Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/4965—Non-condensed pyrazines
  • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
  • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/04—Centrally acting analgesics, e.g. opioids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/12—Antihypertensives
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
  • C07C271/06—Esters of carbamic acids
  • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
  • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
  • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
  • C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
  • C07D233/90—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
  • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
  • C07D241/04—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D269/00—Heterocyclic compounds containing rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms according to more than one of groups C07D261/00 - C07D267/00
  • C07D269/02—Heterocyclic compounds containing rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms according to more than one of groups C07D261/00 - C07D267/00 having the hetero atoms in positions 1 and 3
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D271/00—Heterocyclic compounds containing five-membered rings having two nitrogen atoms and one oxygen atom as the only ring hetero atoms
  • C07D271/02—Heterocyclic compounds containing five-membered rings having two nitrogen atoms and one oxygen atom as the only ring hetero atoms not condensed with other rings
  • C07D271/10—1,3,4-Oxadiazoles; Hydrogenated 1,3,4-oxadiazoles
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
  • C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
  • C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D307/38—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
  • C07D307/52—Radicals substituted by nitrogen atoms not forming part of a nitro radical
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
  • C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
  • C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D307/38—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
  • C07D307/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
  • C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
  • C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D307/56—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
  • C07D307/68—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D309/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings
  • C07D309/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
  • C07D309/04—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings having no double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
  • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
  • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
  • C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
  • C07D333/38—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
  • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
  • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
  • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
  • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
  • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
  • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

• the present invention relates to anti-angiogenic treatments and compounds for use in anti- angiogenic treatments, particularly of conditions characterised by neovascularisation such as. for example, age-related macular degeneration.
• the present invention also relates to treatments of hyperpermeabilily disorders and compounds for use in treating hyperpermeabi lity disorders.
• the present invention also relates to treatments of neuropathic and neurodegenerative disorders and compounds for use in treating neuropathic and neurodegenerative disorders, such as, for example, Alzheimer's disease.
• the present invention also relates to pain treatments, and compounds for use in treating pain.
• the present invention also relates to methods of reducing the risk of pre-eclampsia, and compounds for use in such methods.
• Age-related macular degeneration a disease causing vision loss that affects the central area of the macula, is the leading cause of blindness in people over 50 years of age (Bressler, 2004).
• Exudative AMD is the most severe form of AMD (Ferris et al, 1984) primarily arising from the choroidal circulation beneath the macula and characterized by choroidal neovascularization (CNV).
• CNV choroidal neovascularization
• CNV retinal pigmented epithelium
• VEGF Vascular endothelial growth factor
• VEGF is a complex gene that is alternatively spliced to form a family of multiple isoforms (Leung et al, 1989; Jingjing et al, 1999), each isoform differing in biological property, activity and function (Houck et al , 1991). Most cells commonly express isoforms VHGF n i . VEGF 165 , and VEGFi S9 , whereas VEGF 1 4 5 and VEGF 206 are comparatively rare. The majority of VEGF isoforms contain exons 1 -5 (the exception being VEGF i n (Mineur et al. 2007)) but differing portions of exons 6 and 7 that encode heparin sulphate (HS) binding domains.
• HS heparin sulphate
• VEGF XXX and VEGF xxx b may have separate regulatory pathways.
• anti-angiogenic isoforms such as VEGFi 65 b and VEGF 12 ib have been shown to be potently anti-angiogenic in animal models o retinal and choroidal ncovasc u lari sat i on, following intra-ocular injection (Hua et al 2008), and result in both endothelial and retinal epithelial cell cytoprotection (Magnussen et al 2010).
• the first therapy to be FDA approved for the treatment of neovascular AMD in December 2004 was a VEGF
• pegaptinib dose-dependently reduced the risk of severe visual acuity loss and slowed the progression o neovascular AMD (Gragoudas et al, 2004), but did not result in significant improvement in vision.
• Ranibizumab (Lucentis) a novel humanized anti-VEGF antibody fragment, was FDA approved for the treatment of neovascular AMD.
• Such intraocular injections result in increased intraocular pressure (Good et ah, 2010) and a risk, albeit minor, of endopthalmitis and other severe adverse effects (Jager et ah, 2004).
• bcvici/umab an anti-VEGF antibody from which Lucentis was derived, was shown to bind VEGF 165b with equal potency to VEGF, ⁇ , thus targeting both pro and anti-angiogenic VEGF isoforms (Varey et al 2008).
• WO 2008/1 1077, WO 2009/106855, WO 2010/058227, WO 201 1/036429 and WO 201 1/148200 describe therapeutic and other physiological uses f agents which direct expression in favour of the VEGF xxx b isoforms.
• SRP inhibitors can in principle constitute such agents.
• WO 2005/063293 describes a class of SRPK. inhibitors including SRPIN340 and derivatives and analogues thereof.
• WO 2014/060763 (PCT/GB2013/052716), the contents of which are incorporated herein by reference, describes SRPK inhibitors targeting SRPK l specifically for use as anti-angiogenic agents, neuroprotective agents, agents for use in treating or preventing hyperpermeability disorders, as agents for treating pain, and as agents for reducing the risk of, or treatment of, pre-eclampsia.
• SRPK inhibitors targeting SRPK l specifically for use as anti-angiogenic agents, neuroprotective agents, agents for use in treating or preventing hyperpermeability disorders, as agents for treating pain, and as agents for reducing the risk of, or treatment of, pre-eclampsia.
• agents for directing expression of VEGF xxx b isoforms represents a new era not only in the treatment of, for example, ncovascular AMD, but all other diseases in which VEGF xxx b is implicated.
• the present invention is based in part on new small molecule inhibitors targeting SRPKl specifically for use as anti-angiogenic agents, neuroprotective agents, agents for use in treating or preventing hyperpermeability disorders, as agents for treating pain, and as agents for reducing the risk of. or treatment of, pre-eclampsia.
• the present invention is also based at least in part on the surprising finding that these low molecular weight compounds known to inhibit SRPK1 (e.g. SRPIN340 and derivatives and analogues thereof) could be used topically or in dose-dependent manner to inhibit CNV progression.
• SRPK1 e.g. SRPIN340 and derivatives and analogues thereof
• n 1, 2, 3 or 0;
• Ri I I; a 4- to 8-membered carbocyclic group, which may have one or more substituent; a 4- to 8-membered heterocyclic group comprising one oxygen atom, which may have one or more substituent; a 4- to 8-membered heterocyclic group comprising one nitrogen atom, which may have one or more substituent, a 4- to 8-membered heterocyclic group comprising one nitrogen atom and one oxygen atom which may have one or more substituent; a 4- to 8- membered heterocyclic group comprising two nitrogen atoms which may have one or more substituent; a 4- to 8-rncmbcrcd heterocyclic group comprising three nitrogen atoms which may have one or more substituent, or a condensed aromatic heterocyclic group, which may have one or more substituent;
• X CH, O, NH or N
• Y CH, O, NH or N
• R 2 H; a Ci_ 6 alkyl group; a phenyl group; a 4- to 8-membered heterocyclic group or a condensed aromatic heterocyclic group, each of which may have one or more substituent; for use in dose-dependent treatment or prevention of ocular neovascularisation.
• the dose dependency is preferably a sigmoidal efficacy/dose relationship, e.g. of the type illustrated in Figure 4C of the accompanying drawings.
• ocular neovascularisation includes within its scope diseases and disorders characterised by ocular neovascularisation, including for example choroidal neovascu larisation such as age-related macular degeneration.
• ocular neovascularisation also includes within its scope diseases and disorders characterized by retinal neovascularisation.
• the invention provides a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof for use in the topical treatment or prevention of ocular neovascularisation.
• the first and second aspects of the invention also provide respective methods of treatment or prevention of ocular neovascularisation by administration of the compound of formula (I) to a subject in need of such treatment, and respective uses of a compound of formula (I) in the preparation of a medicament for treatment or prevention of ocular neovascularisation, as a dose-dependent treatment and/or as a topical treatment.
• the compounds used in the present invention enable dose-dependent treatment or prevention of ocular neovascularisation or topical treatment or prevention of ocular neovascu larisation.
• Dose-dependent treatment is not inherently predictable, yet is highly desirable and beneficial for effective treatment.
• R[ is a 4- to 8-rnembcred heterocyclic group comprising one nitrogen atom which may have one or more substituent.
• examples of such compounds include those in which Ri is a 6-membered heterocyclic aromatic group comprising one nitrogen atom, for example a 2- or 3- or 4-pyridyl group.
• R t is a 6-membered heterocyclic aromatic ring comprising two or three nitrogen atoms, for example a pyrimidine ring, a pyridazine ring, a pyrazine ring, or a triazine ring, each of which may have one or more substituent.
• R t is a 6-mcmbered heterocyclic non- aromatic group comprising one nitrogen atom, for example a piperidine group or a pipcrazine group, each of which may have one or more substituent.
• the compounds of Formula (I) include those in which Ri is a nitrogen- containing 6- membered heteroaryl ring as described herein or a phenyl group, with any combination of n, X, Y, Z and R 2 as herein described.
• Ri is a nitrogen-containing 6- membered heteroaryl ring as described herein or a phenyl group
• R 2 is H
• a Ci_ 6 alkyl group is a phenyl group; a 4- to 8-membered heterocyclic group or a condensed aromatic heterocyclic group, each of which may have one or more substituent
• n refers to an alkyl bridging unit between the pipcrazine ring and R,.
• compositions comprising the novel compounds and the use of the novel compounds and pharmaceutical compositions comprising them in anti-angiogenic treatments (including the treatment and prevention of disorders and diseases characterised by abnormal or excessive angiogenesis), treatments of hyperpermeability disorders, treatments of neuropathic and neurodegenerative disorders, treatment of non-inflammatory pain and methods of reducing the risk of pre-eclampsia constitute further aspects of the present invention.
• the present invention also provides (i) methods o treating or preventing disorders and diseases characterised by abnormal or excessive angiogenesis as defined herein; (ii) methods of treating or preventing hypcrpcrnieability disorders as defined herein; (iii) methods of treating or preventing neuropathic and neurodegenerative disorders as defined herein; (iv) methods of treating or preventing non-inflammatory pain; and (v) methods of reducing the risk of pre-eclampsia, comprising administering a compound of Formula (I) to a patient in need thereof.
• is a 2- or 3- or 4-pyridyi group which may have one or more substituent, for example an unsubstituted 3-pyridyl group (namely, where the nitrogen heteroatom of the pyridyl group is meta to the 5-membered ring).
• Ri is a phenyl group which may have one or more substituent, for example a methoxy substituent.
• is a condensed aromatic heterocyclic group which may have one or more substituent.
• R is an indolyl group, an isoindolyl group, a benzimidazolyl group, a quinolyl group or an isoquinolyl group.
• R 2 is a 2- or 3- or 4-pyridyl group which may have one or more substituent, or a pyrimidinyl group.
• the 2- or 3- or 4-pyridyl group which may have one or more substituent may, for example, be an unsubstituted 3-pyridyl group (namely, where the nitrogen hetcroatom of the pyridyl group is meta to the 5-membered ring).
• Ri is a phenyl group which may have one or more substituent, or a condensed aromatic heterocyclic group, wherein the condensed aromatic heterocyclic group may comprise an indolyl group, an isoindolyl group, a benzimidazolyl group, a quinolyl group or an isoquinolyl group.
• R 2 is hydrogen or a C w , alky 1 group, for example a methyl group or an ethyl group.
• R 2 is hydrogen or a C w , alky 1 group, for example a methyl group or an ethyl group.
• n 1
• i is a 2- or 3- or 4-pyridyl substituent
• R 2 is a phenyl substituent or a 2- or 3- or 4-pyridyl substituent, or a phenyl substituent or a 2- or 3- or 4-pyridyl group which may have one or more substituent; and
• their pharmaceutically acceptable salts, solvates, hydrates or prodrugs examples of the compounds of formula (I) include compounds, the formulae of which are shown in Table 1 or Table 2.
• the compounds of Formula (I) and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs may be further characterized by one or more of the following features, which are, whether individually or in any combination, combinable with any o the examples and preferences stated herein, or in WO 2005/063293, for the compounds:
• R 2 may represent a tetrahydropyranyl group, a phenyl group or a 2- or 3- or 4-pyridyl group.
• R t is a 4- to 8-membered earbocyclic group, which may have one or more substituent; a 4- to 8-membered heterocyclic group comprising one oxygen atom which may have one or more substituent or a 4- to 8-membered heterocyclic group comprising one or two nitrogen atoms which may have one or more substituent;
• R 2 is a 2- or 3- or 4-pyridyl ring.
• n is preferred in the order of ( 1 ) to (2) with (2) preferred; Ri is preferred in the order of (3) to (4), with (4) more preferred; X and Y are preferred in the order of (5) to (6), with (6) more preferred; Z is preferred in the order of (7) to (8), with (8) more preferred; R 2 is preferred in the order of (9) to (12), with (12) more preferred.
• the compounds of the present invention are SRPK1 -specific inhibitors and may therefore be used in methods of treating or preventing any disease or condition in which SRPK1 is implicated. Such conditions and treatments will now be described.
• the compounds of the present invention may be used in anti-angiogenic treatments.
• the anti- angiogenic treatment preferably includes the treatment or prevention of any disease or disorder associated with abnormal angiogenesis or abnormal over-production of pro- angiogenic VEGF isolbrms (VEGF XXX ).
• diseases and disorders include, for example, vascular disease (e.g. vasoconstriction and disorders characterised by vasoconstriction, and cardiovascular disease), malignant and benign neoplasia (e.g. angiogenesis-dependent cancers, for example tumorous cancers), tumor metastasis, inflammatory disorders, diabetes, diabetic retinopathy and other complications of diabetes (e.g.
• diabetic ncovascularisation trachoma
• retrolental hyperplasia neo vascular glaucoma
• age-related macular degeneration haemangioma.
• immune rejection of implanted corneal tissue corneal angiogenesis associated with ocular injury or infection, Osier-Webber Syndrome, myocardial angiogenesis. wound granulation, telangiectasia, hemophiliac joints, angiofibroma, telangiectasia psoriasis scleroderma, pyogenic granuloma, rubeosis, obesity, arthritis (e.g.
• the anti-angiogenic treatment according to the present invention may also include non-therapeutic treatments performed on healthy subjects, for example to inhibit vascular development for cosmetic purposes.
• non-therapeutic treatments performed on healthy subjects, for example to inhibit vascular development for cosmetic purposes.
• the compounds of the present invention may be used in the treatment or prevention of ocular neovascularisation, which may include retinal neovascularisation or choroidal neovascularisation or age-related macular degeneration.
• ocular neovascularisation which may include retinal neovascularisation or choroidal neovascularisation or age-related macular degeneration.
• the compounds of the present invention may be used in the treatment or prevention of malignant neoplasias or cancers, for example prostate cancer and breast cancer.
• Microvascular hyperpermeability disorders, disorders of epithelial cell survival and disorders of fenestrations of epithelial filtration membranes may also be used as therapeutic agents in treating other disorders in which the alternatively spliced VEGF xxx b isoform has been implicated.
• VEGF xxx b is active against a range of microvascular hyperpermeability disorders, disorders of epithelial cell survival and disorders of fenestrations of epithelial filtration membranes.
• Examples of such conditions include, for example, proteinuria, uraemia, microalbuminuria, hypoalbuminemia, renal hyperfiltration, nephrotic syndrome, renal failure, pulmonary hypertension, capillary hyperpermeabi 1 ity. microaneurysms, oedema and vascular complications of diabetes.
• vascular complications of diabetes include, for example, diabetic retinopathy, both proliferative and non-proliferative, and diabetic nephropathy.
• vascular complications of diabetes can be associated with either Type I or Type II diabetes.
• the loss of proteins from the blood can lead to further complications, for example thromboses, especially thromboses in the brain, and susceptibility to infections. Loss of natural proteins from the blood can seriously impair the efficacy of cancer therapies.
• the microvascular hyperpermeability disorder may particularly be a renal disorder, for example a permeability disorder of the GFB, for example a permeability disorder of the podocytes.
• disorders where treatment to support epithelial cell survival would be effective are as follows: acute pulmonary fibrotic disease, adult respiratory distress syndrome, adult respiratory distress syndrome, advanced cancer, allergic respiratory disease, alveolar injury, angiogenesis, arthritis, ascites, asthma, asthma or edema following burns, atherosclerosis, autoimmune diseases, bone resorption, bullous disorder associated with subepidermal blister formation including bullous pemphigoid, cardiovascular condition, certain kidney diseases associated with proliferation of glomerular or mesangial cells, chronic and allergic inflammation, chronic lung disease, chronic occlusive pulmonary disease, cirrhosis, corneal angiogenisis, corneal disease, coronary and cerebral collateral vascularization, coronary restenosis, damage following heart disease, dermatitis herpetiformis, diabetes, diabetic nephropathy, diabetic retinopathy, endotoxic shock, erythema multiforme, fibrosis, glomerular nephritis, glomerulonophritis, graft rejection,
• ncovascular glaucoma non-insulin-dependent diabetes mellitus (NIDDM), obliterative bronchiolitis, ocular conditions or diseases, ocular diseases associated with retinal vessel proliferation.
• NIDDM non-insulin-dependent diabetes mellitus
• Osier- Weber-Rendu disease osteoarthritis
• ovarian hyperstimulation syndrome osier- Weber-Rendu disease
• the present invention may be used in the treatment of macular dystrophy.
• macular dystrophy This includes: Stargardt disease/fundus flavimaculatus; Stargardt-like macular dystrophy; Stargardt-like macular dystrophy; Autosomal dominant "bull'seye” macular dystrophy Best macular dystrophy; Adult vitelliform dystrophy; Pattern dystrophy; Doyne honeycomb retinal dystrophy; North Carolina macular dystrophy; Autosomal dominant macular dystrophy resembling MCDR 1 ; North Carolina-like macular dystrophy associated with deafness; Progressive bifocal chorioretinal atrophy; Sorsby's fundus dystrophy; Central areolar choroidal dystrophy; Dominant cystoid macular dystrophy; Juvenile retinoschisis; Occult Macular Dystrophy; Non-familial Occult Macular Dystrophy.
• the disorder may particularly be a disorder of the retinal epithelium, such as geographic atrophy, or age related macular degeneration.
• SRPK1 inhibitors may also be used as therapeutic agents in treating other disorders in which the alternatively spliced VEGF xxx b isoform has been implicated.
• VEGF xxx b has neuroprotective and neuroregenerative effects.
• Neuropathic disorders to be treated or prevented according to the present invention include neuropathic pain and diabetic and other neuropathies.
• Neurodegenerative disorders to be treated or prevented according to the present invention include neurodegeneration of the cognitive and non-cognitive types, neuromuscular degeneration, motor-sensory neurodegeneration. ocular neurodegeneration.
• the activities of the proteins of the VEGF xxx b family are predicted to both actively prevent and actively reverse the conditions and disorders.
• the present invention is also applicable to non-therapeutic treatments of healthy people to adjust or normalise their cognitive function and behaviour, including thinking, memory, learning, concentration and reasoning.
• neuroregeneration can assist in normalising brain neural networks in subjects having psychiatric or behavioural abnormalities, whether or not these are diagnosable as one or more recognised psychiatric condition, the present invention is also applicable to therapeutic treatment of persons having psychiatric disorders and to non-therapeutic treatment of physically healthy people to adjust their cognition and behaviour towards the normal state.
• the present invention provides for the treatment or prevention of: pain (for example, neuropathic pain), dementia, age-related cognitive impairment, Alzheimer ' s disease, senile dementia of the Alzheimer's type (SDAT), Lewy body dementia, vascular dementia, Parkinson's disease, postencephalitic Parkinsonism, depression, schizophrenia, muscular dystrophy including facioscapulohumeral muscular dystrophy (FSH), Duchenne muscular dystrophy, Becker muscular dystrophy and Bruce's muscular dystrophy, Fiichs' dystrophy, myotonic dystrophy, corneal dystrophy, refle sympathetic dystrophy syndrome (RSDSA), neurovascular dystrophy, myasthenia gravis, Lambert Eaton disease, Huntington ' s disease, motor neurone diseases including amyotrophic lateral sclerosis (ALS), multiple sclerosis, postural hypotension, traumatic neuropathy or neurodegeneration e.g.
• pain for example, neuropathic pain
• dementia age-related cognitive impairment
• Cockayne syndrome Down syndrome, corticobasal ganglionic degeneration, multiple system atrophy, cerebral atrophy, olivopontocerebellar atrophy, dentatorubral atrophy, pallidoluysian atrophy, spinobulbar atrophy, optic neuritis, sclerosing pan- encephalitis (SSPE), attention deficit disorder, post-viral encephalitis, post-pol iomyelitis syndrome, Fahr's syndrome.
• SSPE sclerosing pan- encephalitis
• Joubert syndrome Guillain-Barre syndrome, lissencephaly, Moyamoya disease, neuronal migration disorders, autistic syndrome, polyglutainine disease, Niemann-Pick disease, progressive multifocal leukoencephalopathy, pseudotumor cerebri, Refsum disease, Zellweger syndrome, supranuclear palsy, Friedreich ' s ataxia, spinocerebellar ataxia type 2, Rhett syndrome, Shy-Drager syndrome, tuberous sclerosis.
• Pick ' s disease chronic fatigue syndrome, neuropathies including hereditary neuropathy, diabetic neuropathy and mitotic neuropathy, prion-based ncurodegeneration.
• CJD Creutzfeldt- Jakob disease
• variant CJD new variant CJD
• bovine spongiform encephalopathy (BSE) bovine spongiform encephalopathy
• GSS GSS
• FFI. kuru and Alper's syndrome Joseph's disease, acute disseminated encephalomyelitis, arachnoiditis, vascular lesions of the central nervous system, loss of extremity neuronal function
• Charcot-Marie- Tooth disease Krabbe's disease
• leukodystrophies susceptibility to heart failure
• asthma epilepsy
• auditory neurodegeneration macular degeneration
• pigmentary retinitis pigmentary retinitis and glaucoma-induced optic nerve degeneration.
• psychiatric disorders with which the present invention is concerned include, without limitation: anxiety disorders (for example, acute stress disorder, panic disorder, agoraphobia, social phobia, specific phobia, obsessive-compulsive disorder, sexual anxiety disorders, post-traumatic stress disorder, body dysmorphic disorder and generalized anxiety disorder), childhood disorders (for example, attention-deficit hyperactivity disorder (ADHD), Asperger's disorder, autistic disorder, conduct disorder, oppositional defiant disorder, separation anxiety disorder and Tourette's disorder), eating disorders (for example, anorexia nervosa and bulimia nervosa), mood disorders (for example, depression, major depressive disorder, bipolar disorder (manic depression), seasonal affective disorder (SAD), cyclothymic disorder and dysthymic disorder), sleeping disorders, cognitive psychiatric disorders (for example, delirium, amnestic disorders), personality disorders (for example, paranoid personality disorder, schizoid personality disorder, schizotypal personality disorder, antisocial personality disorder,
• SRPK1 inhibitors may also be used as therapeutic agents in treating other disorders in which the alternatively spliced VEGF xxx b isoform has been implicated.
• VEGF xxx b has an analgesic effect on VEGFR2-mediated non-inflammatory pain in mammals.
• VEGFR2-mediated non-inflammatory pain to be treated or prevented according to the present invention includes non-inflammatory neuropathic and nociceptive pain where the VEGFR2 receptor is involved in the cause or transmission of the pain.
• the compounds according to the present invention are predicted to have activity against non-inflammatory allodynia and pain (antiallodynic and analgesic activity). Pain states of this type include chronic pain, whether of the intermittent or constant form.
• Such pain states may include, for example, low back pain, neuralgia, atypical pains such as atypical facial pain, pain exhibited post-surgery, post-injury (for example, after surgery or injury causing nerve damage) or in association with cancer or with cancer therapy such as cytotoxic or radiation therapy, or neuropathy associated with diabetes (diabetic neuropathy, insulin neuritis) or other systemic or autoimmune disease or pathology, or the treatment thereof, alcoholism or HIV infection, ageing associated neuropathy, or neuropathy of unknown origin.
• the activities of the proteins of the VEGFR2 agonists for example the VEGF xxx b family, are predicted to both actively prevent and actively reverse VEGFR2-mediated non-inflammatory pain.
• the compounds used in the present invention may be employed in association with one or more different pain treatment agent for the purpose of normalising the sensitivity towards pain of the subject treated (or being co-treated) with the said one or more different pain treatment agent.
• normalising means moving the subject ' s pain sensitivity towards normal levels, and may include enhancement of the sensitivity if the one or more different pain treatment agent causes an excessive reduction in feeling or in sensitivity towards pain.
• the one or more different pain treatment agent may be selected from pain treatment agents currently known or yet to be devised. Such selection will be well within the skill of the person of ordinary skill in this art. Such combination treatments can enable fine control of pain sensitivity in subjects and minimisation of overall side effects according to the particular condition and needs of the sub ject.
• SRPK1 inhibitors may also be used as therapeutic agents in treating other disorders in which the alternatively spliced VEGF xxx b i so form has been implicated.
• VEGF xxx b levels in pregnant female mammals increase the risk of the female mammal developing prc-cclampsia.
• compounds of the present invention may be used to increase VEGF xxx b levels in a pregnant female mammal so as to reduce the risk of pre-eclampsia the female mammal developing pre-eclampsia or a complication linked thereto, or of a fetus of the female mammal developing a fetal or neonatal deficiency linked to maternal pre-cclampsia.
• Pre-eclampsia in humans can develop as early as 20 weeks of gestation.
• Pre-eclampsia that develops before about 34 weeks of gestation is normally referred to as “early pre-eclampsia” or “early-onset pre-eclampsia”.
• Pre-eclampsia that develops after about 34 weeks of gestation is normally referred to as “late pre-eclampsia” or “late-onset pre-eclampsia”.
• pre-eclampsia can be categorised as "severe pre-eclampsia” according to criteria established by the United Kingdom Royal College of Obstetricians and Gynaecologists. Under these criteria, a patient with "severe pre-cclampsia” will have systolic blood pressure (BP) greater than 169 mniHg or diastolic BP greater than 109 mmllg with proteinuria greater than 1 g/24h; or will show occurrence of HELLP syndrome (haemolysis, elevated liver enzymes and low platelet count).
• BP systolic blood pressure
• pre-eelampsia For further details on pre-eelampsia, and methods to reduce the risk of a pregnant female mammal developing developing pre-eclampsia or a complication linked thereto, or of a fetus of the female mammal developing a fetal or neonatal deficiency linked to maternal preeclampsia, see WO 201 1/036429, the contest of which are incorporated herein by reference.
• Compounds of the present invention are as defined by Formula (1) and have been shown to be inhibitors of one or both of the kinases SRPK1, and SRPK2, and thus useful in treatments as described herein.
• the compounds of the present invention may be synthesised by any known method. Suitable methods as disclosed in WO 2005/063293 may be adapted as required. An exemplary synthesis for compound 12 is described below in the Examples.
• the compounds of the present invention may, if desired, be co-administered with one or more additional active agent, for example one or more agent selected from, but not limited to, cholinesterase inhibitors, dopamine agonists (e.g. L-dopa), COMT inhibitors, MAO-B inhibitors, anti-cholinergics, acetylcholine agonists, serotonin agonists, AMPA receptor agonists, GAB A receptor agonists, NMDA receptor agonists, ⁇ -adrcnoccptor agonists, digoxin, dobutamine, anti-inflammatories, neurotrophic factors, statins, adenosine A 2a receptor antagonists, aldose reductase inhibitors, immunomodulators, cannabinoid agonists. interferon or tricyclic anti-depressants.
• dopamine agonists e.g. L-dopa
• COMT inhibitors e.g. L-dopa
• Ci_ 6 alkyl group refers to a linear or branched alkyl group comprising one to six carbon atoms, which is a monovalent group derived by removing an arbitrary hydrogen atom from an aliphatic hydrocarbon consisting of one to six carbons.
• the C,_ 6 alkyl group includes, for example, a methyl group, an ethyl group, a 1 -propyl group, a 2-propyl group, a 2-methyl-l -propyl group, a 2-methyl-2-propyl group, a 1 -butyl group, a 2-butyl group, a 1 - pentyl group, a 2-pentyl group, a 3-pentyl group, a 2-methyl- l -butyl group, a 3-methyl- l - butyl group, a 2-methyl-2-butyl group, a 3-methyl-2-butyl group, a 2,2-dimetliyl-l -propyl group, a 1 -liexyl group, a 2-hexyl group, a 3-hexyl group, a 2-methyl- l -pentyl group, a 3- methyl- 1 -pentyl group, a 4-methyl-
• heterocycle or “heterocyclic group” refers to an aromatic or non-aromatic ring that may comprise double bonds within the ring, wherein at least one, for example one or two, of the atoms constituting the ring are heteroatoms;
• n i t rogen-con taini ng heterocycle or "heterocyclic group comprising one or more nitrogen atoms” refers to an aromatic or non-aromatic ring that may comprise double bonds within the ring, wherein at least one, for example one or two, of the atoms constituting the ring are nitrogen atoms;
• oxygen-containing heterocycle or “heterocyclic group comprising one or more oxygen atoms” refers to an aromatic or non-aromatic ring that may comprise double bonds within the ring, wherein at least one. for example one or two, of the atoms constituting the ring are oxygen atoms;
• heteroatom refers to a sulfur atom, an oxygen atom, or a nitrogen atom
• nitrogen-containing 5- to 10-membered heteroaryl ring or “nitrogen-containing 5- to 10- membcred heteroaromatic group” refers to an aromatic ring in which five to ten atoms constitute the ring, wherein at least one of the atoms constituting the ring is a nitrogen atom, and one or more heteroatoms other than nitrogen atoms may further be comprised.
• the nitrogen-containing 5- to 10-membered heteroaryl ring includes, for example, a pyridine ring, an indole ring, an isoindole ring, a pyridazine ring, a pyrimidine ring, a pyra/.ine ring, a quinoline ring, an isoquinoline ring, and a benzimidazole ring;
• nitrogen-containing 5- to 10-membered heteroaryl group refers to a mono-or divalent group derived by removing one or two arbitrary hydrogen atoms from the above-defined "5- to 10- membered heteroaryl ring".
• the nitrogen-containing 5- to 10-membered heteroaryl group includes, for example, a pyridyl group, an indolyl group, an isoindolyl group, a pyridazinyl group, a pyrimidinyl group, a pyrazinyl group, a quinolyl group, an isoquinolyl group, and a benzimidazolyl group;
• nitrogen-containing 4- to 8-mcmbcred heteroaryl ring or “nitrogen-containing 4- to 8- membered heteroaromatic group” refers to an aromatic ring in which four to eight atoms constitute the ring, wherein at least one of the atoms constituting the ring is a nitrogen atom, and one or more heteroatoms other than nitrogen atoms may further be comprised.
• the nitrogen-containing 4- to 8-membered heteroaryl ring includes, for example, a pyridine ring, a pyridazine ring, a pyrimidine ring, and a pyrazine ring;
• nitrogen-containing 4- to 8-membered heteroaryl group refers to a mono-or divalent group derived by removing one or two arbitrary hydrogen atoms from the above-defined "4- to 8- membered heteroaryl ring".
• the nitrogen-containing 4- to 8-mcmbercd heteroaryl group includes, for example, a pyridy l group, a pyridazinyl group, a pyrimidinyl group, and a pyrazinyl group;
• oxygen-containing 5- to 10-mcmbered heteroaryl ring refers to an aromatic ring in which five to ten atoms constitute the ring, wherein at least one of the atoms constituting the ring is an oxygen atom, and one or more heteroatoms other than oxygen atoms may further be comprised.
• the oxygen-containing 5- to 10-membered heteroaryl ring includes, for example, a pyran ring;
• oxygen-containing 4- to 8-membered heteroaryl group refers to an aromatic ring in which four to eight atoms constitute the ring, wherein at least one of the atoms constituting the ring is an oxygen atom, and one or more heteroatoms other than oxygen atoms may further be comprised, for example a pyranyl group;
• one or two of the atoms constituting the ring are heteroatoms
• one to three carbonyl groups may be comprised in the ring:
• the group is monocyclic.
• the 4- to 8-membered non-aromatic heterocyclic ring is preferably a nitrogen-containing 4- to 8-membered heterocyclic ring that comprises nitrogen atoms as heteroatoms.
• the 4- to 8-membered non-aromatic heterocyclic ring includes, for example, an azetidine ring, a pyrrolidine ring, a piperidinc ring, an azepane ring, an azocine ring, a tetrahydrofunm ring, a tetrahydropyran ring, a morpholine ring, a thiomorpholine ring, a piperazine ring, a thiazolidine ring, a dioxane ring, an imidazoline ring, and a thiazoline ring.
• the "4- to 8-membered heterocyclic ring" preferably includes a pyrrolidine ring, a piperidinc ring, a morpholine ring, and a piperazine ring;
• 4- to 8-membered non-aromatic heterocyclic group refers to a mono- or divalent group derived by removing one or two arbitrary hydrogen atoms from the above-defined "4- to 8- membered non-aromatic heterocyclic ring".
• the 4- to 8-membered heterocyclic group includes, for example, an azetidinyl group, a pyrrolidinyl group, a piperidinyl group, an azepanyl group, an azocanyl group, a tetrahydrofuryl group, a tetrahydropyranyl group, a morpholinyl group, a thoimorpholinyl group, a piperazinyl group, a thia/.olidinyl group, a dioxanyl group, an imidazolyl group, and a thiazolyl group;
• Condensed aromatic hctcrocycle refers to a ring structure in which the heterocyclic moiety is fused, for example ortho-condensed, with an aromatic ring, such as a benzene ring.
• the heterocyclic moiety is an above-defined hctcrocycle.
• Condensed aromatic heterocyclic group refers to a ring structure in which the heterocyclic moiety is fused, for example ortho-condensed, with a ring, for example an aromatic ring, such as benzene ring.
• the heterocyclic moiety is an above-defined heterocyclic group.
• the condensed aromatic heterocyclic group includes, for example, an indolyl group, an indolinyl group, an isoindolyl group, an isoindolinyl group, and a 1 ,2,3,4-tetrahydroquinolinc.
• "4- to 8-membered carbocyclic ring” refers to a saturated or unsaturated carbon ring, for example a furan ring, a tetrahydrofuran ring, a pyran ring or a tetrahydropyran ring;
• “4- to 8-membered carbocyclic group” refers to a saturated refers to a mono- or divalent group derived by removing one or two arbitrary hydrogen atoms from the above-defined 4- to 8-membered carbocyclic ring.
• “4- to 8-membered carbocyclic group” may refer to a furanyl group, a tetrahydrofuranyl group, a pyranyl group or a tetrahydropyranyl group;
• halogenated Q.,, alkyl group refers to a group in which at least one arbitrary hydrogen atom in the above-defined "Ci_6 alkyl group” is replaced with an above-defined "halogen atom”.
• the halogenated Ci_6 alkyl group includes, for example, a trifluoromethyl group, a difluoromethyl group, and a monofluoromethyl group.
• the phrase "may have one or more substituent” means that a certain group or compound may optionally have an arbitrary selection or combination of one or more substituent at substitutable positions.
• the substituents can include, for example, atoms or groups selected from one or more of: halogen, hydroxy!, hydroxymethyl, hydroxyethyl, mercapto, nitro, cyano, formyl, carboxyl.
• Ci_6 alkyl for example methyl
• Ci_6 alkoxy for example, methoxy
• Q.6 thioalkyl for example thiomethyl
• Ci ⁇ alkoxycarbonyl C « alkylsul onyl.
• C2-6 alkenyl group refers to a linear or branched alkenyl group comprising two to six carbons.
• the C2-6 alkenyl group includes, for example, a vinyl group, an allyl group, a 1-propenyl group, a 2-propenyl group, a 1-butenyl group, a 2-butenyl group, a 3- butenyl group, a pentenyl group, and a hexenyl group;
• C 2 -6 alkynyl group refers to a linear or branched alkynyl group comprising two to six carbons.
• the C 2 -e alkynyl group includes, for example, an ethynyl group, a 1 - propynyl group, a 2-propynyl group, a butynyl group, a pentyiiyl group, and a hexynyl group.
• Ci_6 alkoxy group refers to an oxy group to which the above-defined “Ci_ 6 alkyl group” is Jinked.
• the Ci_ 6 alkoxy group includes, for example, a methoxy group, an ethoxy group, a 1 -propv oxy group, a 2-propyIoxy group, a 2-metfiyl- 1 -propyloxy group, a 2-methyl- 2-propyloxy group, a 1 -butyloxy group, a 2-butyloxy group, a 1 -pentyloxy group, a 2- pentyloxy group, a 3-pentyloxy group, a 2-methyl- 1 -butyloxy group, a 3 -methyl- 1 -butyloxy group, a 2-melhyl-2-butyloxy group, a 3-methyl-2-butyloxy group, a 2.2-dimethyl- l - propy loxy group, a 1 -hexyl
• the "Ci_ 6 alkylthio group” includes, for example, a methy lth io group, an ethylthio group, a 1 -propylthio group, a 2-propylthio group, a but lthio group, and a pentylthio group;
• "Ci_6 alkoxycarbonyl group” refers to a carbonyl group to which the above-defined "Ci_ 6 alkoxy group” is linked.
• the Ci_ 6 alkoxycarbonyl group includes, for example, a methoxy carbonyl group, an ethoxy carbonyl group, a 1 -propyloxycarbonyl group, and a 2- propyloxycarbonyl group;
• Ci.6 alkylsulfonyl group refers to a sulfonyl group to which the abov e-defined "Ci -6 alkyl group” is linked.
• the Ci -6 alkylsulfonyl group includes, for example, a methy lsulfonyl group, an ethylsulfonyl group, a 1 -propylsulfonyl group, and a 2- propylsul fonyl group.
• halogen atom refers to a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom;
• C 6 -io aryl group refers to an aromatic cyclic hydrocarbon group comprising six to ten carbon atoms.
• the C ⁇ -w ary l group includes, for example, a phenyl group, a 1 -naphthy group, and a 2-naphthyl group;
• Salt is not particularly limited, so long as it is a pharmaceutical acceptable salt which is formed with a compound according to the present invention.
• Such salts include, for example, inorganic acid salts, organic salts, inorganic base salts, organic base salts, and acidic or basic amino acid salts.
• examples of preferable inorganic acid salts include: hydrochloride, hydrobromate, sulfate, nitrate, and phosphate.
• preferable organic salts include: acetate, succinate, fumarate, maleate, tartrate, citrate, lactate, stearate, benzoate, mcthanesulfonate, and p-toluene sulfonate.
• preferable inorganic base salts include: alkali metal salts, such as sodium salts and potassium salts; alkali earth metal salts, such as calcium salts and magnesium salts; aluminium salts; and ammonium salts.
• preferable organic base salts include: diethylamine salts, diethanol amine salts, meglumine salts, and N,N'-dibenzylethylenediamine salts.
• preferable acidic amino acid salts include: aspartate and glutamate.
• preferable basic amino acid salts include: arginine salts, lysine salts, and ornithine salts.
• the compounds of the present invention When left in air, the compounds of the present invention sometimes absorb moisture, and are sometimes attached to absorbed water or converted to hydrates. Such hydrates are also included in the present invention.
• compounds of the present invention are sometimes converted into solvates, absorbing some other solvents. Such solvates are also included in the present invention.
• Any organic solvent may in principle be used to prepare a solvate of the compounds of the present invention.
• a solvate can include also water together with the one or more organic solvent.
• the solvent may be selected from ketones, alcohols, ethers, esters, aromatic solvents, and, where possible, mixtures thereof with each other, with other organic solvents and/or with water.
• prodrug forms of the compounds of formula (I) may be used in the present invention.
• “Pharmaceutically acceptable prodrugs” means those prodrugs of the compounds which are, within the scope of sound medical and vetinary judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds.
• prodrug means compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example by hydrolysis in blood. Functional groups which may be rapidly transformed, by metabolic cleavage, in vivo form a class of groups reactive with the carboxyl group.
• prodrugs Because of the ease with which the metabolically cleavable groups of the compounds are cleaved in vivo, the compounds bearing such groups act as pro-drugs. A thorough discussion of prodrugs is provided in the following: Design of Prodrugs, H. Bundgaard, ed. , Elsevier, 1985; Methods in Enzymology, K. Widder et al, Ed., Academic Press, 42, p. 309-396,1985; A Textbook of Drug Design and Development, Krogsgaard-Larsen and H. Bundgaard, ed. , Chapter 5; Design and Applications of Prodrugs p. 1 13- 191. 1991 ; Advanced Drug Delivery Reviews, H. Bundgard. 8, p.
• the compound according to the present invention may be administered in the form of a composition comprising the active agent and any suitable additional component.
• the composition may, for example, be a pharmaceutical composition (medicament), suitably for topical administration (e.g. as eyedrops or cream or lotion), or parenteral administration (e.g. injection, implantation or infusion).
• the composition may alternatively, for example, be a foodstuff, food supplement, beverage or beverage supplement.
• composition means a composition comprising an active agent and comprising additionally one or more pharmaceutically acceptable carriers.
• the composition may further contain ingredients selected from, for example, diluents, adjuvants, excipicnts, vehicles, preserving agents, fillers, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavouring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents and dispersing agents, depending on the nature of the mode of administration and dosage forms.
• compositions may take the form, for example, of tablets, dragees, powders, elixirs, syrups, liquid preparations including suspensions, sprays, inhalants, tablets, lozenges, emulsions, solutions, cachets, granules, capsules and suppositories, as well as liquid preparations for injections, including liposome preparations.
• Techniques and formulations generally may be found in Remington, The Science and Practice of Pharmacy, Mack Publishing Co., Easton, PA, latest edition.
• Liquid form preparations include solutions, suspensions, and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection or topical administration. Liquid preparations can also be formulated in solution in aqueous polyethylene glycol solution. Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for either topical, oral or parenteral administration. Such liquid forms include solutions, suspensions, and emulsions. These particular solid form preparations are most conveniently provided in unit dose form and as such are used to provide a single liquid dosage unit.
• liquid form preparations intended to be converted to liquid form may contain, in addition to the active material, flavourings, colourants, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilising agents, and the like.
• the liquid utilized for preparing the liquid form preparation may be water, isotonic water, ethanol, glycerine, propylene glycol, and the like as well as mixtures thereof. Naturally, the liquid utilized will be chosen with regard to the route of administration, for example, liquid preparations containing large amounts of ethanol are not suitable for topical or parenteral use.
• the composition may be in a formulation intended for topical application.
• the formulation may be a gelling formulation to control release and therefore availability of the active agent following topical application.
• the formulation may contain one or more gelling agents, for example hydroxypropyl mcthylcellulose.
• the formulation may contain one or more surfactants, for example a non-ionic liquid polymer, examples of which include Tyloxapol, and the Pluronics® poloxamcrs from BASF.
• the formulation may contain one or more solubilizers, for example dextrose or sorbitol.
• the formulation may contain one or more antimicrobial or antiseptic agents, for example benzalkonium chloride.
• the aforementioned named gelling agents, surfactants, solubilizers and antimicrobial agents are listed purely by way of example and it will be appreciated that other agents to perform these functions are known.
• the dosages may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. General ly, treatment is initiated with the smaller dosages which are less than the optimum dose of the compound. Thereafter the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired.
• the dosage regime for administration of the active agent may, for example, comprise a total dose of up to 1 pg, for example up to 500 ng, for example up to 50ng, for example less than 20ng of active agent in a dosing period ranging, for example, between 1 and 14 days. For example, a total dose of less than 18 ng, 1 7 ng, 16 ng, 15, ng, 14 ng, 13 ng, 12 ng, 1 1 ng or 10 ng may be administered.
• the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof may be administered in a therapeutically effective amount.
• a therapeutically effective amount o a compound of Formula (I) for topical administration for treatment of CNV may be at least about 5 ⁇ g/10 pi of delivery vehicle.
• a therapeutically effective amount may be at least about 100 pg mL, for example at least about 200 pg mL, at least about 300 pg/mL, at least about 400 pg'mL. at least about 500 pg/mL, at least about 600 pg/mL.
• a therapeutically effective amount may be at least about 1 mg/mL, for example at least about 2 mg/mL, at least about 3 mg/mL, at least about 4 mg/mL, at least about 5 mg/mL.
• a therapeutically effective amount may be less than about 5 mg/mL, for example less than about 4 mg/mL, less than about 3 mg/mL, less than about 2 mg/mL, less than about 1 mg/mL.
• the therapeutically effective amount may be administered daily, for a dosing period ranging, for example, between 1 and 14 days.
• the therapeutically effective amount may be a total daily dosage which may be divided and administered in portions during the day, for example twice daily.
• a therapeutically effective amount of a compound of Formula (I) for anti-angiogcnic treatment of a mammalian subject or for use in treating or preventing microvascular hy perpermeabi 1 i ty disorders, or in regulating the pro-angiogenic pro-permeability properties of VEGF XXX isoforms, or in supporting epithelial cell survival without increased permeability, or in reducing the nature (for example the number density and/or size) of fenestrations of epithelial filtration membranes, or for use in treating or preventing neuropathic and neurodegenerative disorders, or for use as a neuroprotective or neuroregenerative agent in vivo or in vitro, or for use in treating or preventing VEGFR2 -mediated non-inflammatory pain, or for use in reducing the risk of a female mammal developing pre-ec!ampsia or a complication linked thereto, or of a fetus of the female mammal developing a fetal or neonatal
• the therapeutically effective amount may be less than about 100 mg/kg, for example less than about 90 mg/kg, less than about 80 mg/kg, less than about 70 mg/kg, less than about 60 mg/kg, less than about 50 mg/kg, less than about 40 mg/kg, less than about 30 mg/kg, or less than about 20 mg/kg, for example less than about 10 mg/kg. less than about 5 mg/kg.
• the expression ''treating or preventing refers to all forms of healthcare intended to remove or avoid the disorder or to relieve its symptoms, including preventive, curative and palliative care, as judged according to any of the tests available according to the prevailing medical and psychiatric practice.
• An intervention that aims with reasonable expectation to achieve a particular result but does not always do so is included within the expression "treating or preventing " .
• An intervention that succeeds in slowing or halting progression of a disorder is included within the expression '"treating or preventing".
• Certain neurological and psychiatric disorders are considered as "spectrum" conditions, in which individuals may exhibit some or all of a range of possible symptoms, or may exhibit only a mild form of the disorder. Furthermore, many neurological and psychiatric conditions are progressive, starting with relatively mildly abnormal symptoms and progressing to more severely abnormal symptoms.
• the present invention includes the treatment and prevention of all neurological and psychiatric conditions of whatever type and stage.
• non-therapeutic method refers particularly to an intervention performed on an individual who is neurologically or psychologically within the normal range, to normalise or enhance or improve a function of the neurological or psychological kind.
• a neurological function that may suitably be treated non-therapeutically may include, for example, cognition (including thinking, reasoning, memory, recall, imagining and learning), concentration and attention, particularly towards the milder end of the scale of conditions, and mild abnormal behavioural or personality traits.
• a psychological function that may suitably be treated non-therapeutically may include, for example, human behaviour, mood, personality and social function, for example grief, anxiety, depression, moodiness, moroseness, teenage moods, disrupted sleep patterns, vivid dreaming, nightmares, and sleepwalking.
• Normal The expression • 'normalise' " and analogous terms used herein refers particularly to a physiological adjustment towards a condition characteristic of general normal neurological or psychiatric health, whether or not a condition is actually reached that would be characterised as normal. Mammals
• the present invention is also useful in a range of mammals.
• mammals include non-human primates (e.g. apes, monkeys and lemurs), for example in zoos, companion animals such as cats or dogs, working and sporting animals such as dogs, horses and ponies, farm animals, for example pigs, sheep, goats, deer, oxen and cattle, and laboratory animals such as rodents (e.g. rabbits, rats, mice, hamsters, gerbils or guinea pigs).
• rodents e.g. rabbits, rats, mice, hamsters, gerbils or guinea pigs.
• the disorder or function to be treated is exclusive to humans, then it will be understood that the mammal to be treated is a human. The same applies respectively to any other mammalian species if the disorder or function to be treated is exclusive to that species.
• Figure 1 shows the activity of compounds 12 to 14 (termed SPHINX31, SPHINX32 and SPHINX33 respectively) of Formula (I) against SRPK1;
• Figure 2 shows the effects of SPHINX31 relative to reference compounds SPHINX and SPHINX7 on SRSF1 phosphorylation in SRPK1 de-repressed (DDS) cells;
• Figure 3 shows the dose response curve for SPHINX31 relative to reference compounds SPHINX and SPHINX7 in DDS cells;
• Figures 4A and 4B show the effects of SPHINX31 in a laser-induced mouse CNV model relative to reference compounds SRPIN340 and SPHTNX7;
• Figure 4C shows the dose response curve for SPHINX 31 in the laser-induced mouse CNV model;
• Figure 4D shows images of fluorescein angiography showing representative lesion sizes in the laser-induced mouse CNV model after treatment with SPHTNX31 or reference compound SPHINX7;
• Figure 5 shows the hERG inhibition profile of SPHINX31 .
• Figure 6 shows the TREEspotTM results of a kinome screen of SPHINX31 against all known kinases using the DiscoverX K NOMEscan ® binding affinity assay;
• Figure 7 shows the equivalent binding affinity calculated using differential scanning fluorimetry;
• Figure 8 shows that MCI salts of compounds of the invention salts dose-dependently inhibit SRPK1 activity in vitro and as efficiently as unconjugated SPHINX compounds;
• Figure 9 shows that SPHINX31 also inhibits SRSF l phosphorylation in PC-3 cells (at ⁇ ⁇ SPHINX31);
• Figures 1 OA and 10B show that SPHINX3 1 blocks nuclear localisation of SRSFl in PC-3 cancer cells ( Figure 10a) and MDA-MB-231 cancer cells ( Figure 10b)
• Figure 1 1 shows that SPHINX compounds dose depcndently increase expression of anti- angiogenic VEGF ⁇ b in Denys Drash (DDS) podocytes and normal podocytes;
• DDS Denys Drash
• Figure 12 shows that SPHINX compounds increase expression of anti-angiogenic VEGF 165 b in MCF7 breast cancer cells (left hand chart) and MDA-MB-231 breast cancer cells (right hand chart);
• Figure 13 shows that SPHINX compounds dose dependent!y increase expression of anti- angiogenic VEGFi f b in RPE cells;
• Figure 14A shows a permeability chamber used to test whether compounds of the invention can penetrate through the sclera. Pazaponib was used as a control;
• Figure 14B shows the concentration of compounds in the lower chamber ("vitreous") after 0, 4 or 24 hours in rabbit eye tissue;
• Figure 14C shows retinal concentration after 4 hrs as a percentage of applied concentration in rabbit eye tissue;
• Figure 14D shows concentration of compounds in sclera tissue after 24 hrs as a percentage of applied concentration in porcine eye tissue;
• Figure 14E shows concentration of compounds in RPE/choroid tissue after 24 hrs as a percentage of applied concentration in porcine eye tissue;
• Figure 14F shows concentration of compounds in retinal tissue after 24 hrs as a percentage of applied concentration in porcine eye tissue;
• Figure 14G shows concentration of compounds in the lower chamber (“vitreous") after 24 hrs as a percentage of applied concentration in porcine eye tissue:
• Figure 15 shows substantial accumulation of compound in the retina of a mouse 24 hours after addition of SPHINX31 ;
• Figure 16 shows greater retinal penetration for SPUINX3 1 relative to pazopanib in in vivo rabbit studies
• Figure 1 7 shows that SPHINX31 binds to melanin substantially less than pazopanib does
• Figure 1 8 shows the stability of compounds in human plasma relative to propantheline bromide
• Figure 19 shows that neither SPHINX3 1 nor its metabolite SPHINX46 induce genotoxicity in an Ames test;
• Figure 20 shows Ganzfeld ERG recordings in mice taken 24 h after treatment with control or SPHINX31 at 2 ng/ml by topical eye drop administration;
• Figure 21 shows Ganzfeld ERG recordings in mice taken 24 h after treatment with control or SPHINX31 at 2 n /ml by topical eye drop administration;
• Figure 22a shows the effects of SPHINX compounds on RPE cells expressing genes with alternative splicing, which are not linked to SRPKl ; and Figure 22b shows the effects of SPHINX compounds on RPE expressing genes with alternative splicing, which are linked to
• Figure 23 shows the effect of SPHINX compounds on MKNK2, an RPE expressed gene with SRPKl dependent alternative splicing.
• Figure 23 A shows the alternative splicing of the MKNK2 gene; and
• Figures 23 B and 23 C show the effect of SPHINX compounds on this alternative splicing.
• Trifluoroacetic acid (21 .5 mL, 280.96 mmol) was added dropwise to a solution of Boc- piperazine (3) (2.98 g, 10.76 mmol) in dichloromethane (21 .5 ml .) at 0°C (ice). The solution was stirred at 0°C for 10 min then the cold bath was removed and the solution was stirred at room temperature for 4 hours. The solution was neutralised to pH 9 using saturated aqueous sodium bicarbonate solution. The dichloromethane layer was removed and the remaining aqueous solution was extracted with dichloromethane (x 2).
• a flask was charged w ith ester 8 (2.17 g, 10.58 mmol), 4-pyridinyIboronic acid (11) (1.00 g. 8.14 mmol), PdCl 2 (PPh 3 ) 2 (0.29 g, 0.4 1 mmol), 2 M aqueous sodium carbonate solution ( 1 0.2 ml,, 22.4 mmol) and 1 ,2-dimethoxyethane (81 mL).
• the flask was freeze-pump-thawed (x 3), backfi lled with argon and heated at reflux for 17 hours. The solution was cooled to room temperature and the DME was removed under reduced pressure.
• the pH of the residue was adjusted to pH I using 2 M aqueous hydrochloric acid solution.
• the solution was extracted with dichloromethane ( ⁇ 3).
• the dichloromcthane extracts were discarded.
• the remaining aqueous solution was neutralised to pl f 9 using solid sodium bicarbonate and extracted with ethyl acetate (x 3).
• the respective organic extracts were combined and washed with water and brine, then dried (Na2SC> ).
• the solvent was removed under reduced pressure to afford the product 10 as a white solid (1.41 g. 85 %), which was of sufficient purity to use in the next step, with all analytical data matching that reported in the literature (H. Y. Fu, I I. Doucct, Eur. J. Org. Chem.
• the mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with dichloromethane (x 3). The organic extracts were combined and washed with water and brine, then dried (NaiSCXt). The solvent was removed under reduced pressure to afford the product as a white solid (0.19 g. 67 %), which was of sufficient purity to use in the next step. On occasion when the crude product was impure it could be purified by flash chromatography on deactivated silica gel. eluting with 5 % methanol/ethyl acetate to afford the product.
• Candidate compounds were screened by the Kinase-Glo assay (Promega; Koresawa and Okabe, 2004), the results of which are shown in Table 1 and Table 2.
• a reaction buffer containing 9.6mM MOPS pH7 and 0.2nM EDTA pH8 was added to ⁇ SRSF1 RS peptide (NH 2 - SPSYGRSRSRSRSRSRSRSRSNSRSRSY-OH (SEQ ID NO: 1)) and 0.1 ⁇ -g of purified SRPK1 kinase.
• Candidate compounds were serially diluted from 10 ⁇ -0.5 ⁇ and added to the reaction mixture, wells with omitted SRPK 1 kinase and omitted compounds were also added as controls.
• DDS Denys Drash Syndrome
• WT 1 mutant that fails to repress S PK 1 expression
• Animals were culled on cither day 4 or day 14 and eyes were either unfixed for retinal dissection and protein extraction, or fixed and enucleated and choroids stained for isoIectin-B4 and examined, or imaged by fluorescein angiography.
• Isoform specific ELISA was performed as described in Varey et al (2008) and Carter et al (2014).
• Scleral permeability was measured using a modified Ussing chamber assembly in isotonic assay buffer (pH 7.4). Rabbit or porcine excised eye tissues were mounted in the chambers such that the episcleral side faced the donor chamber and the retinal side faced the receiver chamber. The chambers were filled with equal volumes of assay buffer, with (donor side) or without (receiver side) 1 ig ml compound. After 4 or 24 hours tissue was removed from the chamber and the receiver side ("vitreous") sampled. Tissue was dissected into sclera, ehoroid/RPE, and retina and homogenised. A tracer (SPHINX7) was added, and tissue extracted by acetonitrile extraction as described in Gammons et al (2013). Compounds were then analysed by mass spectrometry as described in Gammons et al (2013).
• Rabbits were treated thrice daily for six days, with 50 g SPHINX3 1 in one eye and 5( ⁇ ig pazopanib as 200 ⁇ eye drops. Rabbits were killed 12 hours after the last eye drop, blood and liver taken, and the retina dissected from the choroid/sclera, incisions made, and laid out flat, and photographs taken. Both eye compartments were dissected into 17 different areas. All samples were weighted. Compound was extracted by reverse phase extraction from the retina and choroid/sclera samples and the liver and plasma as above, and amount determined by mass spectrometry in different areas of the eye, in the blood and in the liver. Amounts per gram of tissue were calculated for SPHINX31 and pazopanib for each sample, and averaged.
• mice were treated for six days with 2pg per eye SPHINX31 as eye drops, and ERG carried out using a Micron IV Ganzfield ERG system as recommended by the manufacturers instructions.
• SRPK 1 inhibitors To identify novel SRPK 1 inhibitors, a range of inhibitors were screened in an in vitro kinase assay (Promega; Koresawa and Okabe, 2004). The previously identified SRPK inhibitors SPHINX and SPHINX7 were used as positive controls for the identification of novel candidates. Kinase assays showed that compounds 12 to 14 in Table 1 (termed SPHINX31- 33 respectively) had a 10-20 fold increase in potency compared with the previously reported compounds, resulting in IC 50 values of 3.2-17nM ( Figure 1). A structure activity relationship study to identify the mechanism and potential for new compounds was undertaken, with new compounds generated with structures shown in Table 2. Additional activity was reached with these compounds down to sub iiM potencies for compound 61 .
• a kinome screen of SPHINX31 against all known kinases using substrate DiscovcrX binding affinity assay demonstrated that the only other kinases that showed binding were the closely related Clkl and Clk4, which showed 27% and 14% binding at ⁇ ⁇ ( Figure 6).
• SPHINX31 had a binding affinity 44 x greater for SRPK1 ( ⁇ 12.8°C) than SRPK2 (( ⁇ ' ⁇ 6.7°C) or Clkl ( ⁇ 6.7°C) and 88 fold greater than Clk4 ( ⁇ 5.7°C).
• SPHINX3 1 could access the retina of an animal with a large eye a rabbit was exposed to 150 g per day SPI IINX 1 or pazopanib (Figure 16). After 6 days the animal was killed and the eyes harvested. Individual sections of sclera and retina from the rear half of the eye were then assayed for pazopanib or SPHINX31. Retinal penetration was seen for both pazopanib and SPHINX3 1. but the concentrations for SPHINX3 1 were 1 Ox the IC 50 for the compound whereas for pazopanib the concentration was similar to the IC 50 .
• Novel drug candidates are typically screened for an ability to inhibit the 'hERG' potassium channel, due to the established association between pharmacological blockade of hERG channels and drug- induced long QT syndrome and torsades de pointes arrhythmia (Han cox et al, 2008; Gintant, 2008). SPHINX did not inhibit hERG, as has previously been described (Gammons et al., 2013).
• Plasma level of all compounds of Formula (I) tested during topical local application to the eye were extremely low (below detection level of IpM) and consequently substantial hERG channel block in the heart is unlikely to occur during in vivo use of these compounds as eye drops.
• SRPIN340 and SPHINX do not inhibit hERG suggests that it is possible to have significant pharmacological actions against SRPK 1 without substantial hERG activity, (Gammons et al. 2013).
• SPHTNX3 1 which inhibited hERG with an IC 50 of 0.3 ⁇ , 100-fold higher than its IC 50 value against SRPK1 (3.2 nM) (Figure 5).
• ERG amplitude over time following stimulation with green light (Fig 20A) and UV light (Fig 20B) at 3.756 cd.s.nr.
• ERG amplitude at different intensities of green (Figs 20C, 20 E) or UV (Figs 20D, 20F) are shown for the A wave (Figs 20C. 20D) and B wave ( Figs 20E, 20F).
• the ratio of A-wave: B-wave was unaffected by SPHINX31 treatment (Fig 20G, Fig 20H ).
• eyes were enucleated, dissected, homogenised and spiked with SPHINX7 to measure extraction efficiency, then analysed by mass spectrometry ( Fig 201, Fig 20J).
• SPFIINX3 1 was detected in the retina (0. 165 % of applied eye drop dose when normalised to extraction efficiency) and choroid (0.0175% of applied dose). SPFIINX3 1 levels in control eyes are shown for comparison as background levels.
• Photopic ERG recordings were taken to isolate cone responses from rod responses following 10 minutes light adaptation and with continuous background stimulation with white light at an intensity of 30 cd.m.s 2 .
• VEGF vascular endothelial growth factor
• '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 VEGF 183, a novel spliced variant of vascular endothelial growth factor', lovs, 40(3), 752-759. Houck, K. A..
• 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(1 1), 2422-2427.
• VEGF Vascular Endothelial Growth Factor
• VEGI ' -A The carboxyl terminus of VEGI ' -A is a potential target for anti-angiogenic therapy.

Landscapes

• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Health & Medical Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Life Sciences & Earth Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Medicinal Chemistry (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• General Chemical & Material Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Epidemiology (AREA)
• Neurosurgery (AREA)
• Neurology (AREA)
• Biomedical Technology (AREA)
• Cardiology (AREA)
• Heart & Thoracic Surgery (AREA)
• Urology & Nephrology (AREA)
• Ophthalmology & Optometry (AREA)
• Pain & Pain Management (AREA)
• Hospice & Palliative Care (AREA)
• Psychiatry (AREA)
• Endocrinology (AREA)
• Reproductive Health (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Heterocyclic Compounds Containing Sulfur Atoms (AREA)
• Furan Compounds (AREA)
• Plural Heterocyclic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Priority Applications (8)

Applications Claiming Priority (2)

Related Child Applications (2)

Publications (1)

Family

ID=50928932

Family Applications (1)

Country Status (8)

Cited By (8)

Families Citing this family (2)

Citations (2)

Family Cites Families (7)

• 2014
  • 2014-04-17 GB GBGB1406956.1A patent/GB201406956D0/en not_active Ceased
• 2015
  • 2015-04-17 ES ES15718262T patent/ES2836105T3/es active Active
  • 2015-04-17 JP JP2017505735A patent/JP6579714B2/ja active Active
  • 2015-04-17 CN CN201580031650.5A patent/CN107001344B/zh active Active
  • 2015-04-17 AU AU2015248656A patent/AU2015248656B2/en active Active
  • 2015-04-17 US US15/304,678 patent/US9695160B2/en active Active
  • 2015-04-17 WO PCT/GB2015/051172 patent/WO2015159103A1/en not_active Ceased
  • 2015-04-17 EP EP15718262.7A patent/EP3131886B1/en active Active
• 2017
  • 2017-05-31 US US15/609,596 patent/US9796707B2/en active Active
  • 2017-09-19 US US15/708,315 patent/US9932330B2/en active Active

Patent Citations (2)

Non-Patent Citations (6)

Also Published As

Similar Documents

Legal Events