Classifications

• • 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
  • C07D233/64—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 substituted hydrocarbon radicals attached to ring carbon atoms, e.g. histidine
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
  • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
  • C07D263/30—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D263/32—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having two or three 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
• • 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
  • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
  • A61K31/42—Oxazoles
  • A61K31/421—1,3-Oxazoles, e.g. pemoline, trimethadione
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0048—Eye, e.g. artificial tears
• • 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
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
  • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
  • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
  • C07D213/06—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom containing only hydrogen and carbon atoms in addition to the ring nitrogen atom
  • C07D213/16—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom containing only hydrogen and carbon atoms in addition to the ring nitrogen atom containing only one pyridine ring
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
  • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
  • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
  • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
  • C07D213/36—Radicals substituted by singly-bound nitrogen atoms
  • C07D213/40—Acylated substituent nitrogen atom
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
  • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
  • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
  • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
  • C07D213/54—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
  • C07D213/56—Amides

Definitions

• the present invention relates to compounds for use as therapeutically active substances in the treatment and / or prevention of neuroretinal diseases, and in particular in the treatment and / or prevention of neuroretinal diseases leading to photoreceptor loss or degeneration of the outer retina .
• the main feature of neurodegenerative diseases is an increasing loss of nerve cells, resulting in various neurological symptoms.
• the diseases can arise in different periods of life, which proceed diffusely or generalized and produce specific patterns of damage.
• retinitis pigmentosa retinitis pigmentosa
• retinopathia pigmentosa The chief function of the retina is transduction of light into nervous impulses by the rods and the cones.
• Retinitis pigmentosa is a chronic retinal degeneration where the deterioration is accompanied by abnormal deposits of pigment in the rods of the retina. The disease causes a progressive decrease in peripheral vision leading to malfunction of the side vision. Eventually, the person with retinitis pigmentosa can see only straight ahead so that the patient experiences a condition known as "tunnel vision".
• WO 2016/073931 discloses a method for the treatment of retinitis pigmentosa in a human that comprises administering to the human a therapeutically effective amount of N- acetylcysteine amide (NACA) which reduces cone cell death in the eye.
• NACA N- acetylcysteine amide
• EP 2 734 202 discloses a pharmaceutical composition containing 4- bromo-iV- (imidazolidin-2-ylidene) -1 H- benzimidazol-5-amine as active ingredient for modulating the alpha 2 adrenergic receptors. It was shown that said compound reduced and protected the retina from the damage caused by blue light.
• US 2015/290215 discloses a composition comprising clozapine, n-desmethyl clozapine, olanzapine or derivatives thereof for treating a retinal disorder, which is caused by oxidative stress .
• US 2016/0213671 relates to a pharmaceutical composition for the treatment or prophylaxis of a neurodegenerative disease, which is not based on a protein-folding disorder comprising as the active agent an inhibitor of the valosin-containing protein (VCP inhibitor) .
• WO 2014/079850 discloses both substituted heterocyclic compounds which were believed to stimulate adult neuronal stem cells and that said compounds may be used for a plurality of different diseases.
• neuronal stem cells have the ability to differentiate into several cell types, it cannot be predicted whether said new cell types can be stimulated by the same compounds.
• a significant number of compounds which stimulate neuronal stem cells have no or only a weak activity with regard to other cell types such as retinal precursor cells.
• US 6,117,675 discloses stem cells isolated from the retina of mammals and retinal cells differentiated from these stem cells and a method for obtaining cells from a retinal pigment epithelial layer of a mammal.
• precursor cells encompasses in this context any form of proliferative and non-proliferative cells such as stem cells per se and progenitor cells that can give rise to further differentiated functional tissues of the eye.
• precursor cells include in particular retinal precursor cells .
• the problem of the present invention is therefore to provide new compounds, which stimulate the proliferation of retinal precursor cells.
• A is a 5-oxazolyl residue or a pyridine-4-yl residue
• Ri is selected from the group consisting of fluoro and chloro ;
• R 2 , R 3 , R 4 , R 5 and R 6 of the phenyl ring B are independently from each other selected from the group consisting of hydrogen, a linear or branched alkyl having 1 to 4 carbon atoms, trifluoromethyl, 2, 2, 2-trifluoroethyl, methylsulfany1, ethylsulfany1, methylsulfonyl, ethylsulfonyl, difluoromethoxy, trifluoromethoxy, fluoro, bromo, chloro, methoxy, ethoxy, propoxy, butoxy, hydroxy and amino; and at least two of R 2 , R.3, R.4, Rs and R6 are hydrogens, with the proviso that if Ri is chloro, R5 is not methoxy.
• pharmaceutically acceptable salt stands for therapeutically active, non-toxic acid salt forms, which the compound according to the present invention is able to form.
• alkyl refers to a straight or branched hydrocarbon chain containing 1 to 4 of carbon atoms.
• alkyl as used herein include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert- butyl.
• the residue A may be a 5-oxazolyl group of the formula (II)
• residue A may be a pyridine group of the formula (III) wherein denotes the point of attachment to the remainder of the molecule.
• the phenyl ring B in the compound of the present invention is monosubstituted or disubstituted, but it is also possible that all of R 2 , R. 3 , R. 4 , Rs and R 6 are hydrogen.
• the term "monosubstituted” means that one of R 2 , R 3 , R 4 , R 5 and Rg is not hydrogen.
• the term "disubstituted” means that two of R 2 , R 3 , R 4 , R 5 and R 6 are not hydrogens.
• Ri is chloro. Said compounds show an outstanding biological activity.
• Ri is a residue as defined above and the phenyl ring B is not substituted, that is, all of R 2 , R 3 , R 4 , R 5 and R 6 are hydrogens.
• R ! is a residue as defined above and the phenyl ring B is monosubstituted, that is, one of R 2 , R 3 , R 4 , R 5 and R 6 is not hydrogen .
• R 2 is preferably selected from the group consisting of methyl, trifluoromethyl, methylsulfanyl, methylsulfonyl, difluoromethoxy, fluoro, bromo, chloro, methoxy, and ethoxy, most preferably difluoromethoxy and chloro, and R 2 , R 3, R 4 , R 5 and R 6 are hydrogen.
• R 2 , R 3, R 4 , R 5 and R 6 are hydrogen.
• R 2 is hydrogen and one of R 3 R 4 , R 5 and R 6 is preferably selected from the group consisting of trifluoromethyl, difluoromethoxy, methoxy, preferably trifluoromethyl and difluoromethoxy .
• the phenyl ring B is disubstituted, that is, two of R 2 , R 3 , R 4 , Rs and R 6 are not hydrogens.
• the disubstitution may be an ortho, meta or para substitution.
• R 2 is selected from the group consisting of fluoro, bromo and chloro
• one of R 3 , R 4 or R 5 is selected from the group consisting of fluoro, bromo and chloro.
• the two residues which are different from hydrogen may be the same or different from each other.
• R 2 is chloro and R 5 is fluoro resulting in a para-substitution, or both R 2 and R 4 are fluoro resulting in a meta-substitution.
• the compound of formula (I) is selected from the group consisting of compounds of the formula (I), wherein A, R lf R 2 , R 3 , R 4 , R 5 and R 6 are
• the compound of formula (1) showed an increase of cell proliferation of 103%, the compound of formula (10) of 58%, compound of formula (4) of 57%, compound of formula (9) of 49%, compound of formula (3) of 43%, and compound of formula (2) of 42%.
• the present invention relates to a pharmaceutical composition
• a pharmaceutical composition comprising a pharmaceutically acceptable carrier and/or adjuvant; and a compound of the formula (la) or a pharmaceutically acceptable salt thereof, wherein
• A is a 5-oxazolyl residue or a pyridine-4-yl residue
• Ri' is selected from the group consisting of methoxy, hydrogen, fluoro and chloro;
• R 2 , R 3 , R 4 , R 5 and Rg of the phenyl ring B are independently from each other selected from the group consisting of hydrogen, a linear or branched alkyl having 1 to 4 carbon atoms, trifluoromethyl, 2, 2, 2-trifluoroethyl, methylsulfanyl, ethylsulfanyl, methylsulfonyl, ethylsulfonyl, difluoromethoxy, trifluoromethoxy, fluoro, bromo, chloro, methoxy, ethoxy, propoxy, butoxy, hydroxy and amino; and at least two of R 2 , R 3 , R 4 , R 5 and R 6 are hydrogens, with the proviso that if Ri' is hydrogen or methoxy, A is a pyridine-4-yl residue. as a therapeutically active substance.
• the definition of the compound of formula (la) differs from the definition of the compound of formula (I) in that Ri' is selected from the group consisting of methoxy, hydrogen, fluoro and chloro instead of Ri that was only selected from the group consisting of fluoro and chloro.
• prevention refers to the prevention or reduction of signs and symptoms associated with neuroretinal diseases, in particular of primary neuroretinal diseases leading to photoreceptor loss or degeneration of the photoreceptor layer of the retina in subjects who are at risk for developing the disease. In these subjects a predisposing factor may be retained, but the signs and/or symptoms of the disease do not occur or take significantly longer to develop. Further, it also includes the prevention of a further deterioration of the symptoms once the disease has occurred .
• composition means a composition that is suitable for administering to human patients for the treatment of diseases. Said pharmaceutical composition efficiently stimulates proliferation, migration or both proliferation and migration of endogenous retinal precursor cells in a patient.
• the pharmaceutical composition comprises a pharmaceutically acceptable carrier and/or adjuvant; and a compound of the formula (I) as defined above, and in particular a compound of formula (I) as disclosed in Table 1 and/or Table 2.
• the pharmaceutical composition comprises the compound of formula (la), wherein R lf R 2 , R 3 , R 4 , R 5 and R 6 are
• the compounds according to the present invention and the compositions according to the present invention stimulate the proliferation of retinal precursor cells.
• they are suitable in the treatment and/or prevention of neuroretinal diseases, in particular of primary neuroretinal diseases leading to photoreceptor loss or degeneration of the photoreceptor layer of the retina.
• Compounds and compositions according to the present invention are suitable for the use in the treatment and/or prevention of a disease selected from the group consisting of inherited retinal dystrophies including retinitis pigmentosa (RP) , including syndromic and non-syndromic forms, X-chromosome linked, recessive, dominant and sporadic forms, rod-cone dystrophies, Usher's syndrome, Stargardt's disease, cone-rod dystrophies, cone dystrophies, achromatopsia, blue cone monochromacy, enhanced S-cone syndrome, rod dystrophies, choroideremia, Leber's congenital amaurosis, juvenile X-chromosome linked retinoschisis (JXLR), fundus albipunctatus , retinitis punctata albescens, fleck retina of Kandori, bietti crystalline retinal dystrophy, fenestrated sheen macular dystrophy, adult-onset foveomacular vit
• the compound of the present invention is used in the treatment of retinitis pigmentosa (RP) , including syndromic and non-syndromic forms, X-chromosome linked, recessive, dominant and sporadic forms .
• RP retinitis pigmentosa
• Compounds and compositions according to the present invention are suitable for the use in the treatment and/or prevention of acquired degeneration selected from the group consisting of crystalline maculopathy (drug-related, hyperoxaluria, cystinosis, Sj ogren-Larsson syndrome), west African crystalline maculopathy, solar retinopathy, talc retinopathy, diabetic retinopathy, sickle cell retinopathy, macular telangectasia, eales disease, retinal detachment, retinal dialysis, peripheral retinoschisis.
• crystalline maculopathy drug-related, hyperoxaluria, cystinosis, Sj ogren-Larsson syndrome
• west African crystalline maculopathy solar retinopathy
• talc retinopathy diabetic retinopathy
• sickle cell retinopathy macular telangectasia
• eales disease retinal detachment
• retinal dialysis peripheral retinoschisis
• Compounds and compositions according to the present invention are suitable for the use in the treatment and/or prevention of vascular related retinal degeneration selected from the group consisting of central/branch retinal artery occlusion (CRAO/BRAO) , central/branch retinal vein occlusion (CRVO/BRVO) , haemorrhagic occlusive retinal vasculitis (HORV) .
• CRAO/BRAO central/branch retinal artery occlusion
• CRVO/BRVO central/branch retinal vein occlusion
• HORV haemorrhagic occlusive retinal vasculitis
• Compounds and compositions according to the present invention are suitable for the use in the treatment and/or prevention of drug-induced maculopathies selected from the group consisting of chloroquine, hydroxychloroquine, phenothiazine, quinine sulfate, thioridazine, clofazimine, cholopromazine, deferoxamine, chloroquine-derivatives , cisplatin, carmustine, chlofazimine and vigabatrin as well as crystal-induced maculopathies including tamoxifen, talc, canthaxanthine, methoxyflurane, nitrofurantoin, cystoid macular edema (CME) including Epinephrine, latanoprost and nicotinic acid.
• drug-induced maculopathies selected from the group consisting of chloroquine, hydroxychloroquine, phenothiazine, quinine sul
• Compounds and compositions according to the present invention are suitable for the use in the treatment and / or prevention of infectious and/or inflammatory eye diseases selected from the group consisting of progressive outer retinal necrosis (PORN) , acute retinal necrosis (ARN) , CMV- retinitis, Sarcoidosis, acute syphilitic posterior placoid chorioretinitis, tuberculosis chorioretinitis, toxoplasmic retinochoroiditis , posterior Uveitis and retinal vasculitis, intermediate uveitis, pars planitis +/- CME, enophthalmitis (anterior and/or posterior) , posterior scleritis and masquerade syndromes.
• PORN progressive outer retinal necrosis
• ARN acute retinal necrosis
• CMV- retinitis CMV- retinitis
• Sarcoidosis acute syphilitic posterior placoid chorioretinitis
• Compounds and compositions according to the present invention are suitable for the use in the treatment and / or prevention of white dot syndromes selected from the group consisting of multifocal choroiditis and panuveitis (MCP) , punctate inner choroidopathy (PIC) , birdshot retinochoroidopathy, acute macular neuroretinopathy (AMN) and acute zonal occult outer retinopathy (AZOOR) .
• MCP multifocal choroiditis and panuveitis
• PIC punctate inner choroidopathy
• APN acute macular neuroretinopathy
• AZOOR acute zonal occult outer retinopathy
• the compound or the composition according to the present invention can be administered to a patient, either alone or in combination with one or more additional therapeutic agents.
• "Patient” as used herein includes mammals such as humans, non-human primates, rats, mice, rabbits, hares, dogs, cats, horses, cows and pigs, preferably human.
• the pharmaceutical composition according to the present invention may comprise one or more additional therapeutic agents .
• controlled release pharmaceutical compositions refers to any composition or dosage form, which comprises the compound of the present invention and which is formulated to provide a longer duration of pharmacological response after administration of the dosage form than is ordinarily experienced after administration of a corresponding immediate release composition comprising the same drug in the same amount. Controlled release may be extended up to several months depending on the matrix used. Preferably, the release of the compound according to the present invention takes place over a period of up to 12 months, most preferably over a period of up to 6 months. Such a controlled release formulation results in an increased patient comfort and in significant lower costs.
• the matrix material used for a pharmaceutical composition according to the present may comprise hydrophobic release controlling agents. It is preferably selected from but not limited to polyvinyl acetate dispersion, ethyl cellulose, cellulose acetate, cellulose propionate (lower, medium or higher molecular weight) , cellulose acetate propionate, cellulose acetate butyrate, cellulose acetate phthalate, cellulose triacetate, poly (methyl methacrylate) , poly (ethyl methacrylate) , poly (butyl methacrylate) , poly (isobutyl methacrylate), and poly (hexyl methacrylate), poly (isodecyl methacrylate), poly (lauryl methacrylate), poly (phenyl methacrylate) , poly (methyl acrylate) , poly (isopropyl acrylate), poly (isobutyl acrylate), poly (octadecyl acrylate) , waxes such
• the compound of the invention can be delivered to the eye through a variety of routes, including but not limited to topical application to the eye or by intraocular injection into, for example, the vitreous or subretinal (interphotoreceptor) space; locally by insertion or injection into the tissue surrounding the eye; systemically through an oral route or by subcutaneous, intravenous or intramuscular injection; or via catheter or implant. Most preferably, the compound of the present invention is delivered by intraocular injection.
• the compound of the invention can be administered prior to the onset of the condition to prevent its occurrence, such as during eye surgery, immediately after the onset of the pathological condition, or during the occurrence of an acute or protracted condition.
• the compound according to the present invention may be incorporated in any pharmaceutically acceptable dosage form, such as for example, liquids, including solutions, suspensions and emulsions, tablets, suppositories, pills, capsules, powders or the like, preferably dosage forms suitable for single administration of precise dosages, or sustained release dosage forms for continuous controlled administration. Most preferred are liquids.
• Liquid pharmaceutically administrable dosage forms can be for example a solution, a suspension or an emulsion, preferably a solution comprising a compound of the present invention and optional pharmaceutical adjutants in a carrier, such as for example, water, saline, aqueous dextrose, glycerol, ethanol, DMSO and the like, to thereby form a solution or suspension.
• a carrier such as for example, water, saline, aqueous dextrose, glycerol, ethanol, DMSO and the like.
• the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like. Typical examples of such auxiliary agents are sodium acetate, sorbitan monolaurate, triethanolamine, sodium acetate and triethanolamine oleate.
• the present invention also relates to a method of treating a neuroretinal disease that leads to photoreceptor loss or outer-retina degeneration, comprising administering a compound of formula (la) or a pharmaceutically acceptable salt thereof to a patient having the retinal disease so as to be delivered to an eye of the patient in an amount effective to treat the retinal disease.
• a compound of formula (la) is defined above in detail.
• the compounds of formula (I) may be prepared by methods described below, together with synthetic methods known in the art of organic chemistry, or modifications that are familiar to those of ordinary skill in the art.
• the starting materials used herein are available commercially or may be prepared by routine methods known in the art, such as those methods described in standard reference books such as "Compendium of Organic Synthetic Methods, Vol. I-X1N” (published with Wiley-lnterscience, ISSN: 1934-4783).
• Preferred methods include, but are not limited to, those described below.
• HPLC high-pressure liquid chromatography
• Analytical ultra-performance liquid chromatography used in the following examples and preparations was effected according to the following method unless modified in specific examples.
• a Chromegabond WR C18 (3 cm x 3.2 mm, 3m) column operated with a flow rate of 1.5 mL/min.
• mobile phases 0.02% TFA in water (mobile phase C) and 0.02% TFA in CH 3 CN (mobile phase D) were used in a gradient starting at 90% C and 10% D, changed to 10% C and 90% D in 3.0 min, then to 90% C and 10% D in 4.0 min, which was held constant up to 5.1 min .
• R1 is as described in formula I, R are hydroxy groups or R together with the boron atom form a 4, 4,5,5- tetramethyl-1 , 3 , 2-dioxaborolane group .
• Compounds of general formula IVa may be prepared from compounds of general formulae VI and VII in the presence of a palladium catalyst such as tetrakis ( triphenylphosphin) palladium ( 0 ) and a base such as potassium carbonate or other Suzuki-Miyaura coupling reaction conditions known to chemists skilled in the art of organic synthesis.
• a palladium catalyst such as tetrakis ( triphenylphosphin) palladium ( 0 )
• a base such as potassium carbonate or other Suzuki-Miyaura coupling reaction conditions known to chemists skilled in the art of organic synthesis.
• Compounds of general formula IVb may be prepared by reduction of the nitro group in compounds of general formula X using procedures known to chemists skilled in the art.
• Compounds of general formula X may be prepared from aldehydes of general formula VIII by reaction in the presence of a reagent such as isocyanomethane ) sulfonyl-4- methylbenzene (IX) in the presence of a base such as potassium carbonate.
• reaction mass was purified by preparative HPLC to yield N- ( 3-chloro-4- (oxazol-5-yl ) phenyl ) -2- ( 2 , 4- difluorophenyl ) acetamide (Compound (4)) (57 mg, 32%) .
• reaction mass was purified by preparative HPLC to yield N- (3-chloro-4- (oxazol-5-yl) phenyl) -2- (2- chloro-5-fluorophenyl ) acetamide (Compound (5)) (34 mg,
• Trifluoromethyl-phenyl ) -acetic acid (136 mg, 0.67 mmol) in DMF (1 mL) were added DIPEA (0.26 mL) and HATU (392 mg, 1.03 mmol) at rt and the reaction was stirred for 16h at rt .
• reaction mass was purified by preparative HPLC to yield N- ( 3-chloro-4- (oxazol-5- yl) phenyl) -2- (3- (trifluoromethyl) phenyl) acetamide
• reaction mass was purified by preparative HPLC to yield 2— ( 3— (difluoromethoxy) phenyl) -N- (3-fluoro-4- (oxazol-5- yl ) phenyl ) acetamide (Compound (13)) (104 mg, 25%) .
• UPLC Rt :
• reaction mass was purified by preparative HPLC to yield 2- ( 2-chlorophenyl ) -IV- ( 3- fluoro-4- (oxazol-5-yl ) phenyl ) acetamide (Compound (17)) (123 mg, 44%).
• UPLC Rt 1.50 min; MS: 331.2 (M+H).
• curved or angled micro-dissecting scissors were used to cleave any remaining extraocular muscle tissue, the optic nerve, and cut the eyeball into symmetrical halves; beginning and finishing the cut from the hole left by the optic nerve.
• the two eye halves were peeled apart.
• the lens, optic nerve, and vitreous were separated from the eye shells and the eye shells were transferred into a new petri dish (also containing cold, sterile aCSF) .
• CE ciliary epithelium
• eye shells were oriented with the cornea on the right and retinal pigmented epithelium (RPE) on the left.
• a pair of straight forceps were used to pin down the eye shell on the RPE side while a scalpel blade was inserted between the CE and the iris, using pressure to slice the iris/cornea side off from the rest of the shell.
• the scalpel was run along the border between the CE and the RPE to obtain the CE isolated as a thin strip of tissue.
• the CE strips were then transferred to a 35mm dish containing 2mL of dispase solution (Sigma; T1005) and incubated for 10 minutes at 37°C.
• the strips were transferred from dispase into a 35mm dish containing 2mL of sterile filtered kynurenic acid, trypsin and hyaluronidase solution and incubated at 37°C for 10 minutes.
• the dish was returned to the dissecting scope, and the CE strips were pinned down with straight, non- serrated forceps, while non-serrated curved forceps were used to scrape the CE off from the underlying sclera.
• the bare scleral strips were then discarded, such that only the CE cells remained in the enzyme solution.
• the cells and enzyme solution were transferred to a 15mL tube and triturated approximately 45 times to break apart the tissue. The 15mL tube/cell suspension was centrifuged for 5 minutes at 1500 rpm.
• the supernatant was gently aspirated from the resulting pellet using a fire-polished, cotton-plugged glass pipette and 2mL of trypsin inhibitor solution was added to the pellet. Using a small borehole, fire-polished, cotton- plugged glass pipette, the sample was triturated approximately 45 times until it was a single-cell suspension. The 15mL tube/cell suspension was centrifuged for 5 minutes at 1500 rpm. The supernatant was gently aspirated from the resulting pellet and l-2mL of SUM with
• FGF2 and heparin plating media was added.
• the cells and media were mixed to ensure a uniform cell suspension and a lOuL sample was taken and cell density was determined.
• the cells were then seeded and cultured at 10c/pL in culture- treated plates or flasks. After one week, roughly 1 in 500 cells proliferated to form free-floating, clonal spheres greater than 80pm in diameter.
• Human-derived spheres were passaged using the kynurenic acid, trypsin, hyaluronidase enzyme solution with the addition of collagenase I (0.5mg/mL), collagenase II (0.5mg/mL) and elastase (O.lmg/mL) .
• Mouse-derived spheres were passaged using hyaluronidase (0.67mg/mL), collagenase I (0.5mg/mL), and collagenase II (0.5mg/mL) dissolved in Accustase solution (Sigma; SCR005) .
• Spheres were collected en masse from culture plates or flasks, transferred into one or more 50mL tubes and centrifuged for 5 minutes at 1500rpm. The supernatant was gently aspirated from the pellet and 2- 5mL of enzyme solution was added to the pellet and mixed thoroughly. The 2-5mL enzyme and sphere suspension was transferred to a 15mL tube and laid horizontally on an automated rocker at 37°C for 45 minutes. After incubation, the enzyme solution with spheres was triturated approximately 45 times to mechanically dissociate the spheres. The cell suspension was centrifuged for 5 minutes at 1500 rpm. The supernatant was gently aspirated and l-2mL of trypsin inhibitor solution was added to the pellet and triturated approximately 45 times.
• the cell suspension was centrifuged for 5 minutes at 1500 rpm. The supernatant was gently aspirated from the resulting pellet and l-2mL of SFM with FGF2 and heparin (plating media) was added. The cells and media were mixed to ensure a uniform cell suspension and a lOuL sample was taken and cell density was determined from that sample. The remaining cells were then seeded and cultured at 10c/pL in prepared 96-well or 24-well plates with 0.1% DMSO or a selected concentration of drug in 0.1% DMSO. Cells were grown for one week and then live stained for nuclei (Hoechst 33258; 10pg/mL) .
• an actin-green fluorescent protein (GFP) transgenic mouse strain FVB . Cg-Tg (CAG-EGFP) B5Nagy/J
• FVB . Cg-Tg CAG-EGFP
• B5Nagy/J actin-green fluorescent protein
• calcein AM ThermoFisher C3100MP; 2mM
• cell number comparisons were made based on nuclei and calcein fluorescence-based quantification.

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