1-Methyl-1H-pyrazol-3-yl derivatives for the treatment of angiogenic diseases

Abstract

Abnormal angiogenesis or pro-angiogenic VEGF, especially in or on the eye xxx Anti-angiogenic treatment of conditions associated with abnormal overproduction of isoforms, and compounds for use in anti-angiogenic treatment, are described.

Description

[Technical Field]

[0001] The present invention relates to the treatment of abnormal angiogenesis or pro-angiogenic VEGF, particularly in or on the eye. xxx The present invention relates to the anti-angiogenic treatment of conditions associated with the abnormal overproduction of isoforms, such as ocular neovascularization, choroidal neovascularization, age-related macular degeneration and diabetic retinopathy, and to compounds for use in the anti-angiogenic treatment.

[0002] The present invention also relates to methods for treating intraocular hyperpermeability disorders, such as diabetic macular edema, and to compounds for use in treating hyperpermeability disorders.

[0003] The present invention also relates to methods of treating or preventing ocular degeneration, such as geographic atrophy or glaucoma, and compounds for use in such methods. [Background technology]

[0004] Diabetic macular edema (DMO, also known as DME) and age-related macular degeneration (AMD) are vision-decreasing diseases affecting the central region of the macula and are the leading cause of blindness in high-income countries (Bressler, 2004). DMO results from increased expression of proangiogenic isoforms of vascular endothelial growth factor (VEGF), which disrupts the inner and outer blood-retinal barrier (Perrin et al., 2005). This leads to the proliferation of new blood vessels, leakage of fluid and proteins from blood vessels into the retina, and increased fluid transport through the retinal pigment epithelium, resulting in retinal edema and vision loss. Wet AMD (also known as wet AMD or wAMD) is the most severe form of AMD (Ferris et al., 1984). It primarily arises from the choroidal circulation below the macula and is characterized by choroidal neovascularization (CNV). CNV is the abnormal proliferation of new blood vessels from the choroid into the retinal pigment epithelium (RPE) (Patz et al., 1977), which is thought to result in leakage of blood and serous fluid beneath and through the RPE, ultimately leading to loss of photoreceptors, retinal detachment, and vision loss due to macular scar compaction (Fine et al., 2000; Campochiaro et al., 2006). Vascular endothelial growth factor (VEGF), a key factor in angiogenesis and vascular leakage (Dvorak et al., 1995), increases during the progression of DMO and CNV (Spilsbury et al., 2000; Anderson et al., 2002; Das et al., 2003), making it a lead therapeutic target for the treatment of exudative AMD.

[0005] VEGF is a complex gene that is alternatively spliced ​​to form a family of isoforms (Leung et al., 1989; Jingjing et al., 1999). Each isoform differs in biological properties, activity, and function (Houck et al., 1991). Most cells express VEGF 121 , VEGF 165 , and VEGF 189 isoforms are commonly expressed, but VEGF 145 and VEGF 206are relatively rare. Most VEGF isoforms contain exons 1-5 (with the exception of VEGF 111 (Mineur et al., 2007)), which contains distinct portions of exons 6 and 7 that encode the heparin sulfate (HS)-binding domain.

[0006] In 2002, differential splicing of exon 8 was demonstrated from the proximal splice site (PSS) to the distal splice site (DSS) 66 bases downstream (Bates et al., 2002; Woolard et al., 2004). Alternative splicing in this region gives rise to a second family of isoforms (VEGFs) notable for their anti-angiogenic (Perrin et al., 2005) and anti-permeability properties. xxx b) was generated. WO 03 / 012105, the entire contents of which are incorporated herein by reference, describes alternatively spliced ​​isoforms and their therapeutic importance.

[0007] During pathological vascular growth, pro-angiogenic, pro-permeability isoforms are selectively increased ( Bates et al., 2002 ; Perrin, 2005 ; Varey et al., 2008 ; Pritchard-Jones et al., 2007 ), and VEGF xxx and VEGF xxx b may have separate regulatory pathways. 165 b and VEGF 121 These antiangiogenic isoforms, such as b, are potently antiangiogenic and have been shown to inhibit VEGF-mediated vascular permeability after intraocular injection in animal models of retinal and choroidal neovascularization (Hua et al., 2008; Ved et al., 2016), resulting in protection of endothelial, retinal epithelial, and neuronal cells (Beazley Long 2015; Magnussen et al., 2010). Splicing switching from proangiogenic, propermeability isoforms to antiangiogenic, antipermeability, cyto-, and neuroprotective isoforms may offer a potential therapeutic approach for patients with ocular diseases in which vascular leakage, vascular proliferation, or neuronal or epithelial cell degeneration are key drivers of pathology.

[0008] The first therapy approved by the FDA for the treatment of neovascular AMD in December 2004 was VEGF. 165 , VEGF 189 and VEGF 206 The first known treatment was the specific aptamer pegaptanib sodium (Macugen®). In clinical trials, pegatinibs dose-dependently reduced the risk of severe vision loss and slowed the progression of neovascular AMD, but did not significantly improve vision. In 2006, ranibizumab (Lucentis®), a novel humanized anti-VEGF antibody fragment, was approved by the FDA for the treatment of neovascular AMD. This approval was based on the results of three clinical trials, which showed that approximately 95% of patients treated monthly with Lucentis® (0.5 mg) maintained their vision (defined as a loss of less than 15 letters), and 40% or less improved their vision (defined as a gain of 15 or more letters) after one year, compared with 11% in the sham-treated group (Rosenfeld et al., 2006; Brown et al., 2006; Brown et al., 2009). Current treatment regimens require monthly intraocular injections of Lucentis (Brown et al., 2009; Schmidt-Erfuth et al., 2011). These intraocular injections increase intraocular pressure (Good et al., 2010) and carry the risk of mild endophthalmitis and other serious side effects (Jager et al., 2004). Furthermore, the anti-VEGF antibody bevicizumab (Avastin®), from which Lucentis® is derived, inhibits VEGF. 165 VEGF, which targets both pro- and anti-angiogenic VEGF isoforms with efficacy comparable to that of 165b (Varey et al., 2008). All of these treatments require periodic injections of drugs into the vitreous of patients with the disease. These invasive, uncomfortable, and potentially damaging procedures are necessary because we have not yet been able to develop anti-angiogenic agents that target VEGF and can penetrate the RPE and other retinal tissues without injection. The development of localized treatments that arise from solving the problem of how to deliver molecules to the back of the eye would be an important novel approach that could provide substantial benefit to patients with these and other retinal or ocular neovascularization / hyperpermeability diseases.

[0009] Because both antiangiogenic and angiogenic isoforms of VEGF originate from the same gene, regulation of the isoform family is a result of alternative splicing regulation. We have identified part of the pathway that controls VEGF splicing at the proximal splice site and shown that the RNA-binding protein SRSF1 (Nowak et al., 2008; Amin et al., 2011) and its kinase SRPK1 (Sanford et al., 2005) are key components required for cells to decide to use the proximal splice site and thus generate the proangiogenic isoform of VEGF (Nowak et al., 2008; Nowak et al., 2010). Knockdown of SRPK1 potently reduced VEGF-mediated angiogenesis in tumors in vivo, and inhibition of SRPK1 reduced angiogenesis in vivo (Amin et al., 2011).

[0010] WO 2008 / 110777, WO 2009 / 106855, WO 2010 / 058227, WO 2011 / 148200, and WO 2019 / 064512 (the disclosures of which are incorporated herein by reference) disclose VEGF xxx Therapeutic and other physiological uses of agents that favorably induce expression of the b isoform are described. SRPK inhibitors could, in principle, constitute such agents. WO 2005 / 063293 describes a class of SRPK inhibitors that includes SRPIN340 and its derivatives and analogues.

[0011] WO 2014 / 060763 describes SRPK inhibitors that target SRPK1, in particular for use as anti-angiogenic agents, neuroprotective agents, agents for use in the treatment or prevention of hyperpermeability disorders, agents for the treatment of pain, and agents for reducing the risk of or treating pre-eclampsia. VEGF xxx The development of drugs that induce the expression of the b isoform is expected to be useful not only for the treatment of neovascular AMD, but also for the treatment of VEGF xxx This represents a new era in SRPK1 and all other diseases involving SRPK1. However, to achieve this, it will be necessary to develop molecules that are both potent SRPK1 inhibitors and penetrate the eye. Summary of the Invention [Problem to be solved by the invention]

[0012] The present invention is based on our discovery that novel small molecule inhibitors surprisingly and unexpectedly possess high ocular permeability as topical therapeutic agents while maintaining SRPK1 inhibitory activity, particularly for use as topical anti-angiogenic agents, neuroprotective agents, agents for the treatment or prevention of hyperpermeability disorders, and agents for the treatment or prevention of ocular fibrosis. The present invention is also based, at least in part, on the surprising discovery that these small molecular weight compounds can be used topically to inhibit the progression of CNV and the expression of angiogenic (but not anti-angiogenic) VEGF. [Means for solving the problem]

[0013] In a first aspect, the present invention provides a compound of formula (I) where X is CF3, methyl, Cl or cyclopropyl and n=1 or 2: [ka] or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.

[0014] The present invention also provides a compound of formula (I) for use in the treatment or prevention of ocular neovascularization.

[0015] The compounds of formula (I) and their pharmaceutically acceptable salts, solvates, hydrates or prodrugs are novel compounds in themselves (as well as their uses in the prevention and treatment described herein) and they form an aspect of the present invention.

[0016] It is surprising and unexpected that the compounds used in the present invention have systemic ocular permeability that allows for effective treatment or prevention of ocular neovascularization, or localized treatment or prevention of ocular neovascularization, in eyes from animals that have similar characteristics to the human eye in terms of size, thickness, content, and function.

[0017] Pharmaceutical compositions comprising the Novel Compounds and the use of the Novel Compounds and pharmaceutical compositions comprising them in antipermeability and / or antiangiogenic therapy (including the treatment and prevention of disorders and diseases characterized by aberrant or excessive ocular angiogenesis or permeability), treatment of ocular hyperpermeability disorders, treatment of ocular neuropathies and neurodegenerative disorders, treatment of ocular epithelial degenerative disorders constitute further aspects of the present invention.

[0018] Accordingly, the present invention also provides (i) methods for treating or preventing disorders and diseases characterized by abnormal or excessive ocular angiogenesis as defined herein; (ii) methods for treating or preventing ocular hyperpermeability disorders as defined herein; (iii) methods for treating or preventing ocular neurodegenerative disorders as defined herein; and (iv) methods for treating or preventing ocular epithelial degenerative disorders, comprising administering a compound of Formula (I) to a patient in need thereof. In some embodiments, for any of the methods described herein, the compound of Formula (I) is administered to a patient in need thereof in a therapeutically effective amount.

[0019] Certain compounds of formula (I) and preferred exemplary subclasses of compounds of formula (I) may be specifically mentioned for use in the present invention.

[0020] Examples of compounds of formula (I) that may be mentioned include: n=1 and X is CF3 or Cl; and Included are those where n=2 and X is CF3 or Cl.

[0021] The compounds of the present invention are specific inhibitors of SRPK1 and therefore may be used in methods for treating or preventing any disease or condition of the eye in which SRPK1 is involved. Such conditions and treatments are described below.

[0022] antiangiogenic treatment The compounds of the present invention may be used in ocular anti-angiogenic therapy. The anti-angiogenic therapy preferably involves the treatment of aberrant angiogenesis or pro-angiogenic VEGF isoforms (VEGF xxx The present invention also includes the treatment or prevention of any disease or disorder associated with abnormal overproduction of vasculovascular endothelial cells (vasculovascular endothelial cells). Such diseases and disorders include, for example, diabetic retinopathy, trachoma, retrolental hyperplasia, neovascular glaucoma, age-related macular degeneration, hemangioma, corneal angiogenesis associated with ocular injury or infection, and proliferative diabetic retinopathy. Anti-angiogenic treatments according to the present invention may also include non-therapeutic treatments performed in healthy individuals to inhibit blood vessel development, for example, for cosmetic purposes. For further details regarding diseases and disorders associated with abnormal angiogenesis and anti-angiogenic treatments, see WO 2008 / 110777, the contents of which are incorporated herein by reference.

[0023] In particular, the compounds of the present invention may be used to treat or prevent ocular neovascularization, which may include, but is not limited to, retinal or choroidal neovascularization, diabetic retinopathy, or age-related macular degeneration. Additionally, the compounds of the present invention may be used to treat or prevent malignant ocular neoplasms or cancers, such as uveal melanoma.

[0024] Microvascular hyperpermeability disorder, impaired epithelial cell viability The compounds of the present invention as SRPK1 inhibitors inhibit the activity of alternatively spliced ​​VEGF xxx It may also be used as a therapeutic agent in the treatment of other diseases in which the VEGF b isoform is involved. For example, in WO 2010 / 058227, the contents of which are incorporated herein by reference, xxx b has been shown to be active against a variety of disorders of microvascular hyperpermeability, disorders of epithelial cell viability, and disorders of epithelial filtering membrane fenestrations.

[0025] Microvascular hyperpermeability, VEGF xxx Dysregulation of the pro-angiogenic and pro-permeability properties of isoforms, impaired epithelial cell viability and permeability, and / or impaired fenestration properties (e.g., number density and / or size) of epithelial filtering membranes underlie many serious ocular diseases.

[0026] Examples of such conditions include, for example, proliferative and non-proliferative diabetic retinopathy, diabetic macular edema, exudative age-related macular degeneration, and retinal vein occlusion (central and branch).

[0027] Examples of disorders that would benefit from treatments that support epithelial cell viability include:

[0028] Age-related macular degeneration (AMD) (wet or dry), central serous chorioretinopathy, cystoid macular edema, diabetic retinopathy, proliferative diabetic retinopathy, diabetic macular edema, iris neovascularization, retinopathy of prematurity, central and branch retinal vein occlusion, inflammatory / infectious retinal neovascularization / edema (e.g., posterior uveitis, sarcoid, toxoplasmosis, histoplasmosis, Vogt-Koyanagi-Harada disease, chronic uveitis, tuberculosis, syphilis, punctate and multifocal endometriosis), retinoblastoma, ocular melanoma, ocular tumor, retinal detachment, myopic neovascularization, angiodal streak, Eales disease, ischemic retinopathies (retinal artery occlusion, Takayasu's, carotid artery occlusion), choroidal rupture, or any combination thereof. In a most preferred embodiment, the back of the eye condition is age-related macular degeneration (AMD).

[0029] The present invention may be used in the treatment of macular dystrophies, including Stargardt disease / fundus citrate, Stargardt-like macular dystrophy, autosomal dominant "bull eye" macular dystrophy, Best macular dystrophy, adult vitelliform dystrophy, Pattern dystrophy, Doyne honeycomb retinal dystrophy, North Carolina macular dystrophy, MCDR1-like autosomal dominant macular dystrophy, North Carolina-like macular dystrophy with hearing loss, progressive bifocal choroidal atrophy, Sorsby fundus dystrophy, central areolar choroidal dystrophy, dominant cystoid macular dystrophy, juvenile retinal sequestration, occult macular dystrophy, and non-familial occult macular dystrophy.

[0030] The disorder may in particular be a disorder of the retinal epithelium, such as geographic atrophy, or age-related macular degeneration. For further details regarding disorders of microvascular hyperpermeability, disorders of epithelial cell viability, disorders of epithelial filtering membrane fenestration, and their treatment, see WO 2010 / 058227, the contents of which are incorporated herein by reference.

[0031] active compound The compounds of the present invention are as defined by formula (I) and have been shown to be inhibitors of the kinase SRPK1 and therefore VEGF xxx The compounds of the present invention are useful in treating diseases described herein in which SRPK1 and / or SRPK1 have been shown to be involved. The compounds of the present invention may be SRPK1-specific inhibitors.

[0032] The compounds of the invention may be synthesized by any known method. Exemplary synthetic methods are described in the Examples below.

[0033] Concurrent medication The compounds of the present invention may, if desired, be administered with one or more additional active agents, such as, but not limited to, one or more agents selected from a cholinesterase inhibitor, a dopamine agonist (e.g., L-dopa), a COMT inhibitor, an MAO-B inhibitor, an anticholinergic, an acetylcholine agonist, a serotonin agonist, an AMPA receptor agonist, a GABA receptor agonist, an NMDA receptor agonist, a b-adrenoceptor agonist, digoxin, dobutamine, an anti-inflammatory agent, a neurotrophic factor, a statin, an adenosine A2a receptor antagonist, an aldose reductase inhibitor, an immunomodulator, a cannabinoid agonist, an interferon, or a tricyclic antidepressant.

[0034] definition In the definition of formula (I) herein, The term "salt" is not particularly limited as long as it is a pharmaceutically acceptable salt formed by the compound according to the present invention. Examples of such salts include inorganic acid salts, organic acid salts, inorganic base salts, organic base salts, and acidic or basic amino acid salts. Preferred examples of inorganic acid salts include hydrochloride, hydrobromide, sulfate, nitrate, and phosphate. Preferred examples of organic acid salts include acetate, succinate, fumarate, maleate, tartrate, citrate, lactate, stearate, benzoate, methanesulfonate, and p-toluenesulfonate.

[0035] Preferred examples of inorganic base salts include alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, aluminum salts, and ammonium salts. Preferred examples of organic base salts include diethylamine salts, diethanolamine salts, meglumine salts, and N,N'-dibenzylethylenediamine salts.

[0036] Examples of preferred acidic amino acid salts include aspartate and glutamate, and examples of preferred basic amino acid salts include arginine, lysine, and ornithine salts.

[0037] When left in the air, the compounds of the present invention may absorb moisture and may adhere to the absorbed moisture or convert to a hydrate. In some embodiments, the hydrate may include a hemihydrate. Such hydrates are also included in the present invention.

[0038] Furthermore, the compounds of the present invention may absorb some other solvents and be converted into solvates, and such solvates are also included in the present invention.

[0039] To prepare solvates of the compounds of the present invention, in principle any organic solvent can be used.

[0040] The solvate may also include water together with one or more organic solvents.

[0041] Thus, for example, the solvent may be chosen from ketones, alcohols, ethers, esters, aromatic solvents, and possibly mixtures thereof with one another, other organic solvents and / or with water.

[0042] Pharmaceutically acceptable prodrug forms of compounds of formula (I) may be used in the present invention. "Pharmaceutically acceptable prodrug" refers to a prodrug of a compound, and possibly a zwitterionic form of the compound, that is suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., within the scope of sound medical and veterinary judgment, commensurate with a reasonable benefit / risk ratio, and is effective for its intended use. The term "prodrug" refers to a compound that is rapidly transformed in vivo, for example, by hydrolysis in blood, to yield the parent compound of the above formula. Functional groups that can be rapidly transformed in vivo by metabolic cleavage form a class of groups reactive with carboxyl groups. Compounds bearing metabolically cleavable groups act as prodrugs because such groups are readily cleaved in vivo.A thorough discussion of prodrugs is provided in the following, which are incorporated herein by reference: Design of Prodrugs, H. Bundgaard, ed., Elsevier, 1985; Methods in Enzymology, K. Widder et al., Ed., Academic Press, 42, pp. 309-396, 1985; A Textbook of Drug Design and Development, Krogsgaard-Larsen and H. Bundgaard, ed., Chapter 5; Design and Applications of Prodrugs p. 113-191, 1991; Advanced Drug Delivery Reviews, H. Bundgaard, 8, pl-38, 1992; Journal of Pharmaceutical Sciences, 77, p. 285, 1988; Chem. Pharm. Bull., N. Nakeya et al., 32, p. 692, 1984; Prodrugs as Novel Delivery Systems, T. Higuchi and V. Stella, Vol. 14 of the ACSSymposium Series, and Bioreversible Carriers in Drug Design, Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, 1987.

[0043] Compositions and Dosages The compounds according to the invention may be administered in the form of a composition comprising the active agent and any suitable additional ingredients, for example a pharmaceutical composition (medicament) suitable for topical administration (e.g. as eye drops or a cream or lotion).

[0044] The term "pharmaceutical composition" or "medicament" in the context of the present invention refers to a composition comprising an active agent and further comprising one or more pharmaceutically acceptable carriers. Depending on the nature of the dosage and administration form, the composition may further comprise components selected from, for example, diluents, adjuvants, excipients, vehicles, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavorings, flavoring agents, antibacterial agents, antifungal agents, lubricants, and dispersing agents. The composition may be in the form of, for example, a liquid formulation, including a suspension, powder, emulsion, solution, cachet, granule, and liposomal formulation. Techniques and formulations may generally be found in Remington, The Science and Practice of Pharmacy, Mack Publishing Co., Easton, PA, latest edition.

[0045] Liquid form preparations include solutions, suspensions, and emulsions. For example, water or water-propylene glycol solutions for topical administration may be mentioned. Liquid preparations can also be formulated in solution in aqueous polyethylene glycol solution.

[0046] Also included are solid form preparations intended to be converted to liquid form preparations for topical administration immediately prior to use. Such liquid forms include solutions, suspensions, and emulsions. These particular solid form preparations are most conveniently provided in unit dosage form and, as such, are used to provide a single liquid dosage unit. Alternatively, sufficient solid may be provided so that, after conversion to liquid form, multiple individual liquid doses can be obtained by measuring a predetermined volume of the liquid form preparation using a syringe, teaspoon, or other volumetric container or device. Solid form preparations intended to be converted to liquid form may contain, in addition to the active ingredient, flavors, colorants, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like. Liquids utilized to prepare liquid form preparations may be water, isotonic water, ethanol, glycerin, propylene glycol, and the like, as well as mixtures thereof.

[0047] The composition may be a formulation intended for topical application. The formulation may be a gelling formulation to control the release and therefore availability of the active agent after topical application. The formulation may include one or more gelling agents, such as hydroxypropyl methylcellulose. The formulation may include one or more surfactants, such as non-ionic liquid polymers, examples of which include Tyloxapol and Pluronics® poloxamers from BASF. The formulation may include one or more solubilizing agents, such as glucose or sorbitol. The formulation may include one or more antimicrobial agents or preservatives, such as benzalkonium chloride. The aforementioned gelling agents, surfactants, solubilizing agents, and antimicrobial agents are provided purely by way of example, and it will be understood that other agents that perform these functions are known.

[0048] Dosage of the active agent (e.g., a compound of formula (I)) may vary depending on the needs of the patient, the nature, severity and extent of the condition, the age and condition of the patient, the compound used, and other factors known to those skilled in the art.

[0049] In some instances, treatment is initiated with a dosage lower than the optimal dose of the compound. The dosage is then increased by small increments until the optimal effect under the circumstances is reached. For convenience, the total daily dose may be administered in divided doses throughout the day, if desired. For example, the total daily dose may be administered two, three, or four times daily. The total daily dose may be administered for a period ranging from 1 to 14 days, or longer if necessary. For example, for long-term treatment of chronic eye disorders, the total daily dose may be administered for a period of at least 2 years, such as at least 3 years, for example at least 4 years, for example at least 5 years, for example at least 10 years, for example at least 15 years, or for example at least 20 years.

[0050] A dosage regimen for administration of an active agent may, for example, comprise a total daily dose of up to 30 mg, such as up to 20 mg, for example up to 10 mg, for example up to 500 μg, such as up to 400 μg, for example up to 300 μg, such as up to 200 μg, for example up to 100 μg, such as up to 50 μg, for example up to 20 μg, for example 10 μg of active agent.

[0051] A dosage regimen for administration of the active agent may, for example, comprise a total daily dose of at least 10 μg, such as at least 20 μg, for example at least 50 μg, such as at least 60 μg, for example at least 100 μg, such as at least 200 μg, for example at least 300 μg, such as at least 400 μg, for example at least 500 μg, such as at least 1 mg, for example at least 10 mg, such as at least 20 mg, for example at least 30 mg of active agent.

[0052] The compounds of formula (I), or pharmaceutically acceptable salts, solvates, hydrates, or prodrugs thereof, may be administered in a therapeutically effective amount. As used herein, the term "therapeutically effective amount" refers to that amount of active compound or pharmaceutical agent that elicits the biological or pharmacological response in a tissue system, animal, or human that is desired by a researcher, veterinarian, physician, or other clinician, including alleviation of the symptoms of the disease or disorder being treated.

[0053] A therapeutically effective amount of a compound of Formula (I) for topical administration to treat CNV may be at least about 5 μg / 10 μL of delivery vehicle. Alternatively, the therapeutically effective amount may be at least about 100 μg / mL, such as at least about 200 μg / mL, at least about 300 μg / mL, at least about 400 μg / mL, at least about 500 μg / mL, at least about 600 μg / mL, at least about 700 μg / mL, at least about 800 μg / mL, at least about 900 μg / mL, or at least about 1000 μg / mL. Alternatively, the therapeutically effective amount may be at least about 1 mg / mL, such as at least about 1.5 mg / mL, for example at least about 2 mg / mL, at least about 3 mg / mL, at least about 4 mg / mL, or at least about 5 mg / mL. Alternatively, the therapeutically effective amount may be less than about 5 mg / mL, e.g., less than about 4 mg / mL, less than about 3 mg / mL, less than about 2 mg / mL, less than about 1.5 mg / mL, or less than about 1 mg / mL. The therapeutically effective amount may be administered daily, for example, for a dosing period ranging from 1 to 14 days. The therapeutically effective amount may be administered daily for life, for example, in the long-term treatment of chronic eye disorders. The therapeutically effective amount may be a total daily dose that may be administered in portions throughout the day, e.g., twice daily or four times daily.

[0054] "Treatment or prevention" As used herein, the expression "treatment or prevention" and similar terms refer to all forms of medical care intended to eliminate or prevent a disorder or alleviate its symptoms, including preventive, curative, and palliative care, as determined according to any of the tests available in accordance with common medical and psychiatric practice. Interventions undertaken with a reasonable expectation of achieving a particular result, but not always, are included within the expression "treatment or prevention." Interventions that are successful in slowing or halting the progression of a disorder are included within the expression "treatment or prevention."

[0055] As used herein, unless otherwise indicated, the terms "treat," "treatment," and the like include the management and care of a subject or patient, preferably a mammal, more preferably a human, for the purpose of combating a disease, condition, or disorder, and includes the administration of a compound of the invention to prevent the onset of symptoms or complications, alleviate symptoms or complications, slow the progression of the disease or disorder, or eliminate the disease, condition, or disorder. The term "treat" or "treatment" further includes (a) inhibiting the disease state, i.e., arresting its development, and / or (b) alleviating the disease state, i.e., causing regression of the disease state.

[0056] As used herein, "prevention" includes the prophylactic treatment of an asymptomatic disease state in a mammal, particularly a human, with the goal of reducing the likelihood of the occurrence of the clinical disease state. Patients are selected for prophylactic treatment based on factors known to increase their risk of suffering from the clinical disease state compared to the general population. As used herein, "prophylaxis" refers to the protective treatment of a disease state to reduce and / or minimize the risk of the disease state and / or reduce the risk of recurrence by administering to a patient a therapeutically effective amount of at least one compound of the present invention or a pharmaceutically acceptable salt, hydrate, or solvate thereof. Patients may be selected for prophylaxis treatment based on factors known to increase the risk of suffering from a clinical disease state compared to the general population. For prophylaxis treatment, the clinical disease state may not yet be present. "Prophylaxis" treatment is divided into (a) primary prevention and (b) secondary prevention. Primary prevention is defined as treatment that reduces or minimizes the risk of a disease state in patients who have not yet exhibited a clinical disease state, while secondary prevention is defined as treatment that minimizes or reduces the risk of recurrence or a second occurrence of the same or similar clinical disease state.

[0057] "Susceptible to" As used herein, the phrase "susceptible" and similar terms refer to an individual who is at higher than normal risk of developing a medical or psychiatric disorder or personality change, particularly as assessed using known risk factors for the individual or disorder. Such individuals may be classified as having a substantial risk of developing one or more particular disorders or personality changes, to the extent that, for example, medications may be prescribed and / or special dietary, lifestyle, or similar recommendations may be made to the individual.

[0058] mammalian In addition to being useful for treating humans, the present invention is also useful in a variety of mammals, including, for example, non-human zoo primates (e.g., apes, monkeys, and lemurs), companion animals such as cats or dogs, working and sport animals such as dogs, horses, and ponies, farm animals such as pigs, sheep, goats, deer, oxen, and cattle, and laboratory animals such as rodents (e.g., rabbits, rats, mice, hamsters, gerbils, or guinea pigs).

[0059] It will be understood that the mammal to be treated is a human when the disorder or function to be treated is limited to humans, the same applies to other mammalian species, respectively, when the disorder or function to be treated is exclusive to that species.

[0060] Embodiments of the present invention will now be described, purely by way of example, with reference to the accompanying drawings, in which: [Brief explanation of the drawings]

[0061] [Figure 1] 1 shows the permeability of Compound 1 and other compounds as measured by the method described in the Ocular Permeability section. [Figure 2] The amount of compound not bound to melanin is shown. [Figure 3a]1 shows the penetration of Compound 1 into the retina compared to a chemically similar compound (N-(2-(4-((1H-pyrazol-3-yl)methyl)piperazin-1-yl)-5-(trifluoromethyl)-phenyl)-5-(tetrahydro-2H-pyran-4-yl)furan-2-carboxamide, designated Compound R) and pazopanib. [Figure 3b] 1 shows the distribution of Compound 1 through various ocular tissues. [Figure 4] 1 shows the relationship between ex vivo permeability and the amount of compound found in the retina for Compound 1, Compound R, and SPHINX31 (WO 2015 / 159103). [Figure 5] ELISA shows that compound 1 switches alternative splicing and reduces the expression of the VEGF-A165a isoform in retinal pigment epithelial cell lines. [Figure 6a] 1 shows the results of Compound 1 administered every other day for 28 days on VEGF expression in diabetic rats as measured by Western blot. [Figure 6b] The figure shows that total VEGF was reduced. [Figure 6c] This shows that VEGF-A165b was increased. [Figure 6d] 1 shows that the ratio of VEGF-A165b to total VEGF was increased by Compound 1. [Figure 7a] Fluorescein angiography images are shown. [Figure 7b] The same data are shown graphically demonstrating that Compound 1 has the same anti-angiogenic activity on lesion size as the reference compound in a laser-induced mouse model of CNV. [Figure 8a] 1 shows the amount of Compound 1 found in the retinas of cynomolgus monkeys treated with 0.5 mg / mL, 1.0 mg / mL, and 1.5 mg / mL eye drops every other day for 3 weeks. [Figure 8b] Color intensity indicating the amount of Compound 1 in different tissues within the eye. [Figure 9]1 shows the concentration of Compound 1 in water after a single ocular instillation of Compound 1 at different doses. [Figure 10a] Figure 1 shows changes in retinal VEGF levels in monkeys treated with Compound 1 as eye drops (top: Pan VEGF-A165, bottom: VEGF-A165b). [Figure 10b] Western blot band intensities are shown relative to the mean value of untreated for each blot (treated, N=3 per group, N=6 eyes per group; untreated, N=6 mice, N=12 eyes, mean±SEM), (Two-Way ANOVA p<0.001, FDR post hoc BKY test **=p<0.01, ***=p<0.001 compared to VEGF165b, ##=p<0.01, ###=p<0.001 compared to untreated). [Figure 10c] The ratio of panVEGF to VEGF365b for each sample is shown, with a decrease representing a switch to a less angiogenic state (One way ANOVA p<0.01, **=p<0.01 compared to untreated) (Holm Sidak post hoc). DETAILED DESCRIPTION OF THE INVENTION

[0062] method synthesis The compounds of the present invention can be prepared based on any synthetic method known to those skilled in the art, such as those described in WO 2015 / 159103 and WO 2017 / 064512. Specifically, compounds 1 to 3 were prepared according to the following exemplary methodology.

[0063] compound 1 tert-Butyl 4-(2-nitro-4-(trifluoromethyl)phenyl)piperazine-1-carboxylate [ka] A suspension of tert-butyl piperazine-1-carboxylate (2.90 g, 15.5 mmol), 1-chloro-2-nitro-4-(trifluoromethyl)benzene (3.99 g, 15.5 mmol), and sodium carbonate (4.10 g, 38.66 mmol) in DMF (25 mL) was heated at 110 °C for 3 h. The resulting reaction mixture was cooled to room temperature, quenched with cold water (100 mL), and extracted with ethyl acetate (3 times). The organic extracts were combined, washed with brine, and dried over anhydrous NaSO. The solvent was removed under reduced pressure to give the title product as an orange liquid (5.4 g, 92%), which had all analytical data consistent with the required structure and was sufficiently pure for use in the next step.

[0064] 1 LCMS:[M + -56]320.01m / z, purity 98.37%.

[0065] tert-Butyl 4-(2-amino-4-(trifluoromethyl)phenyl)piperazine-1-carboxylate [ka] Under a nitrogen atmosphere, hydrazine hydrate (17.87 g, 357.5 mmol) was added dropwise to a solution of piperazine (5.4 g, 14.3 mmol), iron(III) chloride (0.46 g, 2.8 mmol), and charcoal (2.7 g) in methanol (50 mL) at 0 °C. The resulting reaction mixture was heated to reflux for 1 h. After the reaction mixture was cooled to room temperature, it was filtered through a short pad of Celite and eluted with ethyl acetate. The organic filtrate was concentrated under reduced pressure. The residue was diluted with water (50 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure. The resulting crude material was triturated with n-pentane (2 × 10 mL) to give the title product as an off-white solid (3.5 g, 71%).

[0066] 1 LCMS:[M + -56]290.01m / z, purity 98.80%.

[0067] N-(2-(piperazin-1-yl)-5-(trifluoromethyl)phenyl)-5-(tetrahydro-2H-pyran-4-yl)furan-2-carboxamide [ka] Under a nitrogen atmosphere, a 2 M solution of trimethylaluminum in toluene (8.7 mL, 17.37 mmol) was added dropwise to a solution of tert-butyl 4-(2-amino-4-(trifluoromethyl)phenyl)piperazine-1-carboxylate (2 g, 5.8 mmol) in dichloromethane (8 mL) at 0 °C. The reaction mixture was stirred at room temperature for 1 hour, and then a solution of 5-(tetrahydro-2H-pyran-4-yl)-2-fluorate (1.22 g, 5.8 mmol) in methyldichloromethane (5 mL) was added dropwise at room temperature. The resulting reaction solution was stirred at room temperature for an additional 15 hours. To quench the reaction, a saturated aqueous solution of Rochelle's salt was added dropwise at room temperature, and the solution was stirred at room temperature for an additional 15 minutes. The reaction mixture was diluted with saturated aqueous sodium bicarbonate (100 mL) and extracted with dichloromethane (3 times). The organic extracts were combined, washed with water and brine, and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure to give the title product as a pale yellow solid (1.8 g, 73%).

[0068] 1 H-NMR(DMSO-d6)δ1.65-1.76(m,2H),1.94(dd,J=2,12.4Hz,2H),2.85-2.88(m,4H),2.97-2.99(m,4H),3.01-3.09(m,1H),3.45-3.51(m,2) LCMS:[MH] + 424.22m / z, purity 43.56%.

[0069] N-(2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)piperazin-1-yl)-5-(trifluoromethyl)phenyl)-5-(tetrahydro-2H-pyran-4-yl)furan-2-carboxamide [ka] A solution of piperazine (0.10 g, 0.23 mmol) and 1-methyl-1H-pyrazole-3-carbaldehyde (0.026 g, 0.23 mmol) in methanol (5 mL) was stirred at ambient temperature under a nitrogen atmosphere for 30 minutes. Sodium cyanoborohydride (0.44 g, 0.71 mmol) was then added portionwise to the reaction mixture at 0 °C. The resulting reaction mixture was allowed to stir at room temperature for 2 hours. The reaction mixture was then diluted with saturated sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined, washed with brine, and dried over anhydrous NaSO. The solvent was removed under reduced pressure. The resulting crude material was purified by preparative HPLC using 10 mM ammonium bicarbonate in water and acetonitrile as the mobile phase to give the title product (Compound 1) as an off-white solid (0.05 g, 40.91%).

[0070] Mp: 148-150°C; 1 H NMR(400MHz,DMSO-d6)δ1.71-1.82(m,2H),1.97-2.00(m,2H),2.65-2.67(m,4H),2.92(br s,4H),3.06-3.12(m,1H),3.47-3.54(m,4H),3.78(s,3H),3.98(d,J=9.6Hz,2H),6.15(s,1H),6 .47(d,J=3.2Hz,1H),7.25(d,J=3.6Hz,1H),7.47(s,2H),7.62(s,1H),8.61(s,1H),9.47(s,1H). HPLC purity: 100% MS (ESI-MS): m / z C 26 H 31 F3N5O3[MH] + Calculated value: 518.24, Measured value: 518.12

[0071] compound 2 tert-Butyl 4-(2-nitro-4-(trifluoromethyl)phenyl)-1,4-diazepane-1-carboxylate [ka] A suspension of 1-chloro-2-nitro-4-(trifluoromethyl)benzene (2.25 g, 9.99 mmol), tert-butyl 1,4-diazepane-1-carboxylate (2.0 g, 9.99 mmol), and solid sodium carbonate (3.18 g, 29.96 mmol) in anhydrous DMF (30 mL) was heated to 110 °C for 17 h. The resulting reaction mixture was cooled to room temperature and filtered through a short pad of Celite, eluting with ethyl acetate. The organic filtrate was concentrated under reduced pressure. The residue was diluted with water (100 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined and dried over anhydrous NaSO. The solvent was removed under reduced pressure. The resulting crude material was purified by column chromatography on silica using 90% ethyl acetate in hexane as the eluent to give the title product (3.78 g, 97%) as a pale yellow solid.

[0072] 1 H NMR(400MHz,DMSO-d6)δ1.12-1.20(m,9H),1.80(br s,2H),3.12-3.24(m,2H),3.30-3.38(m,2H,combined with residual moisture of DMSO),3.50(t,J=6Hz,2H),3.6 1-3.69(m,2H),7.35-7.40(m,1H),7.72(d,J=8.8Hz,1H),8.02-8.03(m,1H);LCMS:[M + -56]334.08m / z, 100% purity. 1-(2-nitro-4-(trifluoromethyl)phenyl)-1,4-diazepane [ka] Under a nitrogen atmosphere at 0°C, 4N HCl in dioxane (25 mL) was added dropwise to a solution of tert-butyl 4-(2-nitro-4-(trifluoromethyl)phenyl)-1,4-diazepane-1-carboxylate (3.78 g, 9.71 mmol) in dioxane (10 mL). The resulting reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was poured into a saturated solution of sodium bicarbonate (100 mL). The product was extracted with ethyl acetate (3 times). The organic extracts were combined, washed with water and brine, and dried over anhydrous NaSO. The solvent was removed under reduced pressure to give the title product as an orange liquid (2.85 g, quantitative), which was used directly in the next step without further purification.

[0073] 1 H NMR(400MHz,DMSO-d6)δ1.79-1.81(m,2H),2.78(t,J=5.2Hz,2H),2.97(t,J=5.2Hz,2H),3.19(d,J=4.8Hz,2H ),3.46(t,J=5.2Hz,2H),7.36(d,J=9.2Hz,1H),7.72(dd,J=2.4,9.2Hz,1H),8.04(d,J=1.6Hz,1H);LCMS:[MH] + 289.96m / z, purity 99.97%.

[0074] 1-((1-methyl-1H-pyrazol-3-yl)methyl)-4-(2-nitro-4-(trifluoromethyl)phenyl)-1,4-diazepane [ka] To a solution of 1-(2-nitro-4-(trifluoromethyl)phenyl)-1,4-diazepane (1.4 g, 4.84 mmol) and 1-methyl-1H-pyrazole-3-carbaldehyde (5) (0.639 g, 5.81 mmol) in 1,2-dichloroethane (25 mL) was added sodium sulfate (0.343 g, 2.42 mmol), followed by glacial acetic acid (0.581 g, 9.68 mmol). The resulting reaction mixture was stirred at ambient temperature under a nitrogen atmosphere for 3 hours. Sodium triacetoxyborohydride (1.54 g, 7.26 mmol) was then added in portions to the reaction mixture at 0 °C. The resulting reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was then diluted with saturated aqueous sodium bicarbonate (100 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined, washed with brine, and dried over anhydrous NaSO. The solvent was removed under reduced pressure. The resulting crude material was purified by flash chromatography on silica using (4% methanol in chloroform) as the eluent to give the title product as an orange liquid (1.80 g, 97%).

[0075] 1 H NMR(400MHz,CDCl3)δ2.01(quin,J=5.2Hz,2H),2.74(t,J=5.2Hz,2H),2.84(t,J=4.8Hz,2H),3.37(t,J=5.6Hz,2H),3.47(t,J=4.4Hz,2H),3.67 LCMS:[MH] + 384.14m / z, purity 99.35%.

[0076] 2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepan-1-yl)-5-(trifluoromethyl)aniline [ka] Under a nitrogen atmosphere at 0°C, hydrazine hydrate (5.88 g, 117.38 mmol) was added dropwise to a solution of 1-((1-methyl-1H-pyrazol-3-yl)methyl)-4-(2-nitro-4-(trifluoromethyl)phenyl)-1,4-diazepane (1.80 g, 4.70 mmol), iron(III) chloride (0.125 g, 0.94 mmol), and charcoal (0.2 g) in methanol (30 mL). The resulting reaction mixture was heated to reflux for 30 minutes. After cooling to room temperature, the reaction mixture was filtered through a short pad of Celite and eluted with ethyl acetate. The organic filtrate was concentrated under reduced pressure. The residue was diluted with water (50 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure. The resulting crude material was purified by flash chromatography on silica using (4% methanol in chloroform) as the eluent to give the title product as a colorless liquid (1.4 g, 84%).

[0077] 1 H NMR(400MHz,DMSO-d6)δ1.81(quin,J=5.6Hz,2H),2.71-2.75(m,4H),3.01-3.07(m,4H),3.58(s,2H),3.77(s,3H),5.07(br s,2H),6.14(d,J=2Hz,1H),6.80(dd,J=1.6,8Hz,1H),6.93(d,J=2Hz,1H),7.04(d,J=8Hz,1H),7.58(d,J=2Hz,1H);LCMS:[MH] + 354.22m / z, 100% purity.

[0078] N-(2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepan-1-yl)-5-(trifluoromethyl)phenyl)-5-(tetrahydro-2H-pyran-4-yl)furan-2-carboxamide [ka] To a solution of 2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepan-1-yl)-5-(trifluoromethyl)aniline (0.2 g, 0.56 mmol) and 5-(tetrahydro-2H-pyran-4-yl)-2-furoic acid (0.133 g, 0.67 mmol) in tetrahydrofuran (8 mL) was added triethylamine (0.114 g, 1.13 mmol) and T3P (0.179 g, 0.56 mmol) at room temperature under a nitrogen atmosphere. The resulting reaction mixture was heated to reflux for 4 hours. The reaction mixture was cooled to room temperature, diluted with water (100 mL), and extracted with ethyl acetate (3 times). The organic extracts were combined and dried over anhydrous Na2SO4. The organic solvent was removed under reduced pressure. The resulting crude material was purified by preparative HPLC using isopropyl alcohol:methanol (70:30) and n-heptane as the mobile phase to give the title product (compound 2) as a brown semi-solid (0.059 g, 19.61%).

[0079] 1 H NMR(400MHz,DMSO-d6)δ1.64-1.74(m,2H),1.90-1.93(m,4H),2.77-2.79(m,4H) ,2.98-3.04(m,1H),3.13-3.16(m,4H),3.42-3.47(m,2H),3.58(s,2H),3.77(s, 3H),3.91(d,J=9.6Hz,2H),6.13(d,J=2Hz,1H),6.43(d,J=3.2Hz,1H),7.24(d,J =3.6Hz,1H),7.38-7.46(m,2H),7.59(d,J=1.6Hz,1H),8.35(s,1H),9.55(s,1H). HPLC purity: 100% MS (ESI-MS): m / z C 27 H 33 F3N5O3[MH] + Calculated value: 532.25, Measured value: 532.08

[0080] compound 3 tert-Butyl 4-(4-chloro-2-nitrophenyl)-1,4-diazepane-1-carboxylate [ka] A suspension of 1-bromo-4-chloro-2-nitrobenzene (2.36 g, 9.98 mmol), tert-butyl 1,4-diazepane-1-carboxylate (2.0 g, 9.98 mmol), and solid sodium carbonate (4.14 g, 29.94 mmol) in anhydrous DMF (30 mL) was heated to 110 °C for 17 h. The resulting reaction mixture was cooled to room temperature and filtered through a short pad of Celite, eluting with ethyl acetate. The organic filtrate was concentrated under reduced pressure. The residue was diluted with water (100 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure. The resulting crude material was purified by column chromatography on silica using 100% chloroform as the eluent to give the title product as an orange liquid (3.20 g, 57%).

[0081] 1 H NMR(400MHz,CDCl3)δ1.39-1.48(m,9H),1.95(br s,2H),3.21-3.43(m,4H),3.51-3.57(m,4H),7.10(d,J=8.8Hz,1H),7.37(dd,J=2.8,9.2Hz,1H),7.69(d,J=2.8Hz,1H);LCMS:[M + -56]299.87m / z, 100% purity.

[0082] 1-(4-chloro-2-nitrophenyl)-1,4-diazepane [ka] Under a nitrogen atmosphere at 0 °C, 4N HCl in dioxane (20 mL) was added dropwise to a solution of tert-butyl 4-(4-chloro-2-nitrophenyl)-1,4-diazepane-1-carboxylate (2.20 g, 6.18 mmol) in dioxane (10 mL). The resulting reaction mixture was stirred at room temperature for 15 hours. After completion of the reaction, the reaction mixture was poured into a saturated solution of sodium bicarbonate (100 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined, washed with water and brine, and dried over anhydrous NaSO. The solvent was removed under reduced pressure to give the title product as a yellow solid (1.7 g, quantitative), which was used directly in the next step without further purification.

[0083] 1 H NMR(400MHz,DMSO-d6)δ2.73(t,J=5.2Hz,2H),2.88(t,J=4.8Hz,2H),3.10(t,J=4.8Hz,2H),3.36(t ,J=5.6Hz,2H),7.23(d,J=9.6Hz,1H),7.49(dd,J=2.8,9.2Hz,1H),7.79(d,J=2.8Hz,1H);LCMS:[MH] + 255.96m / z, 100% purity.

[0084] 1-(4-chloro-2-nitrophenyl)-4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepane [ka] To a solution of 1-(4-chloro-2-nitrophenyl)-1,4-diazepane (0.5 g, 1.96 mmol) and 1-methyl-1H-pyrazole-3-carbaldehyde (0.258 g, 2.35 mmol) in 1,2-dichloroethane (12 mL) was added sodium sulfate (0.138 g, 0.97 mmol) followed by glacial acetic acid (0.234 g, 3.91 mmol) at room temperature. The resulting reaction mixture was stirred at ambient temperature under a nitrogen atmosphere for 6 hours. Sodium triacetoxyborohydride (0.621 g, 2.93 mmol) was then added in portions to the reaction mixture at 0 °C. After stirring the resulting reaction mixture at room temperature for 17 hours, the reaction mixture was diluted with saturated aqueous sodium bicarbonate (100 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined, washed with brine, and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure. The resulting crude material was purified by flash chromatography on silica using (2% methanol in chloroform) as the eluent to give the title product as an orange liquid (0.78 g, quantitative).

[0085] 1 H NMR(400MHz,CDCl3)δ1.70(br s,2H),3.10(br s,4H),3.27-3.30(m,4H),3.92(s,3H),3.97(br s,2H),6.46(br s,1H),7.09(d,J=9.2Hz,1H),7.38-7.45(m,2H),7.70-7.72(m,1H);LCMS:[MH] + 350.03 m / z, purity 98.32%.

[0086] 5-chloro-2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepan-1-yl)aniline [ka] Under a nitrogen atmosphere at 0°C, hydrazine hydrate (2.79 g, 55.74 mmol) was added in several portions to a solution of 1-(4-chloro-2-nitrophenyl)-4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepane (0.78 g, 2.23 mmol), iron(III) chloride (0.072 g, 0.44 mmol), and charcoal (0.1 g) in methanol (30 mL). The resulting reaction mixture was heated to reflux for 30 minutes. After cooling to room temperature, the reaction mixture was filtered through a short pad of Celite and eluted with ethyl acetate. The organic filtrate was concentrated under reduced pressure. The residue was diluted with water (50 mL) and extracted with ethyl acetate (3 times). The organic extracts were combined and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure. The resulting crude material was purified by flash chromatography on silica using (2% methanol in chloroform) as the eluent to give the title product as a pale brown liquid (0.5 g, 70%).

[0087] 1 H NMR(400MHz,DMSO-d6)δ1.82(br s,2H),2.78(br s,4H),2.96(t,J=5.6Hz,4H),3.64(br s,2H),3.78(s,3H),5.03(br s,2H),6.18(br s,1H),6.48(d,J=2.4,8.4Hz,1H),6.65(d,J=2.4Hz,1H),6.90(d,J=8.4Hz,1H),7.61(br s,1H);LCMS:[MH] + 320.07m / z, 100% purity.

[0088] N-(5-chloro-2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepan-1-yl)phenyl)-5-(tetrahydro-2H-pyran-4-yl)furan-2-carboxamide [ka] To a solution of 5-chloro-2-(4-((1-methyl-1H-pyrazol-3-yl)methyl)-1,4-diazepan-1-yl)aniline (0.2 g, 0.62 mmol) in dichloromethane (8 mL) was added dropwise a 2.0 M solution of trimethylaluminum (0.93 mL, 1.88 mmol) in toluene under a nitrogen atmosphere at 0 °C. After stirring this mixture at room temperature for 1 hour, a solution of methyl 5-(tetrahydro-2H-pyran-4-yl)-2-furoate (0.131 g, 0.62 mmol) in dichloromethane (3 mL) was added dropwise at room temperature under a nitrogen atmosphere. The resulting reaction mixture was stirred at room temperature for 21 hours. The resulting reaction mixture was diluted with saturated aqueous sodium bicarbonate (50 mL) and extracted with dichloromethane (3 times). The organic extracts were combined, washed with brine, and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure. The resulting crude material was purified by preparative HPLC using 0.1% formic acid in water and acetonitrile as the mobile phase to give the title product (compound 3) as a brown solid (0.035 g, 11.24%).

[0089] Mp: 148-150℃; 1 H NMR(400MHz,DMSO-d6)δ1.67-1.71(m,2H),1.87-1.93(m,4H),2.76(br s,2H),2.83(t,J=5.6Hz,2H),3.01-3.04(m,5H),3.44(td,J=8,11.6Hz,2H),3.60(br s,2H),3.77(s,3H),3.91(dd,J=2,11.6Hz,2H),6.14(d,J=2Hz,1H),6.44(dd,J=0.8,3.6Hz,1H),7.13(dd,J=2.8, 8.4Hz,1H),7.23(d,J=3.6Hz,1H),7.33(d,J=8.8Hz,1H),7.60(d,J=2Hz,1H),8.29(d,J=2.4Hz,1H),9.61(s,1H). HPLC purity: 98.70% MS (ESI-MS): m / z C 26 H 33 ClN5O3[MH] + Calculated value: 498.23, Measured value: 498.12

[0090] In vitro kinase assay Kinase assays 33 Radioactive filter binding assays with P ATP were performed by the MRC Dundee Kinase Centre (Hastie, et al 2006. Nat Protoc. 2006;1(2):968-71; Bain, et al 2007. Biochem J. 2007 Dec 15;408(3):297-315).

[0091] In vivo angiogenesis assay: Laser-induced choroidal neovascularization (CNV) protocol Six- to eight-week-old female C57 / B6 mice were anesthetized by intraperitoneal injection of a mixture of 50 mg / kg ketamine and 0.5 mg / kg medetomidine. Pupils were immediately dilated by topical application of a dilating agent, such as 5% phenylephrine hydrochloride and 1% tropicamide. Using a green Merilas 532α laser (450 mW, 130 ms), four photocoagulated lesions were created in each eye, one to two papillary diameters apart, between "large" retinal vessels in a peripapillary distribution, with clear, avascular space. Only clean laser lesions with subretinal air bubbles at the time of treatment were included in the study. Immediately after laser photocoagulation, animals received topical instillation of 2 μg / mL, 0.2 μg / mL, or 0.066 μg / mL of the candidate compound or a control eye drop formulation twice daily as indicated (10 μL, with the eye held for 30 seconds to prevent the animal from wiping off the drop). Experiments were performed with an initial eye drop formulation containing 1% hydroxypropylmethylcellulose, 0.2% tyloxapol, 3.4% dextrose, 0.006% benzalkonium chloride, and 0.025% ethylenediaminetetraacetic acid in PBS, and 1% DMSO, and the data were confirmed with a second eye drop formulation containing 7% polyoxyl 40 stearate and 4% dextrose in PBS.

[0092] One week later, mice were anesthetized with an intraperitoneal injection of a mixture of 50 mg / kg ketamine and 0.5 mg / kg medetomidine. Pupils were immediately dilated by topical application of dilating agents such as 5% phenylephrine hydrochloride and 1% tropicamide. Mice were then given an intraperitoneal injection of sodium fluorescein (10%). Phase-contrast and green fluorescent fundus images were taken using an angiographic microscope and camera, focusing on each lesion. Mice were sacrificed using schedule 1 methods, and eyes were either left unfixed for retinal detachment and protein extraction or fixed and enucleated, and the choroids were stained and examined.

[0093] VEGF 165 a VEGF ELISA with capture antibody In a 96-well clear microplate (High Sensitivity ThermoImmunon or Costar 9018), 10 μg / ml LVEGF was added per well. xxx b or 0.25 μg / mL anti-hVEGF 165The plates were coated with 100 μL of a. The plates were sealed with parafilm and incubated overnight on a shaker at room temperature. Each well was aspirated and washed twice with wash buffer (200 μL PBS-Tween 0.05%) for a total of three times. After the final wash, the remaining wash buffer was removed by inverting the plate and blotting it onto a clean paper towel. The plate was blocked by adding 100 μL of reagent diluent (1% BSA / PBS) to each well and incubated for 2 hours on a shaker at room temperature. The aspirate / wash cycle was repeated. 100 μL of standard or sample in 1% BSA / PBS was added to each well, covered with parafilm, and incubated for 2 hours at room temperature. The aspirate / wash cycle was repeated, and 100 μL of 100 ng / mL detection antibody (BAF293) diluted in reagent diluent was added to each well. The wells were covered with parafilm and incubated for 2 hours at room temperature. After repeated aspiration / washing, 100 μL of working dilution of Streptavidin-HRP (1:200 dilution) was added to each well. The plate was covered and incubated at room temperature for 30 minutes. The plate was washed, and 100 μL of substrate solution (1:1 A:B from DY999) was added to each well and incubated at room temperature for 20-60 minutes. 50 μL of stop solution (1 M HCl) was added to each well. The optical density of each well was immediately measured using a microplate reader set at 450 nm.

[0094] Melanin binding assay Test compounds (1 μg / mL) in 1% DMSO in PBS were incubated with 1 mg / mL melanin for 1 hour at 37°C. The solution was then spun at 15 kg for 15 minutes, the supernatant was collected, and the compounds were extracted into methanol and quantified by mass spectrometry.

[0095] Rabbit pharmacokinetic study Rabbits were treated with a single 50 μL instillation of 80 μg / mL (maximum solubility limit) pazopanib and 500 μg / mL Compound R in an initial eye drop formulation containing 1% hydroxypropylmethylcellulose, 0.2% tyloxapol, 3.4% dextrose, 0.006% benzalkonium chloride, and 0.025% ethylenediaminetetraacetic acid in PBS, plus 1% DMSO, or with a single 50 μL instillation of 500 μg / mL Compound 1 in an eye drop formulation containing 7% polyoxyl 40 stearate and 4% dextrose in PBS. No differences were observed between the eye drop formulations in separate studies. After instillation, rabbits were sacrificed at the indicated time points, bled, and harvested. The retinas were dissected from the choroid and sclera, dissected, and laid flat. The retina, RPE / choroid, and sclera compartments were dissected into seven distinct regions. All samples were weighed. Compounds were extracted from retina and choroid / sclera samples, as well as plasma, by reverse-phase extraction, as described above, and determined by mass spectrometry in different regions of the eye and in blood. For each sample, the amounts per milligram of tissue for Compound R, Compound 1, and pazopanib were calculated and averaged.

[0096] Scleral permeability was measured using a modified Ussing chamber assembly in an initial eye drop formulation containing 1% hydroxypropylmethylcellulose, 0.2% tyloxapol, 3.4% dextrose, 0.006% benzalkonium chloride, and 0.025% ethylenediaminetetraacetic acid in PBS (pH 7.4), and 1% DMSO. Porcine excised eye tissue was placed in the chamber with the episcleral side facing the donor chamber and the retinal side facing the receiver chamber. The donor side was filled with 10 μg / mL of compound, and the receiver side was filled with an equal volume of 10 μg / mL of compound-free eye drop formulation. After 24 hours, the tissue was removed from the chamber, and the receiver side ("vitreous") was sampled. The tissue was disaggregated into sclera, choroid / RPE, and retina and homogenized. Tracer (SPHINX7; WO 2015 / 159103) was added and tissues were extracted by acetonitrile extraction as described by Batson et al. (2017), after which compounds were analyzed by mass spectrometry as described by Batson et al. (2017).

[0097] Pharmacokinetic study in non-human primates Cynomolgus monkeys were treated every other day for 20 days by instilling 35 μL of Compound 1 at 0.5 mg / mL, 1.0 mg / mL, or 1.5 mg / mL in an eye drop solution containing 7% poly-oxyl-40-stearate and 4% dextrose in PBS. Blood and aqueous samples were collected 1, 4, 8, 10, and 14 hours after the eye drop administration on Day 1, and at the end of the study. One hour after the final eye drop on Day 21, animals were euthanized, and ocular tissues and blood were collected. Retinas were detached from the choroid and sclera, dissected, and laid flat. The retina, RPE / choroid, and sclera ocular compartments were dissected into seven different regions. All samples were weighed. Compounds were extracted from the retina and choroid / sclera samples, as well as plasma, by reverse-phase extraction, as described above, and determined in different eye regions and blood by mass spectrometry. For each sample, the amount per milligram of tissue or nM of aqueous solution and blood was calculated and averaged.

[0098] Retinal sections sampled from half of the eye were homogenized in NP40 lysis buffer as described by Gammons et al., 2013. Extracts were then incubated with rabbit anti-panVEGF (Santa Cruz A20sc-152; 1:500) or mouse anti-VEGF (Santa Cruz A20sc-152; 1:500) for 1 hour. 165 b (MAB3045; R&D; 1:500) or

[0099] result The following examples demonstrate the present invention: The compounds of the present invention are potent SRPK1 inhibitors and have high ocular permeability.

[0100] We have high efficacy (IC 50 <10 -8 We have previously determined that it is possible to generate SRPK1 inhibitors that have a potent SRPK1 activity (M) and are antiangiogenic in a mouse model of choroidal neovascularization, and that can penetrate through the sclera to the choroid and retinal pigment epithelium layers of rabbits. Maximizing the intraocular permeability of molecules is important for the development of effective topical therapeutics. The compounds of the present invention have improved properties over the compounds described in WO 2014 / 060763 and WO 2017 / 064512, and are useful for the topical treatment of ocular neovascularization and hyperpermeability disorders that depend on the overexpression of antiangiogenic VEGF isoforms. [Example]

[0101] transparency To determine whether compounds can penetrate the sclera of larger animals, pig sclera were clamped between two chambers, and the eye drop formulation was added to the lower chamber, while the compound was added to the upper chamber. After 24 hours, the fluid (vitreous) from the lower chamber and retinal tissue were separated, and the compounds were purified by methanol or acetonitrile extraction and HPLC. This demonstrated the feasibility of producing compounds with both high permeability and high potency. In particular, the novel compounds of the present invention (shown for compounds 1-3) exhibited surprisingly high permeability, far exceeding that of SPHINX31 (Figure 1; Table 1), while still retaining high potency (Table 2). This was particularly surprising, given that chemically similar compounds lacking the methyl group on the pyrazole, such as compound R described above, exhibited significantly lower permeability (Figure 1). Furthermore, the compounds of the present invention also have substantially greater penetration than compounds previously shown to inhibit VEGF signaling through VEGFR2 but which failed in clinical trials due to lack of exposure (pazopanib, regorafenib, LHA510), and greater penetration than compounds that are effectively used as topical eye drops but act on the anterior segment (indomethacin, celecoxib) - see Figure 1. [Table 1] Melanin binding has been proposed to be important for compound bioavailability in the eye. The melanin binding of the compounds of the present invention was measured and determined to be significantly less than previous SRPK1 inhibitors (SPHINX31; WO 2015 / 159103 and Compound R) and previous VEGFR2 inhibitors that have been tried as anti-angiogenic agents (Figure 2).

[0102] To determine whether compounds can access the RPE in animals with large eyes, rabbits were exposed to 500 μg / mL of compound by eye dropwise instillation, and concentrations were measured in the retina, sclera, vitreous, and other ocular tissues (cornea, lens, and RPE / choroid) (Figure 3a). After 1, 4, 12, or 24 hours, the animals were sacrificed and the eyes were harvested. Individual sections of the cornea (100), lens (102), and vitreous (110), as well as sections of the sclera (108), RPE / choroid (106), and retina (104) from the posterior portion of the eye, were then assayed for compound. Retinal penetration was observed for compounds of the present invention. Results obtained for Compound 1 are shown in Figures 3a and 3b. Permeability was superior to that of previously studied SRPK1 inhibitors, such as SPHINX31, and had a sustained permeability (e.g., 4 hours) superior to that of chemically similar molecules, such as Compound R (Figure 4).

[0103] SRPK1 selectivity The inhibitory activity of the compounds of the present invention against SRPK1 was measured and the data are shown in Table 2. All compounds were potent inhibitors of SRPK1 and had high selectivity for SRPK1 when tested against a panel of kinases at 1 μM. [Table 2]

[0104] Compound 1 switches expression to the anti-angiogenic isoform in human cells. To determine whether compound 1 can switch the splicing of VEGF isoforms, VEGF levels in retinal pigment epithelial cells were measured by isoform-specific ELISA. Figure 5 shows that treatment with compound 1 dose-dependently suppressed the pro-angiogenic VEGF-A activity. 165 a Shown to decrease the expression of isoforms.

[0105] Compound 1 switches expression away from pro-angiogenic VEGF isoforms in a rat diabetic retinopathy model. To determine whether Compound 1 could effectively switch VEGF splicing in a diabetic model, we used the STZ model of diabetes in Norway Brown rats. The diabetic animals were treated with Compound 1 eye drops twice daily for 4 weeks, then sacrificed and their retinas were detached. Protein was extracted and VEGF-A was analyzed. 165 6a to 6d show that total VEGF was increased in diabetic animals compared with healthy rats, but VEGF-A was not. 165 b was decreased, which was reversed by treatment with Compound 1.

[0106] Compound 1 inhibits choroidal neovascularization in vivo. We previously demonstrated that SRPK1 inhibition by SPHINX31 is antiangiogenic in a mouse model of choroidal neovascularization, with maximal efficacy at 2 μg / mL as an eye drop. These compounds are relatively lipophilic and have high intraocular permeability. Therefore, we tested the effects of compound 1 as an eye drop in this same model. Compound 1 significantly inhibited choroidal neovascularization at 0.066 μg / mL (Figure 7b).

[0107] Compound 1 is systemic at effective doses in non-human primates. To determine whether Compound 1 could penetrate the primate retina, cynomolgus monkeys were dosed with 0.5, 1.0, or 1.5 mg / mL of Compound 1 twice daily for 21 days. Figure 8 shows that significant concentrations of Compound 1 were observed in the retina, choroid, vitreous, and other ocular tissues of the primate retina, at concentrations far higher than those required for efficacy based on mouse models (Figures 8a and 8b). Water droplets and plasma were collected during treatment, and Compound 1 in the fluid was measured. Significant concentrations were observed at all three doses, with a calculated half-life in water of 2 hours after the 1.5 mg / mL dose (Figure 9). To determine whether these doses were sufficient to inhibit angiogenic VEGF-A isoform expression, retinal tissue was treated with either pan-VEGF or VEGF-A. 165Fig. 10a shows that retinal VEGF levels were altered in monkeys treated with Compound 1 by eye drop administration, and Fig. 10b shows that total VEGF levels were reduced and VEGF-A levels were increased after 3 weeks of every other day administration. 165 b shows that levels do not decrease but increase at 1 mg / ml, and Figure 10c shows that the ratio of angiogenic to anti-angiogenic VEGF-A is decreased in the retinas of monkeys treated with Compound 1.

[0108] References Bressler, S., Bressler, NM, Clemons, T., Ferris, FL, Milton, RC, Klien, R., Klien, B. and Age-Related Eye Dis Study, G. (2004)'Ocular risk factors for developing neovascular AMD in the fellow eyes of patients with unilateral neovascular AMD', Investigative Ophthalmology & Visual Science, 45, U924-U924. Ferris, FL, Fine, SLand Hyman, L. (1984)'Age-related macular degeneration and blindness due to neovascular maculopathy', Archives of Ophthalmology, 102(11), 1640-1642. Patz, A., Fine, SL, Finkelstein, D. and Yassur, Y. (1977) 'Diseases of macula - diagnosis and management of choroidal neovascularization', Transactions American Academy of Ophthalmology and Otolaryngology, 83(3), 468-475. Fine,S.L.,Berger,J.W.,Maguire,M.G.and Ho,A.C.(2000)‘Drug therapy: Age-related macular degeneration’,New England Journal of Medicine,342(7),483-492. Campochiaro,P.A.,Nguyen,Q.D.,Shah,S.M.,Klein,M.L.,Holz,E.,Frank,R.N.,Saperstein,D.A.,Gupta,A.,Stout,J.T.,Macko,J.,DiBartolomeo,R.and Wei,L.L.(2006)‘Adenoviral vector-delivered pigment epithelium-derived factor for neovascular age-related macular degeneration: Results of a phase I clinical trial’,Human Gene Therapy,17(2),167-176. Dvorak,H.F.,Brown,L.F.,Detmar,M.and Dvorak,A.M.(1995)‘Vascular-permeability factor vascular endothelial growth-factor,microvascular hyperpermeability,and angiogenesis’,American Journal of Pathology,146(5),1029-1039. Spilsbury,K.,Garrett,K.L.,Shen,W.Y.,Constable,I.J.and Rakoczy,P.E.(2000)‘Overexpression of vascular endothelial growth factor(VEGF)in the retinal pigment epithelium leads to the development of choroidal neovascularization’,American Journal of Pathology,157(1),135-144. Anderson,D.H.,Mullins,R.F.,Hageman,G.S.and Johnson,L.V.(2002)‘Perspective - A role for local inflammation in the formation of drusen in the aging eye’,American Journal of Ophthalmology,134(3),411-431. Das,A.,Fanslow,W.,Cerretti,D.,Warren,E.,Talarico,N.and McGuire,P.(2003)‘Angiopoietin / Tek interactions regulate MMP-9 expression and retinal neovascularization’,Laboratory Investigation,83(11),1637-1645. Leung,D.W.,Cachianes,G.,Kuang,W.J.,Goeddel,D.V.and Ferrara,N.(1989)‘Vascular endothelial growth-factor is a secreted angiogenic mitogen’,Science,246(4935),1306-1309. Jingjing,L.,Xue,Y.,Agarwal,N.and Roque,R.S.(1999)‘Human Muller cells express VEGF183,a novel spliced variant of vascular endothelial growth factor’,Iovs,40(3),752-759. Houck,K.A.,Ferrara,N.,Winer,J.,Cachianes,G.,Li,B.and Leung,D.W.(1991)‘The vascular endothelial growth-factor family - identification of a 4th molecular-species and characterization of alternative splicing of rna’,Molecular Endocrinology,5(12),1806-1814. Mineur,P.,Colige,A.C.,Deroanne,C.F.,Dubail,J.,Kesteloot,F.,Habraken,Y.,Noel,A.,Voo,S.,Waltenberger,J.,Lapiere,C.M.,Nusgens,B.V.and Lambert,C.A.(2007)‘Newly identified biologically active and proteolysis-resistant VEGF-A isoform VEGF111 is induced by genotoxic agents’,Journal of Cell Biology,179(6),1261-1273. Tischer,E.,Gospodarowicz,D.,Mitchell,R.,Silva,M.,Schilling,J.,Lau,K.,Crisp,T.,Fiddes,J.C.and Abraham,J.A.(1989)‘Vascular endothelial growth-factor - a new member of the platelet-derived growth-factor gene family’,Biochemical and Biophysical Research Communications,165(3),1198-1206. Neufeld,G.,Cohen,T.,Gengrinovitch,S.and Poltorak,Z.(1999)‘Vascular endothelial growth factor(VEGF)and its receptors’,Faseb Journal,13(1),9-22. Bates,D.O.,Cui,T.G.,Doughty,J.M.,Winkler,M.,Sugiono,M.,Shields,J.D.,Peat,D.,Gillatt,D.and Harper,S.J.(2002)‘VEGF(165)b,an inhibitory splice variant of vascular endothelial growth factor,is down-regulated in renal cell carcinoma’,Cancer Research,62(14),4123-4131. Woolard,J.,Wang,W.Y.,Bevan,H.S.,Qiu,Y.,Morbidelli,L.,Pritchard-Jones,R.O.,Cui,T.G.,Sugiono,M.,Waine,E.,Perrin,R.,Foster,R.,Digby-Bell,J.,Shields,J.D.,Whittles,C.E.,Mushens,R.E.,Gillatt,D.A.,Ziche,M.,Harper,S.J.and Bates,D.O.(2004)‘VEGF(165)b,an inhibitory vascular endothelial growth factor splice variant: Mechanism of action,in vivo effect on angiogenesis and endogenous protein expression’,Cancer Research,64(21),7822-7835. Perrin,R.M.,Konopatskaya,O.,Qiu,Y.,Harper,S.,Bates,D.O.and Churchill,A.J.(2005)‘Diabetic retinopathy is associated with a switch in splicing from anti- to pro-angiogenic isoforms of vascular endothelial growth factor’,Diabetologia,48(11),2422-2427. Varey,A.H.R.,Rennel,E.S.,Qiu,Y.,Bevan,H.S.,Perrin,R.M.,Raffy,S.,Dixon,A.R.,Paraskeva,C.,Zaccheo,O.,Hassan,A.B.,Harper,S.J.and Bates,D.O.(2008)‘VEGF(165)b,an antiangiogenic VEGF-A isoform,binds and inhibits bevacizumab treatment in experimental colorectal carcinoma: balance of pro- and antiangiogenic VEGF-A isoforms has implications for therapy’,British Journal of Cancer,98(8),1366-1379. Pritchard-Jones,R.O.,Dunn,D.B.A.,Qiu,Y.,Varey,A.H.R.,Orlando,A.,Rigby,H.,Harper,S.J.and Bates,D.O.(2007)‘Expression of VEGF(xxx)b,the inhibitory isoforms of VEGF,in malignant melanoma’,British Journal of Cancer,97(2),223-230. Hua,J.,Spee,C.,Kase,S.,Rennel,E.S.,Magnussen,A.L.,Qiu,Y.,Varey,A.,Dhayade,S.,Churchill,A.J.,Harper,S.J.,Bates,D.O.and Hinton,D.R.(2010)‘Recombinant Human VEGF(165)b Inhibits Experimental Choroidal Neovascularization’,Investigative Ophthalmology & Visual Science,51(8),4282-4288. Magnussen,A.L.,Rennel,E.S.,Hua,J.,Bevan,H.S.,Long,N.B.,Lehrling,C.,Gammons,M.,Floege,J.,Harper,S.J.,Agostini,H.T.,Bates,D.O.and Churchill,A.J.(2010)‘VEGF-A(165)b Is Cytoprotective and Antiangiogenic in the Retina’,Investigative Ophthalmology & Visual Science,51(8),4273-4281. Rosenfeld,P.J.,Rich,R.M.and Lalwani,G.A.(2006)‘Ranibizumab: Phase III clinical trial results’,Ophthalmology clinics of North America,19(3),361-72. Brown,D.M.,Kaiser,P.K.,Michels,M.,Soubrane,G.,Heier,J.S.,Kim,R.Y.,Sy,J.P.,Schneider,S.and Grp,A.S.(2006)‘Ranibizumab versus verteporfin for neovascular age-related macular degeneration’,New England Journal of Medicine,355(14),1432-1444. Brown,D.M.,Michels,M.,Kaiser,P.K.,Heier,J.S.,Sy,J.P.and Ianchulev,T.(2009)‘Ranibizumab versus Verteporfin Photodynamic Therapy for Neovascular Age-Related Macular Degeneration: Two-Year Results of the ANCHOR Study’,Ophthalmology,116(1),57-65. Schmidt-Erfurth,U.,Eldem,B.,Guymer,R.,Korobelnik,J.-F.,Schlingemann,R.O.,Axer-Siegel,R.,Wiedemann,P.,Simader,C.,Gekkieva,M.,Weichselberger,A.and Grp,E.S.(2011)‘Efficacy and Safety of Monthly versus Quarterly Ranibizumab Treatment in Neovascular Age-related Macular Degeneration: The EXCITE Study’,Ophthalmology,118(5). Good,T.J.and Kahook,M.Y.(2010)‘The role of endothelin in the pathophysiology of glaucoma’,Expert Opinion on Therapeutic Targets,14(6),647-654. Jager,R.D.,Aiello,L.P.,Patel,S.C.and Cunningham,E.T.(2004)‘Risks of intravitreous injection: A comprehensive review’,Retina-the Journal of Retinal and Vitreous Diseases,24(5),676-698. Nowak,D.G.,Amin,E.M.,Rennel,E.S.,Hoareau-Aveilla,C.,Gammons,M.,Damodoran,G.,Hagiwara,M.,Harper,S.J.,Woolard,J.,Ladomery,M.R.and Bates,D.O.(2010)‘Regulation of Vascular Endothelial Growth Factor(VEGF)Splicing from Pro-angiogenic to Anti-angiogenic Isoforms a novel therapeutic strategy for angiogenesis’,Journal of Biological Chemistry,285(8),5532-5540. Amin,E.M.,Oltean,S.,Hua,J.,Gammons,M.V.R.,Hamdollah-Zadeh,M.,Welsh,G.I.,Cheung,M.-K.,Ni,L.,Kase,S.,Renne,E.S.,Symonds,K.E.,Nowak,D.G.,Royer-Pokora,B.,Saleem,M.A.,Hagiwara,M.,Schumacher,V.A.,Harper,S.J.,Hinton,D.R.,Bates,D.O.and Ladomery,M.R.(2011)‘WT1 Mutants Reveal SRPK1 to Be a Downstream Angiogenesis Target by Altering VEGF Splicing’,Cancer Cell,20(6),768-780. Sanford,J.R.,Ellis,J.D.,Cazalla,D.and Caceres,J.F.(2005a)‘Reversible phosphorylation differentially affects nuclear and cytoplasmic functons of splicing factor 2 / alternative splicing factor’,Proceedings of the National Academy of Sciences of the United States of America,102(42),15042-15047. Nowak,D.G.,Woolard,J.,Amin,E.M.,Konopatskaya,O.,Saleem,M.A.,Churchill,A.J.,Ladomery,M.R.,Harper,S.J.and Bates,D.O.(2008)‘Expression of pro- and anti-angiogenic isoforms of VEGF is differentially regulated by splicing and growth factors’,Journal of Cell Science,121(20),3487-3495. Doukas,J.,Mahesh,S.,Umeda,N.,Kachi,S.,Akiyama,H.,Yokoi,K.,Cao,J.,Chen,Z.,Dellamary,L.,Tam,B.,Racanelli-Layton,A.,Hood,J.,Martin,M.,Noronha,G.,Soll,R.and Campochiaro,P.A.(2008)‘Topical administration of a multi-targeted kinase inhibitor suppresses choroidal neovascularization and retinal edema’,Journal of Cellular Physiology,216(1),29-37. Fukuhara,T.,Hosoya,T.,Shimizu,S.,Sumi,K.,Oshiro,T.,Yoshinaka,Y.,Suzuki,M.,Yamamoto,N.,Herzenberg,L.A.and Hagiwara,M.(2006)‘Utilization of host SR protein kinases and RNA-splicing machinery during viral replication’,Proceedings of the National Academy of Sciences of the United States of America,103(30),11329-11333. Rennel,E.S.,Regula,J.T.,Harper,S.J.,Thomas,M.,Klein,C.and Bates,D.O.(2011)‘A Human Neutralizing Antibody Specific to Ang-2 Inhibits Ocular Angiogenesis’,Microcirculation,18(7). Aubol,B.E.,Chakrabarti,S.,Ngo,J.,Shaffer,J.,Nolen,B.,Fu,X.D.,Ghosh,G.and Adams,J.A.(2003)‘Processive phosphorylation of alternative splicing factor / splicing factor 2’,Proceedings of the National Academy of Sciences of the United States of America,100(22),12601-12606. Velazquez-Dones,A.,Hagopian,J.C.,Ma,C.T.,Zhong,X.Y.,Zhou,H.L.,Ghosh,G.,Fu,X.D.and Adams,J.A.(2005)‘Mass spectrometric and kinetic analysis of ASF / SF2 phosphorylation by SRPK1 and Clk / Sty’,Journal of Biological Chemistry,280(50),41761-41768. Ngo,J.C.K.,Chakrabarti,S.,Ding,J.H.,Velazquez-Dones,A.,Nolen,B.,Aubol,B.E.,Adams,J.A.,Fu,X.D.and Ghosh,G.(2005)‘Interplay between SRPK and Clk / Sty kinases in phosphorylation of the splicing factor ASF / SF2 is regulated by a docking motif in ASF / SF2’,Molecular Cell,20(1),77-89. Xu,J.,Dou,T.,Liu,C.,Fu,M.,Huang,Y.,Gu,S.,Zhou,Y.and Xie,Y.(2011)‘The evolution of alternative splicing exons in vascular endothelial growth factor A’,Gene,487(2). Caires,K.C.,de Avila,J.M.,Cupp,A.S.and McLean,D.J.(2012)‘VEGFA Family Isoforms Regulate Spermatogonial Stem Cell Homeostasis in Vivo’,Endocrinology,153(2). Zhao,M.,Shi,X.,Liang,J.,Miao,Y.,Xie,W.,Zhang,Y.and Li,X.(2011)‘Expression of pro- and anti-angiogenic isoforms of VEGF in the mouse model of oxygen-induced retinopathy’,Experimental Eye Research,93(6),921-926. Harris,S.,Craze,M.,Newton,J.,Fisher,M.,Shima,D.T.,Tozer,G.M.and Kanthou,C.(2012)‘Do Anti-Angiogenic VEGF(VEGF xxx b)Isoforms Exist? A Cautionary Tale’,Plos One,7(5). McFee,R.M.,Rozell,T.G.and Cupp,A.S.(2012)‘The balance of proangiogenic and antiangiogenic VEGFA isoforms regulate follicle development’,Cell and Tissue Research,349(3). Ishida,S.,Usui,T.,Yamashiro,K.,Kaji,Y.,Amano,S.,Ogura,Y.,Hida,T.,Oguchi,Y.,Ambati,J.,Miller,J.W.,Gragoudas,E.S.,Ng,Y.S.,D’Amore,P.A.,Shima,D.T.and Adamis,A.P.(2003)‘VEGF( 164 )-mediated inflammation is required for pathological,but not physiological,ischemia-induced retinal neovascularization’,Journal of Experimental Medicine,198(3),483-489. Geroski,D.H.and Edelhauser,H.F.(2000)‘Drug delivery for posterior segment eye disease’,Investigative Ophthalmology & Visual Science,41(5),961-964. Keyt,B.A.,Nguyen,H.V.,Berleau,L.T.,Duarte,C.M.,Park,J.,Chen,H.and Ferrara,N.(1996)‘Identification of vascular endothelial growth factor determinants for binding KDR and FLT-1 receptors - Generation of receptor-selective VEGF variants by site-directed mutagenesis’,Journal of Biological Chemistry,271(10),5638-5646. Stalmans,I.,Ng,Y.S.,Rohan,R.,Fruttiger,M.,Bouche,A.,Yuce,A.,Fujisawa,H.,Hermans,B.,Shani,M.,Jansen,S.,Hicklin,D.,Anderson,D.J.,Gardiner,T.,Hammes,H.P.,Moons,L.,Dewerchin,M.,Collen,D.,Carmeliet,P.and D’Amore,P.A.(2002)‘Arteriolar and venular patterning in retinas of mice selectively expressing VEGF isoforms’,Journal of Clinical Investigation,109(3). Gammons,M.V.,Dick,A.D.,Harper,S.J.,Bates,D.O.(2013)SRPK1 Inhibition Modulates VEGF Splicing to Reduce Pathological Neovascularization in a Rat Model of Retinopathy of Prematurity Invest.Ophthalmol.Vis.Sci.vol.54(8)5797-5806. Gammons,M.V.,Fedorov,O.,Ivison,D.,Du,C.,Clark,T.,Hopkins,C.,Hagiwara,M.,Dick,A.D.,Cox,R.,Harper,S.J.,Hancox,J.C.and Bates,D.O.(2013)Topical Antiangiogenic SRPK1 Inhibitors Reduce Choroidal Neovascularization in Rodent Models of Exudative AMD Invest.Ophthalmol.Vis.Sci.54(9)6052-6062. Federov O,Niesen FH,Knapp S.Kinase Inhibitor Selectivity Profiling Using Differential Scanning Fluorimetry.In: Kuster B,ed.Kinase Inhibitors: Methods and Protocols: Springer,2011:109-18. Carter JG,Gammons MV,Damodaran G,Churchill AJ,Harper SJ,Bates DO.(2015)The carboxyl terminus of VEGF-A is a potential target for anti-angiogenic therapy.Angiogenesis 18(1),23-30. Chomczynski,P.,and Sacchi,N.Single-step method of RNA isolation by acid quanidinium thiocyanate phenol chloroform extraction.Anal.Biochem.,162: 156-159,1987. Batson,J.,Toop,H.D.,Redondo,C.,Babaei-Jadidi,R.,Chaikuad,A.,Wearmouth,S.F.,Gibbons,B.,Allen,C.,Tallant,C.,Zhang,J.,Du,C.,Hancox,J.C.,Hawtrey,T.,Da Rocha,J.,Griffith,R.,Knapp,S.,Bates,D.O.,Morris,J.C.,Development of Potent,Selective SRPK1 Inhibitors as Potential Topical Therapeutics for Neovascular Eye Disease,ACS Chem.Biol.2017,12,825-832.

Claims

1. When X is CF3 and n=1, the compound of formula (I): 【Chemistry 1】 or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

2. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of ocular neovascularization.

3. The treatment or prevention of ocular neovascularization may involve the induction of abnormal angiogenesis or pro-angiogenic VEGF. xxx 3. A compound for use according to claim 2, including the treatment or prevention of eye diseases associated with abnormal overproduction of isoforms.

4. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of age-related macular degeneration.

5. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of diabetic macular oedema.

6. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of diabetic retinopathy.

7. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of retinal vein occlusion.

8. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of retinopathy of prematurity.

9. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of ocular neurodegenerative disorders.

10. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of ocular degenerative diseases such as geographic atrophy.

11. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of ocular hyperpermeability disorders.

12. 10. A compound of formula (I) as defined in claim 1 for use in the treatment or prevention of ocular epithelial degenerative disorders.

13. A compound for use according to any one of claims 2 to 12, including topical treatment or prevention.

14. 10. A pharmaceutical composition comprising a compound of claim 1, optionally one or more other active ingredients, and a pharmaceutically acceptable carrier.

15. 10. A pharmaceutical composition in a form suitable for intraocular injection, comprising a compound according to claim 1, optionally one or more other active ingredients, and a pharmaceutically acceptable carrier.

16. 10. A pharmaceutical composition in a form suitable for topical administration to the eye, comprising a compound of claim 1, optionally one or more other active ingredients, and a pharmaceutically acceptable carrier.

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