Peripheral retina and peripheral choroid samples were only assayed for groups 1-8, 12, 14, and 16 per study protocol.

[0543] Rabbit aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid samples were analyzed. Calibration curves were prepared in a control matrix to determine the concentration of Compound-I in various tissues.

[0544] Example 6

[0545] Five-day PK results with topical ocular Compound-I in a cyclodextrin-based solution

[0546] Ocular pharmacokinetics were calculated following ocular dosing of various dosing regimens of topical ophthalmic solutions of Compound-I containing hydroxypropyl-β-cyclodextrin ("HDβCD") in Dutch striped rabbits. Nine (9) different topical ophthalmic solutions with four different doses of Compound-I were administered once daily (qd) or twice daily (bid) for 4 or 5 consecutive days. The study design consisted of forty (40) rabbits, each receiving one of four (4) Compound-I dose strengths of 30 μL bilateral topical ophthalmic doses using a positive displacement pipette.

[0547] All doses were administered within ±1 hour of the scheduled dose time, except for Groups 1, 4, and 10 on Day 1. The first dose was administered at 12:00 PM on Day 1, and the second dose (for Groups 1 and 10) was administered approximately 4 hours later. This was due to delayed arrival of the formulation. All other dosing for these groups was scheduled. On Day 1, Groups 4-8 and 11-12 received one dose (qd) for four (4) days. On Days 1 to 4, Groups 1-3 and 9-10 received bid dosing, approximately 8 hours apart, for four (4) days.

[0548] For all groups, ocular sampling was performed 1 hour after the first daily dose on Day 5, except for Groups 5b and 6b, where ocular sampling was performed 24 hours after the first daily dose on Day 4.

[0549] For all animals, blood samples for plasma collection were obtained just prior to scheduled euthanasia. Aqueous humor, cornea, central and peripheral retina, and central and peripheral choroid samples were collected. Cornea, central retina, and central choroid samples from groups 1-12 were assayed. Aqueous humor, peripheral retina, and peripheral choroid samples were not assayed.

[0550] Example 7

[0551] Concentration of Compound-I in various intraocular fluids and tissues (in μM)

[0552] Ocular solutions of compound-I containing cyclodextrin were prepared and tested in different animal groups. After 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) containing 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.

[0553] The central choroidal concentration of Compound-I was 0.259 μΜ to 0.769 μΜ. See Table 10A. The central retinal concentration of Compound-I was 0.0531 µM-0.124 µM. See Table 10A. The aqueous humor concentration of Compound-I was 0.00313 µM-0.00656 µM. See Table 10A. And the corneal concentration of Compound-I was 6.49 µM-30 µM. See Table 10A. The cyclodextrin used to prepare the solutions was KLEPTOSE® HPB or CAPTISOL®. See Table 10B.

[0554] Example 8

[0555] Ocular Toxicology Studies

[0556] The ocular toxicity of dose limitation is characterized by the cornea and conjunctiva results in Dutch striped rabbits and beagles. These ocular results from repeated dose toxicology studies with compound-I ophthalmic gel drops are based on clinical ophthalmology and histopathological evaluations and are limited to conjunctival hyperemia, chemosis, hyperemia and secretions, corneal opacification and epithelial erosion and keratoconjunctivitis. No adverse changes involving the deeper structures (iris, lens, ciliary body, retina, choroid, sclera) or optic nerve of the eye are observed. During the full-field electroretinogram performed in rabbits, retinal function was normal in all test articles and vehicle treatment groups.

[0557] The goals of the exploratory ocular toxicology studies are to identify: a) a well tolerated topical ocular formulation, and b) a topical ocular formulation that can achieve targeted therapeutic concentrations of Compound-I in the central choroid.

[0558] In Dutch striped rabbits, mean Compound-I ocular tissue concentrations following twice daily topical administration of a 0.3% Compound-I ophthalmic gel drop formulation (with or without benzalkonium chloride) were highest in the cornea (236-260 μM) >> peripheral choroid (2.79-4.10 μM), central choroid (0.340-0.496 μM), peripheral retina (0.150-0.309 μM), and aqueous humor (0.0197-0.0395 μM).

[0559] In Dutch striped rabbits, the Tris-based suspension formulation was well tolerated, with clinical ophthalmological examinations revealing a marked absence of corneal findings with only a few occasional episodes of mild conjunctivitis. In addition, the eyes of animals receiving a 0.3% w / v Tris-based suspension of Compound-I twice daily for 30 days were considered normal during microscopic evaluation. For twice daily topical administration of a 0.3% Compound-I Tris-based suspension (with or without benzalkonium chloride), the mean Compound-I ocular tissue concentrations evaluated 1 hour ± 15 minutes after the first daily topical ocular dose on Day 30 were highest in the cornea (2.69-3.10 μM), followed by the peripheral choroid (0.781-1.21 μM), central choroid (0.303-0.319 μM), peripheral retina (0.0819-0.0868 μM), central retina (0.0495-0.0592 μM) and aqueous humor (0.00127-0.00145 μM).

[0560] A cyclodextrin-based solution using hydroxypropyl-β-cyclodextrin (HP-β-CD, KLEPTOSE® HPB) was well tolerated when applied topically twice daily for 30 days in Dutch striped rabbits at 0.1% Compound-I (2.1% HP-β-CD), twice daily at 0.2% Compound-I (4.21% HP-β-CD), once or twice daily at 0.4% Compound-I (8.41% HP-β-CD), and once or twice daily at 0.6% Compound-I (up to 12.62% HP-β-CD). In addition, in a similar repeated dosing study, a cyclodextrin-based solution of 0.4% w / v Compound-I in Kleptose® HPB, Kleptose® HP, or Captisol® was well tolerated when administered twice daily for 24 hours. In any study, no significant ocular toxicity associated with Compound-I or vehicle treatment was found during clinical ophthalmologic or microscopic examinations.

[0561] In the 24-day study, ocular tissue concentrations of eyes treated with a cyclodextrin-based solution of Compound-I were evaluated at 1 hour ± 15 minutes after the first daily topical ocular dose on Day 24 and were, in descending order, highest in the cornea (6.49-30 μΜ) >> central choroid (0.212-0.769 μΜ) > central retina (0.0531-0.124) > aqueous humor (0.002-0.007).

[0562] In summary, dose-limiting corneal toxicity was observed with the compound-I ophthalmic gel drop formulation. The ophthalmic gel drops allow the corneal concentration of compound-I to be five to fifteen times that of a cyclodextrin-based solution, and the corneal concentration of compound-I to be fifty to one hundred times that of a Tris-based suspension. Tris-based suspensions and cyclodextrin-based solutions of compound-I were well tolerated with no evidence of obvious ocular toxicity. The well-tolerated dose levels of cyclodextrin-based solutions or Tris-based suspensions of compound-I ranged from 0.005% to about 5.0% w / v when applied once or twice daily for at least 30 days. When using three formulations at equimolar doses, cyclodextrin-based solutions also provide the highest central choroidal concentration of compound-I, and reach or exceed the target therapeutic concentration.

[0563] Example 9

[0564] Phase I Protocol for a Dose Escalation Study in Patients with Neovascular AMD

[0565] Phase I study is a 12-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 neovascular age-related macular degeneration (AMD). Up to a total of 60 patients were treated 1 to 2 times a day with topical ocular administration of compound-I ophthalmic solution for three months, with three dose-escalating monotherapy groups and one adjuvant therapy group planned, using Lucentis® plus a single intravitreal injection of the maximum tolerated monotherapy dose (15 patients per treatment group). Patients who met the pre-specified visual acuity and CNV damage criteria confirmed by an independent reading center were allowed to stop topical ocular administration and receive standard of care treatment at the same time.

[0566] Compound-1 ophthalmic solutions for clinical studies were manufactured in at least 3 dosage strengths ranging from 0.1%-1.0% (as Compound-1). Strengths used for GLP batches were 0% (placebo), 0.1%, 0.3%, 0.6%, and 1.0% Compound-1 HCl. Up to 2 to 3-fold incremental doses (approximately ½ log unit steps) were administered to subsequent cohorts.

[0567] Example 10

[0568] A physically and chemically stable "non-gel", "non-viscous" uniform ophthalmic solution topical preparation at a drug strength of 0.1-1.0% (1 to 10 mg / ml) was prepared by measuring the effects of Compound-I concentration, cyclodextrin complex concentration, pH and tonicity on the solubility and stability of Compound-I. A suitable buffer system prevents the effects of pH drift on stability at a concentration of less than 1 mg / mL. Both phosphate and tromethamine (Tris) were evaluated as buffers. Sodium chloride was used to adjust tonicity.

[0569] The product quality attributes are shown in Table 14.

[0570] Table 14: Product quality attributes

[0571] Solvents Solubility (mg / mL) Color appearance of the preparation Clear and colorless, not noticeable to the eyes pH pH 5.5-7.0 Turbidity clarify Viscosity Free-flowing, water-like and filterable Tension Isotonic settlement none Mixed endpoint Clear and colorless, not noticeable to the eyes Solubility Solubility ≥6mg / ml

[0572] Preparation

[0573] The formulations outlined in Tables 12 and 13 were prepared using the general procedures listed.

[0574] The preparation is made up to volume with water for injection and stirred at 500 rpm for 30 minutes. Final pH is checked and adjusted to the target range with NaOH or HCl. About 5 ml aliquots are filtered directly into translucent 5 ml LDPE bottles while continuously stirring at a constant speed by means of a Watson Marlow Pumpsil D tube that is assembled to a Flexicon filler and connected to a 0.2 micron PVDF capsule filter. Samples are stored at 2-8 ° C until all sample preparations are complete. All samples will be submitted to analysis for storage and testing.

[0575] Embodiment 11

[0576] EGFR tyrosine phosphorylation assay in corneal epithelial cells (hTCEpi cells) to determine the activity of compounds of formula I or II EGFR activity

[0577] 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 compounds of Formula I or II.

[0578] The hTCEpi cells of 35 mm dishes were serum starved, then pretreated with different concentrations of Formula I or II compounds (e.g., Compound-I) or control AG1478 (EGFR kinase inhibitor) for 30 minutes, then treated with EGF (10, 50, 100 ng / ml) for 15 minutes. For each EGF concentration (10, 50, 100 ng / ml), six concentrations of Formula I or II compounds, such as 0, 1 μM, 3 μM, 10 μM, 30 μM, 100 μM or 1 μM AG1478 were used as controls. Additional control runs were without Formula I or II compounds or without EGF. All experiments were repeated three times. Cells were then harvested and immunoblotted for determination of phosphorylated EGFR (tyrosine 1068 and tyrosine 1045) and total EGFR.

[0579] EGFR phosphorylation serves as a readout of receptor activity. Figure 1 As shown in, high concentrations of Formula I or II compounds can inhibit ligand-stimulated EGFR tyrosine phosphorylation, and increasing the concentration of Formula I or II compounds reduces EGFR tyrosine phosphorylation at two sites. Increasing the concentration of EGF overcomes the EGFR inhibition caused by Formula I or II compounds and corneal adversity ( Figure 1 ). The EGFR-specific inhibitor AG1478 has an IC 50 = 3 nM, and at 1 μM, AG1478 completely blocked EGFR phosphorylation. It is estimated that 100 μM of Formula I or II compound inhibits EGFR phosphorylation by about 75-80%.

[0580] Example 12

[0581] EGFR tyrosine phosphotransferase in corneal epithelial cells (hTCEpi cells) using a compound of Formula I or II and a second active agent Acidification assay

[0582] An EGFR tyrosine assay in corneal epithelial cells was performed to determine whether vitamin K, niacin or niacinamide can prevent the compounds of Formula I or II from inhibiting EGFR and whether EGFR activity is increased.

[0583] hTCEpi cells in 35 mm dishes were serum starved and then pretreated with vitamin K (50 μM), niacin (10 μM) or nicotinamide (10 μM) for 4 hours, followed by treatment with different concentrations of Formula I or II compounds (e.g., Compound-I) for 30 minutes. The cells were then incubated with EGF (50 ng / ml) for 15 minutes. The cells were harvested and the cell lysates were immunoblotted for determination of phosphorylated EGFR (tyrosine 1068 and tyrosine 1045) and total EGFR concentrations. Figure 2 As shown in, vitamin K, niacin or niacinamide overcome the corneal adversity (EGFR inhibition) mediated by the compounds of formula I or II. In addition, Figure 2 Vitamin K3 (menadione) was shown to be most effective in attenuating the inhibition of EGFR tyrosine phosphorylation mediated by compounds of Formula I or II.

[0584] Example 13

[0585] Cell migration / proliferation (in vitro wound healing) assay in corneal epithelial cells (hTCEpi cells): Formula I or II Effects of compounds and EGF

[0586] A cell migration / proliferation assay in corneal epithelial cells was performed to determine cell migration / proliferation in the presence of varying concentrations of a compound of Formula I or II and EGF.

[0587] hTCEpi cells were plated with silicone plugs. The cells were then serum starved and pretreated with various concentrations of Formula I or II compounds or control compound AG1478 for 30 minutes. The silicone plugs were then removed to produce a cell-free area. The cells were photographed and then treated with EGF (10, 50 or 100 ng / ml) or VEGF (10 ng / ml) together with Formula I or II compounds or AG1478 for 16 hours. The cells were photographed again and cell migration was quantified from microphotographs. All experiments were repeated three times.

[0588] For each EGF concentration or VEGF, four concentrations of Formula I or II compound (0, 3 μM, 10 μM, 30 μM) or 3.2 μM AG1478 (EGFR kinase inhibitor) were used. Additional control runs included no Formula I or II compound and no EGF.

[0589] The data allow determination of in vitro measures of whether compounds of Formula I or II prevent corneal epithelial wound healing (cell migration and proliferation), and whether higher levels of EGF overcome recep...

Claims

1. A topical ophthalmic suspension formulation comprising: (a) a first active agent of formula II: or a pharmaceutically acceptable salt thereof; (b) carboxymethyl cellulose or its salts; (c) a second active agent, or a pharmaceutically acceptable salt thereof, wherein the second active agent is vitamin K3; and (d) a pharmaceutically acceptable excipient selected from one or more of the following: polysorbate 80, poloxamer, povidone, hypromellose and tyloxapol, and combinations thereof; wherein the first active agent or a pharmaceutically acceptable salt thereof is present at 0.005% to 5.0% w / v.

2. The formulation of claim 1, further comprising tromethamine.

3. The formulation of claim 2 comprising 0.2% to 1.0% w / v tromethamine.

4. The formulation of claim 1, further comprising 1% to 2.0% w / v glycerol.

5. The formulation of claim 3, wherein the pharmaceutically acceptable salt of the first active agent is a hydrochloride salt provided by the formula: (Compound-I).

6. The formulation of claim 1 comprising 0.005%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0% or 5.0% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

7. The formulation of claim 1, further comprising hydrochloric acid or sodium hydroxide.

8. The formulation of claim 1 comprising 0.1% to 1.0% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

9. The formulation of claim 1 comprising 0.2% to 1.0% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

10. The formulation of claim 1 comprising 0.1%, 0.2%, 0.3% or 0.4% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

11. The formulation of claim 1 comprising 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1.0% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

12. The formulation of claim 1 comprising 0.2% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

13. The formulation of claim 1 comprising 1.0% w / v of the first active agent or a pharmaceutically acceptable salt thereof.

14. The formulation of claim 1, wherein the second active agent or a pharmaceutically acceptable salt thereof is present in an amount less than 10 μM.

15. The formulation of claim 1, wherein the second active agent or a pharmaceutically acceptable salt thereof is present in an amount of 1 μM, 2 μM, 3 μM, 4 μM or 5 μM.

16. The formulation of claim 1, wherein the second active agent or a pharmaceutically acceptable salt thereof is present in an amount of 1 μM.

17. The formulation of claim 1, further comprising a stabilizer for the second active agent or a pharmaceutically acceptable salt thereof.

18. The formulation of any one of claims 1-17, wherein the formulation has a pH of 6.

19. The formulation of claim 1 comprising 0.005%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0% or 5.0% w / v of the first active agent or a pharmaceutically acceptable salt thereof, and 0.3% - 1.0% w / v of tromethamine.

20. The formulation of claim 1, comprising: (a) Compound I ； (b) salts of carboxymethyl cellulose; (c) the second active agent, or a pharmaceutically acceptable salt thereof, wherein the second active agent is vitamin K3; (d) 0.2% to 1.0% w / v tromethamine; (e) Poloxamers; (f) 1% to 2.0% w / v glycerol; and (g) having a pH of 6.0 at or below 40°C.

21. The formulation of claim 1, comprising: (a) 0.1% to 2.0% w / v of a compound of formula II, or a pharmaceutically acceptable salt thereof ； (b) carboxymethyl cellulose or its salts; (c) the second active agent, or a pharmaceutically acceptable salt thereof, wherein the second active agent is vitamin K3; (d) 0.6% w / v tromethamine; (e) 2.0% w / v glycerol; (f) 0.2% w / v hydroxyethyl cellulose; and (g) 0.08% w / v Hydroxypropyl methylcellulose.

22. Use of a formulation according to any one of claims 1 to 21 for the preparation of a medicament suitable for administration into the posterior segment of the eye and / or for the treatment and / or amelioration of a posterior segment disease of the eye selected from the group consisting of diabetic retinopathy, age-related macular degeneration, pathological choroidal neovascularization, pathological retinal neovascularization, uveitis, retinal vein occlusion, ocular trauma, surgically induced edema, surgically induced neovascularization, cystic macular edema, ocular ischemia, retinopathy of prematurity, Coat's disease, sickle cell retinopathy and neovascular glaucoma.

23. The use according to claim 22, wherein the diabetic retinopathy is background diabetic retinopathy, proliferative diabetic retinopathy or diabetic macular edema.

24. The use according to claim 22, wherein the age-related macular degeneration is neovascular age-related macular degeneration, dry age-related macular degeneration or geographic atrophy.

25. Use according to claim 24, wherein the neovascular age-related macular degeneration is wet or exudative age-related macular degeneration.

26. The use according to claim 22, wherein the retinal vein occlusion is a central or branch occlusion.

27. The use according to claim 22, wherein the posterior segment disease of the eye is age-related macular degeneration.

28. The use according to claim 22, wherein the posterior segment disease of the eye is diabetic retinopathy.

29. The use according to claim 22, wherein the posterior segment disease of the eye is cystic macular edema.

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