EA045318B1 - COMPOUNDS FOR THE TREATMENT OF EYE DISEASES ASSOCIATED WITH EXCESSIVE VASCULARIZATION - Google Patents

Description

Field of technology to which the present invention relates

The present invention relates to compounds and compositions for the treatment of a disease or disorder associated with excessive vascularization of the eye, such as, for example, corneal neovascularization, iris neovascularization, ciliary body neovascularization, corneal pannus, uveal neovascularization, retinal neovascularization, wet age-related degeneration macula, proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy.

BACKGROUND OF THE INVENTION

Laquinimod and tashinimod.

Laquinimod and tashinimod, second-generation quinoline-3-carboxamides, are developed as oral immunomodulators for the treatment of relapsing multiple sclerosis (MS) and chemotherapy-naïve metastatic prostate cancer (mCRPC), respectively.

The efficacy and safety of laquinimod have been evaluated in phase 1 to 3 clinical studies and have a clinical safety profile well established over more than 14,000 patient-years of daily doses of up to 0.6 mg administered to patients with relapsed MS. Data from the MS clinical trial program showed a consistent clinical reduction in the annualized relapse rate, which is a widely used endpoint for MS relapse. Treatment with laquinomod also resulted in some indications of progressive disability.

The efficacy and safety of taschinimod were assessed in phase 2 and 3 global randomized placebo-controlled studies. Disease progression (the primary endpoint) was significantly delayed with tashinimod treatment in the phase 2 and 3 studies.

Eye disorders.

Many eye diseases and disorders have no early symptoms. They may be painless and the patient may not notice changes in their vision until the disease is quite advanced. Therefore, it is important to prevent, treat and/or delay the progression of such diseases or disorders.

Age-related macular degeneration (AMD) is a debilitating disease that affects individuals over 60 years of age. It is the leading cause of irreversible severe vision loss in developing countries. The disease damages acute and central vision. Central vision is necessary for seeing objects clearly and for performing common daily tasks such as reading and driving. AMD affects the macula, the central part of the retina that allows the eye to see fine detail. There are two forms of AMD, wet and dry.

Dry AMD occurs when the spot thins over time as part of the aging process, gradually clouding central vision. The dry form is more common, accounting for 70-90% of AMD cases, and progresses more slowly than the wet form. Over time, as the function of the spot weakens, central vision in the affected eye is gradually lost. Dry AMD usually affects both eyes. One of the most common early signs of dry AMD is drusen.

The exudative (wet) or neovascular form of AMD—wet AMD—leads to vision loss due to abnormal growth of blood vessels (choroidal neovascularization) in the choriocapillaris through Bruch's membrane, which ultimately leads to leakage of blood and protein below the macula. Bleeding, leaking, and scarring of these blood vessels, if left untreated, can lead to retinal pigment epithelial detachment and permanent photoreceptor damage and rapid vision loss.

Brief Disclosure of the Present Invention

As noted above, treatment of an eye disease or eye disorder associated with excess vascularization, such as wet age-related macular degeneration, is urgently needed. Such treatment can prevent or reduce damage to ocular tissues such as the macula, which will significantly improve the prognosis for subjects suffering from such eye diseases or ocular disorders.

The present disclosure relates to a composition containing a compound of formula (I) for use in the treatment of an ocular disease or ocular disorder associated with excessive vascularization of the eye. The authors of the present disclosure unexpectedly found that treatment of induced vascularization of ocular tissues resulted in reduced vascularization of the cornea or choroid. More specifically, the present inventors have surprisingly found that compositions containing the compounds of the present invention suppress choroidal neovascularization in a rat model of laser-induced choroidal neovascularization. The present inventors also discovered an unexpected effect in the treatment of growth factor-stimulated neovascularization in a mouse model where compositions containing compounds of the present invention were able to reduce the area of vascularization. These findings have led to the development of a new treatment pathway for diseases and disorders associated with excess vascularization of the eye, which may lead to a better prognosis for patients suffering from diseases such as

- 1 045318 as corneal neovascularization, iris neovascularization, ciliary body neovascularization, corneal pannus, choroidal neovascularization, retinal neovascularization, wet age-related macular degeneration, proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy.

Thus, the present disclosure relates to a composition containing a compound of any of formulas (I) to (IX), for use in the treatment of an eye disease or ocular disorder.

One object of the present disclosure is a composition containing a compound of formula (IX)

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

One object of the present invention is a composition containing a compound of formula (I)

Where

R ¹ represents chlorine, R ² represents ethyl and R ³ represents hydrogen,

R ¹ represents methoxy, R ² represents methyl and R ³ represents trifluoromethyl,

R ¹ is chlorine, R ² is hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy, R ² represents hydrogen and R ³ represents trifluoromethyl;

or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

One particular subject of the present disclosure is a composition containing a compound of formula (II), formula (III), formula (IV), or formula (V)

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or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

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A particular subject of the present disclosure is a composition containing a compound selected from the group consisting of the following: laquinimod, 5-chloro-^ethyl-4-hydroxy^-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2- dihydroquinoline-3-carboxamide, 5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-M-ethyl-4-hydroxy-2-oxo -M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-M-ethyl-4,8-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide, 5-chloro-M-ethyl-4,7-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide, 5-chloro-M-ethyl-4,6-dihydroxy-1 -methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide, 5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-M-vinyl-1,2-dihydroquinoline- 3-carboxamide, 5-chloro-M-ethyl-4-hydroxy-M-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide, ]H-ethyl-4-hydroxy- 1 -methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5'-chloro-1-ethyl-1'-methyl-2'H-spiro[ind oline-3,3'- quinoline]-2,2',4'(1'H)-trione, tashinimod, 4-hydroxy-5-methoxy-M-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2 -dihydroquinoline-3carboxamide,

4,5-dihydroxy-N, 1-dimethyl-2-okco-N-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3 carboxamide,

4-hydroxy-5-methoxy-1-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,6-dihydroxy-5-methoxy-M,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide,

4,7-dihydroxy-5-methoxy-M,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihyde roquinoline-3-carboxamide and

4,8-dihydroxy-5-methoxy-K,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide, or a pharmaceutically acceptable salt thereof, for use in the treatment of ocular disease or eye disorder.

One object of the present disclosure is a method of treating an eye disease or ocular disorder, wherein said method comprises administering a composition containing a compound of formula (IX)

R ⁶

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

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R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof.

One object of the present disclosure is the use of a compound of formula (IX)

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of an eye disease or eye disorder.

Description of the figures

Fig. 1. Photograph of a mouse eye with clearly visible vascularity. The overlay figure shows how to measure the mean vessel length (VL) from the limbal vessels towards the pellet and continuous circular zone (on the clock = CH).

Fig. 2. Suppression of corneal vascularization induced by VEGF. Laquin. = laquinimod. Avastin 5 mg/kg leads to 100% suppression (reference). Dose 0.5 mg/kg laquinimod bid. does not lead to suppression. Dose 2.5 mg/kg laquinimod bid. results in 46% suppression. Dose 0.5 mg/kg laquinimod qid. results in 8% suppression. Dose 2.5 mg/kg laquinimod qid. results in 50% suppression.

Fig. 3. Suppression of corneal vascularization induced by bFGF. Laquin. = laquinimod. Sutent 40 mg/kg results in 80% suppression. Dose 0.5 mg/kg laquinimod big. results in 11% suppression. Dose 2.5 mg/kg laquinimod bid. results in 35% suppression. Dose 0.5 mg/kg laquinimod qid. results in 37% suppression. Dose 2.5 mg/kg laquinimod qid. results in 56% suppression.

Fig. 4. Normalized effect size for the average vessel length. The data were normalized so that the value for the diluent group corresponded to a mean effect size of 0% and for the sulforaphane (positive control) group a mean effect size of 100%. The data for Eylea support a larger effect size (effect size 18%). For the group receiving study compound ABR215174 (effect size 88.2%), the effect size was significantly greater than for the group receiving diluent. Data for the test compound ABR215062 support an increase in effect size (effect size 53%).

Detailed Disclosure of the Present Invention

Definitions

The term C1-C4 alkyl means a fragment containing or consisting of 1, 2, 3 or 4 carbon atoms and a number of hydrogen atoms. Examples of C1-C4 alkyl groups are methyl, ethyl, vinyl, isopropyl, n-propyl, n-butyl, t-butyl, isobutyl or sec-butyl.

Laquinimod or ABR-215062 means a chemical compound of formula (II)

Tashinimod means a chemical compound of formula (III)

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ABR-215174 means carboxylic acid phenylamide, i.e. a compound of formula (IV)

CF3 formula (III).

5-chloro-4-hydroxy-1-methyl-2-oxo-1,2-dihydroquinoline-3-

formula (IV).

ABR-215691 means 4-hydroxy-5-methoxy-1-methyl-2-oxo-N-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide, i.e. the compound of formula (V)

CF ₃ formula (V).

Vascularization and neovascularization refer to the process by which new blood vessels are formed. Vascularization and neovascularization are used interchangeably in the present invention.

As used in the present invention, ocular vascularization is synonymous with ocular neovascularization.

Excessive vascularization means a phenomenon in which vascularization occurs to a degree that is detrimental to the normal functioning of the affected tissue. Such excess vascularization occurs during or as a consequence of ocular diseases or ocular disorders such as corneal neovascularization, iris neovascularization, ciliary body neovascularization, corneal pannus, uveal neovascularization, proliferative diabetic retinopathy, retrolental fibroplasia, ischemic retinopathy, retinal neovascularization and wet form age-related macular degeneration.

As used herein, the terms ocular disease or ocular disorder associated with excessive vascularization of the eye and ocular disease or ocular disorder associated with ocular vascularization mean any ocular disease or ocular disorder that those skilled in the art consider to be caused and/or caused by vascularization of one or more more ocular tissue, for example, in which said vascularization is detrimental to the normal functioning of the affected tissue. Such diseases or disorders can lead to vision loss.

Treatment generally means eliminating, preventing, limiting and slowing, stopping or reversing the progression or severity of an eye disease or eye disorder.

When applied to vascularization, the term grade refers to the severity of vascularization. This degree of vascularization can be determined by various measurable parameters, such as the area of vascularization, the number of vessels in the vascularized area, the length of the vessels in the vascularized area, or the thickness of the vessels in the vascularized area.

Laquinimod diluent means the diluent used for laquinimod. The diluent does not contain laquinimod.

VEGF stands for vascular endothelial growth factor. In mammals, the VEGF family includes five members, namely, VEGF-A, VEGF-B, VEGF-C, VEGF-D and placental growth factor (PGF). VEGF stimulates cellular responses by binding to the VEGF receptor (VEGFR).

bFGF stands for basic fibroblast growth factor. Compounds and compositions for use

One embodiment of the present disclosure is a composition containing a compound of formula (I)

Where

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R ¹ represents chlorine, R ² represents ethyl and R ³ represents hydrogen,

R ¹ represents methoxy, R ² represents methyl and R ³ represents trifluoromethyl,

R ¹ is chlorine, R ² is hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy, R ² represents hydrogen and R ³ represents trifluoromethyl;

or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

Another embodiment of the present disclosure is a composition containing a compound of formula (I), wherein R ¹ is chlorine, R ² is ethyl and R ³ is hydrogen, for use in treating an eye disease or ocular disorder. The common name for the compound where R ¹ is Cl, R ² is ethyl and R ³ is H is laquinimod.

Another embodiment of the present disclosure is a composition containing a compound of formula (I), wherein R ¹ is methoxy, R ² is methyl, and R ³ is trifluoromethyl, for use in the treatment of an eye disease or ocular disorder. The common name for the compound where R ¹ is methoxy, R ² is methyl and R ³ is trifluoromethyl is taschinimod.

One embodiment of the present invention is a method of treating an eye disease or ocular disorder, said method comprising administering to a subject in need thereof a composition containing a therapeutically effective amount of a compound of formula (I)

R ¹ OH

R ³ formula (I), where

R ¹ represents chlorine, R ² represents ethyl and R ³ represents hydrogen,

R ¹ represents methoxy, R ² represents methyl and R ³ represents trifluoromethyl,

R ¹ is chlorine, R ² is hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy, R ² represents hydrogen and R ³ represents trifluoromethyl;

or a pharmaceutically acceptable salt thereof.

In one embodiment, the present disclosure relates to the use of a compound of formula (I)

R ¹ OH

R ³ formula (I), where

R ¹ represents chlorine, R ² represents ethyl and R ³ represents hydrogen,

R ¹ represents methoxy, R ² represents methyl and R ³ represents trifluoromethyl,

R ¹ is chlorine, R ² is hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy, R ² represents hydrogen and R ³ represents trifluoromethyl;

or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of an eye disease or eye disorder.

The present invention also discloses metabolites of laquinimod and tashinimod. Laquinimod and tashinimod are metabolized after administration to a subject. Some metabolites, such as those disclosed in the present invention, have therapeutic activity. One embodiment of the present disclosure is a composition containing laquinimod, taquinimod, or an active metabolite thereof, for use in treating an ocular disease or ocular disorder. In one embodiment of the present disclosure, laquinimod or its active metabolite is administered to a person in need of

- 7 045318 to this subject. In another embodiment, tashinimod or its active metabolite is administered to a subject in need thereof.

Metabolites of laquinimod include those formed by hydroxylation of quinoline at various positions, demethylation of quinoline, deethylation of aniline, and hydroxylation of aniline at the para position. Specific examples of laquinimod metabolites include

5-chloro-X-ethyl-4-hydroxy-X-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-X-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-X-ethyl-4-hydroxy-2-oxo-X-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-X-ethyl-4,8-dihydroxy-1-methyl-2-oxo-X-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-X-ethyl-4,7-dihydroxy-1-methyl-2-oxo-X-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-X-ethyl-4,6-dihydroxy-1-methyl-2-oxo-X-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-X-phenyl-X-vinyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-X-ethyl-4-hydroxy-X-(4 -hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

X-ethyl-4-hydroxy-1-methyl-2-oxo-X-phenyl-1,2-dihydroquinoline-3-carboxamide and

5'-chloro-1-ethyl-G-methyl-2'H-spiro[indoline-3,3'-quinoline]-2,2',4'(HN)-trione.

A preferred embodiment of the present disclosure is laquinimod, a metabolite of 5-chloro-4-hydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinoline-3-carboxamide (phenylamide 5-chloro4-hydroxy-1-methyl- 2-oxo-1,2-dihydroquinoline-3-carboxylic acid, ABR-215174).

Metabolites of tashinimod include those formed by demethylation of aniline, demethylation of quinoline at the N atom, and demethylation of quinoline at the O atom. Specific examples of metabolites of tashinimod include

4-hydroxy-5-methoxy-M-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,5-dihydroxy-N, 1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3 carboxamide and

4-hydroxy-5-methoxy-1-methyl-2-oxo-X-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,6-dihydroxy-5-methoxy-X,1-dimethyl-2-oxo-X-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide,

4,7-dihydroxy-5-methoxy-X,1-dimethyl-2-oxo-X-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide and

4,8-dihydroxy-5-methoxy-X,1-dimethyl-2-oxo-X-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide.

A preferred embodiment of the present disclosure is the metabolite of taschinimod, 4-hydroxy-5-methoxy-1-methyl-2-oxo-N-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3-carboxamide.

In a preferred embodiment of the present disclosure, the compound is selected from the group consisting of the following: laquinimod, tashinimod, 5-chloro-4-hydroxy-1-methyl-2-oxo1,2-dihydroquinoline-3-carboxylic acid phenylamide (compound of formula (IV) ) and 4-hydroxy-5-methoxy-1-methyl2-oxo-N-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3-carboxamide (compound of formula (V)).

One embodiment of the present disclosure is a composition containing a compound selected from the group consisting of: laquinimod,

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5-chloro-N-ethyl-4-hydroxy-H-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-M-ethyl-4-hydroxy-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-M-ethyl-4,8-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-M-ethyl-4,7-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-L1-ethyl-4,6-dihydroxy-1-methyl-2-oxo-L1-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-M-vinyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-M-ethyl-4-hydroxy-M-(4 -hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

M-ethyl-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5'-chloro-1-ethyl-G-methyl-2'H-spiro[indoline -3,3'-quinoline]-2,2',4'(GN)-trione, tashinimod,

4-hydroxy-5-methoxy-M-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,5-dihydroxy-N, 1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3 carboxamide,

4-hydroxy-5-methoxy-1-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,6-dihydroxy-5-methoxy-bT,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide,

4,7-dihydroxy-5-methoxy-L'G,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide and

4,8-dihydroxy-5-methoxy-L'G,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3-carboxamide, or a pharmaceutically acceptable salt thereof, for use in treating an eye disease or eye disorder, wherein said eye disease or eye disorder is associated with excessive vascularization of the eye.

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One embodiment of the present disclosure is a method of treating an eye disease or ocular disorder, said method comprising administering to a subject in need thereof a composition containing a therapeutically effective amount of a compound selected from the group consisting of: laquinimod,

5-chloro-X-ethyl-4-hydroxy-X-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-M-ethyl-4-hydroxy-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-X-ethyl-4,8-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-M-ethyl-4,7-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-M-ethyl-4,6-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-M-vinyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-X-ethyl-4-hydroxy-X-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

X-ethyl-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5'-chloro-1-ethyl-G-methyl-2'H-spiro[indoline-3,3'-quinoline]-2,2',4'(1'H)-trione, tashinimod,

4-hydroxy-5-methoxy-M-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,5-dihydroxy-N, 1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3 carboxamide,

4-hydroxy-5-methoxy-1-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,6-dihydroxy-5-methoxy-T4,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide,

4,7-dihydroxy-5-methoxy-N,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide and

4,8-dihydroxy-5-methoxy-N,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide, or a pharmaceutically acceptable salt thereof.

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One embodiment of the present disclosure is the use of a compound selected from the group consisting of the following: laquinimod, 5-chloro-^ethyl-4-hydroxy^-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline -3-carboxamide, 5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-M-ethyl-4-hydroxy-2-oxo- M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-M-ethyl-4,8-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide, 5 -chloro-M-ethyl-4,7-dihydroxy-1 -methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide, 5-chloro-M-ethyl-4,6-dihydroxy-1 - methyl 2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide, 5-chloro-4-hydroxy-1 - methyl-2-oxo-T4-phenyl-T4-vinyl-1,2-dihydroquinoline-3 -carboxamide, 5-chloro-M-ethyl-4-hydroxy-M-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

M-ethyl-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide, 5'-chloro-1-ethyl-G-methyl-2'H-spiro[indoline -3,3'-quinoline]-2,2',4'(1'H)-trione, tashinimod,

4-hydroxy-5-methoxy-M-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,5-dihydroxy-N, 1-dimethyl-2-oxo-N-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3 carboxamide,

4-hydroxy-5-methoxy-1-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,6-dihydroxy-5-methoxy-M,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide,

4,7-dihydroxy-5-methoxy-T4,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihyde roquinoline-3-carboxamide and

4,8-dihydroxy-5-methoxy-M,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide, or a pharmaceutically acceptable salt thereof for the preparation of a medicinal product for treatment eye disease or eye disorder.

One embodiment of the present disclosure is a composition containing a compound of formula (VI)

R ⁶

Where

R ¹ represents chlorine,

R ² represents ethyl or hydrogen,

R ³ represents hydrogen or hydroxy group,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

One embodiment of the present disclosure is a composition containing a compound of formula (VII)

Where

R ¹ represents a methoxy group or a hydroxy group,

R ² represents methyl or hydrogen,

R ³ represents trifluoromethyl,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

One embodiment of the present disclosure is a composition containing a compound of formula (VIII)

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy, and

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine, or a pharmaceutically acceptable salt thereof, for use in the treatment of an eye disease or ocular disorder.

Composition containing a compound of formula (IX)

R ⁶

formula (IX) where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

One embodiment of the present disclosure is a method of treating an eye disease, said method comprising administering a composition containing a therapeutically effective amount of a compound of formula (VI), formula (VII), formula (VIII), formula (IX) disclosed in the present invention , or a pharmaceutically acceptable salt thereof.

One embodiment of the present disclosure is the use of a compound of formula (VI), formula (VII), formula (VIII), or formula (IX) disclosed in the present invention, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of an eye disease or ocular disorder .

Excessive vascularization of eye tissue

The present disclosure relates to the treatment of an eye disease or eye disorder associated with excessive vascularization of the eye. This vascularization may occur in response to the eye being exposed to external stimuli, such as excessive stress. Vascularization can also occur as a natural result of aging.

Vascularization of some ocular tissues can be harmful to vision. The eye is made up of many different tissues, such as the cornea, iris, ciliary body, choroid, retina, or macula. Each of these tissues can undergo vascularization.

One embodiment of the present invention is a compound for treating a subject where the subject suffers from vascularization of the cornea, iris, ciliary body, choroid, retina, or maculum.

One embodiment of the present invention is a compound for treating a subject where the subject suffers from vascularization of tissue within the eye, such as the cornea, iris or ciliary body. It is important for vision to function that the cornea is transparent. Therefore, vascularization of the cornea is detrimental to the subject's vision. Thus, a preferred embodiment of the present disclosure is a composition containing a compound according to the present disclosure for the treatment of corneal neovascularization.

In one embodiment of the present disclosure, the eye disease or eye disorder is selected from the group consisting of the following: corneal neovascularization, iris neovascularization, ciliary body neovascularization, corneal pannus, uveal neovascularization, retinal neovascularization, wet age-related macular degeneration, proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy.

In one embodiment of the present disclosure, the eye disease or eye disorder is associated with excessive vascularization of the eye. In one embodiment of the present disclosure, the ocular disease or ocular disorder is corneal neovascularization, iris neovascularization, ciliary body neovascularization, corneal pannus, uveal neovascularization, retinal neovascularization, wet age-related macular degeneration, proliferative diabetic retinopathy, retrolental fibroplasia, or ischemic retinopathy. associated with excessive vascularization of the eye.

Retinopathy is a disorder of the retina that can cause vision impairment. Retinopathy may be a retinal vascular disease or damage to the retina caused by abnormal blood flow. Thus, one embodiment of the present invention is a compound for treating a subject where the subject suffers from proliferative diabetic retinopathy, retrolental fibroplasia or ischemic retinopathy. Diabetes is a common cause of retinopathy and diabetic retinopathy is one of the leading causes of blindness in working-age people. Thus, one embodiment of the present disclosure is a composition containing a compound according to the present disclosure for treating a subject where the subject suffers from proliferative diabetic retinopathy.

One embodiment of the present invention is a compound for treating a subject where the subject suffers from vascularization of tissue of the back of the eye, such as the choroid, retina or macula. The purpose of the retina is to perceive light that is focused by the lens, convert the light into nerve signals, and send these signals to the brain for visual recognition. Therefore, any retinal disorder such as vascularization may affect the subject's vision. Thus, a preferred embodiment of the present disclosure is a composition containing a compound of the present disclosure for treating a subject suffering from retinal neovascularization.

The macula is the central part of the retina. The spot provides high-resolution central color vision, which is possible in good lighting. Damage to the spot results in loss of central vision, which can severely impact the subject's ability to read and recognize faces. Thus, the most preferred embodiment of the present disclosure is a composition containing a compound according to the present disclosure for treating a subject suffering from macular vascularization. Macular vascularization is also known as the wet form of age-related macular degeneration. Thus, the most preferred embodiment of the present disclosure is a composition containing a compound according to the present disclosure for treating a subject suffering from the wet form of age-related macular degeneration. In one embodiment of the present disclosure, the wet form of age-related macular degeneration is associated with excessive vascularization of the eye.

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In a preferred embodiment of the present disclosure, the term ocular diseases or ocular disorders associated with excessive vascularization of the eye does not include uveitis or conjunctivitis.

Administration of the disclosed compositions together with VEGF inhibitors.

Vascular endothelial growth factor (VEGF) stimulates the formation of blood vessels. VEGF inhibitors have the ability to reduce tissue vascularization by binding to VEGF. Alternatively, VEGF inhibitors may affect VEGF activity by binding to the VEGF receptor.

The compositions of the present disclosure can be administered in combination with a VEGF inhibitor for the treatment of an ocular disease or ocular disorder, such as those associated with excessive vascularization of the eye. Such combination treatment may be more effective for treating an ocular disease or ocular disorder than the compositions of the present disclosure and/or the VEGF inhibitor alone. Thus, in one embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with administration of a VEGF inhibitor.

VEGF inhibitors include antibodies, antibody-derived fragments, recombinant proteins and recombinant fusion proteins such as aflibercept, ranibizumab, bevacizumab, brolucizumab, abicipar pegol, conbercept and faricimab. Thus, in one embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of aflibercept, ranibizumab, bevacizumab, brolucizumab, abicipar pegol, conbercept, or faricimab. In a preferred embodiment, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of aflibercept, ranibizumab, bevacizumab or brolucizumab.

The recombinant protein aflibercept (Eylea) has affinity for VEGF-A, VEGF-B and PGF. This VEGF inhibitor has been shown to be effective in the treatment of wet AMD. In one preferred embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of aflibercept.

The monoclonal antibody fragment ranibizumab has affinity for VEGF-A. This antibody fragment is known to be effective in the treatment of wet AMD. In one embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of ranibizumab.

Bevacizumab (Avastin) is an IgG1-based antibody that binds VEGF-A. Bevacizumab has been shown to be effective for the treatment of wet AMD. Thus, in one embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of bevacizumab.

Brolucizumab is an sc antibody fragment that has affinity for VEGF-A. In one embodiment of the present disclosure, an ocular disease or eye is treated by administering a composition according to the present disclosure in combination with the administration of bevacizumab.

Abicipar pegol peptide is a known VEGF-A inhibitor. In one embodiment of the present disclosure, an eye disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of abicipar pegol.

Conbercept is a recombinant fusion protein and has an affinity for VEGF-A. Thus, in one embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a composition according to the present disclosure in combination with the administration of conbercept.

Bispecific monoclonal antibodies faricimab modulate the activity of angiopoietin-2 and VEGF-A. Thus, in one embodiment of the present disclosure, an ocular disease or ocular disorder is treated by administering a compound of the present disclosure in combination with faricimab.

In one embodiment of the present disclosure, the composition of the present disclosure contains or is administered in combination with an angiogenesis inhibitor. In a preferred embodiment of the present disclosure, the angiogenesis inhibitor is aflibercept.

In one embodiment of the present disclosure, a composition containing a compound of the present disclosure may be administered to a subject in need thereof by topical, oral, intravitreal, subconjunctival, retrobulbar, intracameral, or systemic routes.

To treat a disease or disorder that is highly localized to one part of the body, such as an eye disease and/or an eye disorder, it may be useful to administer a drug for the disease or disorder in a manner that ensures that the drug is located primarily at the site of the disease or disorder . Thus, in a preferred embodiment of the present disclosure, a composition containing a compound according to the present disclosure can be administered to a subject in need thereof by the topical, intravitreal, subconjunctival, retrobulbar, or intracameral route.

In one embodiment of the present disclosure, the composition according to the present disclosure is administered by a route that provides the effect of systemic administration of the composition. In another version

- 14 045318 implementation of the present disclosure, the administration of the composition according to the present disclosure is oral administration.

For the treatment of a disease or disorder where the disease or disorder requires frequent dosing of a medicament to a subject, it may be useful to formulate the medicament in a manner that allows for self-administration. One skilled in the art will know which types of formulations are suitable for self-administration.

When used in the present invention, the compositions of the present invention can be administered in combination with a VEGF inhibitor. Thus, in one embodiment, the composition of the present disclosure and the VEGF inhibitor are contained in the same composition and are therefore administered simultaneously.

The combination treatment described in the present invention is not limited to compositions containing a compound according to the present disclosure and a VEGF inhibitor. In contrast, the compound of the present invention and the VEGF inhibitor can be administered in different formulations. The choice of the administered compound of the present disclosure and the VEGF inhibitor as separate formulations may be influenced by the fact that the compound of the present disclosure and the VEGF inhibitor have different preferred modes of administration. For example, it may be useful to administer a compound of the present disclosure frequently, such as daily or once a week, while a VEGF inhibitor may be advantageously administered infrequently, such as once every few months. The choice of the administered compound of the present disclosure and the VEGF inhibitor as separate formulations may be influenced by the fact that the compound of the present disclosure and the VEGF inhibitor are not suitable for the same type of route of administration. For example, a compound of the present disclosure may be particularly suitable for one type of route of administration, such as topical administration, and a VEGF inhibitor may be suitable for a second type of route of administration, such as intravitreal injection. Thus, in one embodiment, a compound of the present disclosure and a VEGF inhibitor are contained in different formulations where the formulations are administered at different frequencies. In another embodiment, the compound of the present disclosure and the VEGF inhibitor are contained in different formulations, where the formulations are administered using different routes of administration.

VEGF inhibitors are typically administered by intravitreal injection to treat ocular diseases or ocular disorders such as those associated with ocular vascularization. Vitreous injections are often performed in a hospital or general practitioner's office. Intravitreal injections for the treatment of diseases associated with vascularization of the eye are usually performed every few months, such as once a month, such as once every 3 months, or once every 6 months. Injections into the vitreous body are performed under local anesthesia. Side effects of intravitreal injection include increased eye pressure, floating cells, inflammation, bleeding, scratched cornea, damage to the retina or surrounding nerves, and infection. In addition, having to visit a general practitioner or hospital every few months for an injection may be inconvenient and subjects receiving intravitreal injections may find the experience unpleasant and difficult. Topical treatment with a compound of the present disclosure before, during, or between intravitreal injections of VEGF inhibitors may prolong the period between additional VEGF injections required. Thus, in one embodiment of the present disclosure, topical treatment of an ocular disease or ocular disorder, such as those associated with excess vascularization of the eye, with a composition containing a compound of the present disclosure results in intravitreal injections of a VEGF inhibitor being required less frequently than when administered only intravitreal injections of a VEGF inhibitor. Additionally, in one embodiment, a compound of the present disclosure and a VEGF inhibitor are contained in different compositions, wherein administration of a composition containing a compound of the present disclosure provides treatment for an ocular disease or ocular disorder and reduces the frequency of required intravitreal injections of the VEGF inhibitor.

Treatment

One embodiment of the present disclosure is a composition containing a compound according to the present disclosure. Another embodiment of the present disclosure is a composition comprising a compound of the present disclosure and a pharmaceutically acceptable excipient.

There are different ways to administer drugs into the eye. For example, drugs can be administered topically to the eye. Thus, in one embodiment, a composition containing a compound of the present disclosure is administered topically to the eye. One skilled in the art will understand what types of routes of administration are suitable for administration to the eye.

In one embodiment of the present disclosure, a composition containing a compound of the present disclosure is administered orally.

One embodiment of the present disclosure is a composition containing a compound of formula (IX)

- 15 045318

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ is methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for use in the treatment of an eye disease or eye disorder, wherein the eye disease or eye disorder is a wet form of age-related macular degeneration.

One embodiment of the present disclosure is a method of treating wet age-related macular degeneration in a subject, said method comprising administering to the subject a composition containing a compound of formula (I)

Where

R ¹ represents chlorine, R ² represents ethyl and R ³ represents hydrogen,

R ¹ represents methoxy group, R ² represents methyl and R ³ represents trifluoromethyl,

R ¹ is chlorine, R ² is hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy, R ² represents hydrogen and R ³ represents trifluoromethyl.

Another embodiment of the present disclosure is a method of treating wet age-related macular degeneration in a subject, said method comprising administering laquinimod to said subject. Another embodiment of the present disclosure is a method of treating a wet form of age-related macular degeneration, the method comprising administering to a subject a therapeutically effective amount of taschinimod. Another embodiment of the present disclosure is a method of treating wet age-related macular degeneration, which method comprises administering to a subject a therapeutically effective amount of ABR-215691. Another embodiment of the present disclosure is a method of treating wet age-related macular degeneration, which method comprises administering to a subject a therapeutically effective amount of ABR-215174.

The compounds disclosed in the present invention can be used for the preparation of drugs. Thus, in one embodiment of the present invention, the compound of formula (I)

Where

R ¹ represents chlorine, R ² represents ethyl and R ³ represents hydrogen,

R ¹ represents methoxy, R ² represents methyl and R ³ represents trifluoromethyl,

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R ¹ is chlorine, R ² is hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy group, R ² represents hydrogen and R ³ represents trifluoromethyl, is used for the preparation of a medicament for the treatment of wet age-related macular degeneration. In another embodiment, laquinimod is used to formulate a medicament for the treatment of wet age-related macular degeneration. In yet another embodiment, tashinimod is used to formulate a medicament for the treatment of wet age-related macular degeneration. In yet another embodiment, ABR215174 is used to formulate a medicament for the treatment of wet age-related macular degeneration. In yet another embodiment, ABR-215691 is used to formulate a medicament for the treatment of wet age-related macular degeneration.

Paragraphs.

1. Composition containing a compound of formula (IX)

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for use in the treatment of an ocular disease or ocular disorder.

2. Composition for use according to paragraph 1, where the compound is a compound of formula (VIII)

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy, and

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine, or a pharmaceutically acceptable salt thereof.

3. Composition for use according to any of the previous paragraphs, wherein the compound is a compound of formula (VII)

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Where

R ¹ represents a methoxy group or a hydroxy group,

R ² represents methyl or hydrogen,

R ³ represents trifluoromethyl,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof.

4. Composition for use according to any of the previous paragraphs, wherein the compound is a compound of formula (VI)

Where

R ¹ represents chlorine,

R ² represents ethyl or hydrogen,

R ³ represents hydrogen or hydroxy group,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof.

5. Composition containing 5'-chloro-1-ethyl-1-methyl-2'H-spiro[indoline-3,3'-quinoline]-2,2',4'(1Ή)trione or a pharmaceutically acceptable salt thereof , for use in the treatment of an eye disease or eye disorder.

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6. A composition for use according to any of the preceding paragraphs, wherein the compound is a compound selected from the group consisting of the following: laquinimod,

5-chloro-X-ethyl-4-hydroxy-N-(4-hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-M-ethyl-4-hydroxy-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide,

5-chloro-M-ethyl-4,8-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-M-ethyl-4,7-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-M-ethyl-4,6-dihydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3 carboxamide,

5-chloro-4-hydroxy-1-methyl-2-oxo-M-phenyl-M-vinyl-1,2-dihydroquinoline-3-carboxamide, 5-chloro-M-ethyl-4-hydroxy-M-(4 -hydroxyphenyl)-1-methyl-2-oxo-1,2-dihydroquinoline-3 carboxamide,

M-ethyl-4-hydroxy-1-methyl-2-oxo-M-phenyl-1,2-dihydroquinoline-3-carboxamide, tashinimod,

4-hydroxy-5-methoxy-M-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,5-dihydroxy-N, 1-dimethyl-2-oxo-X-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3 carboxamide,

4-hydroxy-5-methoxy-1-methyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2-dihydroquinoline-3carboxamide,

4,6-dihydroxy-5-methoxy-X,1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide,

4,7-dihydroxy-5-methoxy-M,1-dimethyl-2-oxo-X-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide and

4,8-dihydroxy-5-methoxy-M)1-dimethyl-2-oxo-M-(4-(trifluoromethyl)phenyl)-1,2dihydroquinoline-3-carboxamide, or a pharmaceutically acceptable salt thereof.

7. Composition for use according to any of the previous paragraphs, wherein the compound is a compound of formula (I)

R ¹ OH

R ³ formula (I), where

R ¹ is chlorine, R ² is ethyl or hydrogen and R ³ is hydrogen, or

R ¹ represents methoxy, R ² represents methyl or hydrogen and R ³ represents trifluoromethyl;

or a pharmaceutically acceptable salt thereof.

8. A composition for use according to any of the preceding paragraphs, wherein the compound is laquinimod or a pharmaceutically acceptable salt thereof.

9. A composition for use according to any of the preceding paragraphs, wherein the compound is tasquinimod or a pharmaceutically acceptable salt thereof.

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10. Composition for use according to any of the previous paragraphs, wherein the compound is a compound of formula (IV)

or a pharmaceutically acceptable salt thereof.

11. Composition for use according to any of the previous paragraphs, wherein the compound is a compound of formula (V)

or a pharmaceutically acceptable salt thereof.

12. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is selected from the group consisting of the following: corneal neovascularization, iris neovascularization, ciliary body neovascularization, corneal pannus, uveal neovascularization, retinal neovascularization, wet age-related macular degeneration, proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy.

13. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is selected from the group consisting of the following: corneal neovascularization, iris neovascularization, ciliary body neovascularization, and corneal pannus.

14. A composition for use according to any of the preceding paragraphs, wherein the ocular disease or ocular disorder is corneal neovascularization.

15. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is selected from the group consisting of proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy.

16. A composition for use according to any of the preceding paragraphs, wherein the eye disease or eye disorder is proliferative diabetic retinopathy.

17. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is retrolental fibroplasia.

18. A composition for use according to any of the preceding paragraphs, wherein the eye disease or eye disorder is ischemic retinopathy.

19. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is selected from the group consisting of the following: uveal neovascularization, retinal neovascularization, and wet age-related macular degeneration.

20. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is neovascularization of the choroid.

21. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is retinal neovascularization.

A composition for use as defined in any of the preceding paragraphs, wherein the eye disease or eye disorder is a wet form of age-related macular degeneration.

22. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is not uveitis or conjunctivitis.

23. A composition for use as defined in any of the preceding paragraphs, wherein the ocular disease or ocular disorder is associated with excessive vascularization of the eye.

24. A composition for use as defined in any of the preceding paragraphs, wherein the composition contains or is administered in combination with an angiogenesis inhibitor, such as aflibercept.

25. A composition for use as defined in any of the preceding paragraphs, wherein the composition is administered in combination with one or more VEGF inhibitors.

26. A composition for use as defined in any of the preceding paragraphs, wherein the composition is administered in combination with a single VEGF inhibitor.

27. A composition for use as defined in any of the preceding paragraphs, wherein the composition further comprises one or more VEGF inhibitors.

28. A composition for use as defined in any of the preceding paragraphs, wherein the composition further comprises one VEGF inhibitor.

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29. A composition for use as defined in any of the preceding paragraphs, wherein the VEGF inhibitor is selected from the group consisting of the following: aflibercept, ranibizumab, bevacizumab, brolucizumab, abicipar pegol, conbercept and faricimab.

30. A composition for use as defined in any of the preceding paragraphs, wherein the VEGF inhibitor is selected from the group consisting of aflibercept, ranibizumab, bevacizumab and brolucizumab.

31. The composition for is aflibercept.

32. The composition for is ranibizumab.

33. Composition for is bevacizumab.

34. Composition for is brolucizumab.

35. Composition for is abicipar pegol.

36. The composition for is conbercept.

37. Composition for use for any use for any use for any use for any use for any use for any use for any of the previous paragraphs, from the previous paragraphs, from the previous paragraphs, from the previous paragraphs, from the previous paragraphs, from the previous paragraphs, from the previous paragraphs, where a VEGF inhibitor where a VEGF inhibitor where a VEGF inhibitor where a VEGF inhibitor where a VEGF inhibitor where a VEGF inhibitor where the VEGF inhibitor is faricimab.

38. A composition for use according to any of the preceding paragraphs, wherein the composition contains or is administered in combination with at least one pharmaceutically acceptable excipient.

39. A composition for use as defined in any of the preceding paragraphs, wherein the route of administration is local, oral, intravitreal, subconjunctival, retrobulbar, intracameral or systemic.

40. Composition for use according to any of the previous paragraphs, where the route of administration is local.

41. A composition for use according to any of the previous paragraphs, where the route of administration is oral.

42. A composition for use according to any of the previous paragraphs, wherein the route of administration is intravitreal administration.

43. Composition for use according to any of the previous paragraphs, where the route of administration is subconjunctival.

44. Composition for use according to any of the previous paragraphs, where the route of administration is retrobulbar.

45. Composition for use according to any of the previous paragraphs, where the route of administration is intracameral.

46. Composition for use according to any of the previous paragraphs, where the route of administration is systemic.

47. A method of treating wet age-related macular degeneration, said method comprising administering to a subject in need thereof a composition containing a therapeutically effective amount of a compound of formula (IX)

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen,

- 21 045318 or a pharmaceutically acceptable salt thereof.

48. Use of a compound of formula (IX)

R ⁶

Where

R ¹ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ² is selected from the group consisting of the following: hydrogen and C1-C4 alkyl such as methyl, ethyl or vinyl,

R ³ is selected from the group consisting of the following: hydrogen, hydroxy, methyl, methoxy, fluorine, chlorine, bromine, trifluoromethyl and trifluoromethoxy,

R ⁴ is selected from the group consisting of the following: hydrogen, fluorine and chlorine, provided that

R ⁴ is selected from the group consisting of fluorine and chlorine only if R ³ is selected from the group consisting of fluorine and chlorine,

R ⁵ represents hydrogen or hydroxy and

R ⁶ represents methyl or hydrogen, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of an eye disease or eye disorder.

Example 1. Suppression of choroidal neovascularization in a rat model of laser-induced choroidal neovascularization.

Study design.

Forty-eight (48) pigmented Brown Norway rats were divided into six (6) groups of eight (8) animals. Choroidal neovascularization was induced using a 532 nm argon laser photocoagulator (six (6) 75 μm spots at 150 mW for 0.1 s) in the right eyes on day 0. The test agent was administered three times daily by instillation or twice daily. day by oral administration from day 0 (D0) immediately after ChNV induction until day 21 (the last day of the study). The control agent (diluent) was instilled three times daily and the reference agent (dexamethasone in olive oil) was administered by daily oral administration from day 0 immediately after induction of neovascularization until day 21. Newly formed vessels at the fundus were examined in the right eyes on days 14 and 21 using Heidelberg's Retinal Angiograph (HRA). Lesion size was determined from flat-mounted choroidal preparations labeled with Isolectin-B4 at the end of the in vivo period.

Induction of neovascularization.

On day 0, animals were anesthetized by intramuscular injection of a mixture of xylazine (5 mg/kg) and ketamine (25 mg/kg). The pupils of the right eyes were dilated by instilling one drop of 0.5% tropicamide. Six (6) choroidal burns (spot diameter 75 μm) were then made using a slit lamp with a contact lens around the optic disc between the branches of the main vessel using an argon laser photocoagulator (532 nm; 150 mW; duration 0.1 s). The formation of a bubble during laser exposure confirmed the rupture of Bruch's membrane.

Route and method of administration.

Study, control, or reference agents were administered by topical instillation (10 μl each) or orally (1 ml/kg) from day 0 (immediately after induction) to day 21 (end of study). The study and control agents were administered three times a day (in the case of instillation) and twice a day (in the case of oral administration). The reference agent was administered once daily orally.

Body mass.

The body weights of the animals were recorded before the start of the study and then once a week.

Fluorescein angiography.

Fluorescein angiography was performed on days 14 and 21 using HRA (Heidelberg's Retinal Angiograph). After anesthesia (with the same mixture used to induce ChNV) and dilation of the pupils, 250 μl/100 g (body weight) of 10% fluorescein sodium salt solution was administered by subcutaneous injection using a 26-G insulin syringe and photographs of fluorescein were recorded 10 min after dye injections.

Fluorescein angiography study.

Fluorescein leakage on angiograms was determined by two investigators in a blinded manner and scored as follows:

Indicator 0, no leakage;

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Indicator 1, weak staining;

Indicator 2, moderate staining;

Indicator 3, strong staining.

If two scores for a particular lesion did not match, the highest score was used for analysis.

Data processing.

Statistical analysis using the Mann-Whitney U test or appropriate statistical model was performed on lesion scores visualized by HRA for each animal and on scores for flat-mounted preparations.

Animal behavior and mortality.

The general behavior and appearance of all animals were examined and reported in the raw data. No specific sign was detected and the general behavior and appearance of the animals were normal. All animals were alive before scheduled euthanasia.

Animal body weight.

Animal body weights were recorded before induction and treatment (baseline) and then once a week. The body weights of the animals in the initial state were in the normal range: 166-202 g (minimum - maximum, n=48). On day 21, no significant difference was found between the study and control agents. Animals treated with the reference drug (dexamethasone) experienced a 21% loss in body weight during the study. This loss is an expected side effect of an orally administered corticosteroid.

Angiography studies.

Fluorescein angiography (FA) evaluation is based on the fluorescence intensity of each lesion. For each treatment group, results are presented as the average for the group at that time point. Table 1 shows FA data recorded for 10 min on days 14 and 21 (n=8 animals per group, right eyes). Statistical analysis was performed on the median of individual intensity scores using the Kruskal-Wallis test followed by the Mann-Whitney U multiple comparison test, which was used to compare each study, control, or reference group.

Table 1

FA study for treatment group

Treatment Route of administration Administration mode Moment of time Average Fluorescein Leakage Rate Average value (n=nb studied spots/48) % reduction compared to the corresponding thinner Laquinimod Burying 5% 3x/day Day 14 0.7 (n = 38) 0% Day 21 0.4 (n = 45) 63% 1% 3x/day Day 14 1.1 (n = 36) <0% Day 21 1.2 (n = 46) <0% 0.2% Zx/day Day 14 0.7 (n = 35) 0% Day 21 0.9 (n = 42) 18% Per os 1 mg/kg 2x/day Day 14 0.8 (n = 42) <0% Day 21 0.9 (n = 38) 18% Laquinimod diluent Burying 3x/day Day 14 0.7 (n = 35) - Day 21 1.1 (n = 38) - Dexamethasone Orally 0.5 mg/kg Day 14 0.7 (n = 48) 71% 1 x/day Day 21 0.8 (n = 48) 73%

Conclusion.

On day 14 and day 21, 63% and 66% of the stains tested showed leakage in the diluent- and product-treated animals, respectively, indicating formation and persistence

ChNV. However, daily topical administration of laquinimod effectively reduces vascular leakage and this shows that laquinimod effectively counteracts the manifestation of ChNV.

Example 2 Suppression of VEGF- and bFGF-induced neovascularization.

Method for determining the area of neovascularization.

The area of neovascularization can be determined by the formula:

where the length of the vessel (VL) and the continuous circular zone (on the clock =CH) are determined as shown in Fig. 1.

Laquinomod treatment of VEGF-induced corneal vascularization.

Hydron pellets for inducing vascularization were prepared from a stimulant (VEGF) and a coupling agent (sucralfate).

6- to 8-week-old female CR C57BL/6 mice were prepared for surgery by anesthetizing with 90 mg/kg pentobarbital, ip (intraperitoneal). Corneal vascularization was induced by placing the pellet in a corneal pocket cut into the eye. Monitor closely for signs of eye irritation or infection.

Doses of laquinimod in DI (deionized) water, diluent (DDW = double distilled water) and Avastin (positive control) were administered directly into the eye containing the pellet. Vascularization was determined on day 8. The treatment regimen is given in Table. 2. The results are shown in table. 3 and in fig. 2.

table 2

Treatment regimens for inducing corneal vascularization via VEGF followed by laquinimod

Group η Treatment mode 1 Treatment mode 2 Means ng/animal Path Scheme Means µg/animal Path Scheme 1 8 VEGF 129 Wed qdxl - - - - 2 4 VEGF 129 Wed qdxl avas. 5* ip qdx7 3 4 VEGF 129 Wed qdxl veh. - local qdx7 first day 2 doses 4 8 VEGF 129 Wed qdxl laqui. 10 local bidx7 first day 1 dose 5 8 VEGF 129 Wed qdxl laqui. 50 local bidx7 first day 1 dose 6 8 VEGF 129 Wed qdxl laqui. 10 local qidx7 first day 2 doses 7 8 VEGF 129 Wed qdxl laqui. 50 local qidx7 first day 2 doses

*mg/kg av=in the corneal pocket bid=twice a day qd=daily qid=four times a day laqui.=laquinimod veh.=diluent αvas.=avastin

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Table 3

Treatment results for VEGF-induced corneal vascularization

Group η Pellet facilities Area vasc. Area as a percentage of the positive control area 1 8 VEGF 1.47 ±0.09 - 2 4 VEGF 0.00±0.00 0% 3 4 VEGF 1.63 ±0.13 111% 4 8 VEGF 1.49 ±0.13 101% 5 7 VEGF 0.79 ± 0.06 54% 6 8 VEGF 1.35 ±0.09 92% 7 8 VEGF 0.74 ±0.11 50%

vasc.=vascularization

Treatment of vascularization induced by VEGF is characterized by a dose-dependent effect after administration of laquinimod: low dose (0.5 mg/kg, bid, group 4) does not lead to any suppression. Increasing the frequency of administration resulted in only slight suppression (0.5 mg/kg, qid, group 6, 8% suppression). Increasing dose results in significant suppression of vascularization, with low (2.5 mg/kg, bid, group 5, 46% suppression) and high (2.5 mg/kg, qid, group 7, 50% suppression) doses at day.

Laquinomod treatment of bFGF-induced corneal vascularization.

Hydron pellets for inducing vascularization were prepared from a stimulant (bFGF) and a coupling agent (sucralfate).

6- to 8-week-old female CR C57BL/6 mice were prepared for surgery by anesthetizing with 90 mg/kg pentobarbital, ip. Corneal vascularization was induced by placing the pellet in a corneal pocket cut into the eye. Monitor closely for signs of eye irritation or infection.

Doses of laquinimod in DI (deionized) water, diluent (DDW = double distilled water) and Avastin (positive control) were administered directly into the eye containing the pellet. Vascularization was determined on day 6. The treatment regimen is given in Table. 4. The results are shown in table. 5 and fig. 3.

Table 4

Treatment regimens for inducing corneal vascularization with bFGF followed by laquinimod

Group η Treatment mode 1 Treatment mode 2 Means ng/animal Path Scheme Means µg/animal Path Scheme 1 8 bFGF 200 SR qdxl - - - - 2 6 bFGF 200 SR qdxl sutent 5* po qdx5 3 4 bFGF 200 SR qdxl veh. - local qdx5 first day 2 doses 4 6 bFGF 200 SR qdxl laqui. 10 local bidx5 first day 1 dose 5 6 bFGF 200 Wed qdxl laqui. 50 local bidx5 first day 1 dose 6 6 bFGF 200 SR qdxl laqui. 10 local qidx5 first day 2 doses 7 6 bFGF 200 Wed qdxl laqui. 50 local qidx5 first day 2 doses

*mg/kg laqui.=laquinimod veh.=diluent

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Table 5

Treatment results for bFGF-induced corneal vascularization

Group η Pellet products Area vase. Area as a percentage of the positive control area 1 8 bFGF 3.54 ±0.10 - 2 6 bFGF 0.72 ± 0.08 20% 3 4 bFGF 3.54 ±0.15 100% 4 6 bFGF 3.16 ±0.06 89% 5 6 bFGF 2.30 ± 0.23 65% 6 6 bFGF 2.25 ±0.16 63% 7 6 bFGF 1.56 ±0.15 44%

vasc.=vascularization

Conclusion.

Treatment of vascularization induced by bFGF is characterized by a dose-dependent effect after administration of laquinimod: a low dose (0.5 mg/kg, bid, group 4) leads to a suppression of vascularization by 11% vascularization, and a higher dose (2.5 mg/kg). kg, bid, group 5) leads to suppression of vascularization by 35%. Increasing the frequency of administration resulted in further increases in suppression, with many low doses (0.5 mg/kg, qid, group 6) resulting in suppression (37%) similar to suppression at the few high doses (group 5). Many daily high doses (2.5 mg/kg, qid, group 7) resulted in even greater suppression (56%) of vascularization.

Example 3 Laquinimod and ABR-215174 influence LPS-activated microglia-induced microvascular endothelial cell tube formation in the human retina.

Techniques

The study included the following experimental groups.

Group 1: Control (diluent, 0.1% DMSO).

Group 2: Sulforaphane (10 µM, positive control, antiangiogenic effects).

Group 3: Aflibercept (Eylea 40 μg/ml, positive control, partial antiangiogenic effects).

Group 4: ABR-215174 (0.1 µM).

Group 5: ABR-215062 (10 µM).

Human retinal microvascular endothelial cells (HRMECs) were purchased from Neuromics (Cat# HEC09, Lot# 2872) and grown according to the manufacturer's instructions in EndoGrowth media (Cat# EKG001, Lot# EKG0011902269) supplemented with endothelial growth factor (Cat# EKG001, Lot# EGK00125) in T25 vessels coated with AlphaBiocoat at 37°C 5% CO ₂ .

Primary microglia from human brain were purchased from Celprogen (Cat# 3708901, Lot# 1614454-01) and grown in complete microglia culture media with antibiotics (Cat# M37089-01, Lot# 2010089205-03) supplemented with 10% standard fetal stock. bovine serum (Neuromics, Lot# 042P20) in poly-L-lysine (PLL, 50 mg/ml) coated T25 vials at 37°C 5% CO2.

Human microglia were seeded onto PLL-coated cell culture inserts (Sarstedt, Cat# 83,3932,040) at 103,000 cells/ ^cm2 . Microglia were treated with test compounds, aflibercept, and vehicle for 24 h prior to activation with lipopolysaccharide (LPS) using the following concentrations in complete microglia culture media: Vehicle (0.1% DMSO) Aflibercept (Eylea®, 40 g/mL), 0. 1% DMSO ABR-215174 (0.1 µM), 0.1% DMSO ABR-215062 (10 µM), 0.1% DMSO.

The coculture technique for microglia and HRMECs was modified based on the protocol described in Ding et al. 2018 (Ding X, Gu R, Zhang M, Ren H, Shu Q, Xu G, Wu H. Microglia enhanced the angiogenesis, migration and proliferation of co-cultured RMECs. BMC Ophthalmol. 2018,18(1):249. doi : 10.1186/sl2886-018-0886-z. PMID: 30223824; PMCID: PMC6142340) and Ji Cho et al. 2019 (Ji Cho M, Yoon SJ, Kim W, Park J, Lee J, Park JG, Cho YL, Hun Kim J, Jang H, Park YJ, Lee SH, Min JK. Oxidative stress-mediated TXNIP loss causes RPE dysfunction. Exp Mol Med 2019 Oct 15;51(10): 1-13. doi: 10.1038/s12276-019-0327-y. PMID: 31615975; PMCID: PMC6802648). Microglia were activated with LPS (100 ng/ml) in complete microglia culture media without FBS and co-treated with freshly prepared test compounds, Eylea and diluent for 24 h. HRMECs were incubated in Endo-Growth basal media for 24 h for coculture.

HRMECs were seeded into Matrigel®-coated 24-well plates (42,000 cells/ ^cm2 ) and simultaneously treated with freshly prepared test compounds, aflibercept and sulforaphane in Endo-Growth Basic Media:

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Thinner (0.1% DMSO)

Sulforaphane (10 M), 0.1% DMSO

Aflibercept (Eylea®, 40 mcg/ml), 0.1% DMSO

ABR-215174 (0.1 µM), 0.1% DMSO

ABR-215062 (10 µM), 0.1% DMSO

Media in microglial inserts containing LPS were replaced with the corresponding freshly prepared test compounds, aflibercept and sulforaphane in Endo-Growth master media. Inserts containing activated microglia were transferred into 24-well plates containing HRMECs. Co-cultures were incubated at +37°C 5% CO ₂ , stained with calcein-AM (5 μM) for 30 min and visualized using a fluorescence microscope (Leica Thunder 3D Tissue Imager, Leica Microsystems).

Images were analyzed using AngioTool software (Zudaire E, Gambardella L, Kurcz C, Vermeren S (2011) A Computational Tool for Quantitative Analysis of Vascular Networks. PLOS ONE 6(11): e27385. https://doi.org/10.137 I/journal, pone.0027385) for ImageJ (NIH public domain). The total tubule area, tubule length, density, lacunarity, and branching index (number of junctions and end points) were quantified.

Images were analyzed using AngioTool software (NIH, Bethesda, MD; publicly available) and the following characteristics were determined: average vessel length, total vessel length, vessel area, percent vessel area, total number of connections, density of connections, total number of terminals points and average lacunarity. Raw data were presented in mm or µm (and ^mm2 or ^µm2 ) for length and area. Raw data for all characteristics were presented and analyzed using ordinary one-way ANOVA with Dunnett's post hoc multiple comparison test.

Effect sizes were calculated by subtracting the mean of the vehicle group from each value, then normalizing the values to those of the sulforaphane group (positive control), so that the mean of the sulforaphane group corresponds to the maximum effect size (100%) and for the group receiving the diluent, corresponds to no effect (0%). Effect size data were analyzed using the nonparametric Kruskal-Wallis test with Dunn's post hoc multiple comparison test.

Results and conclusions

The difference between the normalized effect sizes for mean vessel length for the sulforaphane and vehicle groups reached statistical significance (Figure 4). For the group receiving test compound ABR215174 (effect size 88.2%), the effect size for mean vessel length was significantly greater than for the diluent group. The effect sizes for the Eylea group (effect size 18%) and the study compound ABR215062 group (effect size 53%), although not statistically significant, support an increase in effect size for mean vessel length.

Since sulforaphane is a known and common compound for use as a positive control to prevent angiogenesis, these results indicate that the test compounds ABR215174 and ABR215062 can also be used to prevent angiogenesis.

Example 4 Additive effects for LPS-activated microglia-induced microvascular endothelial cell tube formation in the human retina when combining laquinimod or ABR-215174 with an angiogenesis inhibitor such as aflibercept (Eylea®) versus monotherapy.

Microglia, especially activated microglia, play an important role in angiogenesis and maintaining homeostasis of vascular function in retinal microvessels (Ding et al 2018). It is contemplated that coculture of human retinal microvascular endothelial cells (HRMEC's) and human brain microglia can be used to evaluate the effects of a compound of the present disclosure, such as laquinimod, taquinimod, ABR-215174 or ABR-215691, on neovascularization. You can use a technique such as the one described below.

Techniques

This study can be carried out essentially as described in Example 3.

Study of tube formation.

Tube formation assays were performed using wells of a 24-well Matrigel-coated plate or a 96-well plate as described by Ding et al. 2018 (Ding et al. BMC Ophthalmology (2018) 18:249): 50 µL/well of Matrigel was applied to the wells of a 96-well plate at 37°C for 30 min. After co-culture with microglia for 24 h, HRMECs were seeded at 1.5 x 10 ⁴ cells/well in 100 μl of medium. After a certain period of time, for example equal to 4 hours, the formation of tubes was observed and they were photographed using a microscope (Leica Microsys-

Claims (16)

themes). Images were analyzed using the Angiotool plugin (Zudaire et al., 2011, PLoS one, 6, 11, e27385) for ImageJ (NIH public domain). The total tubule area, tubule length, density, lacunarity, and branching index (number of junctions and end points) were quantified. Results and conclusions It is believed that when a compound of the present disclosure, such as laquinimod, 1 and 10 μM or ABR-215174, 0.01 and 0.1 μM, is combined with aflibercept (30 nM), there is a clear additive effect compared to when each compound is used in as monotherapy. CLAIM 1. Use of a composition containing a compound of formula (I) Where R ¹ is chlorine, R ² is ethyl or hydrogen, and R ³ is hydrogen, or a pharmaceutically acceptable salt thereof, for the treatment of an ocular disease or ocular disorder selected from the group consisting of the following: corneal neovascularization, iris neovascularization, neovascularization ciliary body, corneal pannus, uveal neovascularization, retinal neovascularization, wet age-related macular degeneration, proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy. 2. Use according to claim 1, wherein the compound is laquinimod or a pharmaceutically acceptable salt thereof. 3. Use according to claim 1, wherein the compound is a compound of formula (IV) or a pharmaceutically acceptable salt thereof. 4. The use of any of the preceding claims, wherein the ocular disease or ocular disorder is selected from the group consisting of the following: corneal neovascularization, iris neovascularization, ciliary body neovascularization, and corneal pannus. 5. Use according to claim 4, wherein the eye disease or eye disorder is corneal neovascularization. 6. Use according to any one of claims 1 to 3, wherein the eye disease or eye disorder is selected from the group consisting of the following: proliferative diabetic retinopathy, retrolental fibroplasia and ischemic retinopathy. 7. Use according to claim 6, wherein the eye disease or eye disorder is proliferative diabetic retinopathy. 8. Use according to claim 6, wherein the eye disease or eye disorder is retrolental fibroplasia. 9. Use according to claim 6, wherein the eye disease or eye disorder is ischemic retinopathy. 10. Use according to any one of claims 1 to 3, wherein the eye disease or ocular disorder is selected from the group consisting of the following: uveal neovascularization, retinal neovascularization and wet age-related macular degeneration. 11. Use according to claim 10, wherein the ocular disease or ocular disorder is neovascularization of the choroid. 12. Use according to claim 10, wherein the eye disease or eye disorder is retinal neovascularization. 13. Use according to claim 10, wherein the eye disease or eye disorder is a wet form of age-related macular degeneration. 14. Use as claimed in any of the preceding claims, wherein the composition contains or is administered in combination with an angiogenesis inhibitor such as aflibercept. 15. Use according to any of the previous claims, wherein the composition contains or is administered in combination with a VEGF inhibitor. 16. Use according to claim 15, wherein the VEGF inhibitor is selected from the group consisting of the following: afli- -