CN1997359A - Management of ophthalmologic disorders, including macular degeneration - Google Patents

Management of ophthalmologic disorders, including macular degeneration Download PDF

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CN1997359A
CN1997359A CN 200580011513 CN200580011513A CN1997359A CN 1997359 A CN1997359 A CN 1997359A CN 200580011513 CN200580011513 CN 200580011513 CN 200580011513 A CN200580011513 A CN 200580011513A CN 1997359 A CN1997359 A CN 1997359A
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chemical compound
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hydrogen
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R·R·兰多
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Harvard College
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Harvard College
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Abstract

A drug may be used in the preparation of a medicament for the treatment or prevention of an ophthalmologic disorder, wherein the drug inihibits, antagonizes, or shortcircuits the visual cycle at a step of the visual cycle that occurs outside a disc of a rod photoreceptor cell.

Description

Comprise the processing of the ophthalmic diseases of degeneration of macula
The cross reference of related application
[0001] the application requires the U.S. Provisional Patent Application serial number 60/545,456 of submission on February 17th, 2004; The U.S. Provisional Patent Application serial number 60/567,604 that on May 3rd, 2004 submitted to; With the rights and interests of the U.S. Provisional Patent Application serial number of submitting on June 9th, 2,004 60/578,324, all these documents all are attached to herein by reference in full.
The research of relevant federal government-funded or the statement of exploitation
[0002] national health institute (National Institutes of Health) subsidizes number (Grant No.) R01-EY-04096 for the application's subject research provides the part support.Therefore, U.S. government has some right to the application's theme.
Foreword
[0003] in industrialized country, senile ophthalmic is the health problem that increases day by day.Senile degeneration of macula (AMD) influences the millions of crowd in the whole world, and it is visual deterioration and a blind main cause in the old group.In this disease, vision in daytime (vision of cone domination) is degenerated in time, because concentrate on the cone photoreceptors death of foveal region of retina.This disease incidence rate by rise in 50 years old age colony's less than 10% 75 years old surpass 30%, and crossed and should incidence rate continue to rise after the age.In this seizure of disease and the retinal pigment epithelium (RPE) and complicated toxicity biochemical on every side and the lipofuscin among the RPE accumulate relevant.Accumulating of these retinal toxicity mixture is one of known most important risk factor in the AMD etiology
[0004] RPE forms retinal blood barrier part, and also supports to comprise shaft-like and photoreceptor cell function taper.In other activity, RPE engulfs the consumption acromere (spent outer segments) of staff cell usually.In the degeneration of macula of some form at least, the accumulating part of lipofuscin in RPE is because this phagocytosis.Retinal toxicity chemical compound (retinotoxic compound) forms in the dish (disk) of shaft-like photoreceptors acromere.Therefore, the retinal toxicity chemical compound in the dish is brought into RPE, and they further weaken the acromere phagocytosis there, and cause the RPE apoptosis.The photoreceptor cell death that comprises the essential pyramidal cell of vision in daytime then loses RPE and supports.
[0005] one of retinal toxicity chemical compound that forms in the dish of shaft-like acromere is the inferior retinyl (retinylidene) of N--N-retinyl ethanolamine (A 2E), it is the important component of retinal toxicity lipofuscin.Under the normal condition, A 2E forms in dish, but amount very little thus it does not weaken RPE function when engulfing.But under some pathological conditions, so many A 2E can accumulate in dish, consequently RPE " poisoning " when acromere is engulfed.
[0006] A 2E is produced by alltrans-retinal, and alltrans-retinal is one of rod-vision circulation intermediate.At normal vision cycle period (summary is seen Fig. 1), alltrans-retinal produces in rod cell acromere dish.Alltrans-retinal can react with the PHOSPHATIDYL ETHANOLAMINE (PE) of dish film component, forms the inferior retinyl-PE of N-.Rim albumen (RmP), ATP are arranged in the film of rod cell acromere dish in conjunction with box (cassette) transport protein, then with alltrans-retinal and/or the inferior retinyl of N--PE transhipment placing and be transported in the rod cell acromere dish kytoplasm.Environment there helps hydrolyzing N-Ya retinyl-PE.Alltrans-retinal is reduced to alltrans-retinol in the rod cell kytoplasm.Alltrans-retinol passes rod cell acromere plasmalemma and enters extracellular space then, is absorbed by retinal pigment epithelium (RPE).Alltrans-retinol is converted into 11-cis-retinal by series reaction, and it returns photoreceptors and continues visual cycle (visual cycle).
[0007] still, remove alltrans-retinal in the dish by prevention, the defective among the RmP can upset this process.(incidence rate that influences the child usually is 1/10,000 to be called the recessive form of degeneration of macula of Shi Tajiate disease (Stargardt ' s disease); Have 25,00 individual influenced in the U.S.), the gene of coding RmP, abcr undergos mutation, and transport protein is a non-functional.Therefore, the inferior retinyl-PE of alltrans-retinal and/or N-is trapped in the dish.Then the inferior retinyl-PE of N-can with another alltrans-retinal molecular reaction, form the inferior retinyl of N--N-retinyl ethanolamine (A 2E); This process is summarized among Fig. 2.Form some A as mentioned above, even in normal condition 2E; But because the cause of defective transport protein is worked as A 2When the precursor of E is accumulated, just produce A in a large number in dish 2E, thereby can cause degeneration of macula.
[0008] degeneration of macula of other form also can be produced by the pathology that causes lipofuscin to be accumulated.The gene that coding speciality chain fatty acid-4 extends, the sudden change of elovl4 causes the dominant form of Shi Tajiate disease, be called the autosomal dominant macula lutea atrophy that chromosome 6-connects (ADMD, OMIM#600110).
[0009] even the prophylactic treatment of couple AMD is arranged, also be few, and treatment is intervened only effective to some not really common disease form.
General introduction
[0010] disclosure relates to the processing degeneration of macula, prevents that more particularly the retinal toxicity chemical compound is in retinal pigment epithelium and the compositions of accumulating, system and method on every side.
[0011] in one embodiment, prevention and minimizing A 2E accumulating in rod cell acromere dish.Discovery can reduce A by the medicine that limits visual cycle 2E produces in dish.Available accomplished in many ways should restriction.In a method, medicine can effectively shorten generation A 2The circuit (circuit) of the visual cycle part of E precursor alltrans-retinal.In another method, medicine can suppress the concrete ladder (step) in the synthetic necessary visual cycle of alltrans-retinal.In another method, medicine can stop midbody product (retinyl ester) to combine with some chaperone in the retinal pigment epithelium.
[0012] in one embodiment, the method for treatment or prevention patient degeneration of macula can comprise that giving the patient shortens the medicine that occurs in the visual cycle circuit on the outer visual cycle ladder of shaft-like photoreceptor cell dish.In another embodiment; the method of treatment or prevention patient degeneration of macula can comprise that giving the patient suppresses and/or disturb at least a lecithin retinol acyltransferase, RPE65, the medicine of 11-cis-retinol dehydrogenase and isomery hydrolytic enzyme (isomerohydrolase).
[0013] in another scheme again, identifies that the method for degeneration of macula medicine can comprise suffering from or have patient's candidate medicine that the degeneration of macula risk takes place, and measure the retinal toxicity chemical compound that is accumulated in patient's retinal pigment epithelium.
[0014] estimates to use multiple medicine.In certain embodiments, the visual cycle inhibitor comprises that retinoid is like thing.In other embodiments, the medicine of shortening visual cycle circuit comprises aromatic amine and hydrazine.
The accompanying drawing summary
[0015] Fig. 1 represents visual cycle.
[0016] Fig. 2 represents A 2E's is synthetic.
[0017] Fig. 3 represents to shorten the intervention of visual cycle circuit (circuiting).
[0018] Fig. 4 A-C lists relevant alltrans-tretinoin and the bonded data of RPE65.
[0019] Fig. 5 A-C lists relevant 13-cis-tretinoin and the bonded data of RPE65.
[0020] Fig. 6 A-C list relevant N-(4-hydroxy phenyl) look yellow amide (retinamide) (4-HPR) with the bonded data of RPE65.
[0021] Fig. 7 lists between relevant alltrans-tretinoin and the Palmic acid alltrans-retinyl ester the competitive bonded data with RPE65.
[0022] Fig. 8 lists relevant alltrans-tretinoin (atRA), 13-cis-tretinoin (13cRA) and N-(4-hydroxy phenyl) and looks yellow amide (4-HPR) to 1]-data of cis-retinol biosynthesis influence.
[0023] Fig. 9 A1, A2, B1 and B2 list relevant alltrans-retinol and Palmic acid alltrans-retinyl ester and the bonded data of pure sRPE65.Fig. 9 C lists relevant vitamin A and the bonded data of sRPE65.Fig. 9 D lists the binding constant of the various binding partners of measurement.
[0024] Figure 10 A-C lists the palmitoylation data of mRPE65 in the relevant body.
[0025] Figure 11 A-D lists the data that relevant mRPE65 and sRPE65 transform mutually.
[0026] Figure 12 A-C lists the palmitoylation data of relevant 11-cis-retinol.
[0027] Figure 13 A and B represent how described adjusting composition can regulate and control flowing of retinoid in the vision (retinoids).
[0028] Figure 14 A-18B lists the data that act in the body of relevant short circuit (short circuit) medicine.
[0029] Figure 19-24 lists the data of effect in relevant enzyme inhibitor and/or the RPE65 antagonist body.
[0030] Figure 25 lists the relevant lower body profile that exists at aromatic amine and becomes A 2The data of E.Describe in detail
[0031] 1. overview
[0032] disclosure provides by preventing or reducing A2E accumulates in rod cell acromere dish Process composition and the method for macular degeneration. Can be present in rod cell acromere dish by minimizing In alltrans-retinene amount prevent or reduce A2E accumulates. In a method, can give Give the medicine that suppresses the one or more enzymatic ladders in the visual cycle, to reduce alltrans-look The generation of yellow aldehyde. In another method, can impel 11-cis-retinene among the RPE Isomery turns to the medicine of alltrans-retinene, thereby minimizing is returned the acromere dish and turned to by isomery The amount of the 11-cis-retinene of alltrans-retinene.
[0033] 2. definition
[0034] for ease of understanding, before further setting forth the exemplary embodiment, will be used for Some term in specification of the present invention, embodiment and the appended claims concentrates on this. Should treat these definition according to disclosure remainder and those skilled in the art's understanding.
[0035] article used herein " " refers to 1 of article or more than 1 (namely Refer at least 1) grammar object. As an example, " composition " expression a kind of composition or a kind of Above composition.
[0036] term " access to plant " is the technical term of generally acknowledging, comprises any suitable increasing Add or medical treatment device that maintenance and anatomic region approach. The technician of medical science and surgical field is familiar with Such device. Access to plant can be pin, conduit, intubate, trocar, tubing, isocon, Drainage tube or endoscope, for example otoscope, electric nasopharyngoscope, an organ mirror or be applicable to hinge area Any other endoscope in territory, or be applicable to and enter or remain positioned in advance the dissection of selecting Any other medical treatment device in the zone.
[0037] when relating to the chemical compound use, term " biocompatible chemical compound " and " biocompatibility " are technical known.For example, the biocompatible chemical compound comprises itself neither to host (for example animal or human) toxigenicity, also not according to the chemical compound of the speed degraded (if degradation) that produces monomer subunit or oligomer subunit or other By-product Toxicology concentration in the host.In certain embodiments, biodegradation is usually directed to the degraded of chemical compound in organism, for example is degraded to the monomer whose subunit, and known single subunit is nontoxic certainly.But the intermediate oligomerization product that this degraded produces may have different toxicology properties, or biodegradation may relate to the generation compound molecule but not oxidation or other biochemical reaction of monomer subunit.Therefore, in certain embodiments,, can determine to be intended to use in vivo, for example grow or be expelled to the toxicology characteristic of the biodegradable compound among the patient through behind one or more oxicity analysis.Need not any compositions have 100% purity, but just can think biocompatible; Certainly, but only to need an above-mentioned compositions be biocompatible.Therefore, this compositions can comprise 99%, 98%, 97%, 96%, 95%, 90%, 85%, 80%, 75% or even still less the biocompatible chemical compound that for example comprises described chemical compound and other material and excipient herein, but and biocompatible still.
[0038] but for determining that whether chemical compound or other material are biocompatible, may must carry out oxicity analysis.This type of mensuration is well-known in the art.An example of this type of mensuration is can be in the following manner, and with the cancerous cell of living, for example the GT3TKB tumor cell carries out this mensuration: under 37 ℃, sample is degraded until observing degraded fully in 1M NaOH.Use 1M HCl neutralization solution then.The catabolite sample of the various concentration of about 200 μ L is placed 96 hole tissue culturing plates, by 10 4/ hole density, inoculation gastric carcinoma cells (GT3TKB).With catabolite sample and GT3TKB cell incubation together 48 hours.Can relative growth % to the mapping of the degraded sample concentration in tissue culture hole expression measurement result.In addition, also can for example implant assessing compound and preparation, can not cause significant the stimulation or the inflammation level at subcutaneous implant site to prove conclusively them by the in vivo test of knowing in subcutaneous rat.
[0039] term " biodegradable " " be (art-recognized) well known in the art, comprise chemical compound, compositions and the preparation that will degrade when using, for example described herein those.The common difference with nonbiodegradable chemical compound of biodegradable chemical compound is that the former can be degraded in use.In certain embodiments, this purposes relates to purposes in the body, therapy in the body for example, and in some other embodiment, this purposes relates to external purposes.Generally speaking, relate to the composition subunit of biodegradable degradation for it owing to the degraded of biological degradability (biodegradability), or chemical compound digestion, for example be littler subunit by biochemical process digestion.In certain embodiments, can differentiate two kinds of dissimilar biodegradations usually.For example, a kind of biodegradation type may relate to key (no matter covalent bond still is other key) fracture of chemical compound.In this biodegradation, produce monomer or oligomer usually, more generally, by the one or more substituent bond fission that connects chemical compound this biodegradation takes place.On the contrary, the biodegradation of another kind of type may relate in the side chain or with the key (no matter covalent bond still is other) that side chain is connected with chemical compound and rupturing.For example, other chemical part that can discharge therapeutic agent or be connected with chemical compound by biodegradation as side chain.In certain embodiments, when using chemical compound, biodegradations one or another kind of or two types all may take place usually.Term used herein " biodegradation " comprises the biodegradation of two kinds of general types.
[0040] degradation speed of biodegradable compound depends in part on multiple factor usually, they comprise the degree of cross linking of chemical characteristic, molecular weight, crystallinity, biological stability and this chemical compound of the key of being responsible for any degraded, physical characteristic, shape and the size of implant, and mode of administration and position.For example molecular weight is big more, and degree of crystallinity is high more and/or biological stability is high more, and the degraded of any biodegradable compound is slow more usually.Term " biodegradable " " should comprise and be called (bioerodible) of biological etch " can " material and process,
[0041] in certain embodiments, if biodegradable compound also contains therapeutic agent or other material relevant with it, the biodegradable speed of this chemical compound can characterize by the rate of release of this type of material.In this case, biodegradation rate may not only depend on the chemical characteristic and the physical features of this chemical compound, but also depends on the characteristic of mixing any this type of material wherein.
[0042] in certain embodiments, compound formulation in the application of needs, biodegradation in the acceptable time limit.In certain embodiments, for example in vivo in the therapy, be 25-37 ℃, during the physiological liquid of pH6-8 when being exposed to temperature, this degraded usually occur in approximately less than 5 years, 1 year, half a year, 3 months, 1 month, 15 days, 5 days, 3 days or even 1 day during in.In other embodiments, chemical compound was degraded during a few weeks longer at about 1 hour, and this depends on the application of needs.
[0043] term " comprises ", " containing " " comprise ", " containing " " have " " having " and use by the open implication that comprises, and means to comprise other key element.The term " for example " that this paper refers to use " as " be nonrestrictive, and only be used for illustration purpose." comprise " and " including but not limited to " is used interchangeably.
[0044] term " drug delivery device " is a term well known in the art, is meant that suitable drugs is applied to any medical treatment device of target organ or anatomic region.Even this term comprises this device itself of design and is not formulated as and comprises compositions, also can transport or finish compositions is instiled to those devices of target organ or anatomic region.As an example, transmit the pin or the conduit of compositions and be interpreted as drug delivery device by inserting anatomic region or blood vessel or other structure relevant with anatomic region.As another example, have the stent that is included in its material or is coated on its lip-deep compositions or isocon or conduit and be interpreted as drug delivery device.
[0045] when using relevant therapeutic agent or other material, term " slow release " is well known in the art.For example, but compare in the large bolus injection administration type of synchronization biological utilisation with the material that wherein makes all amounts, this compositions of passing h substance in time can show slow release characteristic.For example, in specific embodiments, when contacting with the body fluid that comprises blood, tissue fluid, lymph etc., chemical compound substrate (by this paper or method known to those skilled in the art preparation) can (be compared with large bolus injection agent release) in the time that continues or prolong can experience progressively degraded (for example by hydrolysis), follows simultaneously to discharge any material that mixes wherein.This release can cause prolonging any therapeutic agent that mixes that transmits the treatment effective dose.In following some embodiment that more elaborates, slow release can be different.
[0046] term " delivery agent " is a term well known in the art, comprises impelling the molecule that transmits therapeutic agent or other material in cell.The example of delivery agent comprises: sterol (for example cholesterol) and lipid (for example cationic lipid, virion virosome or liposome).
[0047] term used herein " or " be interpreted as the expression " and/or ", have in addition in the literary composition offer some clarification on except.
[0048] phrase " gastrointestinal tract external administration " and " parenteral " are terms well known in the art, comprise that non-intestinal and local mode of administration for example inject, include but not limited in intravenous, intramuscular, the pleura, in the blood vessel, in the pericardium, in the intra-arterial, sheath, in the capsule, interior, intracardiac, the intradermal of eye frame, intraperitoneal, under trachea, subcutaneous, epidermis, under the intraarticular, capsule, under the arachnoidea, in the spinal column and breastbone inner injection and infusion.
[0049] term " treatment " is the recognized techniques term, comprises that inhibition diagnosed disease of patient, obstacle or the disease of suffering from disease, obstacle or disease, for example hinders its development; And alleviate disease, obstacle or disease, for example cause disease, obstacle and/or disease to disappear.Treatment disease or disease comprise at least a symptom of alleviating disease specific or disease, and both having made does not influence its pathophysiology basis.
[0050] term " prevention " is the recognized techniques term, comprise stoping susceptibility to disease, obstacle and/or disease but N goes out ill patient that disease, obstacle or disease take place that the prevention state relevant with disease is included in to be diagnosed out after the disease but stoped this state generation before this condition diagnosing goes out.
[0051] term " fluid " is the recognized techniques term, is meant wherein atom or molecule, as free-moving toward each other non-solid matter in gas or liquid.If unfettered when using, flowing material can be by the space that offers it, for example comprises that the surface of cutting part or the shape of the dead space that rim strip (flap) stays flow.Flowing material can be embedded or be expelled to spatial finite part, but its inflow space more most of or it is whole then.Such material can be described as " flowable ".This term is well known in the art, and comprises the fluid composition that for example can be ejected into the position; With the manually-operated outfit fluid composition of the injector to inject of No. 23-gage needles for example; Or the fluid composition that transmits by conduit.Can be by pipe perfusion, extruding, be passed to the position that needs at room temperature for high viscosity " gel sample " material or with enough injection pressures are provided, advance high viscosity substance by transmission system, for example the material of the arbitrary commercially available obtainable injection device injection of pin or conduit is also included within term " flowable " scope.When the chemical compound itself that uses can flow, the compositions that comprises it did not need to comprise the biocompatible solvent that it is distributed in bodily cavity.Certainly, the delivery system of the flowability that available dependence material is natural is passed to easy liquidation compound in the bodily cavity, so that it is applied to the tissue surface that needs.For example, if flowable, injectable contains compound compositions, and the injection back forms interim bio-mechanical barrier with bag quilt or encapsulate internal or tissue, or the coating of available its preparation solid implanting device.In some cases, flowable compositions has the ability that occupies its spatial form of containing in a period of time under body temperature.
[0052] as generally acknowledging in the art, viscosity herein is interpreted as fluidic internal friction power or when flowing material experience is out of shape, the flow resistance that shows.Can pass through the viscosity of the molecular-weight adjusting chemical compound of chemical compound, other method that changes the physical characteristic of particular compound is conspicuous for the technical staff who only has common experimental technique.The molecular weight of the chemical compound that uses can differ widely, and depending on needs rigid solid state (higher molecular weight) still to need fluid state (lower molecular weight).
[0053] phrase " pharmaceutically acceptable " is well known in the art.In certain embodiments, this term is included in the rational medical judgment scope, be applicable to the tissue of contact humans and animals, do not have over-drastic toxicity, stimulation, anaphylaxis or other problem or complication, with rational interests/risk than the compositions that matches, chemical compound and other material and/or dosage form.
[0054] phrase " pharmaceutically acceptable carrier " is well known in the art, comprise for example pharmaceutically acceptable material, compositions or solvent, for example liquid or solid filler, diluent, excipient, solvent or encapsulate material, they participate in another organ or the part that health was transported or be transported to any compositions by the organ or the part of health.With other components compatibility of this compositions and to the patient on the harmless meaning, every kind of carrier must be " acceptable ".In certain embodiments, pharmaceutically acceptable carrier is pyrogen-free.Some example that can be used as these materials of pharmaceutically acceptable carrier comprises: (1) sugar: for example lactose, dextrose plus saccharose; (2) starch: for example corn starch and potato starch; (3) cellulose and derivant thereof, for example sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) tragacanth gum powder; (5) Fructus Hordei Germinatus; (6) gelatin; (7) Pulvis Talci; (8) excipient: for example cocoa butter and suppository wax; (9) oil: for example Oleum Arachidis hypogaeae semen, Oleum Gossypii semen, sunflower oil, Oleum sesami, olive oil, Semen Maydis oil and soybean oil; (10) dihydroxylic alcohols, for example propylene glycol; (11) polyhydric alcohol: for example glycerol, sorbitol, mannitol and Polyethylene Glycol; (12) esters: for example ethyl oleate and ethyl laurate; (13) agar; (14) buffer agent: for example magnesium hydroxide and aluminium hydroxide; (15) alginic acid; (16) apirogen water; (17) isotonic saline solution; (18) ringer's solution; (19) ethanol; (20) phosphate buffer; (21) other is used for the nontoxic compatibility material of pharmaceutical formulation.
[0055] term " pharmaceutically acceptable salt " is well known in the art, comprises the compositions of nontoxic relatively, inorganic and organic acid addition salt, and these compositionss include but not limited to therapeutic agent, excipient, other material etc.The example of pharmaceutically acceptable salt comprises by mineral acid, for example deutero-those acid of hydrochloric acid and sulphuric acid and by organic acid, for example ethyl sulfonic acid, benzenesulfonic acid, right-deutero-those acid such as toluenesulfonic acid.The example that is used to form the suitable inorganic base of salt comprises hydroxide, carbonate and the bicarbonate of ammonia, sodium, lithium, potassium, calcium, magnesium, aluminum, zinc etc.Also available suitable organic base forms salt, and these organic bases comprise nontoxic and those alkali that have the intensity of enough this type of salt of formation.In order to illustrate, the kind of this type of organic base can comprise one, two or trialkylamine, for example methylamine, dimethylamine and triethylamine; One, two or the trihydroxy alkylamine, for example one, two and triethanolamine; Aminoacid: for example arginine and lysine; Guanidine; The N-methylglucosamine; N-methyl glucoside amine; L-glutaminate; N methyl piperazine; Morpholine; Ethylenediamine; The N-benzyl-1-phenylethylamine; (trihydroxy methyl) aminoethane etc.Referring to, J.Pharm.Sci. for example, 66:1-19 (1977).
[0056] " patient ", " curee " or " host " by this method treatment can refer to people or non-human animal, for example primates, mammal and vertebrates.
[0057] term " prevention or therapeutic " treatment is well known in the art, comprises giving the host one or more these compositionss.If before the clinical manifestation of deleterious disease (for example disease of host animal or other unnecessary state), give this compositions, this treatment is preventative so, it is harmful disease development that it prevents the host, if but after harmful disease performance, gave this compositions, then treatment would be curative (being that book it will reduce, alleviates or stablize already present harmful disease or its side effect).
[0058] term " therapeutic agent ", " medicine ", " medicament " and " bioactive substance " are well known in the art, be included in biology, physiology or pharmaceutically active substance that patient or curee part or whole body work, with treatment disease or disease, for example molecule of degeneration of macula and other medicines.This term includes but not limited to its pharmaceutically acceptable salt and prodrug.This type of medicine can be tart, alkaline or salt; They can be neutral molecule, polar molecule or can be in conjunction with the molecular complex of hydrogen; They can be the prodrugs of the form such as ether, ester, amide of bioactivation when giving patient or curee.
[0059] phrase " treatment effective dose " is the recognized techniques term.In certain embodiments, this term is meant works as and the compounds time spent, may be used on any therapeutic treatment, produces the amount of the therapeutic agent of some required effect with rational interests/risk ratio.In certain embodiments, this term is meant the amount of other target that must or be enough to eliminate, reduce or keep (for example preventing diffusion) tumor or concrete therapeutic scheme.Effective dose can be according to this type of factor, constitutes the seriousness of thing (constructs), patient's body weight or disease or disease and decides as the disease of being treated or disease, the concrete targeting that gives.Those of ordinary skills can rule of thumb determine the effective dose of particular compound, need not do unsuitable experiment.In certain embodiments, the treatment effective dose with therapeutic agent in the body depends on multiple factor probably, and they comprise: the rate of release of medicine in chemical compound substrate, this speed depend in part on the chemistry and the physical characteristic of chemical compound; Medicinal property; Mode of administration and method; With any other material that except that medicine, mixes chemical compound substrate.
[0060] " radiosensitizer " is defined as when treating effective dose, promotes the disease or the treatment of conditions agent of one or more available electromagnetic radiation treatments of treatment.Generally speaking, radiosensitizer is decided to be with electromagnetic radiation and unites use, as the part of preventative or therapeutic treatment.The known suitable radiosensitizer that is used for the therapeutic alliance of this compositions of those skilled in the art.
[0061] " electromagnetic radiation " that is used for this description includes but not limited to have 10 -20The radiation that-10 metric waves are long.The specific embodiments of electromagnetic radiation is used γ-radiation (10 -20-10 -13M), the x-beta radiation (10 -11-10 -9M), ultraviolet (10nm-400nm), visible light (400nm-700nm), infra-red radiation (700nm-1.0mm) and microwave radiation (1mm-30cm).
[0062] phrase " whole body is cheated medicine ", " systematicness is cheated medicine ", " periphery is cheated medicine " and " administration on every side " are well known in the art, are included in the position for the treatment of the position of sickness influence away from being subjected to and give patient's compositions or other material.Directly medicine is given within the focus of the disease of treat, on or near, both made subsequently medicine understand the whole body distribution, can be described as " part ", " locality " or " zone " cheats medicine.
[0063] term " ED 50" be well known in the art.In certain embodiments, ED 50Expression produces its maximum reaction or acts on 50% drug dose, perhaps, produces the dosage of predetermined reaction in 50% experimenter or preparation.Term " LD 50" be well known in the art.In certain embodiments, LD 50Expression makes the lethal drug dose of 50% experimenter.Term " therapeutic index " is a term well known in the art, refers to the therapeutic index of medicine, is defined as LD 50/ ED 50
[0064] when with therapeutic agent and chemical compound, compositions for example disclosed herein is relevant when using, term " mixes " and " encapsulate " is well known in the art.In certain embodiments, these terms comprise mixing, preparing or comprise in addition this medicine mixed and make in the compositions that this medicine slowly discharges by required application purpose.These terms can comprise by its therapeutic agent or other material and mix any way in the chemical compound substrate, these chemical compound substrate for example comprise: chemical compound is a polymer, medicine is connected (by covalent bond or other binding interactions) with the monomer of this polymer, and this monomer is the polymeric part that produces polymer formulations, it is distributed in the whole polymeric matrix, attached to polymeric matrix surface (by covalent bond or other binding interactions), encapsulate etc. in polymeric matrix.Term " mixes " altogether or " encapsulate altogether " is meant in this compositions, and at least a other therapeutic agent or other material are mixed in therapeutic agent or other material.
[0065] more particularly, therapeutic agent or the physical form of other material encapsulate in chemical compound can be decided according to specific embodiment.For example, therapeutic agent or other material be encapsulate in micropill earlier, and this mode by the micropill structure that keeps at least a portion combines with described chemical compound then.Perhaps, therapeutic agent or other material are not enough to mix mutually in the controlled release chemical compound so that it is separated into droplet, but not is dissolved in chemical compound.The disclosure is conceived any type of encapsulate or is mixed, so that the therapeutic agent of any encapsulate or other material slowly discharge, whether encapsulate or any form decision encapsulated form of mixing enough can be accepted any concrete purposes.
[0066] term " bio-compatible plasticizer " is well known in the art, is included in solvable or dispersible material in the controlled release composition disclosed herein, and they increase the elasticity of chemical compound substrate, and are biocompatible in amount ranges.Know suitable plasticizer in the art, they are included in U.S. Patent number 2,784, those disclosed in 127 and 4,444,933.As an example, concrete plasticizer comprises: citric acid acetyl three n-butyl (about 20% weight or still less), acetyl trihexyl citrate (about 20% weight or still less), phthalic acid butyl ester benzyl ester, dibutyl phthalate, dioctyl phthalate, the positive bytyry three own esters of citric acid, dibenzoic diglycol laurate (about 20% weight or still less) etc.
[0067] " micromolecule " is term well known in the art.In certain embodiments, this term is meant molecular weight less than about 2000amu or less than about 1000amu, even less than the molecule of about 500amu.
[0068] term " alkyl " is well known in the art, comprises the radical of saturated aliphatic group, contains straight chained alkyl, branched alkyl, cycloalkanes (alicyclic ring) base, the cycloalkyl of alkyl replacement and the alkyl of cycloalkyl substituted.In certain embodiments, the straight or branched alkyl has about 30 carbon atoms (for example the straight chain of C1-C30, the side chain of C3-C30) still less on its skeleton, perhaps about 20 still less.Equally, cycloalkyl has about 10 carbon atoms of about 3-, 5,6 or 7 carbon of perhaps having an appointment in ring structure in their ring structure.
[0069] unless carbon number indicates in addition, and " low alkyl group " is meant the alkyl of above definition, but in its framing structure, has about 10 carbon of 1-, perhaps have about 6 carbon atoms of 1-.Equally, " low-grade alkenyl " has similar chain length with " low-grade alkynyl ".
[0070] term " aralkyl " is well known in the art, is meant the alkyl that is replaced by aryl (for example aryl or heteroaryl).
[0071] term " alkenyl " and " alkynyl " are well known in the art, are meant similar length and can be substituted by the unsaturated aliphatic group of abovementioned alkyl, but contain at least one two keys or triple bond respectively.
[0072] term " aryl " is well known in the art, be meant and comprise 0-4 heteroatomic 5 yuan, 6 yuan and 7 yuan of monocyclic aryl, for example benzene, naphthalene, anthracene, pyrene, pyrroles, furan, thiophene, imidazoles,  azoles, thiazole, triazole, pyrazoles, pyridine, pyrazine, pyridazine and pyrimidine etc.Have heteroatomic those aryl in the ring structure and also can be described as " fragrant heterocycle " or " heteroaryl ".Aromatic ring can be on one or more ring positions be replaced by above-mentioned this type of substituent group: halogen for example; azide; alkyl; aralkyl; alkenyl; alkynyl; cycloalkyl; hydroxyl; alkoxyl; amino; nitro; sulfydryl; imino group; acylamino-; phosphonate ester (phosphonate); phosphinate (phosphinate); carbonyl; carboxyl; silicyl; ether; alkylthio group; sulfonyl; sulfonamido (sulfonamido); ketone; aldehyde; ester; heterocyclic radical; aryl or heteroaryl moieties;-CF 3,-CN etc.Term " aryl " also comprises the multi-loop system that contains two or more rings, two shared two or more carbon of adjacent ring (these rings are " fused rings ") wherein, wherein at least one ring is an aromatics, and for example other ring can be cycloalkyl, cycloalkenyl group, cycloalkynyl radical, aryl and/or heterocyclic radical.
[0073] the term neighbour, and to being well known in the art, refer to 1 respectively, 2-, 1, the benzene that 3-and 1,4-replace.For example, 1,2-dimethylbenzene and ortho-xylene are synon.
[0074] term " heterocyclic radical ", " heteroaryl " or " heterocyclic group " are well known in the art, are meant 3 yuan-Yue 10 ring structures, and perhaps 3 yuan-Yue 7 yuan of rings, their ring structure comprises 1-4 hetero atom.Heterocycle also can be multi-ring.Heterocyclic radical comprises, thiophene for example, thianthrene, furan, pyrans, isobenzofuran, chromene, xanthene, phenol xanthene (phenoxanthene), the pyrroles, imidazoles, pyrazoles, isothiazole, different  azoles, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, iso-indoles, indole, indazole, purine, quinolizine, different quinolizine, quinoline, 2, the 3-benzodiazine, 1, the 5-benzodiazine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridines, acridine, pyrimidine, phenanthroline, azophenlyene, phenarsazine (phenarsazine), phenothiazine, furazan, fen  piperazine, pyrrolidine, tetrahydrofuran (oxolane), tiacyclopentane (thiolane), the  azoles, piperidines, piperazine, morpholine, lactone, lactams, for example azetidinone (azetidinones) and pyrrolinone (pyrrolidinones), sultam, sultone etc.Heterocycle can be in one or more positions be replaced by above-mentioned this type of substituent group, for example halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfydryl, imino group, acylamino-, phosphonate ester, phosphinate, carbonyl, carboxyl, silicyl, ether, alkylthio group, sulfonyl, ketone, aldehyde, ester, heterocyclic radical, aryl or heteroaryl moieties ,-CF 3,-CN etc.
[0075] term " multi-ring base " and " multi-ring group " are well known in the art, be meant two or more rings (for example cycloalkyl, cycloalkenyl group, cycloalkynyl radical, aryl and/or heterocyclic radical), two shared two or more carbon of adjacent ring wherein, for example these rings are " fused rings ".The ring that connects by non-adjacent atom is called " bridge " ring.Each ring in multi-ring can be replaced by above-mentioned this type of substituent group, for example halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfydryl, imino group, acylamino-, phosphonate ester, phosphinate, carbonyl, carboxyl, silicyl, ether, alkylthio group, sulfonyl, ketone, aldehyde, ester, heterocyclic radical, aryl or heteroaryl moieties ,-CF 3,-CN etc.
[0076] term " carbocyclic ring " is well known in the art, is meant that each atom in its medium ring is the aryl or the non-aryl rings of carbon.
[0077] term " nitro " is well known in the art, is meant-NO 2Term " halogen " is well known in the art, be meant-F ,-Cl ,-Br or-I; Term " sulfydryl " is well known in the art, is meant SH; Term " hydroxyl " expression-OH; Term " sulfonyl " is well known in the art, is meant SO 2 -The corresponding anion of " halogenide " expression halogen, and " pseudohalogen " has the definition that Cotton and Wilkinson's set forth in " Advanced Inorganic Chemistry " the 560th page.
[0078] term " amine " and " amino " are well known in the art, are meant amine unsubstituted and that replace, for example can be by the part of following general formula representative:
[0079] wherein R50, R51 and R52 independently represent hydrogen, alkyl, alkenyl, (CH separately 2) m-R61, or R50 and R51 and connected N atom are formed on the heterocycle that has 4-8 atom in the ring structure together; R61 represents aryl, cycloalkyl, cycloalkenyl group, heterocycle or multi-ring; M is 0 or the integer of 1-8.In other embodiments, R50 and R51 (with optional R52) independent separately represent hydrogen, alkyl, alkenyl or-(CH 2) m-R61.Therefore, term " alkylamine " comprises having amido connected replacement or the alkyl that unsubstituted definition is the same, and promptly at least one among R50 and the R51 is alkyl.
[0080] term " amide groups " is well known in the art, and being meant can be by the part of following general formula representative:
[0081] wherein R50 definition is the same, R54 represent hydrogen, alkyl, alkenyl or-(CH 2) m-R61, wherein m and R61 definition is the same.
[0082] term " acylamino-" is the carbonyl of amino well known in the art-replacement, and comprising can be by the part of following general formula representative:
Figure A20058001151300472
[0083] wherein R50 and R51 definition is the same.Some embodiment of amide of the present invention does not comprise the imidodicarbonic diamide of potentially unstable.
[0084] term " alkylthio group " is meant the same alkyl of definition with connected sulfenyl.In certain embodiments, " alkylthio group " part by the S alkyl ,-the S-alkenyl ,-the S-alkynyl and-S-(CH 2) mA representative the among-R61, wherein m and R61 definition is the same.Representative alkylthio group comprises methyl mercapto, ethylmercapto group etc.
[0085] term " carboxyl " is well known in the art, and comprising can be by this type of part of following general formula representative:
[0086] wherein X50 is a key, or represent oxygen or sulfur, R55 and R56 represent hydrogen, alkyl, alkenyl ,-(CH 2) m-R61 or pharmaceutically acceptable salt, R56 represent hydrogen, alkyl, alkenyl or-(CH 2) m-R61, wherein m and R61 definition is the same.When X50 is an oxygen, when R55 or R56 are not hydrogen, this formula representative " ester ".When X50 is an oxygen, and R55 defines when the same, and the part of indication is a carboxyl herein, especially when R55 is hydrogen, and this formula representative " carboxylic acid ".When X50 is an oxygen, and R56 is when being hydrogen, this formula representative " formic acid esters ".Generally speaking, when the oxygen atom of following formula is replaced by sulfur, this formula representative " thiol carbonyl (thiolcarbonyl) ".When X50 is a sulfur, and R55 or R56 be not when being not hydrogen, this formula representative " thiol ester (thiolester) ".When X50 is a sulfur, and R55 is when being hydrogen, this formula representative " thiol carboxylic acid (thiolcarboxylic acid) ".When X50 is a sulfur, when its R56 is hydrogen, this formula representative " thiol formic acid esters (thiolformate) ".On the other hand, when X50 is a key, and R55 is not when being not hydrogen, following formula representative " ketone " base.When X50 is a key, and R55 is when being hydrogen, following formula representative " aldehyde " base.
[0087] term " carbamoyl " is meant-O (C=O) NRR ', and wherein R and R ' independently are H, aliphatic group, aryl or heteroaryl.
[0088] term " oxo " be meant ketonic oxygen (=O).
[0089] term " oxime " and " oxime ether " are well known in the art, and being meant can be by the part of following general formula representative:
Figure A20058001151300481
[0090] wherein R75 be hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl or-(CH 2) m-R61.When R was H, part was an oxime; When R be alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl or-(CH 2) mDuring-R61, it is " an oxime ether ".
[0091] term " alkoxyl (alkoxyl) " or " alkoxyl (alkoxy) " are well known in the art, are meant the same alkyl of definition with connected oxygen base.Representative alkoxyl comprises methoxyl group, ethyoxyl, propoxyl group, uncle-butoxy etc." ether " is by two covalently bound hydrocarbon of oxygen.Therefore, cause alkyl ether to be or resemble the substituent group of the alkyl of alkoxyl, can by-O-alkyl ,-O-alkenyl, O-alkynyl ,-O-(CH 2) mA representative the among-R61, wherein m and R61 definition is the same.
[0092] term " sulphonic acid ester " is well known in the art, and being meant can be by the part of following general formula representative:
Figure A20058001151300491
[0093] wherein R57 is electron pair, hydrogen, alkyl, cycloalkyl or aryl.
[0094] term " sulfuric ester " is well known in the art, and comprising can be by the part of following general formula representative:
[0095] wherein the R57 definition is the same.
[0096] term " sulfonamido " is well known in the art, and comprising can be by the part of following general formula representative:
Figure A20058001151300493
[0097] wherein R50 and R56 definition is the same.
[0098] term " sulfamoyl " is well known in the art, and being meant can be by the part of following general formula representative:
[0099] wherein R50 and R51 definition is the same.
[0100] term " sulfonyl " is well known in the art, and being meant can be by the part of following general formula representative:
[0101] wherein R58 is one of following group: hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic radical, aryl or heteroaryl.
[0102] term " sulfoxide group (sulfoxido) " is well known in the art, and being meant can be by the part of following general formula representative:
[0103] wherein the R58 definition is the same.
[0104] term " phosphoryl " is well known in the art, can be represented by following formula usually:
[0105] wherein Q50 represents S or O, and R59 represents hydrogen, low alkyl group or aryl.When for example being used to replace alkyl, the phosphoryl of phosphoryl alkyl can be by following general formula representative:
Figure A20058001151300504
[0106] wherein independently Q50 and R59 definition is the same separately, and Q51 represents O, S or N.When Q50 was S, phosphoryl partly was " D2EHDTPA root (phosphorothioate) ".
[0107] term " phosphoramidate (phosphoramidite) " is well known in the art, can be represented by following general formula:
Figure A20058001151300511
[0108] wherein Q51, R50, R51 and R59 definition are the same.
[0109] term " amido phosphonate (phosphonamidite) " is well known in the art, can be represented by following general formula:
Figure A20058001151300512
[0110] wherein Q51, R50, R51 and R59 definition are the same, and R60 represents low alkyl group or aryl.
[0111] can similarly replace alkenyl and alkynyl, for example produce the alkenyl or the alkynyl of amino alkenyl, amino alkynyl, acylamino-alkenyl, acylamino-alkynyl, imino group alkenyl, imino group alkynyl, sulfo-alkenyl (thioalkenyls), sulfo-alkynyl (thioalkynyls), carbonyl-replacement.
[0112] when various expression, for example alkyl, m, n etc. occur in any structure when once above, plan to make its definition be independent of the definition in other place in same structure.
[0113] term " alkane seleno (selenoalkyl) " is well known in the art, is meant the alkyl of the seleno with connected replacement.Exemplary " selenide (selenoethers) " that can replace on alkyl be selected from-the Se-alkyl ,-the Se-alkenyl ,-the Se-alkynyl and-Se-(CH 2) mOne of-R61, m and R61 definition are the same.
[0114] term triflyl, tosyl, mesyl and nonaflyl are well known in the art, refer to trifyl, ptoluene-sulfonyl, mesyl and nine fluorine butane sulfonyls respectively.Term triflate, tosylate, mesylate and nonaflate are well known in the art, refer to triflate, right-tosylate, methanesulfonates and nine fluorine fourth sulphonic acid ester functional groups respectively and contain the molecule of described group.
[0115] abbreviation Me, Et, Ph, Tf, Nf, Ts and Ms difference represent methylidene, ethyl, phenyl, trifyl sulfonyl, nine fluorine fourth sulfonyls, ptoluene-sulfonyl and mesyl.The catalogue of more fully abridging that the common organic chemist in this area uses is published in the first phase of every volume Journal of Organic Chemistry; This catalogue is published in the table that title is Standard List of Abbreviations usually.
[0116] some contained chemical compound can exist with concrete geometry or stereoisomeric forms in any ratio in the present composition.In addition, polymer of the present invention also can be an optical activity.The present invention includes all these compounds that fall in the scope of the invention, comprise cis and transisomer, R-and S-enantiomer, diastereomer, (D)-isomer, (L)-isomer, its racemic mixture and other mixture thereof.Other asymmetric carbon atom can be present in substituent group, for example in the alkyl.All these type of isomers and composition thereof are all planned to be included in the present invention.
[0117] if for example need the concrete enantiomer of The compounds of this invention, can derive by asymmetric synthesis or by chiral auxiliary prepares it, and the non-enantiomer mixture that obtains is separated, and prothetic group is dissociated, and obtains the pure enantiomer that needs.Perhaps, to contain basic functionality for example amino when molecule, or acidic functionality forms diastereoisomeric salt with suitable optically active acid or alkali for example during carboxyl, split the diastereomer that so forms by fractional crystallization well known in the art or chromatography method subsequently, reclaim pure enantiomer subsequently.
Be appreciated that [0118] " replacement " or " being substituted " comprises that precondition in secret is, this replacement meets the atom of replacement and the valency of substituent permission, replace to produce stable chemical compound with this, for example for example conversion by rearrangement, cyclization, elimination or other reaction of its unautogenous experience.
[0119] substituent group of the organic compound of all permissions also planned to comprise in term " replacement ".In aspect widely, the substituent group of permission includes acyclic and ring-type, side chain or unbranched, carbocyclic ring and heterocycle, aromatics and the non-aromatic substituent of organic compounds.The illustrative substituent group comprises for example mentioned above those.The substituent group of permission can be a kind of and multiple identical or different substituent group of the organic compound that suits.In the present invention, hetero atom, for example nitrogen can have the hydrogen substituent group and/or the substituent group of any permission that meets the hetero atom valency of described organic compound herein.The present invention does not plan to be subjected to the restriction of substituent any way of the permission of organic compound.
[0120] 3. compositions
[0121] as mentioned above, can be present in the mode of the amount of the alltrans-retinal in the rod cell photoreceptors acromere dish, by disturbing visual cycle treatment or prevention degeneration of macula by minimizing.Can ignore the retinal toxicity chemical compound that produces by the awl cell, because rod cell is represented 95% of all photoreceptorss.
[0122] Fig. 1 explains mammiferous visual cycle.In the visual cycle process, be called the 11-cis-retinal of rhodopsin and opsin's a series of biochemical ladder of complex by causing by absorbing light.Each ladder of this circulation occurs in different places.According to Fig. 1 explanation, light absorption to the opsin dissociate and the initial ladder of the formation of alltrans-retinal occurs in the dish of bar photoreceptor cell acromere.Alltrans-retinal is reduced to alltrans-retinol and occurs in the Cytoplasm of rod cell, and all the other ladders of regeneration 11-cis-retinol occur in the retinal pigment epithelium (RPE).
[0123] designs at least two kinds and main prevent the method that alltrans-retinal is accumulated in dish.In one approach, can suppress one or more enzymatic ladders or the chaperone in visual cycle in conjunction with ladder, to block the approach of synthetic alltrans-retinal.In another approach, part visual cycle " short circuit ", promptly the early stage intermediate in the circulation splits into stagewise intermediate of two or more later stages in the visual cycle, so that these circulation ladders are shunted, and alltrans-retinal precursor is not in dish simultaneously.
[0124] A. enzyme inhibitor
[0125] can realize the retinoid flux of restricted passage visual cycle by any crucial biochemical reaction that suppresses visual cycle.By this mode, each ladder of circulation has potential specificity (addressable).Therefore, available inhibition enzymatic ladder " stops " visual cycle among the RPE, thereby alltrans-retinal is discharged from dish.
[0126] also tends to suppress other ladder of visual cycle.For example, as shown in Figure 1, when alltrans-retinal and derivant thereof are returned RPE, comprise that several enzymes of LRAT (lecithin retinol acyltransferase), 11-cis-retinol dehydrogenase and IMH (isomery hydrolytic enzyme) act on them.In addition, chaperone RPE65 combines with retinyl ester, those typical hydrophobic compounds can be utilized, to be processed as 11-cis-retinol by IMH.These enzymes and chaperone can become inhibition and/or interferential target.
[0127] in certain embodiments; the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), the inhibitor of 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT), or the antagonist of chaperone retinal pigment epithelium (RPE65) has the structure of formula I representative:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X is-O-,-N (R a)-,-C (R b) p-or-S-;
Z be alkyl, haloalkyl ,-(CH 2CH 2O) pR bOr-C (=O) R b
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl or haloalkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0128] in certain embodiments, the antagonist of the inhibitor of the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT) or chaperone retinal pigment epithelium (RPE65) has the structure of formula II representative:
Figure A20058001151300551
Wherein n is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) p-;
Z does not exist or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-(CH 2CH 2O) pR b,
-C(＝O)R b，-C(＝O)CH 2F，-C(＝O)CHF 2，-C(＝O)CF 3，-C(＝O)CHN 2，-C(＝O)OR b，
-C (=O) CH 2OC (=O) R b,-C (=O) C (=C (R b) 2) R b, Or
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0129] in certain embodiments, the antagonist of the inhibitor of the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT) or chaperone retinal pigment epithelium (RPE65) has the structure of formula III representative:
Figure A20058001151300561
Wherein
N is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-CR b(OR b)-,-CR b(N (R a) 2)-,-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) p-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-(CH 2CH 2O) pR b,-C (=O) R b,-C (=NR a) R b,-C (=NOR b) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
 represents singly-bound or trans double bond.
[0130] in certain embodiments, the antagonist of the inhibitor of the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT) or chaperone retinal pigment epithelium (RPE65) has the structure of formula VI representative:
Wherein, the implication of following group when at every turn occurring is independently,
R 1Be hydrogen, alkyl, aryl and aralkyl;
X be alkyl, alkenyl ,-C (R b) 2-,-C (=O)-,-C (=NR a)-,-C (OH) R bOr-C (N (R a) 2) R b-;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bIt is hydrogen or alkyl.
[0131] in certain embodiments; the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), the inhibitor of 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT), or the antagonist of chaperone retinal pigment epithelium (RPE65) has the structure of formula I representative:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X is-O-,-N (R a)-,-C (R b) p-or-S-;
Z be alkyl, haloalkyl ,-(CH 2CH 2O) pR bOr-C (=O) R b
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl or haloalkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0132] in certain embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id representative:
Figure A20058001151300581
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, the impure aromatic ene base, the hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino (acylamino), acylamino-(amido), alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical (sulfonate), sulfate radical (sulfate), sulfonamido, sulfamoyl (sulfamoyl), sulfonyl and sulfoxide group (sulfoxido);
R 4Do not exist, or hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
Y is-C (R b) 2-or-C (=O)-;
X is-O-,-N (R a)-,-C (R b) 2-or-S-;
Z be alkyl, haloalkyl or-C (=O) R b
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl or haloalkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0133] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, wherein R 1It is methyl.
[0134] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein n is 0.
[0135] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein n is 1.
[0136] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein Y is-CH 2-.
[0137] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein X is-O-.
[0138] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, wherein X be-N (H)-.
[0139] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein Z is-C (=O) R b
[0140] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein Z is-C (=O) R bR bIt is haloalkyl.
[0141] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein Z is an alkyl.
[0142] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, and wherein Z is a haloalkyl.
[0143] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, wherein R 3Be hydrogen.
[0144] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ia, Ib, Ic or Id, wherein R 4Be hydrogen, methyl or do not exist.
[0145] in certain embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih representative:
Figure A20058001151300601
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
X is-O-,-N (Ra)-,-C (R b) 2-or-S-;
Z be alkyl, haloalkyl or-C (=O) R b
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bBe hydrogen, alkyl or haloalkyl.
[0146] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein n is 0.
[0147] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein n is 1.
[0148] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein X is-O-.
[0149] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, wherein X be-N (H)-.
[0150] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein Z is-C (=O) R b
[0151] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein Z is-C (=O) R bR bIt is haloalkyl.
[0152] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein Z is an alkyl.
[0153] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein Z is a haloalkyl.
[0154] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, wherein R 3Be hydrogen.
[0155] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein X is-O-; Z is an alkyl.
[0156] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, and wherein X is-O-; Z is a haloalkyl.
[0157] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, wherein X be-N (H)-; Z is an alkyl.
[0158] in other embodiments, the inhibitor of isomery hydrolytic enzyme (IMH) has the structure of formula Ie, If, Ig or Ih, wherein X be-N (H)-; Z is a haloalkyl.
[0159] in one embodiment, the inhibitor of isomery hydrolytic enzyme (IMH) is bromoacetic acid 11-cis-retinyl ester (cBRA):
Figure A20058001151300611
[0160] in certain embodiments; the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), the inhibitor of 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT), or the antagonist of chaperone retinal pigment epithelium (RPE65) has the structure of formula II representative:
Figure A20058001151300621
Wherein
N is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) p-;
Z does not exist, or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-(CH 2CH 2O) pR b,
-C(＝O)R b，-C(＝O)CH 2F，-C(＝O)CHF 2，-C(＝O)CF 3，-C(＝O)CHN 2，-C(＝O)OR b，
-C (=O) CH 2OC (=O) R b,-C (=O) C (=C (R b) 2) R b, Or
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0161] in certain embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
R 4Do not exist or hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
Y is-C (=O)-or-C (R b) 2-;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) 2-;
Z does not exist or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-C (=O) R b,-C (=O) CH 2F ,-C (=O) CHF 2,-C (=O) CF 3,-C (=O) CHN 2,-C (=O) CH 2OC (=O) R b,-C (=O) OR b,
-C (=O) C (=C (R b) 2) R b,
Figure A20058001151300641
Or
Figure A20058001151300642
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0162] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein n is 0.
[0163] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein n is 1.
[0164] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, wherein R 1Be hydrogen or methyl.
[0165] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, wherein R 3Be hydrogen.
[0166] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, wherein R 4Be hydrogen or methyl.
[0167] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein Y is-CH 2-
[0168] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein X is-O-.
[0169] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein X is-NH-.
[0170] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein X is-C (R b) 2-.
[0171] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, wherein X be-C (=O)-.
[0172] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein Z is an alkyl.
[0173] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIa, IIb, IIc or IId representative, and wherein Z is a haloalkyl.
[0174] in certain embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative,
Figure A20058001151300651
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) 2-;
Z does not exist or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-C (=O) R b,-C (=O) CH 2F ,-C (=O) CHF 2,-C (=O) CF 3,-C (=O) CHN 2,-C (=O) CH 2OC (=O) R b,
-C (=O) OR b,-C (=O) C (=C (R b) 2) R b, Or
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl.
[0175] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein n is 0.
[0176] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein n is 1.
[0177] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, wherein R 1Be hydrogen or methyl.
[0178] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, wherein R 3Be hydrogen.
[0179] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, wherein R 4Be hydrogen or methyl.
[0180] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein X is-O-.
[0181] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein X is-NH-.
[0182] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein X is-CH 2-.
[0183] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, wherein X be-C (=O)-.
[0184] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein Z is an alkyl.
[0185] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein Z is a haloalkyl.
[0186] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein Z is-C (=O) R b
[0187] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein X is-O-; Z is-C (=O) R b
[0188] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein X is-CH 2-; Z is-C (=O) R b
[0189] in other embodiments, the inhibitor of lecithin retinol acyltransferase (LRAT) has the structure of formula IIe, IIf, IIg or IIh representative, and wherein X is-NH-; Z is-C (=O) R b
[0190] in one embodiment, the inhibitor of lecithin retinol acyltransferase (LRAT) is a 13-demethyl-13,14-dihydro-alltrans-retinyl (retunyl) trifluoro-acetate (RFA):
Figure A20058001151300671
[0191] in one embodiment, the inhibitor of lecithin retinol acyltransferase (LRAT) is alpha-brominated acetic acid alltrans-retinyl ester.
[0192] in certain embodiments, the antagonist of the inhibitor of the inhibitor of the inhibitor of isomery hydrolytic enzyme (IMH), 11-cis-retinol dehydrogenase, lecithin retinol acyltransferase (LRAT) or chaperone retinal pigment epithelium (RPE65) has the structure of formula III representative:
Figure A20058001151300672
Wherein n is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-CR b(OR b)-,-CR b(N (R a) 2)-,-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) p-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-(CH 2CH 2O) pR b,-C (=O) R b,-C (=NR a) R b,-C (=NOR b) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
 represents singly-bound or trans double bond.
[0193] in certain embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative:
Figure A20058001151300681
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
Y is-C (=O)-,-CR b(OR b)-,-CR b(N (R a) 2)-or-C (R b) 2-;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) 2-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-C (=O) R b,-C (=NR a) R b,-C (=NOH) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl;
P is 0-10; With
 represents singly-bound or trans double bond.
[0194] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein n is 0.
[0195] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein n is 1.
[0196] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, wherein R 1Be hydrogen or methyl.
[0197] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, wherein R 3Be hydrogen.
[0198] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, wherein R 4Be hydrogen or methyl.
[0199] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein X is-O-.
[0200] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein X is-NH-.
[0201] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein X is-C (R b) 2-.
[0202] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, wherein X be-C (=O)-.
[0203] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein Z is an alkyl.
[0204] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III a, IIIb, IIIc or IIId representative, and wherein Z is a haloalkyl.
[0205] in certain embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) 2-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-C (=O) R b,-C (=NR a) R b,-C (=NOH) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
P is 0-10.
[0206] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein n is 0.
[0207] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein n is 1.
[0208] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, wherein R 1Be hydrogen or methyl.
[0209] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, wherein Y be-C (=O)-.
[0210] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein Y is-CH 2-.
[0211] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein Z is-C (=O) R b
[0212] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein Z is-CH (OH) R b-.
[0213] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein Z is CH (NH) R b
[0214] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein Z is an alkyl.
[0215] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula III e, IIIf, IIIg or IIIh representative, and wherein Z is a haloalkyl.
[0216] in-individual embodiment, retinal pigment epithelium (RPE65) antagonist is 13-cis-tretinoin (isoretinoin, ACCUTANE ):
[0217] in certain embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative:
Figure A20058001151300712
Wherein, the implication of following group when at every turn occurring is independently,
N is 1,2,3 or 4;
Y is-C (R b) 2-or-C (=O)-;
X is-O-,-NR a-,-C (R b) 2-or-C (=O)-;
Z is-C (=O) R b,-OR b,-N (R b) 2, alkyl or haloalkyl;
R aBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl.
[0218] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Y is-CH 2-.
[0219] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein X is-O-.
[0220] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Z is-C (=O) R bR bIt is alkyl.
[0221] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Z is an alkyl.
[0222] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Y is-CH 2-; X is-O-; Z is-C (=O) R bR bIt is alkyl.
[0223] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Y is-CH 2-; X is-O-; Z is an alkyl.
[0224] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Y is-CH 2-; X is-C (=O)-; Z is an alkyl.
[0225] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula IV representative, and wherein Y is-CH 2-; X is-C (=O)-; Z is-N (R b) 2R bIt is alkyl.
[0226] in one embodiment, retinal pigment epithelium (RPE65) antagonist is Palmic acid geraniol ester (K D=301nM):
[0227] in one embodiment, retinal pigment epithelium (RPE65) antagonist is Palmic acid farnesyl bromide (K D=63nM)
Figure A20058001151300722
[0228] in one embodiment, retinal pigment epithelium (RPE65) antagonist is Palmic acid geranyl geraniol ester (K D=213nM):
[0229] in one embodiment, retinal pigment epithelium (RPE65) antagonist is geranyl palmityl ether (K D=416nM):
[0230] in one embodiment, retinal pigment epithelium (RPE65) antagonist is farnesyl-palmityl ether (K D=60nM):
[0231] in one embodiment, retinal pigment epithelium (RPE65) antagonist is geranyl geranyl palmityl ether (K D=195nM):
[0232] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound:
[0233] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (K D=96nM):
[0234] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound:
Figure A20058001151300737
[0235] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (K D=56nM):
Figure A20058001151300741
[0236] in one embodiment, retinal pigment epithelium (RPE65) antagonist is the farnesyl-octanone:
Figure A20058001151300742
[0237] in one embodiment, retinal pigment epithelium (RPE65) antagonist is an octyl group method acid imide (farnesimide):
Figure A20058001151300743
[0238] in one embodiment, retinal pigment epithelium (RPE65) antagonist is a palmityl method acid imide:
[0239] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (K D=56nM):
Figure A20058001151300745
[0240] in certain embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative:
Figure A20058001151300746
Wherein, the implication of following group when at every turn occurring is independently,
N is 1,2 or 3;
Y is-C (R b) 2-,-C (=O)-or-CH (OH)-;
X is-O-,-NR a-or-C (R b) 2-;
Z is-C (=O) R b, hydrogen ,-(CH 2CH 2O) pR b, alkyl or haloalkyl;
R aBe hydrogen, alkyl, haloalkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
P is 1-10.
[0241] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein Y is-CH 2-.
[0242] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, wherein Y be-C (=O)-.
[0243] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, wherein Y be-CH (OH)-.
[0244] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein X is-O-.
[0245] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein X is-NR a-.
[0246] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein X is-C (R b)-.
[0247] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein Z is an alkyl.
[0248] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein Z is-C (=O) R bR bIt is alkyl.
[0249] in other embodiments, retinal pigment epithelium (RPE65) antagonist has the structure of formula V representative, and wherein Z is-(CH 2CH 2O) pR bAnd R bIt is alkyl.
[0250] in one embodiment, retinal pigment epithelium (RPE65) antagonist is β-ionoacetyl cetylate (K D=153nM):
Figure A20058001151300751
[0251] in one embodiment, retinal pigment epithelium (RPE65) antagonist is β-ionoacetyl palmityl ether (K D=156nM):
[0252] in one embodiment, retinal pigment epithelium (RPE65) antagonist is Palmic acid retinyl ester (4a; K D=47nM):
[0253] in one embodiment, retinal pigment epithelium (RPE65) antagonist is caproic acid retinyl ester (4b; K D=235nM):
Figure A20058001151300763
[0254] in one embodiment, retinal pigment epithelium (RPE65) antagonist is the valeric acid retinyl ester:
[0255] in one embodiment, retinal pigment epithelium (RPE65) antagonist is acetic acid retinyl ester (4c; K D=1,300nM):
Figure A20058001151300765
[0256] in one embodiment, retinal pigment epithelium (RPE65) antagonist is palmityl retinyl ether (4d, K D=25nM):
Figure A20058001151300766
[0257] in one embodiment, retinal pigment epithelium (RPE65) antagonist is hexyl retinyl ether (K D=151nM):
[0258] in one embodiment, retinal pigment epithelium (RPE65) antagonist is methyl retinyl ether (K D=24nM):
[0259] in one embodiment, retinal pigment epithelium (RPE65) antagonist is retinyl [2-(2 '-methoxyl group) ethyoxyl] ethylether (K D=486nM):
Figure A20058001151300773
[0260] in one embodiment, retinal pigment epithelium (RPE65) antagonist is:
[0261] in one embodiment, retinal pigment epithelium (RPE65) antagonist is that the N-palmityl is looked yellow acid imide (retinimide) (K D=40nM):
Figure A20058001151300775
[0262] in one embodiment, retinal pigment epithelium (RPE65) antagonist is N, and the N-dimethyl is looked yellow acid imide (K D=3,577nM):
Figure A20058001151300776
[0263] in one embodiment, retinal pigment epithelium (RPE65) antagonist is that the N-tert-butyl group is looked yellow acid imide (K D=4,321nM):
Figure A20058001151300781
[0264] in one embodiment, retinal pigment epithelium (RPE65) antagonist is palmityl retinol (K D=170nM):
Figure A20058001151300782
[0265] in one embodiment, retinal pigment epithelium (RPE65) antagonist is the methyl retinol:
[0266] in one embodiment, retinal pigment epithelium (RPE65) antagonist is that palmityl is looked flavone (K D=64nM):
[0267] in one embodiment, retinal pigment epithelium (RPE65) antagonist is the retinyl decanone:
Figure A20058001151300785
[0268] in one embodiment, retinal pigment epithelium (RPE65) antagonist is that methyl is looked flavone (K D=3,786nM):
[0269] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (4e):
[0270] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (4f; K D=64nM)
[0271] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (K D=173nM):
Figure A20058001151300793
[0272] in one embodiment, retinal pigment epithelium (RPE65) antagonist is following chemical compound (K D=3,786nM):
[0273] in one embodiment, retinal pigment epithelium (RPE65) antagonist is:
[0274] in one embodiment, retinal pigment epithelium (RPE65) antagonist is:
[0275] in one embodiment, retinal pigment epithelium (RPE65) antagonist is:
Figure A20058001151300801
[0276] above-mentioned RPE65 agonist compounds and general formula compound, their various substituent group definition and other embodiments also are the LRAT inhibitor, the LRAT inhibitor is attached to herein by reference.
[0277] other RPE65 antagonist and LRAT inhibitor comprise the medicine that suppresses palmitoylation.For example, 2-bromine cetylate suppresses palmitoylation.In certain embodiments, the racemic mixture of 2-bromine cetylate also can be used for suppressing LRAT and/or antagonism RPE65.In other embodiments, also available pure (R)-2-bromine Palmic acid suppresses LRAT and/or antagonism RPE65.In other other embodiment, pure (S)-2-bromine Palmic acid can be used for suppressing LRAT and/or antagonism RPE65.
[0278] in certain embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VI representative:
Figure A20058001151300802
Wherein, the implication of following group when at every turn occurring is independently,
R 1Be hydrogen, alkyl, aryl or aralkyl;
X be alkyl, alkenyl ,-C (R b) 2-,-C (=O)-,-C (=NR a)-,-C (OH) R bOr-C (N (R a) 2) R b-;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bIt is hydrogen or alkyl.
[0279] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VI representative, wherein R 1Be hydrogen.
[0280] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VI representative, and wherein X is-C (R b) 2-.
[0281] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VI representative, wherein X be-C (=O)-.
[0282] in certain embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIa or VIb representative:
Figure A20058001151300811
Wherein, the implication of following group when at every turn occurring is independently,
R 1Be hydrogen, alkyl, aryl or aralkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R 3It is hydrogen or alkyl;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bIt is hydrogen or alkyl; With
 represents singly-bound, cis-double bonds or trans double bond.
[0283] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIa or VIb representative, wherein R 1Be hydrogen.
[0284] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIa or VIb representative, wherein R 2It is alkyl.
[0285] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIa or VIb representative, wherein R 3Be hydrogen or methyl.
[0286] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIc, VId or VIe representative:
Figure A20058001151300821
Wherein, the implication of following group when at every turn occurring is independently,
N is 1-5;
M is 0-30;
R 1Be hydrogen, alkyl, aryl or aralkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R 3It is hydrogen or alkyl;
R 4Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bIt is hydrogen or alkyl.
[0287] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIc representative, wherein R 1Be hydrogen.
[0288] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIc representative, wherein R 4Be hydrogen.
[0289] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIc representative, wherein R 1Be hydrogen; And R 4Be hydrogen.
[0290] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VId representative, and wherein n is 1,2 or 3.
[0291] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VId representative, wherein R 3It is methyl.
[0292] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VId representative, wherein R 1Be hydrogen.
[0293] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VId representative, and wherein n is 1,2 or 3; R 3It is methyl.
[0294] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VId representative, and wherein n is 1,2 or 3; R 3It is methyl; R 1Be hydrogen.
[0295] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIe representative, wherein R 1Be hydrogen.
[0296] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIe representative, and wherein m is 1-10.
[0297] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIe representative, and wherein m is 11-20.
[0298] in other embodiments, 11-cis-retinol dehydrogenase inhibitor has the structure of formula VIe representative, and wherein m is 11-20; And R 1Be hydrogen.
[0299] except other method, also can generate the 11-cis-retinol dehydrogenase inhibitor that has by the structure of formula VIe representative by the several data storehouse method shown in the flow process 1, among other ways:
Figure A20058001151300831
Flow process 1
[0300] in one embodiment, 11-cis-retinol dehydrogenase inhibitor is 13-cis-tretinoin (isoretinoin, ACCUTANE ):
Figure A20058001151300832
[0301] also comprises the pharmaceutically acceptable addition salt and the complex of the chemical compound that following formula provides.Can contain therein in the situation of chemical compound of one or more chiral centres, except that regulation was arranged, Fa Ming chemical compound can be the racemic mixture of single stereoisomers or stereoisomer herein.Also comprise its prodrug, analog and derivant.
[0302] in certain embodiments, can be with two or more enzyme inhibitors and/or the associating of RPE65 binding inhibitors.In certain embodiments, can be with enzyme inhibitor and/or RPE65 binding inhibitors and the associating of short-circuit cycle (short-circuiting) chemical compound.Can select these to unite sequence ladder (promptly a step is right after two steps of another step generation) to suppress visual cycle.
[0303] in certain embodiments, isomery hydrolytic enzyme (IMH) inhibitor can be the chemical compound with structure of general formula 1 representative:
Wherein, the implication of following group when at every turn occurring is independently:
R, R 1, R 2And R 3Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl or heteroarylalkyl;
W and Y are O, NR, R or S;
X is H, alkyl, haloalkyl, aryl or halogenide;
M and n are the integers of 1-6; With
P is the integer of 0-6.
[0304] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be H or Me.
[0305] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein m is 2.
[0306] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein n is 2.
[0307] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein W is O.
[0308] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein W is C.
[0309] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein Y is O.
[0310] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein p is 1.
[0311] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein X is Br.
[0312] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be that H or Me and m are 2.
[0313] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be H or Me, m is 2, and n is 2.
[0314] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be H or Me, m is 2, and n is 2, and W is O.
[0315] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be H or Me, m is 2, and n is 2, and W is that O and Y are O.
[0316] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be H or Me, m is 2, and n is 2, and W is O, and Y is O, and p is 1.
[0317] in another embodiment, the definition that the IMH inhibitor has formula 1 structure and follows, wherein R 2And R 3Be H or Me, m is 2, and n is 2, and W is O, and Y is O, and p is 1, and X is Br.
[0318] in one embodiment, the isomery hydrolase inhibitor is bromoacetic acid 11-cis-retinyl ester (cRBA):
Figure A20058001151300861
[0319] in certain embodiments, the 1MH inhibitor can be a formula 8a chemical compound
Figure A20058001151300862
Wherein, the implication of following group when at every turn occurring is independently:
X is O, S, NR ', CH 2Or NHNR ';
Z is O or NOH;
R 1Be-CH 2F ,-CHF 2,-CF 3,-CH 2N 2,-CH 2C (O) OR ,-OR ' ,-C (O) CHR ' ,-C (NH) CHR ' or-CH=CHR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Figure A20058001151300863
Or
R  is CH 3Or H; With
N is 0,1 or 2;
Wherein  represents singly-bound, cis-double bonds or trans double bond.
[0320] can think that formula 8a chemical compound is irreversible IMH inhibitor because they can with the IMH covalent bond, make its permanent failure.
[0321] in certain embodiments, the IMH inhibitor can be a formula 8a chemical compound, and wherein Z is O.
[0322] in certain embodiments, the IMH inhibitor can be a formula 8b chemical compound,
Figure A20058001151300871
Wherein, the implication of following group when at every turn occurring is independently:
Y is C=O, C=S, C=NR ' or CH 2
R 1Be R ' ,-OR ' or-CN;
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is Or
R " is CH 3Or H; With
N is 0,1 or 2;
Wherein  represents singly-bound, cis-double bonds or trans double bond.
[0323] can think that formula 8b chemical compound is reversible IMH inhibitor,, and not make its permanent inefficacy because they can combine with IMH is non-covalent.
[0324] in certain embodiments, the IMH inhibitor can be a formula 8c chemical compound:
Wherein, the implication of following group when at every turn occurring is independently:
X is O, S, NR ', CH 2Or NHNR ';
Z is O or NOH;
R 1Be-CH 2F ,-CHF 2,-CF 3,-CH 2N 2,-CH 2C (O) OR ,-OR ' ,-C (O) CHR ,-C (NH) CHR ' or-CH=CHR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Or
R  is CH 3Or H; With
N is 0,1 or 2.
[0325] can think that formula 8c chemical compound is irreversible IMH inhibitor because they can with the IMH covalent bond, make its permanent failure.
[0326] in certain embodiments, the IMH inhibitor can be a formula 8c chemical compound, and wherein Z is O.
[0327] in certain embodiments, the IMH inhibitor can be a formula 8d chemical compound,
Wherein, the implication of following group when at every turn occurring is independently:
Y is C=O, C=S, C=NR ' or CH 2
R 1Be R ' ,-OR ' or-CN;
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Figure A20058001151300885
Or
R  is CH 3Or H; With
N is 0,1 or 2.
[0328] can think that formula 8d chemical compound is reversible IMH inhibitor,, and not make its permanent inefficacy because they can combine with IMH is non-covalent.
[0329] in certain embodiments, the LRAT inhibitor can be the chemical compound with structure of formula 2 representatives:
Figure A20058001151300891
Wherein, the implication of following group when at every turn occurring is independently:
R, R 1, R 2And R 3Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl or heteroarylalkyl;
W and Y are O, NR, R or S;
X is H, alkyl, haloalkyl or aryl;
M and n are the integers of 1-6; With
P is the integer of 0-6.
[0330] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following, wherein R 2And R 3Be H or Me.
[0331] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein m is 3.
[0332] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein n is 1.
[0333] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein W is O.
[0334] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein W is C.
[0335] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein Y is O.
[0336] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein p is 0.
[0337] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following thereof, and wherein X is OCF 3
[0338] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following, wherein R 2And R 3Be H or Me, m is 3.
[0339] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following, wherein R 2And R 3Be H or Me, m is 3, and n is 1.
[0340] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following, wherein R 2And R 3Be H or Me, m is 3, and n is 1, and W is O.
[0341] in another embodiment, the LRAT inhibitor has the structure of formula 2 and the definition of following, wherein R 2And R 3Be H or Me, m is 3, and n is 1, and W is that O and Y are O.
[0342] in another embodiment, the LRAT inhibitor has formula structure 2 and definition, wherein R 2And R 3Be H or Me, m is 3, and n is 1, and W is O, and Y is O, and p is 0.
[0343] in another embodiment, the LRAT inhibitor has formula structure 2 and definition, wherein R 2And R 3Be H or Me, m is 3, and n is 1, and W is O, and Y is O, and p is 0, and X is OCF 3
[0344] exemplary L RAT inhibitor is alpha bromoisobutyric acid alltrans-retinyl ester.Another exemplary L RAT inhibitor is a trifluoroacetic acid 13-demethyl-13,14-dihydro-alltrans-retinyl ester (RFA):
[0345] in some embodiments, disturbing the bonded chemical compound of RPE65 can be the chemical compound with structure of formula 3 representatives
Wherein, the implication of following group when at every turn occurring is independently:
R and R 1Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl or heteroarylalkyl;
R 2Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or-CO 2R;
R 3Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or-CH 2OR 4
R 4Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic radical; With
M is the integer of 1-6.
[0346] in another embodiment, the LRAT inhibitor has the structure of formula 3 and the definition of following, wherein R 2Be H, Me or-CO 2H.
[0347] in another embodiment, the LRAT inhibitor has the structure of formula 3 and the definition of following thereof, and wherein m is 4.
[0348] in another embodiment, the LRAT inhibitor has the structure of formula 3 and the definition of following, wherein R 3Be H.
[0349] in another embodiment, the LRAT inhibitor has the structure of formula 3 and the definition of following, wherein R 2Be H, Me or-CO 2H, m are 4.
[0350] in another embodiment, the LRAT inhibitor has the structure of formula 3 and the definition of following, wherein R 2Be H, Me or-CO 2H, m are 4, R 3Be H.
[0351] in certain embodiments, the LRAT inhibitor can be a formula 6a chemical compound:
Wherein, the implication of following group when at every turn occurring is independently:
X is O, S, NR ', CH 2Or NHNR ';
Z is O or NOH;
R 1Be-CH 2F ,-CHF 2,-CF 3,-CH 2N 2, CH 2C (O) OR ,-OR ' ,-C (O) CHR ' ,-C (NH) CHR ' or-CH=CHR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is Or
Figure A20058001151300923
With
N is 1,2 or 3;
Wherein  represents singly-bound, cis-double bonds or trans double bond.
[0352] can think that formula 6a chemical compound is irreversible LRAT inhibitor because they can with the LRAT covalent bond, make its permanent failure.
[0353] in certain embodiments, the LRAT inhibitor can be a formula 6a chemical compound, and wherein Z is O.
[0354] in certain embodiments, the LRAT inhibitor can be a formula 6b chemical compound,
Figure A20058001151300924
Wherein, the implication of following group when at every turn occurring is independently:
Y is C=O, C=S, C=NR ' or CH 2
R 1Be R ' ,-OR ' or-CN;
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is Or
With
N is 1,2 or 3;
Wherein  represents singly-bound, cis-double bonds or trans double bond.
[0355] can think that formula 6c chemical compound is reversible LRAT inhibitor,, and not make its permanent failure because they can combine with LRAT is non-covalent.
[0356] in certain embodiments, the LRAT inhibitor can be a formula 6c chemical compound:
The implication of wherein following group when at every turn occurring is independently:
X is O, S, NR ', CH 2Or NHNR ';
Z is O or NOH;
R 1Be-CH 2F ,-CHF 2,-CF 3,-CH 2N 2,-CH 2C (O) OR ,-OR ' ,-C (O) CHR ' ,-C (NH) CHP ' or-CH=CHR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Figure A20058001151300934
Or
Figure A20058001151300935
With
N is 1,2 or 3.
[0357] can think that formula 6c chemical compound is irreversible LRAT inhibitor because they can with the LRAT covalent bond, make its permanent failure.
[0358] in certain embodiments, the LRAT inhibitor can be a formula 6c chemical compound, and wherein Z is O.
[0359] in certain embodiments, the LRAT inhibitor can be a formula 6d chemical compound:
Figure A20058001151300941
Wherein, the implication of following group when at every turn occurring is independently:
Y is C=O, C=S, C=NR ' or CH 2
R 1Be R ' ,-OR ' or-CN;
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Figure A20058001151300942
Or
With
N is 1,2 or 3.
[0360] can think that formula 6d chemical compound is reversible LRAT inhibitor,, and not make its permanent failure because they can combine with LRAT is non-covalent.
[0361] disturbing an exemplary of the bonded chemical compound of RPE65 is 13-cis-tretinoin (isotretinoin, ACCUTANE ):
[0362] 13-cis-tretinoin is converted into alltrans-tretinoin in vivo, and it is the highly efficient depressor of RPE65 function.
[0363] in certain embodiments, the RPE65 antagonist is the chemical compound with structure of formula 4 representatives:
Figure A20058001151300945
Wherein, the implication of following group when at every turn occurring is independently:
R, R 1, R 2Be H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, alkoxyl, aryloxy group, amino, halo, hydroxyl or carboxyl;
R 3Be alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or ether;
L is H, OH, NH 2, N (R) 2, alkoxyl, aryloxy group, halo, hydroxyl, carboxyl, or two L link together and represent O, S or NR;
X is C (R) 2, O, S or NR; With
M is the integer of 1-6.
[0364] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O.
[0365] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is CH 2
[0366] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is NH.
[0367] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein two L link together and represent O.
[0368] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein two L link together and represent NOH.
[0369] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein L is H, OH or NH 2
[0370] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein each L is H.
[0371] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein m is 4.
[0372] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein m is 3.
[0373] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following, wherein R 2Be H or methyl.
[0374] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following, wherein R 3It is alkyl.
[0375] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following, wherein R 3Be ether.
[0376] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and two L link together and represent O.
[0377] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and each L is H.
[0378] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is NH, and two L link together and represent O.
[0379] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is CH 2, two L link together and represent O.
[0380] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is CH 2, two L link together and represent NOH.
[0381] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and two L link together and represent O, R 2Be H or methyl, m is 4, R 3It is the C15 alkyl.
[0382] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and two L link together and represent O, R 2Be H or methyl, m is 4, R 3It is the C5 alkyl.
[0383] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and two L link together and represent O, R 2Be H or methyl, m is 4, R 3It is methyl.
[0384] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and each L is H, R 2Be H or methyl, m is 4, R 3It is the C15 alkyl.
[0385] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is NH, and two L link together and represent O, R 2Be H or methyl, m is 4, R 3It is the C15 alkyl.
[0386] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is CH 2, two L link together and represent O, R 2Be H or methyl, m is 4, R 3It is the C15 alkyl.
[0387] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and each L is H, R 2Be H or methyl, m is 4, R 3Be ether.
[0388] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is O, and each L is H, R 2Be H or methyl, m is 4, R 3Be-CH 2OCH 2CH 2OCH 2CH 2OC 7H 15
[0389] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is CH 2, two L link together and represent NOH, R 2Be H or methyl, m is 4, R 3It is the C15 alkyl.
[0390] in another embodiment, the RPE65 antagonist has the structure of formula 4 and the definition of following thereof, and wherein X is CH 2, L is H, OH or NH 2, R 2Be H or methyl, m is 4, R 3It is the C15 alkyl.
[0391] in certain embodiments, the RPE65 inhibitor can be a formula 7a chemical compound:
Wherein, the implication of following group when at every turn occurring is independently:
X is O, S, NR ', CH 2Or NHNR ';
Z is O or NOH;
R 1Be-CH 2F ,-CHF 2,-CF 3,-CH 2N 2,-CH 2C (O) OR ,-OR ' ,-C (O) CHR ' ,-C (NH) CHR ' or-CH=CHR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is Or
With
N is 1,2 or 3;
Wherein  represents singly-bound, cis-double bonds or trans double bond.
[0392] can think that formula 7a chemical compound is irreversible RPE65 inhibitor because they can with the RPE65 covalent bond, make its permanent failure.
[0393] in certain embodiments, the RPE65 inhibitor can be a formula 7a chemical compound, and wherein Z is O.
[0394] in certain embodiments, the RPE65 inhibitor can be a formula 7b chemical compound:
Figure A20058001151300983
Wherein, the implication of following group when at every turn occurring is independently:
Y is O, S, NR ', CH 2=O, C=S, C=NR ', CHOR ', CHNR ' R ", CHSR '
Or CH 2
R 1Be R ' ,-OR ' ,-CN or (CH 2CH 2O) mR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R " is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Figure A20058001151300984
Figure A20058001151300985
M is 1,2 or 3; With
N is 1,2 or 3;
Wherein  represents singly-bound, cis-double bonds or trans double bond.
[0395] can think that formula 7b chemical compound is reversible RPE65 inhibitor,, and not make its permanent failure because they can combine with RPE65 is non-covalent.
[0396] in certain embodiments, the RPE65 inhibitor can be a formula 7c chemical compound:
Figure A20058001151300991
Wherein, the implication of following group when at every turn occurring is independently:
X is O, S, NR ', CH 2Or NHNR ';
Z is O or NOH;
R 1Be-CH 2F ,-CHF 2,-CF 3,-CH 2N 2,-CH 2C (O) OR ,-OR ' ,-C (O) CHR ' ,-C (NH) CHR ' or-CH=CHR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Or
With
N is 1,2 or 3.
[0397] can think that formula 7c chemical compound is irreversible RPE65 antagonist because they can with the RPE65 covalent bond, make its permanent failure.
[0398] in certain embodiments, the RPE65 inhibitor can be a formula 7c chemical compound, and wherein Z is O.
[0399] in certain embodiments, the RPE65 inhibitor can be a formula 7d chemical compound:
Wherein, the implication of following group when at every turn occurring is independently:
Y is C=O, C=S, C=NR ', CHOH, CHOR ', NH 2, NHR ', NR ' R ", SH, SR ' or CH 2
R 1Be R ' ,-OR ' ,-CN or-(CH 2CH 2O) mR ';
R ' is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R " is H, alkyl, assorted alkyl, aryl, heteroaryl, aralkyl or heteroarylalkyl;
R is
Figure A20058001151301002
M is 1,2 or 3; With
N is 1,2 or 3.
[0400] B. is used to shorten the circulation compositions
[0401] before 11-cis-retinal leaves RPE, can pass through thermodynamics (thermodynamically) catalysis in RPE, make under 11-cis-retinal and turn to the circuit that alltrans-retinal realizes shortening visual cycle to (downhill) isomery.Fig. 3 explains the intervention of an expection.Can conceive a variety of materials that are fit to this purposes.Generally speaking, suitable medicine comprises anil, promptly contains the phenyl ring of amine side chain.
[0402] shortening the circulation molecule has an effect by elder generation and retinal formation Schiff alkali.After forming Schiff alkali with 11-cis-retinal, isomerization takes place.This is short circuit.
[0403] the short circuit chemical compound also can be caught retinal, so that can not utilize them to form A 2E, its precursor or analog.Availablely catch alltrans-retinal and stop it to form A 2The medicine of E and similar compound and alltrans-retinal form metastable Schiff alkali, and the short circuit medicine combines alltrans-retinal with PHOSPHATIDYL ETHANOLAMINE competitiveness.Then, can make the chemical compound of catching in lysozyme, be decomposed into nontoxic metabolite.The short circuit medicine can be upset visual cycle in one or both approach, promptly by shortening 11-cis-retinal circuit and/or by catching alltrans-retinal.(A 2E has the best features of lipofuscin.Perhaps alltrans-retinal and amine-or even albumen between other is arranged adduct one form Schiff alkali between by active retinal and amine and start their formation).
[0404] though expection arylamine/alltrans-retinal Schiff alkali can not continue to form A 2E sample molecule (because its degraded earlier) stops this situation to take place but can shorten circulation secondary amine medicine by use more reliably.This is because A 2E forms mechanism needs primary amine (two free Hs), because form two new N-alkyl bonds (each alltrans-retinal molecule has), and this situation can not occur in the second month in a season or the tertiary amine raw material.If the short circuit medicine is a secondary amine, it only combines with an alltrans-retinal molecule so, do not reserve and second bonded position of alltrans-retinal, thereby stop in the process shown in Figure 2 and A 2The similar compounds of E thing of the same clan forms.
[0405] the short circuit medicine also provides long term, so that their administration number of times can be seldom.In some cases, may need be administered once in every month.Under other situation, may need to be administered once weekly.The short circuit medicine is effectively exhausted local storage vitamin A within the eye by catching alltrans-retinal.Once medicine reduces the storage of vitamin, visual cycle just suffers damage, and lipofuscin forms and hindered, the purpose of treatment that Here it is.But it is very slow that vitamin A storage storehouse replenishes in eye, so that single gives the short circuit medicine and can keep long-acting.In addition, the short circuit medicine can slowly be removed from eye, so that can utilize the long combination of their types.
[0406] in certain embodiments, the short circuit chemical compound has the structure of formula VII representative:
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or carbonyl;
L is a hydrophobic part, or any two adjacent L are joined together to form condensed aromatic ring or hetero-aromatic ring (for example naphthalene, anthracene, indole, quinoline etc.).
[0407] in certain embodiments, L independently is alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, carbonyl, ether or multi-ring base when occurring at every turn.In certain embodiments, L has formula VIIa:
Wherein, the implication of following group when at every turn occurring is independently:
R ' and X are hydrogen, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, carbonyl, alkoxyl, hydroxyl, mercaptan, alkylthio or amino; With
M is the integer of 1-6.
[0408] in certain embodiments, the short circuit medicine can be represented by following general formula VIIb:
[0409] wherein n is the integer of 1-8.
[0410] in certain embodiments, the short circuit medicine can be represented by following general formula VIIc:
Wherein, the implication of following group when at every turn occurring is independently,
R is H, alkyl or acyl group; With
R ' is alkyl or ether.
[0411] in another embodiment, the short circuit medicine has the structure of formula VIIc and the definition of following thereof, and wherein R is H when occurring for twice.
[0412] in another embodiment, the short circuit medicine has the structure of formula VIIc and the definition of following thereof, and wherein at least one R is an alkyl.
[0413] in another embodiment, the short circuit medicine has the structure of formula VIIc and the definition of following thereof, and wherein at least one R is a methyl.
[0414] in certain embodiments, the short circuit medicine can be represented by following general formula VIId:
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is alkyl or ether.
[0415] in another embodiment, the short circuit medicine has the structure of formula VIId and the definition of following thereof, and wherein R is H when occurring for twice.
[0416] in another embodiment, the short circuit medicine has the structure of formula VIId and the definition of following thereof, and wherein at least one R is an alkyl.
[0417] in another embodiment, the short circuit medicine has the structure of formula VIId and the definition of following thereof, and wherein at least one R is a methyl.
[0418] in certain embodiments, the short circuit medicine can be represented by following general formula VIIe:
Figure A20058001151301032
Wherein, the implication of following group when at every turn occurring is independently:
X be hydrogen or-C (=O) OR ';
R is H, alkyl or acyl group; With
R ' is an alkyl.
[0419] in another embodiment, the short circuit medicine has the structure of formula VIIe and the definition of following thereof, and wherein R is H.
[0420] in another embodiment, the short circuit medicine has the structure of formula VIIe and the definition of following thereof, and wherein at least one R is an alkyl.
[0421] in another embodiment, the short circuit medicine has the structure of formula VIIe and the definition of following thereof, and wherein R is a methyl.
[0422] in certain embodiments, the short circuit medicine can be represented by following general formula VIIf:
Figure A20058001151301041
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is an alkyl.
[0423] in another embodiment, the short circuit medicine has the structure of formula VIAnd if the definition of following thereof, and wherein R is H.
[0424] in another embodiment, the short circuit medicine has the structure of formula VIAnd if the definition of following thereof, and wherein at least one R is an alkyl.
[0425] in another embodiment, the short circuit medicine has the structure of formula VIAnd if the definition of following thereof, and wherein R is a methyl.
[0426] in one embodiment, shortening the circulation medicine is the diamino phenoxy pentane:
[0427] in one embodiment, shortening the circulation medicine is phenetidine (phenetidine):
Figure A20058001151301043
[0428] in one embodiment, shortening the circulation medicine is tricaine (tricaine):
[0429] in one embodiment, shortening the circulation medicine is the 4-butylaniline:
Figure A20058001151301051
[0430] in one embodiment, shortening the circulation medicine is N-methyl-4-butylaniline:
Figure A20058001151301052
[0431] in one embodiment, shortening the circulation medicine is the 3-benzocaine:
[0432] in one embodiment, shortening the circulation medicine is N-methyl-3-benzocaine:
[0433] in one embodiment, shortening the circulation medicine is ethyl 2-aminobenzoate:
Figure A20058001151301055
[0434] in one embodiment, shortening the circulation medicine is N-methyl-ethyl 2-aminobenzoate:
[0435] in certain embodiments, the short circuit medicine can be represented by following general formula VIII:
Figure A20058001151301062
Wherein R ' is hydrogen, alkyl or ether; Or any two adjacent L are joined together to form condensed aromatic ring or hetero-aromatic ring (for example naphthalene, anthracene etc.).
[0436] in certain embodiments, shorten the structure that the circulation medicine has formula IX representative:
ANR 2
IX
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or carbonyl; With
A is optional aryl or the heteroaryl that replaces.
[0437] in certain embodiments, the short circuit medicine can be represented by following general formula X:
AC(＝O)NHNH 2
X
The implication of wherein following group when at every turn occurring is independently:
R ' is hydrogen, alkyl or ether; With
A is optional aryl or the heteroaryl that replaces.
[0438] in certain embodiments, shorten the structure that the circulation medicine can have formula 5 representatives:
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or carbonyl;
L is a hydrophobic part, or any two adjacent L are joined together to form condensed aromatic ring;
With
N is the integer of 0-5.
[0439] in certain embodiments, L independently is alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, carbonyl, ether or multi-ring base when occurring at every turn.In certain embodiments, L has formula 5a:
Figure A20058001151301072
Wherein, the implication of following group when at every turn occurring is independently:
R ' and X are H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, carbonyl, alkoxyl, hydroxyl, mercaptan, alkylthio group or amino;
M is the integer of 1-6; With
P is the integer of 0-5.
[0440] instantiation of the short circuit medicine of Xuan Zeing comprises the diamino phenoxy pentane:
Phenetidine:
Figure A20058001151301074
And tricaine:
[0441] in certain embodiments, the short circuit medicine can be represented by following general formula 5b:
Wherein n is the integer of 1-8.
[0442] in certain embodiments, the short circuit medicine can be represented by following general formula 5c:
Figure A20058001151301083
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is alkyl or ether.
[0443] in another embodiment, the short circuit medicine has the structure of formula 5c and the definition of following thereof, and wherein R is H when occurring for twice.
[0444] in another embodiment, the short circuit medicine has the structure of formula 5c and the definition of following thereof, and wherein at least one R is an alkyl.
[0445] in another embodiment, the short circuit medicine has the structure of formula 5c and the definition of following thereof, and wherein at least one R is a methyl.
[0446] in certain embodiments, the short circuit medicine can be represented by following general formula 5c1:
Figure A20058001151301084
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is alkyl or ether.
[0447] in another embodiment, the short circuit medicine has the structure of formula 5c1 and the definition of following thereof, and wherein R is H when occurring for twice.
[0448] in another embodiment, the short circuit medicine has the structure of formula 5c1 and the definition of following thereof, and wherein at least one R is an alkyl.
[0449] in another embodiment, the short circuit medicine has the structure of formula 5c1 and the definition of following thereof, and wherein at least one R is a methyl.
[0450] in certain embodiments, the short circuit medicine can be represented by following general formula 5d:
Figure A20058001151301091
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is an alkyl.
[0451] in another embodiment, the short circuit medicine has the structure of formula 5d1 and the definition of following thereof, and wherein R is H.
[0452] in another embodiment, the short circuit medicine has the structure of formula 5d and the definition of following thereof, and wherein at least one R is an alkyl.
[0453] in another embodiment, the short circuit medicine has the structure of formula 5d and the definition of following thereof, and wherein R is a methyl.
[0454] in certain embodiments, the short circuit medicine can be represented by following general formula 5d1:
Figure A20058001151301092
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is an alkyl.
[0455] in another embodiment, the short circuit medicine has the structure of formula 5d1 and the definition of following thereof, and wherein R is H.
[0456] in another embodiment, the short circuit medicine has the structure of formula 5d1 and the definition of following thereof, and wherein at least one R is an alkyl.
[0457] in another embodiment, the short circuit medicine has the structure of formula 5d1 and the definition of following thereof, and wherein R is a methyl.
[0458] in certain embodiments, the short circuit medicine can be represented by following general formula 5e:
Figure A20058001151301101
Wherein R ' is alkyl or ether.
[0459] also comprises the pharmaceutically acceptable addition salt and the complex of the chemical compound that following formula provides.Chemical compound can contain in the situation of one or more chiral centres therein, and except that regulation was arranged, Gou Si chemical compound can be the racemic mixture of single stereoisomers or stereoisomer herein.Also comprise its prodrug, analog and derivant.
[0460] in certain embodiments, can unite two or more and shorten the circulation chemical compound.In certain embodiments, can be with enzyme inhibitor and/or RPE65 binding inhibitors and the associating of shortening circulation chemical compound.
[0461] can go up the Pharmaceutical composition that uses in acceptable carrier or excipient preparation the inventive method with one or more physiologys according to a conventional method.Thereby, the physiology of reactive compound and they can be gone up acceptable salt and solvate be mixed with by for example injecting, suck or be blown into the preparation of (through port or nose) or oral, buccal, parenteral or rectally, target cell, the position that exists as the disease cell, i.e. eye or the described chemical compound of retina topical administration.
[0462] chemical compound can be formulated as the multiple administration load that comprises whole body and part or site-specific delivery of drugs.Technology and preparation usually can be at Remmington ' s Pharmaceutical Sciences, Meade Publishing Co., and Easton finds among the PA.For the whole body administration, preferably include intramuscular, intravenous, intraperitoneal and subcutaneous in interior injection.For injection, chemical compound can be mixed with liquid solution, preferably buffer that can be adaptive on the physiology is for example prepared in Hank ' ssolution or the ringer's solution.In addition, chemical compound can be mixed with solid form, face with preceding dissolving again or suspension.Also comprise lyophilized form.
[0463] for oral administration, Pharmaceutical composition can adopt and for example pass through conventional method, uses pharmaceutically acceptable excipient, for example binding agent (for example, pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropyl emthylcellulose); Filler (for example, lactose, microcrystalline Cellulose or calcium hydrogen phosphate); Lubricant (for example magnesium stearate, Pulvis Talci or silica gel); Disintegrating agent (for example, potato starch or primojel); Or the form of tablet, lozenge or the capsule of wetting agent (for example sodium lauryl sulphate) preparation.Can be by method well known in the art with tablet coating.The liquid preparation that is used for oral administration can adopt for example solution, syrup or form of suspension, maybe they can be made and use preceding water or other suitable dissolved desciccate of solvent.Can pass through conventional method, with the suspending agent (for example sorbitol syrups, cellulose derivative or hydrogenation edible fat) for example of acceptable additive pharmaceutically; Emulsifying agent (for example lecithin or arabic gum); Non-water-soluble matchmaker (for example ationd oil, grease, ethanol or fractionated vegetable oil); And antiseptic (for example methyl parahydroxybenzoate or propyl ester or sorbic acid) prepares this type of liquid preparation.If suitable, preparation also can contain buffer salt, correctives, coloring agent and sweeting agent.Can suitably prepare the oral formulations that the controlled release of active ingredient thing is provided.
[0464] for inhalation, can be with the form of chemical compound with aerosol spray, use suitable propellant, for example dichlorodifluoromethane, Arcton 11, dichlorotetra-fluoroethane, carbon dioxide or other suitable gas transmit from pressure packing or aerosol apparatus easily.In the aerocolloidal situation of pressure, can determine dosage unit by the valve that the amount of transmitting metering is provided.Can prepare and contain described chemical compound and suitable powder substrate, for example the capsule of the mixture of powders of lactose or starch and for example be used for the gelatin cartridge case of inhaler or insufflator.
[0465] chemical compound can be mixed with by injection, for example ejection preparation of the parenteral of large bolus injection or continuous infusion.Available additional preservatives is formulated in ejection preparation for example in ampoule or the multi-dose container and prepares unit dosage forms.Compositions can adopt this type of form of oil or water-soluble matchmaker's suspension, solution or emulsion, and can contain formula agent, for example suspending agent, stabilizing agent and/or dispersant.Perhaps, before can being to use, active component uses suitable solvent, for example the powder type of aseptic aqueous solution.
[0466] also chemical compound can be mixed with rectal compositions, for example contain suppository base commonly used, for example the suppository of cocoa butter or glyceride or retention enema.
[0467] except that previous formulations, also chemical compound can be mixed with storage storehouse (depot) preparation.Can give this type of durative action preparation by (for example subcutaneous or intramuscular) implantation or by intramuscular injection.Therefore, for example available suitable macromolecule or hydrophobic substance (for example can accept the emulsion of oil) or ion exchange resin, or slightly soluble derivant, for example slightly soluble salt preparation chemical compound.
[0468] Pharmaceutical composition (comprising cosmetic formulations) can contain about 0.00001-100%, for example the described herein chemical compound of one or more of 0.001-10% or 0.1%-5% weight.
[0469] in one embodiment, described chemical compound is herein mixed contain the topical formulations that is fit to the topical carrier of topical usually and contains any this material known in the art.Can select topical carrier so that the compositions that needs form to be provided, for example ointment, lotion, cream, microemulsion, gel, oil, solution etc., and can contain the material in natural generation or synthetic source.Preferred selected carrier does not have a negative impact to active medicine or other composition of topical formulations.The example that is used for suitable topical carrier herein comprises water, pure and mild other nonpoisonous organic solvent, glycerol, mineral oil, silicone, vaseline, lanoline, fatty acid, vegetable oil, p-Hydroxybenzoate, wax etc.
[0470] preparation can be colorless and odorless ointment, lotion, cream, microemulsion and gel.
[0471] chemical compound can be mixed ointment, it is the semi-solid preparation of substrate with vaseline or other petroleum derivative normally.One skilled in the art will recognize that stand-by concrete ointment base, is the substrate that the optimal drug transmission can be provided and other characteristic that needs and for example property of softening etc. preferably can be provided.When with other carrier or solvent coexistence, ointment base should be inertia, stable, non-irritating and nonsensitized.As setting forth among the Remington ' s that partly quotes at preamble, ointment base can be divided into four classes: oleaginous base; Emulsifiable base; Emulsifying base and water-soluble base.The oiliness ointment base comprises for example vegetable oil, the fat that obtains and the semi-solid hydrocarbon that obtains from oil from animal.But the emulsifying ointment base oil pole that is called the ointment base of absorption again contains less or is not moisture, comprises for example hydroxystearin sulfate, anhydrous lanolin and hydrophilic vaseline.The emulsifying ointment base is Water-In-Oil (W/O) Emulsion or oil-in-water (O/W) Emulsion, for example comprises spermol, glyceryl monostearate, lanoline and stearic acid.(PEGs) prepares exemplary water-soluble ointment base by various molecular weight polyethylene glycol; Details can be again with reference to described in the same Remington ' s.
[0472] chemical compound can be mixed lotion, lotion normally is applied to the preparation that skin surface does not have friction, is that solids comprising active medicine are present in typical liquid or the semi-solid preparation in water or the pure substrate.The normally solid suspension of lotion can contain the oil-in-water liq oily emulsion.Lotion is the preparation of preferably treating the bodice bulk area, because this position is easy to use more multithread body compositions.Usually must grind the insoluble substance in the lotion.It is suspending agent and the fixing chemical compound that keeps active medicine and contact skin, for example methylcellulose, the sodium carboxymethyl cellulose etc. of reaching that forms better dispersion that lotion contains usually.The exemplary lotion preparation of uniting use with the inventive method contains the mixture of propylene glycol and hydrophilic vaseline, for example can be from Beiersdorf, and (Norwalk, Conn.) trade mark of Huo Deing is Aquaphor to Inc RTMThe sort of.
[0473] chemical compound can be mixed cream, cream is oil-in-water or water in oil viscous liquid or semi-solid emulsion normally.The washing of cream base available water, and contain oil phase, emulsifying agent and water.Oil phase is usually by vaseline and aliphatic alcohol, and for example spermol or stearyl alcohol are formed; Although optional, the water volume is usually greater than oil phase, and contains wetting agent usually.Emulsifying agent in the cream as setting forth among the same Remington ' s, normally nonionic, anionic, cationic or zwitterionic surfactant.
[0474] chemical compound can be mixed microemulsion, microemulsion is thermodynamically stable, the isotropism clarification dispersion of two kinds of non soluble liquids normally, for example by stable You Heshui (Encyclopedia of Pharmaceutical Technology (the New York:Marcel Dekker of surfactant molecule interfacial film, 1992), the 9th volume).For the preparation of microemulsion, surfactant (emulsifying agent), cosurfactant (coemulsifier), oil phase and water are essential.Suitable surfactant comprises any surfactant that can be used for preparing emulsion, for example is generally used for preparing the emulsifying agent of cream.Cosurfactant (or " coemulsifier ") is selected from polyglycereol derivant, glycerol derivatives and aliphatic alcohol usually.Although optional, preferred solvent/coemulsifier combination is selected from usually: glyceryl monostearate and Myrj 45; Polyethylene Glycol and palmityl glycol stearate (ethylene glycolpalmitostearate); And Trivent OCG and tricaprin and oleoyl polyethyleneglycol glyceride (macrogolglyceride).Water not only comprises water but also comprises buffer agent, glucose, propylene glycol, Polyethylene Glycol usually; preferred low molecular poly (for example PEG 300 and PEG 400) and/or glycerol etc., and oil phase comprises the mixture of for example fatty acid ester, modified vegetable oil, silicone oil, glycerol one, two and three esters, one and the diester (for example oleoyl polyethyleneglycol glyceride) etc. of PEG usually.
[0475] chemical compound can be mixed gel preparation, said preparation is normally by little inorganic particulate (binary system) or be evenly distributed on the semi-solid systems that suspension that the big organic molecule (single-phase gels) in the whole carrier liquid forms is formed basically.Can be by for example making active medicine, carrier liquid and suitable gellant, for example tragacanth (2-5%), sodium alginate (2-10%), gelatin (2-15%), methylcellulose (3-5%), sodium carboxymethyl cellulose (2-5%), carbomer (0.3-5%) or polyvinyl alcohol (10-20%) mix, and mix until producing the semi-solid product of feature and prepare single-phase gels.Other suitable gellant comprises methyl hydroxylated cellulose, polyoxyethylene-polyoxypropylene, hydroxyethyl-cellulose and gelatin.Although gel generally uses aqueous carrier liquid, pure and mild oil also can be used as carrier liquid.
[0476] various additive-package well known by persons skilled in the art can be contained in preparation, for example in the topical formulations.The example of additive includes but not limited to solubilizing agent, dermal osmosis accelerator, opacifier, antiseptic (for example antioxidant), gellant, buffer agent, surfactant (especially nonionic and amphoteric surfactant), emulsifying agent, softening agent, thickening agent, stabilizing agent, wetting agent, coloring agent, essence etc.Especially preferably include solubilizing agent and/or dermal osmosis accelerator and emulsifying agent, softening agent and antiseptic.Best topical formulations comprises approximately: 2% weight-60 (weight) %, the solubilizing agent and/or the dermal osmosis accelerator of preferred 2% weight-50% weight; 2% weight-50% weight, the emulsifying agent of preferred 2% weight-20% (weight); 2% weight-20% weight softening agent; With 0.01-0.2% weight antiseptic, form the active medicine and the carrier (for example water) of preparation remainder.
[0477] dermal osmosis accelerator is used to impel the intact skin of the active medicine of treatment level by the modest size area.Know suitable promoter in this area, for example comprise: low-level chain triacontanol, for example methanol and 2-propanol; Alkyl methyl sulfoxide, for example dimethyl sulfoxine (DMSO), decyl methyl sulfoxide (C 10MSO) and four decyl methyl sulfoxide; Ketopyrrolidine, for example 2-Pyrrolidone, N-N-methyl-2-2-pyrrolidone N-and N-(hydroxyethyl) ketopyrrolidine; Urea; N, the N-diethyl--toluamide; C 2-C 6Alkanediol; Other solvent, for example dimethyl formamide (DMF), N,N-dimethylacetamide (DMA) and tetrahydrofurfuryl alcohol; The azo-cycle that replaces with 1-heptan-2-ketone, especially 1-n-dodecane basic ring azacyclo-heptan-2-ketone (Laurel nitrogen  ketone (laurocapram); Can be from Whitby Research Incorporated, Richmond, Va. obtains, and trade mark is Azone RTM).
The example of solubilizing agent includes but not limited to following: hydrophilic ether, for example (ethoxydiglycol is with Transcutol for carbitol RTMSell) and the carbitol oleate (with Softcutol RTMSell); Polyethylene castor oil derivant, for example polyoxy 35 castor oils, polyoxy 40 hydrogenation castor oils etc.; Polyethylene Glycol, especially low molecular poly, for example PEG300 and PEG 400 and polyethyleneglycol derivative, for example PEG-8 caprylic/capric glyceride is (with Labrasol RTMSell); Alkyl methyl sulfoxide, for example DMSO; Ketopyrrolidine, for example 2-Pyrrolidone and N-N-methyl-2-2-pyrrolidone N-; And DMA.Multiple solubilizing agent also can be used as absorption enhancer.Single solubilizing agent can be mixed preparation, maybe the mixture of solubilizing agent can be mixed wherein.
[0478] examples of suitable emulsifiers and coemulsifier include but not limited to those emulsifying agents and the coemulsifier of setting forth in relevant microemulsion formulation.Softening agent comprises for example propylene glycol, glycerol, isopropyl myristate, polypropylene glycol-2 (PPG-2) myristyl ether propionic ester etc.
[0479] also other active medicine can be included in the preparation, for example other anti-inflammatory drug, analgesic, antimicrobial drug, antifungal agent, antibiotic, vitamin, antioxidant and in the opacifier preparation sun-proof medicine commonly used, they include but not limited to o-aminobenzoa class, benzophenone (especially benzophenone-3), camphor derivatives, cinnamate derivative (for example octyl methoxycinnamate), dibenzoyl methane (for example butyl methoxydibenzoylmethise), Para-Aminobenzoic (PABA) and derivant thereof, and salicylate (for example ethylhexyl salicylate).
[0480] in some topical formulations, the content of active medicine accounts for about 0.25% weight-75% weight of preparation, preferably accounts for about 0.25% heavy about 1.0% weight-10% weight of preparation.
[0481] the topical skin treating compositions can be packaged in the proper container that is fit to its viscosity and consumer's expection use.For example, lotion or cream can be packaged in the container of aerosol device that bottle or ball applicator or propellant drive or the pump that suitable finger manipulation is installed.When compositions is cream, it simply can be stored in indeformable bottle or squeeze receptacle, for example in pipe or the jar with cover.Also compositions can be included in capsule, for example at U.S. Patent number 5,063, in those capsules of setting forth in 507.Therefore, the hermetic container of acceptable composition on the cosmetics that fill definition herein also is provided.
[0482] in alternate embodiment, be provided for the pharmaceutical formulation of oral or parenteral, in this case, preparation can contain the above-mentioned microemulsion that contains reactive compound, but can contain the alternative pharmaceutically acceptable carrier that especially is fit to oral or parenteral, solvent, additive etc.Perhaps, can be basically by above-mentioned and without polishing, oral administration or parenteral contain the microemulsion of reactive compound.
[0483] can for example use the cell drug of the external processing of described chemical compound by the method afford that gives patient's implant herein, can follow to give for example immunosuppressive drug, for example cyclosporin A.For the rule of pharmaceutical formulation, the reader can be with reference to Cell Therapy:Stem CellTransplantation, Gene Therapy and Cellular Immunotherapy, G.Morstyn﹠amp; W.Sheridan edits, Cambridge University Press, 1996; And Hematopoietic Stem Cell Therapy, E.D.Ball, J.Lister﹠amp; P.Law, Churchill Livingstone, 2000.
[0484] also provides kit herein, for example be used for the treatment of and/or the kit of diagnostic purpose.Kit can comprise one or more described chemical compounds and the optional device that is used to make chemical compound and tissue or cells contacting that comprises herein.Device comprises pin, syringe, stent (stents), resuspension liquid and is used for chemical compound is introduced other device of patient.
[0485] in any previous embodiments, can clearly get rid of 1,5-two (right-aminobenzene 1 oxygen base) pentane.
[0486] in any previous embodiments, can clearly get rid of 11-cis-retinol.
[0487] in any previous embodiments, can clearly get rid of 11-cis-retinal cetylate.
[0488] in any previous embodiments, can clearly get rid of 13-cis-tretinoin (accutane).
[0489] in any previous embodiments, can clearly get rid of 2-bromine Palmic acid.
[0490] in any previous embodiments, can clearly get rid of 3-benzocaine methanesulfonates.
[0491] in any previous embodiments, can clearly get rid of acetaminophen.
[0492] in any previous embodiments, can clearly get rid of amantadine.
[0493] in any previous embodiments, can clearly get rid of alltrans-retinal.
[0494] in any previous embodiments, can clearly get rid of alltrans-tretinoin.
[0495] in any previous embodiments, can clearly get rid of alltrans-retinol (vitamin A).
[0496] in any previous embodiments, can clearly get rid of Palmic acid alltrans-retinyl ester.
[0497] in any previous embodiments, can clearly get rid of ahaline.
[0498] in any previous embodiments, can clearly get rid of cyclohexylamine.
[0499] in any previous embodiments, can clearly get rid of dapson.
[0500] in any previous embodiments, can clearly get rid of the diamino phenoxy pentane.
[0501] in any previous embodiments, can clearly get rid of the m-anthranilic acid ethyl ester.
[0502] in any previous embodiments, can clearly get rid of m-anthranilic acid.
[0503] in any previous embodiments, between can clearly getting rid of-phenetidine.
[0504] in any previous embodiments, can clearly get rid of N-(4-hydroxy phenyl) and look yellow amide (fenretinide (fenretinide)).
[0505] in any previous embodiments, can clearly get rid of N, accelerine.
[0506] in any previous embodiments, can clearly get rid of N, N-dimethyl-right-phenetidine.
[0507] in any previous embodiments, can clearly get rid of methylphenylamine.
[0508] in any previous embodiments, can clearly get rid of N-methyl-right-phenetidine.
[0509] in any previous embodiments, can clearly get rid of neighbour-phenetidine.
[0510] in any previous embodiments, can clearly get rid of right-(positive hexyloxy) aniline.
[0511] in any previous embodiments, can clearly get rid of right-(positive hexyloxy) Benzoylamide.
[0512] in any previous embodiments, can clearly get rid of right-(positive hexyloxy) benzoic acid hydrazides.
[0513] in any previous embodiments, can clearly get rid of right-anisidine.
[0514] in any previous embodiments, can clearly get rid of right-ethyl analine.
[0515] in any previous embodiments, can clearly get rid of right-ethoxybenzene methylamine.
[0516] in any previous embodiments, can clearly get rid of right-thanatol.
[0517] in any previous embodiments, can clearly get rid of phenetidine.
[0518] in any previous embodiments, can clearly get rid of piperidines.
[0519] in any previous embodiments, can clearly get rid of right-n-butoxy aniline.
[0520] in any previous embodiments, can clearly get rid of right-n-butyl aniline.
[0521] in any previous embodiments, can clearly get rid of right-dodecyl aniline.
[0522] in any previous embodiments, can clearly get rid of p-Nitraniline..
[0523] in any previous embodiments, can clearly get rid of sulfbenzamide.
[0524] in any previous embodiments, can clearly get rid of sulfanilamide  azoles (sulfamoxaole).
[0525] in any previous embodiments, can clearly get rid of sulfanilamide.
[0526] in any previous embodiments, can clearly get rid of tricaine.
[0527] in addition, any chemical compound of being attached to herein of list of references also can clearly be got rid of outside any previous embodiments by reference.
[0528] 4. method
[0529] method disclosed herein is used for the treatment of or prevents ophthalmic.Illustrative methods comprises the compositions that the patient treatment of these needs effective dose is arranged, Pharmaceutical composition for example as herein described.There is this patient who needs to know oneself to suffer from the patient that ophthalmic maybe may take place.
[0530] as mentioned above, can give the disclosed compositions of patient so that treatment or prevention degeneration of macula.Can treat other disease, disorder or the disease that are characterised in that the retinal toxicity chemical compound is accumulated in RPE equally.
[0531] in one embodiment, give the medicine that the visual cycle short circuit takes place to make in the visual cycle ladder patient outside shaft-like photoreceptor cell dish.For example as shown in Figure 3, medicine can react with the 11-cis-retinal among the RPE, it is branched to alltrans-retinal, and it rests among the RPE.More particularly, therapeutic agent can react with 11-cis-retinal, forms the intermediate that isomery turns to the alltrans configuration.The alltrans intermediate can discharge this therapeutic agent then, forms alltrans-retinal.Then can be by normal vision circulation residue ladder reprocessing alltrans-retinal among the RPE.Therefore, visual cycle will deteriorate to invalid circulation, and wherein alltrans-retinal is little or have no chance to accumulate in dish.
[0532] in one embodiment, the diagnosable patient who suffers from degeneration of macula can give disclosed medicine then.In another embodiment, can identify patient's (risk factor comprise smoking history, age, women and family history) that the degeneration of macula risk takes place.In another embodiment again, the patient of the Shi Tajiate disease of diagnosable family's form of suffering from degeneration of macula.In certain embodiments, preventability gives medicine.In certain embodiments, the diagnosable patient who before retinal damage is obvious, suffers from disease.For example, can find to carry abcr, the patient of elovl4 mutant gene and/or another gene, therefore the diagnosable patient who before any ophthalmology sign occurs, suffers from the Shi Tajiate disease, or can before recognizing any eyesight influence, the patient find to have patient as the early stage macula lutea change of degeneration of macula indication.In certain embodiments, people patient may know him or she and need treat or prevent degeneration of macula.
[0533] in certain embodiments, can monitor the degree of patient's degeneration of macula.Available several different methods is for example by checking eyes, expansion eye examination, ophthalmofundoscope (fundoscopic inspection, visual discrimination aptitude tests, angiography, luciferin angiography and/or biopsy monitoring patient.Can monitor at different times.For example, can monitor behind patient's medicine giving.Can after first administration 1 day, 1 week, 2 weeks, January, February, June, 1 year, 2 years and/or 5 years, monitor.In certain embodiments, can change drug dose by the monitoring situation.
[0534] in certain embodiments, disclosed method can with other method of treatment or prevention degeneration of macula, for example photodynamic therapy associating.
[0535] in certain embodiments, can treat or prevent the medicine of degeneration of macula for a long time.Can be once a day, every day repeatedly, twice weekly, on every Wendesdays time, once in a week, whenever biweekly, every month once, whenever bimonthly, half a year once, annual and/or give medicine annual twice.
[0536] can give therapeutic agent by above-mentioned number of ways.In certain embodiments, but per os gives tablet, capsule, the medicine of liquid, paste and/or powder type.In certain embodiments, can pass through intraocular injection topical administration medicine.In certain embodiments, but whole body gives support rack type (caged), mask type (masked) medicine, or gives the medicine (for example passing through photodynamic therapy) of other inactivation form of eyes and activity form.In certain embodiments, can give the reservoir type medicine, so that provide long-time, the medicine of a few hours, a few days, several weeks and/or several months slow release for example.
[0537] use therapeutic agent by the treatment effective dose, the treatment effective dose alters a great deal, and depends on employed concrete medicine on very to degree.The time span that the medication amount of mixing compositions also depends on required release mode, the needed drug level of biological effect and the bioactive substance that is used for the treatment of discharges.In certain embodiments, bioactive substance can be mixed with the chemical compound substrate of different load level, in one embodiment, at room temperature not need organic solvent.In other embodiments, compositions can be mixed with microsphere.In certain embodiments, medicine can be mixed with slow release formulation.
[0538] it should be noted that, upset visual cycle to stop A 2E accumulates dark (faint light) vision that may damage the patient and may cause nyctalopia.When suitable, the suitable prevention A that mentions herein 2Some therapeutic agent that E accumulates is used for people or restriction (withheld) really on a small quantity all to be used, because they easily cause nyctalopia.But,, might need the pleased nyctalopia of accepting to a certain degree of patient of this treatment not injure twenty-twenty vision to exchange for owing to recognize that this special nyctalopia cause of disease may become treating and/or preventing property treatment degeneration of macula.This is because above-mentioned visual cycle is moved in the rod cell photoreceptors, and it only moves and operation in the daytime on low illumination level.Therefore, although may to scotopic vision some influence be arranged at night, the visual cycle function reduces seldom influences degeneration of macula.At least some patient and may Most patients may be willing to sacrifice the night vision reduction for reducing cone vision in the daytime probability that they finally lose them.
[0539] Palmitoylation
[0540] in certain embodiments, available LRAT inhibitor is regulated the palmitoylation of RPE65.RPE65 occurs with at least two kinds of forms, film relevant (mRPE65) and solubility (sRPE65).As following more reasoned, mRPE65 is the palmitoylation form of RPE65, and sRPE65 takes off the palmitoylation form.
[0541] the retinoid flux in the visual cycle can be subjected to regulate by the reversibility palmitoylation RPE65 of LRAT.MRPE65 combines with long-chain alltrans-retinyl ester specifically, and is further to process to transfer them in visual cycle.Alltrans-retinyl ester is the substrate of IMH, and this substrate is converted into 11-cis-retinol with them.Need alltrans-retinyl ester that the company of mRPE65 is done in order to transfer these esters.It is not absolute regulating in this article, because in cycle period, the molecule of the mRPE65 that does not infer changes.Report several observations in this article, but relate to this problem and very likely visual cycle is applied adjusting in the RPE65 phase.
[0542] can be summarized as follows about the obvious fact of on the RPE65 level, calling adjusting: (1) mRPE65 and sRPE65 performance different with complementary retinoid in conjunction with behavior.MRPE65 combines with alltrans-retinyl ester specificity, they can be used in IMH processing, and sRPE65 combines with the vitamin A specificity, and it is used to make it can be LRAT.(2) principal mode of isolating RPE65 is sRPE65 rather than mRPE65.(3) state of the palmitoylation of mRPE65 and sRPE65 is different.(4) to transform mutually be harmony to the reversibility between sRPE65 and the mRPE65, and be subjected to the catalysis of LRAT, so that little mRPE65 level variation meeting has the effect of expansion to isomerization.(5) in the presence of LRAT; mRPE65 works as the palmityl donor of 11-cis-retinol; show dual function to mRPE65; be the albumen of association class visual pigment and by reducing the isomerized acry radical donor of mRPE65 horizontal constraints; (6) alltrans-retinyl ester has adverse effect, becomes mRPE65 because they order about sRPE65.
[0543] it is comprehensive to prepare the laboratory observation that the simple working model will carry out herein, becomes the important adjusting composition in the control visual cycle.Figure 13 A-B shows how described adjusting composition may guide the retinoid in the vision to flow.In the dark, when not needing to form vision chromophore 11-cis-retinal, estimate that sRPE65 is the principal mode of RPE65.Make 11-cis-retinol palmitoylation by mRPE65, produce sRPE65, may also will pass through activated palmityl esterase hydrolyzed mRPE65 in the dark.It is quite possible relating to G-albumen coupling incident at this.Light-operated (Light flips) switch (Figure 13 A) is because the rhodopsin photoisomerization in photoreceptors causes vitamin A to flow to RPE.RPE starts the process of chaperone vitamin A to the LRAT of the substrate alltrans-retinyl ester that generates IMH.As this paper to shown in, alltrans-retinyl ester has second effect, orders about sRPE65 and is converted into mRPE65.This process is a harmony, so that little mRPE65 concentration change can have big influence to alltrans-retinyl ester process velocity and isomerization.MRPE65 guiding alltrans-retinyl ester flows to IMH, with its processing, forms 11-cis-retinol there.Once form 11-cis-retinol, by combining with cRALBP, by 11-cis-retinol dehydrogenase oxidation, it can be divided into rhodopsin chromophore 11-cis-retinal subsequently.When the opsin in light can be used as that rhodopsin fades as a result the time, this chromophore flows to photoreceptors.The opsin combines the formation rhodopsin and drives this process with 11-cis-retinal heat release.
[0544] switch cuts out in the dark again because with mRPE65 as acry radical donor with 11-cis-retinol acidylate, form the Palmic acid 11-cis-retinyl ester and the sRPE65 of chromophoric storage form.This process makes system closing, because the latter is the chaperone of vitamin A, is not alltrans-retinyl ester, can not impel IMH to handle.Because of the concertedness of process, little mRPE65 concentration change can have big influence to 11-cis-retinol aggregate velocity again.At the visual cycle run duration, make the also mRPE65 conversion of soluble hypothesis of 11-cis-retinol palmitoylation by mRPE65, although by above prompting, other factors also can impel the mRPE65 hydrolysis.Therefore, the operation of the switch of proposal is very simple: alltrans-retinyl ester level raises and impels the chromophore biosynthesis, because the mRPE65 that regenerates guiding retinoid flows to IMH.11-cis-retinol forms to increase and makes system closing, is converted into sRPE65 because it orders about mRPE65.11-cis-the retinol of known increase is the biosynthetic highly efficient depressor of chromophore in the body, and shown in Figure 12 A, this suppresses to small part because switching effect in this article.At last, should also meet that 11-cis-retinoid regenerates in the dark based on existence of the adjusting composition of switch is the observation of incident very slowly,
[0545] described herein research has than them the influence of vision process (processing) broad sense more.Certainly, except that a mechanism of exploring herein, palmityl switch mechanism can be by multiple signal transduction context operation.On biochemical level, mRPE65 is only relevant with their palmitoylation degree difference in conjunction with the molecular basis of selective difference with the part between the sRPE65.The post translational modification that the representative of albumen palmitoylation is known, its main effect are to increase proteic hydrophobicity, guide them to film, in some cases, also impel protein-protein interaction.Undoubtedly, be converted in the situation of mRPE65 at the sRPE65 that makes of RPE65 palmitoylation mediation, the film guiding is the result.But Bao Dao the palmitoylation that studies show that has two other effects herein.The first, as mentioned above, palmitoylation changes the part binding specificity of modified protein.Whether directly and alltrans-retinyl ester effect the palmityl of mRPE65 thereby impels the combination of these molecules by hydrophobic interaction; Or whether palmitoylation causes the change of albumen structure picture unknown at present.The second, the inventor confirms that also palmitoylation albumen (mRPE65) can bring into play the function of palmityl donor.Set forth the reversibility palmitoylation, this reversibility may have the adjusting importance, and (Houslay 1996; Mumby 1997; Bijlmakers and Marsh 2003; Qanbar and Bouvier 2003).This therein small G-protein be that especially meaningful in the signal transduction process of palmitoylation (Milligan 1995; Morello 1996; Mumby 1997; Resh 1999; Chen andManning 2001; El-Husseini and Bredt 2002; Bijlmakers and Marsh 2003; Qanbar and Bouvier 2003).In these examples, it is believed that, remove the palmityl part by esterase, but the role of the proteic acyl carrier of little G-may not participate in wherein (Mumby1997; Resh 1999; Linder and Deschenes 2003).
[0546] LRAT catalysis mRPE65 and sRPE65 transform mutually, so this enzyme is bifunctional, because its also is responsible for a large amount of synthetic alltrans-retinyl esters in visual cycle.In the research of this paper report, mRPE65 plays the palmityl donor, but not the effect of lecithin.This result because up to now, it is believed that LRAT makes acry radical donor and does the enzyme-specific that is rather narrow (Canada etc., 1990 of acyl acceptor with retinol with lecithin (being DPPC) unexpectedly; Barry etc., 1989; Saari2000).As for the acry radical donor function, PHOSPHATIDYL ETHANOLAMINE and Phosphatidylserine all can not replace lecithin (
Figure A20058001151301241
Deng, 1990).
[0547] LRAT is the basis of the protide that enlarges, wherein many have unknown function (Jahng etc., 2003b).The albumen of unknown function comprises II class tumor inhibitor and EGL-26, a kind of in C.elegans the mediation morphogenetic imaginary enzyme (Hanna-Rose2002; Anantharaman and Arvind 2003).These albumen should be considered as the candidate enzyme of possible palmityl transferring enzyme.Along these clues, notice that enjoyably the special-purpose palmityl transferring enzyme of Shang Weiyou evaluation it is believed that chemistry but not there is (Mumby1997 in the probability of enzymatic palmitoylation; Resh 1999; Linder and Deschenes 2003; Bijlmakersand Marsh 2003).Therefore, can think that herein what identify is the prototype of palmityl transferring enzyme regulator or inhibitor as the regulator of LRAT or the chemical compound of inhibitor, and can be used for regulating other palmityl transferring enzyme of the LRAT apoplexy due to endogenous wind of expansion.
[0548] 5. screening technique
[0549] can identify appropriate drug by multiple screening technique.For example, can suffer from degeneration of macula or the patient who suffers from the degeneration of macula risk is arranged, for example for the animal candidate medicine of the animal model of degeneration of macula and can measure for example A 2The retinal toxicity chemical compound of E.Therefore, will compare the drug identification that can cause the retinal toxicity chemical compound to accumulate minimizing with contrast (not having medicine) is appropriate drug.Perhaps, can analyze, determine whether to exist alltrans-retinal, the inferior retinyl-PE of N-and/or A the photoreceptors dish 2E.The animal model that produces degeneration of macula fast has quite big meaning, because spontaneous degeneration of macula is usually by developing for many years.Several animal models is admissible degeneration of macula model.For example, existing with abcr-/-reject mice as elovl4-/-reject the argumentation that mice is accumulated model as degeneration of macula and/or lipofuscin.In addition, also have lacking mononuclear cell chemoattractant albumen-1 (Ccl-2; Be called MCP-1 again) or the rejecting mice of its homoreceptor, C-C chemokine receptors-2 (Ccr-2) as the argumentation of the acceleration model of degeneration of macula.
[0550] in addition, the external model of visual system can promote to screen the drug research that suppresses or shorten the visual cycle circuit.Can place the solution that contains suitable enzyme to set up external model by intermediate and other the essential cofactor that will select.Perhaps, can use external RPE culture systems.For example, can be by in the solution of the substrate that contains LRAT and LRAT, adding candidate drug test LRAT inhibition and measuring accumulating of expection product.Can imagine the similar system that is used for described other potential inhibition target herein.
[0551] except that referring else, enforcement the inventive method will be used cytobiology, cell culture, molecular biology, genetically modified organism, microbiology, recombinant DNA and the immunology routine techniques in the art technology scope.This type of technology elaborates in the literature.Referring to for example, Molecular Cloning A Laboratory Manual, the 2nd edition, Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press:1989); DNACloning, I and II volume (D.N.Glover edits, 1985); OligonucleotideSynthesis (M.J.Gait edits, 1984); U.S. Patent numbers such as Mullis; 4,683,195; Nucleic Acid Hybridization (B.D.Hames﹠amp; S.J.Higgins edits 1984); Transcription And Translation (B.D.Hames﹠amp; S.J.Higgins edits 1984); Culture Of Animal Cells (R.I.Freshney, Alan R.Liss, Inc., 1987); Immobilized Cells And Enzymes (IRL Press, 1986); B.Perbal, APractical Guide To Molecular Cloning (1984); The treatise, and Methods InEnzymology (Academic Press, Inc., N.Y.); Gene Transfer Vectors ForMammalian Cells (J.H Miller and M.P.Calos edits, and 1987, Cold SpringHarbor Laboratory); Methods In Enzymology, the 154th and 155 volumes editors such as () Wu, Immunochemical Methods In Cell And Molecular Biology (Mayer and Walker edit, Academic Press, London, 1987); Handbook OfExperimental Immunology, I-IV volume (D.M.Weir and C.C.Blackwell, editor, 1986); Manipulating the Mouse Embryo, (Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y., 1986).
Embodiment
[0552] further specify this description by following examples, these embodiment should be nonrestrictive.The list of references content of all references (comprising list of references, patent granted, the disclosed patent application of the document of quoting among the application) is attached to herein by reference.
[0553] Embodiment 1: external
[0554] material: refrigerated no retina cattle optic cup (eyecups) is available from W.L.LawsonCo., Lincoln, NE.Ammonium bicarbonate, BSA, ethylenediaminetetraacetic acid (EDTA), guanidine HCl, imidazoles, DEAE-agarose, phenyl-agarose CL-4B, alltrans-retinol, Palmic acid alltrans-retinyl ester, alpha-cyano-4-hydroxycinnamic acid and Trizma  alkali are available from Sigma-Aldrich.Dithiothreitol, DTT is available from ICN Biomedicals Inc.By the synthetic 11-cis-retinol of following described elsewhere method (Shi etc., 1993) and Palmic acid 11-cis-retinyl ester.Anagrade TMCHAPS and Lauryl.beta.-maltoside (maltoside) are available from Anatrace.HPLC level solvent is available from Sigma-Aldrich Chemicals.Anti-RPE65 (NFITKVNPETLETIK) antibody derives from Genmed Inc, and anti-LRAT antibody is provided by DeanBok professor (University of California at Los Angeles).The rHRPE65 baculovirus is provided by Jian-Xin Ma professor (University of South Carolina).Hank ' sTNM-FH Insect culture medium derives from JRH Biosciences.The sf21 cell is StevenHartison professor's laboratory (Harvard Medical School) experiment reserve.The broad-spectrum protease inhibitor mixture (cocktail) that does not contain EDTA derives from Roche Biosciences.Nickel-NTA resin and tweezer-NTA column spinner is available from Qiagen Inc.Be used for sodium lauryl sulphate-available from Invitrogen according to the precast gel (4-20%) of polyacrylamide gel electrophoresis, prestained benchmark and Magic molecular weight marker.The DEAE agarose is available from AmershamBiosciences.Press buffer at slide-a-lyser TMThe requirement that box Pierce (10 KDa MWCO) spends the night changes buffer by dialysis.Amicon Ultra with Millipore Corp TMCentrifugal filtration instrument (30KDa-termination) concentrates RPE65 solution.Except that referring else, all reagent are AG.
[0555] method
[0556] Purification mRPE65,3RPE65 and rHRPE65: implement purification by aforementioned (Ma etc., 2001).By silver dyeing or coomassie dyeing and Western blotting check these proteic purity (1: 4000 antibody (primary antibody)-1h that comes into being, at room temperature; With 1: 4000 secondary antibody (secondary antibody)-0.5h, at room temperature).
[0557] Purification rCRALBP: implement purification by aforementioned (25).By silver dyeing or coomassie dyeing and Western blotting check these proteic purity (1: 4000 antibody-1h that comes into being, at room temperature; With 1: 4000 secondary antibody-0.5h, at room temperature).
[0558] Fluorescence is in conjunction with mensuration: with the PBS of RPE65,1%CHAPS, pH7.4 solution are used for fluorescence titration research.Lowry method (Lowry etc., 1951) by improvement is measured protein concentration.All titration are all 25 ℃ of operations down.At the 280nm place, the PBS buffer of excited sample carries out fluorescent scanning in the 300-500nm scope.Under 25 ℃,, with the 450 μ L quartz curettes of passage length 0.5cm, measure fluorescence on the Fluoromax 2 with the right angle detection method at JobinYvon Instruments.
[0559] behind 25 ℃ of following balance 10min, measures protein solution fluorescence.Be dissolved in the drips of solution random sample product of dimethyl sulfoxine then with retinoid.In each titration, in 250 μ L protein solutions, add the retinoid of common 0.2 μ L equivalent, mix homogeneously makes its balance 10min then, writes down fluorescence intensity then.Add dimethyl sulfoxine (at every turn adding 0.1%) to fluorescence intensity without any influence.By in order to following equation (Gollapalli etc., 2003), by fluorescence intensity calculations incorporated constant (K D).
P 0 α = R 0 α n ( 1 - α ) - K D n
P wherein 0=total protein concentration, α = F max - F F max - F 0 , The number of n=independence binding site, R 0=add the concentration of fashionable total retinoid, K at every turn D=dissociation constant, F MaxFluorescence intensity during=saturation, and F 0=initial fluorescent intensity.
[0560] Tretinoin (alltrans and 13-cis) and Palmic acid alltrans-retinyl ester are competed Striving property is in conjunction with RPE65:
Carry out the buffer-exchanged experiment, research tretinoin (alltrans and 13-cis) displacement is in conjunction with the ability of Palmic acid alltrans-retinyl ester of RPE65.(pH7.4) the middle 6 μ M tretinoins (alltrans and 13-cis) that add are cultivated 30min down at 4 ℃ for PBS, 1%CHAPS to RPE65 (0.5 μ M).Under 4 ℃, the control sample of the RPE65 of retinoic acid-containing is not cultivated 30min.When cultivating end, with sample and Palmic acid 3(0.65 μ M 20.31Ci/mmol) cultivates 30min to H-alltrans-retinyl ester.When culture period finishes, buffer (PBS-1%CHAPS) is exchanged 10 with Centricon 30KMWCO filter 4Doubly.Sample that record is held back on liquid scintillation counter and buffering liquid stream branch are measured the Palmic acid of holding back 3The amount of H-alltrans-retinyl ester.
[0561] Alltrans-tretinoin (atRA), 13-cis-tretinoin (13cRA) and N-(4- Hydroxy phenyl) looks yellow the influence of amide (4-HPR) to IMH:
In the buffering suspension of 1mL RPE film (100mM Tris pH8.0,76.7 μ g albumen), add 60 μ M or 6 μ M atRA, 13cRA or 4-HPR, at room temperature incubation 15min.At room temperature, the control reaction mixture incubation 15min that also will not have any inhibitor.After the 15min incubation finishes, with alltrans-retinol [11-12- 3H 2] (0.2 μ M) adding reactant mixture (100mM Tris pH8.0,76.7 μ g RPE albumen, 0.2%BSA 100 μ MDPPC, 1mM DTT and 0.2 μ M alltrans-retinol [11-12- 3H 2]) in, incubation 30min at room temperature.After the 30min incubation finishes, with the sample aliquot quencher of reactant, check equivalent adding alltrans-retinol [11,12- 3H 2] and these inhibitor to the effect of LRAT.Afterwards, with this control reaction mixture and atRA (60﹠amp; 6 μ M), 13cRA (60﹠amp; 6 μ M) or 4-HPR (60﹠amp; 6 μ M) incubation 15min.At once with all reactant mixtures and 30 μ M apo-rCRALBP (100mM Tris pH8.0,7.7 μ g RPE albumen, 0.2%BSA100 μ M DPPC, 1mM DTT 30 μ M apo-rCRALBP and 0.2 μ M alltrans-retinol [11-12- 3H 2]) 37 ℃ of following incubations 30 minutes.After this incubation period finishes, add the freezing methanol of 750 μ l, add 100 μ l 1M sodium chloride solution quenchers, 200 μ L reactant mixtures afterwards, add 500 μ l hexanes (containing butylated hydroxytoluene 1mg/mL) and extract retinoid.By aforementioned (27) methods analyst retinoid.The amount of the 11-cis-retinol that forms is measured as IMH is active.All experiments are all done three parts, analyze with the meansigma methods of these measurements.
[0562] makes rHmRPE65's 3H 2Palmitoylation:
With the 6xHis-recombined human film relevant with RPE65 at sf 2Express in the recombinant baculovirus in 1 insect cell.With recombinant baculovirus transfection sf 21 cell subsequently at 25 ℃ of following incubation 8h, adds (0.09 μ M) subsequently 3H 2Palmic acid (0.5mCi/mL).With culture at 25 ℃ of following incubation 48h.Make similar culture (the 0.09 μ M) growth of "dead" Palmic acid in contrast.Express when finishing harvesting under 500xg.Cell is being contained 500mMNaCl-pH8.0, dissolving in the 100mM phosphate buffer of 5mM imidazoles and 6M guanidine HCl.According to manufacturer's explanation, molten born of the same parents' buffer contains an amount of protease inhibitor cocktail.Then with dissolved cell 100, centrifugation cell fragment under the 000xg, by manufacturer explanation, purification on nickel-NTA post.Purifying protein solution is divided into two parts: (1) handles 16h with 0.5M TrispH8.0, and (2) handle 16h with 0.5M azanol pH8.0.Then by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, western blot analysis and radioautographic analysis protein sample.
[0563] MALDI-TOF analyzes the cattle mRPE65 and the sRPE65 of purification:
Voyager-DE STR with Applied Biosystems carries out the MALDI-TOF quality analysis.Purification mRPE65 and sRPE65 as stated above.Make the gel bands of a spectrum that contain pure mRPE65 and the sRPE65 10min that in acetonitrile, dewaters.Ammonium bicarbonate (100mM) solution with dithiothreitol, DTT (10mM) is covered gel strips, reduction albumen 1h under 56 ℃.After being cooled to room temperature, remove the reduction buffer.Carry out gel detergent/dewatering cycle with ammonium bicarbonate/acetonitrile, repeat 3 times, then at 37 ℃, with trypsin 12.5ng/ μ L, 5 μ L/mm 2Gel spends the night) digestion.Centrifugal gel slice is collected supernatant.By under 25 ℃, change a 20mM ammonium bicarbonate and three times 50% acetonitriles and further extract peptide (per 20 minutes replacing solvents).(0.5 μ L is 10mg/mL) as the substrate of each sample (0.5 μ l) with alpha-cyano-4-hydroxycinnamic acid.Press the 20000V accelerating potential and extract the reflector mode operation sample of 200 nanoseconds of time delay.Laser intensity is 1900-2300, collects each spectrographic 100-200 laser exit point.The mass range of gathering is 750-4500Da, and quality lower bound (low mass gate) is 600Da.
[0564] SRPE65 is to the influence of the esterification of tLRAT mediation:
By monitoring tLRAT catalytic by the alltrans-retinol that adds [11,12- 3H 2] retinyl ester that forms of sRPE65 and/or DPPC/ Lauryl.beta.-maltoside measures the LRAT activity.During all of report are studied in this article, and the LRAT (tLRAT) of use truncate (Jahng etc., 2003b).The LRAT of this form has two N and the terminal membrane-spanning domain of C-of the LRAT of truncate, and labelling allows the bacterial expression LRAT and the His (Bok etc., 2003) of purification fully.Never purification is crossed LRAT, can not express in antibacterial.Dynamics research to LRAT and tLRAT shows, their behavior identical (Bok etc., 2003).In present experiment, reactant mixture (0.1mL volume) contains 100mM Tris (pH8.4), 5 μ M tLRAt, 200 μ M DPPC/0.1% Lauryl.beta.-maltosides and/or 0.04 μ M sRPE65,1mM dithiothreitol, DTT and 0.2 μ M alltrans-retinol [11,12- 3H 2], incubation 10min at room temperature.Behind the 10min, with 500 μ L methanol, 100 μ l water and 500 μ L hexane quenchers reaction.Palmic acid alltrans-the retinyl ester that measure to form by positive HPLC [11,12- 3H 2] amount, this amount is as active tolerance.Double is done in each experiment, and the data point of use is the average of these two points.
[0565] Rely on the esterification of the vitamin A of mRPE65 concentration:
By monitoring tLRAT catalysis by add [11,12- 3H 2]-alltrans-retinol and mRPE65 form Palmic acid alltrans-retinyl ester, measure mRPE65 concentration forms speed to Palmic acid alltrans-retinyl ester effect.It should be noted that and to identify the Palmic acid alltrans-retinyl ester that forms by mRPE65 and vitamin A by its mass spectrum and chromatograph character.Reactant mixture (0.1mL volume) contains 100mM Tris (pH8.4), 5 μ M tLRAT 0.3%CHAPS, 1mM dithiothreitol, DTT and 5 μ M alltrans-retinols [11,12- 3H 2] and mRPE65 (0,0.008,0.02,0.028,0.04,0.052,0.06 and 0.08 μ M), at room temperature incubation 10min.Behind the 10min, with 500 μ L methanol, 100 μ L water and 500 μ L hexane quenchers reaction.Palmic acid alltrans-the retinyl ester that measure to form by positive HPLC [11,12- 3H 2] amount, this amount is as active tolerance.By aforementioned, the computational dynamics parameter K M(Kapp) and N (Segal 1993).Each experiment is done three parts, and the data point of use is the meansigma methods of these three points.Standard deviation is represented with error post (error bars).
[0566] The reversible palmitoylation of the vitamin A of RPE65 mediation:
To in the presence of RPE65, the reversibility of the vitamin A palmitoylation by tLRAT mediation is studied.Will be by 100mM Tris pH8.4,0.06 μ M mRPE 65,1mM dithiothreitol, DTT, the reactant mixture incubation 1h that 1mM EDTA 5 μ M tLRAT and 5 μ M alltranses-retinol is formed.Add subsequently 5 μ M alltrans-retinols [11,12- 3H 2] (4.05Ci/mmol).0,2,7,10,20 and 35min after from reactant, shift out sample aliquot, add 500 μ L methanol, 100 μ L H 2O quencher reaction is with 500 μ L hexane extraction.Alltrans-retinyl ester is separated with alltrans-retinol, press the activity specific (Gollapalli and Rando 2003) that preceding method calculates every part of flow point.Each time point is done three times, uses meansigma methods.Standard deviation is represented with the error post.
[0567] External, mRPE65 is converted into sRPE65:
With purification mRPE65 (0.02 μ M) and tLRAT (5 μ M) and alltrans-retinol (0.2 μ M) incubation 2h at room temperature.When reaction finishes,, destroy the endogenous retinoid with UV light (365nm) irradiation reactant mixture 15min.This solution of dialysis in the dialysis buffer liquid that contains 100mM phosphate buffer (pH8.0), 500mM NaCl, 5mM imidazoles and 1%CHAPS.Concentrate the reactant mixture of dialysis then, make it pass through nickel-NTA column spinner, remove the tLRAT of 6xHis labelling.Concentrate flow point, be used for fluorescence as stated above in conjunction with mensuration.Confirm that by Western blotting tLRAT removes (1: 4000 nascent antibody-1h at room temperature analyze and 1: 4000 secondary antibody-0.5h at room temperature analyzes).
[0568] the preferential isomerization of retinol esterification of mRPE65/tLRAT mediation:
By monitoring tLRAT catalysis by the alltrans-retinol that adds [11,12- 3H 2], 11-cis-retinol [15- 3H] and the retinyl ester that forms of mRPE65, measure mRPE65 to processing 11-cis-and the influence of alltrans-retinol.Reactant mixture (0.1mL volume) contain 100mM Tris (pH8.4), 5 μ M tLRAT0.3%CHAPS, the second-rate threitol of 1mM and 0.2 μ M alltrans-retinol [11,12- 3H 2] or 11-cis-retinol [15- 3H] and mRPE65 (0.02 μ M) or 200 μ M/0.4%DPPC/BSA, at room temperature incubation 10min.Behind the 10min, with 500 μ L methanol, l00 μ L water and 500 μ L hexane quenchers reaction.The amount of the Palmic acid retinyl ester that measure to form by positive HPLC, this amount is as active tolerance.Each experiment is done three parts, and the data point of use is the meansigma methods of these three points.Standard deviation is represented with the error post.
[0569] It is suitable to producing 11-that the mRPE65 of 11-cis-retinol mediation takes off palmitoylation The influence of formula-retinol:
In (100mM Tris pH8.0,80 μ g albumen) buffering suspension of 1mL RPE film, add 10 μ M11-cis-retinol, at room temperature incubation 45min.Simultaneously at room temperature incubation does not contain the control reaction mixture 45min of 11-cis-retinol.When the 45min incubation finishes, make reactant mixture be exposed to UV light (354nm), keep 10min, destroy 11-cis-retinoid.Then with alltrans-retinol [11-12- 3H 2] (0.1 μ M) adding reactant mixture (100mM Tris pH8.0,80 μ g RPE albumen 5%BSA and 0.1 μ M alltrans-retinol [11-12- 3H 2], at 37 ℃ of following incubations.0,5,10,15,20,30,45,60,90,120 and 150min after, add 500 μ L methanol successively, 100 μ LH2O quenchers, 100 μ L react sample aliquot, add 500 μ l hexanes (containing the 1mg/mL butylated hydroxytoluene) and extract retinoid.Press preceding method analysis classes visual pigment (Winston and Rando 1998).The amount of the 11-cis-retinol that forms is as the active tolerance of IMH.All experiments are done three parts, with the meansigma methods analysis of these measurements.
[0570] A. blocking RPE65 combines with retinyl ester
[0571] alltrans-tretinoin (atRA), 13-cis-tretinoin (13cRA) and N-(4-hydroxy phenyl) look yellow the influence of amide (4-HPR) to IMH:
In (100mM Tris pH8.0,76.7 μ g albumen) buffering suspension of 1mL RPE film, add 60 μ M or 6 μ M atRA, 13cRA or 4-HPR, at room temperature incubation 15min.At room temperature simultaneously, incubation does not contain the control reaction mixture 15min of any inhibitor.When the 15min incubation finishes, with alltrans-retinol [11-12- 3H 2] (0.2 μ M) adding reactant mixture (100mM Tris pH8.0,76.7 μ g RPE albumen, 0.2%BSAl00 μ MDPPC, 1mM DTT and 0.2 μ M alltrans-retinol [11-12- 3H 2]) in, incubation 30min at room temperature.When incubation finished in 30 minutes, quencher reaction sample aliquot, check equivalent adding alltrans-retinol [11,12- 3H 2] and these inhibitor to the influence of LRAT.Afterwards, with control mixture and atRA (60﹠amp; 6 μ M), 13cRA (60﹠amp; 6 μ M) or 4-HPR (60﹠amp; 6 μ M) incubation 15min.Immediately with all reactant mixtures and 30 μ M apo-rCRALBP (100mM Tris pH8.0,7.7 μ g RPE albumen, 0.2%BSA100 μ M DPPC, 1mM DTT30 μ M apo-rCRALBP and 0.2 μ M alltrans-retinol [11-12- 3H 2]) 37 ℃ of following incubations 30 minutes.When this incubation period finishes, add the freezing methanol quencher 200 μ L reactant mixtures of 750 μ l, add 100 μ L 1M sodium chloride solutions subsequently, add 500 μ L hexanes (containing the 1mg/mL butylated hydroxytoluene) and extract retinoid.By aforementioned (27) analysis classes visual pigment.11-cis-retinol the amount that forms is as the active tolerance of IMH.All experiments are done three parts, with the meansigma methods analysis of these measurements.
[0572] Fig. 4 A, B, C show alltrans-tretinoin and the bonded data of purification RPE65 specificity.Shown in Fig. 4 A, alltrans-tretinoin combines with RPE65 and causes albumen fluorescence to be exponential damping.This decay is followed saturability in conjunction with isothermal line (Fig. 4 B), produces on average in conjunction with K DAbout 109nM (SD=10nM, N=4) (Fig. 4 C).Similar data to 13-cis-tretinoin are listed in Fig. 5 A, B, C, obtain on average in conjunction with K DAbout 195nM (SD=20nM, N=4) (Fig. 5 C).These experimental results show that with the bonded two kinds of tretinoins of RPE65 specificity all have high-affinity.By contrast, under the identical combination condition, Palmic acid alltrans-retinyl ester combines with RPE65, K D=47nM (not produce data).For further estimating binding specificity, to another kind xanthoplane, N-(4-hydroxy phenyl) looks yellow amide (fenretinide):
Study with the binding interactions of RPE65.Estimate that N-(4-hydroxy phenyl) looks yellow amide and RPE65 binding ratio tretinoin is weak, because look yellow the only faint nyctalopia that causes of analog in the amide series.In fact, a little less than N-(4-hydroxy phenyl) looks yellow amide and combines quite with RPE65.Data shown in Fig. 6 A, B, the C obtain on average in conjunction with K DAbout 3547nM (SD=280nM, N=4) (Fig. 6 C).Therefore, it is weak in conjunction with as the hypothesis of expection that the N-that observes (4-hydroxy phenyl) looks yellow amide, and RPE65 is the nyctalopia target.
[0573] B. tretinoin is replaced the Palmic acid alltrans-retinyl ester among the RPE65
[0574] tretinoin (alltrans and 13-cis) and Palmic acid alltrans-retinyl ester is competed in conjunction with RPE65:
Carry out the buffer-exchanged experiment, with the ability of research tretinoin (alltrans and 13-cis) displacement with the bonded Palmic acid alltrans-retinyl ester of RPE65.(pH7.4) the middle 6 μ M tretinoins (alltrans and 13-cis) that add are at 4 ℃ of following incubation 30min for PBS, 1%CHAPS to RPE65 (0.5 μ M).With the RPE65 control sample of retinoic acid-containing not at 4 ℃ of following incubation 30min.After this incubation finishes, with sample and 3H-alltrans-retinyl cetylate (0.65 μ M, 20.31Ci/mmol) incubation 30min.After this incubation period finishes, with Centricon 30KMWCO filter exchange buffering liquid (PBS-1%CHAPS) 10 4Doubly.Sample that record is held back on liquid scintillation counter and buffering liquid stream branch, measurement is held back 3The amount of H-alltrans-retinyl cetylate.
[0575] tretinoin of above report directly in conjunction with research can't determine these molecules whether and alltrans-retinyl ester competition in conjunction with the physiology associated ligands of RPE65.Can pass through 3H-alltrans-retinyl cetylate confirms this problem easily in conjunction with RPE65 in advance, confirms that alltrans-tretinoin competition is in conjunction with (Fig. 7).By earlier with RPE65 and alltrans-tretinoin or 13-cis-tretinoin and (-) tretinoin (contrast) incubation, then with albumen and 3H-alltrans-retinyl cetylate incubation 30min carries out this experiment.Remove excessive retinoid by buffer-exchanged, with liquid scintillation counter record flow point that passes through and the solution that dams.Tables of data photopic vision yellow acid and alltrans-retinyl ester are obviously competed in conjunction with the same binding site on the RPE65.
[0576] C. tretinoin suppresses the RPE65 function
[0577] RPE65 is the chaperone of alltrans-retinyl ester, and is most important to the isomerized transfer of these hydrophobic molecules equally.In present research, with the alltrans-retinol (vitamin A) of bovine retina pigment epithelium cell film system processing adding, to form 11-cis-retinol.Because RPE65 is most important for biosynthesis 11-cis-retinol (4,8,11), its inhibitor is capable of blocking should be synthetic.In experiment shown in Figure 7, alltrans-retinyl ester is enough to make this complex residual in the Centricon column spinner is centrifugal in conjunction with RPE65 stall (off-rate).Alltrans-tretinoin is suitable for (not produce data) equally.This prompting estimates that the inferior ordered pair announcement of incubation RPE65 inhibitor effectively suppresses very important.The precincubation of these films and vitamin A produces alltrans-retinyl ester fast by the quick esterification of LRAT mediation.Synthetic alltrans-retinyl ester and RPE65 strong bonded are processed as 11-cis-retinol by IMH then.60 μ M (unlisted data) alltrans or 13-cis-tretinoin that this system is not subject to incubation suppress.This is the result who estimates, because known tretinoin does not directly suppress IMH.But as shown in Figure 8, produce visibly different result with tretinoin precincubation.In this example, in the presence of tretinoin, the basic inhibition that 11-cis-retinol forms appears, because they enter RPE65.Be enjoyably, observe with N-(4-hydroxy phenyl) and look yellow amide inhibition, prove it is more weak (Fig. 8) basically.This is the result who expects because it to the affinity of RPE65 quite a little less than.
[0578] D. pass through the sRPE65 Stereoselective in conjunction with vitamin A
[0579] as mentioned above, relevant with RPE65 (mRPE65) film combines with Palmic acid alltrans-retinyl ester Stereoselective.In this embodiment, measure retinoid in conjunction with sRPE65 (Gollapalli by the fluorescent method of having set forth, D.R., Maiti, P., Rando, R.R. (2003) RPE65 operates in the vertebrate visual cycle by stereospecificallybinding all-reans-retinyl esters (RPE65 works in conjunction with alltrans-retinyl ester by Stereoselective in the vertebrates visual cycle) .Biochemistry42,11824-30.).Excitation wavelength is 280nm, observes emission and passes the 0.5cm solution layer.Volumetric soiutions is made up of 100mM phosphate buffered salt solution (150mM) pH7.4 and the 1%CHAPS of 0.37 μ MsRPE65.At Fig. 9 A, among the B, show alltrans-retinol (tROL) and Palmic acid alltrans-retinyl ester data in conjunction with purification sRPE65.Fig. 9 A1 shows the emission spectra of the sRPE65 that increases with tROL concentration.Fig. 9 A2 shows titrating linearity square match (squarefit) curve of sRPE65 to tROL.Shown in Fig. 9 A, alltrans-retinol causes albumen fluorescence to be exponential damping in conjunction with sRPE65, and this decay is followed saturated in conjunction with isothermal line, obtains on average in conjunction with K D(Fig. 9 D) about 65nM (Fig. 9 A2).In Fig. 9 B, shown in Palmic acid alltrans-retinyl ester (tRP) have similarity index decay in the albumen fluorescence in conjunction with the data of sRPE65.Fig. 9 B1 represents that the emission spectra of sRPE65 increases with tRP concentration.Fig. 9 B2 shows the titrating linearity square matched curve of sRPE65 to tRP.This decay is followed saturated in conjunction with isothermal line, obtains on average in conjunction with K D(Fig. 9 D) about 1.2 μ M (Fig. 9 B2).In containing the 100mM phosphate buffered solution of 150mM sodium chloride, tROL and rRP and mRPE65 are summarized among Fig. 9 D with the binding constant that contains the sRPE65 of 1%CHAPS.
[0580] E. is when forming alltrans-retinyl ester, and sRPE65 is as the vitamin A chaperone.
[0581] ability of being processed by LRAT by the explanation vitamin shows that the vitamin A in conjunction with sRPE65 is an active metabolite, the only enzyme that becomes known for vitamin A processing metabolic pathway in RPE of LRAT representative.Shown in Fig. 9 C, be the good substrate of the LRAT (tLRAT) of truncate in conjunction with the vitamin A of sRPE65, the LRAT of truncate is the LRAT form that is easy to express, it mechanically (mechanistically) distinguish with LRAT.These studies have shown that the certain bootable vitamin A of sRPE65 to LRAT, thereby vitamin A may have functional importance in conjunction with sRPE65.When not having sRPE65, seldom synthetic Palmic acid alltrans-retinyl ester (Fig. 9 C).Report as Fig. 9 C, in the presence of (1) sRPE65 (0.04 μ M), Lauryl.beta.-maltoside (0.1%) and DPPC (200 μ M), produce tRP, but under (2) Lauryl.beta.-maltoside (0.1%) and DPPC (200 μ M) or (3) sRPE65 (0.04 μ M) individualism, do not produce tRP.All reactant mixtures contain 100mM Tris pH8.4, the second-rate threitol of 1mM, 1mM EDTA, 5 μ M tLRAT and 0.2 μ M tROL.
[0582] The F.sRPE65 palmitoylation
[0583] biochemical relationship between mRPE65 and the sRPE65 is studied the influence of their hydrophobicity post translational modification state.If process is reversible, the seemingly most possible property of S-palmitoylation.By existing with rHRPE65 baculovirus transfection insect cell (sf21) 3H 2Grow in-the Palmic acid, directly test this situation and the mRPE65 that measure to express whether labelling (Ma, J., Zhang with standard mode, J., Othersen, K.L., Moiseyev, G., Ablonczy, Z., Redmond, T.M., Chen, Y., Crouch, R.K. (2001) Expression, purification, and MALDI ahalysis of RPE65 (expression, purification and maldi analysis RPE65) .Invest Ophthalmol Vis Sci.42,1429-35).Figure 10 A shows and exists respectively 3H 2Under the existence of Palmic acid and unlabelled Palmic acid, palmitoylation in the rHmRPE65 body of in the sf21 cell, expressing.L1-4 represents coomassie dyeing gel, and L5-6 represents the autoradiograph of L1-4, and L8 represents the Western blotting of rHmRPE65.In little block diagram A, (L1) expression 14The C molecular weight marker; (L2) expression has in the presence of unlabelled Palmic acid (0.09 μ M) contrast of the purification rHmRPE65 that expresses in the sf21 cell of growth; (L3) expression wherein exists 3H 2Under the existence of Palmic acid (0.09 μ M-0.5mCi/mL), handling the purification rHmRPE65 that expresses in the sf21 cell of 16h with 0.5M Tris pH8.0; (L4) be illustrated in 3H 2Palmic acid (0.09 μ M-0.5mCi/mL) exists down, handles the purification rHmRPE65 that expresses in the sf21 cell of 16h with 0.5M azanol pH8.0 then; The autoradiograph of (L5, L6 and L7) expression L2, L3 and L4.L7 represents the Western blotting with the purification rHmRPE65 of nascent antibody (1: 4000-1h, the room temperature) detection of anti-RPE65.
[0584] shown in Figure 10 A, by adding 3The H-Palmic acid is at the purification mRPE65 of expressed in insect cells labelling really.As expected, the labelling mRPE65 release mark of handling with the azanol that decomposes thioester.For determining to modify in the body position of mRPE65, purification cattle mRPE65 and sRPE65 are carried out the mass spectrum experiment.These sample trypsinizations, and experience mass spectral analysis (Figure 10 B and C).The result show mRPE65 at position C231, C329 and C330 by triple palmitoylations.As if by contrast, data also show sRPE65 palmitoylation not.
[0585] Figure 10 B and C represent the mass spectral analysis of mRPE65 and two kinds of different peptides of sRPE65.Analyze the RPE65 peptide of trypsinization with MALDI-TOF.The peak note is as follows: Figure 10 B; 1378.9Da (aminoacid sequence 223-234; SEIVVQFPCSDR), 1429.4Da (1-14; N-acetyl group-SSQVEHPAGGYKK), 1477-4Da (34-44; IPLWLTGSLLR), 1483.0 Da (114-124; NIFSRFFSYFR), 1616.6Da (223-234, SEIVVQFPC *SDR), 1700.1Da (83-96, FIRTDAYVRAMTEK), 1701.7 (367-381, RYVLPLNID), 1718.7 (83-96, FIRTDAYVRAM#TEK).Figure 10 C, 2770.3 (333-354, GFEFVYNYSYLANLRENWEEVK), 3321.6 (306-332, TSPFNLFHHINTYEDHEFLIVDLCCWK), 3797.8 (306-332, TSPFNLFHHINTYEDHEFLIVDLC *C *WK).C *Expression palmitoylation cysteine, M# represents the oxidation methionine.
[0586] G.mRPE65 and sRPE65 are by the mutual conversion of LRAT
[0587], be how to transform mutually so must understand these two kinds of molecules because mRPE65 and sRPE65 show complementary retinoid binding specificity.What is interesting is most that LRAT can utilize mRPE65 as palmityl donor (Figure 11 A), and this part is transported to vitamin A, produce Palmic acid alltrans-retinyl ester (Figure 11 B, C, D).Figure 11 B represents to rely on the independent mRPE65 (■-) of alltrans-retinol esterification and DPPC (●-) separately.In these experiments,, separate the Palmic acid alltrans-retinyl ester that obtains by HPLC with pure mRPE65, vitamin A and LRAT incubation together.Isolating Palmic acid alltrans-retinyl ester mass spectral analysis shows that it is genuine, so the palmityl part is transported to vitamin A from mRPE65.These data show that also mRPE65 is the efficient palmityl donor more much higher than dipalmitoyl phosphatidyl choline (DPPC), and in the LRAT reaction, DPPC is the standard acry radical donor.Under the same conditions, do not measure tangible DPPC and upgrade (Figure 11 B).Kinetic curve shows sigmoid curve kinetics shown in Figure 11 B, the K of the mRPE65 of calculating M(Kapp) be 0.03 μ M (compare, the value of DPPC is 1.4 μ M).The value that Hill curve (Figure 11 C) provides N is 2.54, points out a more than mRPE65 molecule to participate in the transhipment of palmityl.The sigmoid curve kinetics that observes is pointed out the regulating action of this process, and its slight variation to mRPE65 concentration is reacted.
[0588] determines the reaction reversibility easily.In these experiments (Figure 11 D), with excessive alltrans-retinol with mRPE65 and tLRAT incubation until no longer producing Palmic acid alltrans-retinyl ester.Use subsequently 3The trans retinol of H-is handled.The activity specific of Palmic acid alltrans-retinyl ester raises and in constant Palmic acid alltrans-retinyl ester level, alltrans-retinol activity specific descends and shows the balance (Figure 11 D) of substrate subsequently.Figure 11 D represents that it is the function of time that the activity specific of tRP (■-) and tROL (●-) changes (left y-axle).The total retinyl ester that forms (▲-) (right y-axle) expression ester synthesis reaction saturated.Every kind of reactant contains 100mM Tris pH8.4,0.06 μ M mRPE65,5 μ M tLRAT, the second-rate threitol of 1mM, 1mM EDTA and 10 μ tROL.
[0589] when mRPE65 handled with tLRAT and vitamin A, the palmitoylation RPE65 that takes off of formation showed that retinoid is similar to sRPE65 in conjunction with behavior.With mRPE65 and excessive vitamin A and tLRAT incubation together.After removing retinoid and tLRAT, study sRPE65 and vitamin A and the bonded ability of Palmic acid alltrans-retinyl ester then.Shown in Fig. 2 D, handle mRPE65 with LRAT and vitamin A, mRPE65 is converted into the RPE65 of function combining form, can't offer an explanation with isolating sRPE65 form.
[0590] H. make 11-cis-retinol palmitoylation by mRPE65 and LRAT
[0591] the direct product of IMH effect, 11-cis-retinol also esterified (Figure 12 A).Shown in Figure 12 B, during the retinoid esterification (palmitoylation) of tLRAT/mRPE65 mediation, to compare with vitamin A, 11-cis-retinol is actually better substrate.(1) 11-cis-retinol (2 μ M) and mRPE65 (0.02 μ M).(2) alltrans-retinol (2 μ M) and mRPE65 (0.02 μ M).(3) 11-cis-retinol (2 μ M) and DPPC/BSA (250 μ M/0.4%).(4) alltrans-retinol (2 μ M) and DPPC/BSA (250 μ M/0.4%).All reactant mixtures all contain 100mM Tris pH8.4, the second-rate threitol of 1mM, 1mM EDTA and 5 μ M tLRAT.
[0592] makes 11-cis-retinol palmitoylation by mRPE65; nature mechanism is provided, between the visual cycle on-stream period, upgrades and enforcement control by this mechanism mRPE65; because 11-cis-retinol orders about mRPE65 to sRPE65, effectively close and lead to the biosynthetic approach of chromophore.This directly obtains proof in Figure 12 C, wherein handle the RPE film with 10 μ M11-cis-retinol, orders about mRPE65 and changes to sRPE65.The figure shows the mediation of 11-cis-retinol take off palmitoylation (●-) exist down and do not exist 11-cis-retinol mediation take off palmitoylation (■-) time, rely on [the 11-12-of time 3H 2] 11-cis-retinol generation.Illustration (inset) is represented whole time range.The irradiation under ultraviolet ray sample can destroy 11-cis-retinoid.Then, the sample that will contrast and handle is measured the speed of product 11-cis-retinol of IMH with the vitamin A incubation respectively.With the pretreated sample of 11-cis-retinol, the period of delay (lag period) of performance uniqueness before the synthetic generation of product.
[0593] I. exemplary RPE65 antagonist
[0594] measures the affinity constant (K of several chemical compounds to mice RPE65 αS).The Kd that finds above-mentioned 4a, 4b and 4c chemical compound is respectively 47nM, 235nM and 1300nM.Therefore, adhesion is the function of ester chain length, and promptly chain length is long more, and is strong more and antagonist is strong more to the affinity of RPE65.
[0595] above-listed 4d, 4e and 4f chemical compound also are effective RPE65 antagonisies, K dBe respectively 21nM, 40nM and 64nM.
[0596] The list of references of quoting
[0597] Anantharaman, V., Aravind, L. (2003) .Evolutionary history, structural features and biochemical diversity of the NIpC/P60 superfamilyof enzymes (evolutionary history of NIpC/P60 superfamily enzyme, architectural feature and biochemical multiformity) .Gen.Biol.4, R11.
[0598] Arshavsky, V.Y., Lamb, T.D., and Pugh EN Jr. (2002) Gproteins and phototransduction (G albumen and light transduction) .Annu.Rev.Physiol.64,153-187.
[0599] Barry, R.J., Canada, F.J., and Rando, R.R. (1989) the Solubilization and partial purification of retinyl ester synthetase andretinoid isomerase from bovine ocular pigment epithelium retinyl ester synzyme and the retinoid isomerization of partial purification buphthalmos pigment epithelium cell (dissolving and) .J.Biol.Chem.264,9231-9238.
[0600] Bavik, C.O., Levy, F., Hellman, U., Wemstedt, C., andEriksson, U. (1993) .The retinal pigment epithelial membrane receptor forplasma retinol-binding protein.Isolation and cDNA cloning of the 63-kDa protein (the retinal pigment epithelium membrane receptor of blood plasma retinol binding protein.Separate and cDNA clone 63-kDa albumen) .J.Biol.Chem.26f, 20540-20546.
[0601] Bernstein, P.S., Law, W.C., and Rando, R.R. (1987) .Isomerization of all-trans-retinoids to 11-cis-retinoids in vitro (alltrans retinoid isomery turns to 11-cis-retinoid) .Proc.Natl.Acad.Sci. (USA) 84,1849-1853.
[0602] Bijlmakers, M.J., Marsh, M., (2003) .The on-off story ofprotein palmitoylation (switching process of albumen palmitoylation) .Trends.Cell.Biol.13.32-42.
[0603] Bok, D. (1993) .The retinal pigment epithelium:a versatilepartner in vision (retinal pigment epithelium: the general companion in the vision) .J.Cell.Sci.Suppl.17,189-195.
[0604] Bok, D., Ruiz; A., Yaron, O.; Jahng, W.J., Ray; A.; Xue, L., Rando; R.R. (2003) .Purification and characterization of atransmembrane domain-deleted form of lecithin retinol acyltransferase (purification and characterize the lecithin retinol acyltransferase of membrane spaning domain-disappearance form) .Biochemistry42,6090-8.
[0605] Bridges, C.D.B. (1976) Vitamin A and the role of thepigment epithelium during bleaching and regeneRation of rhodopsin in thefrog eye is (at bleaching of frogeye rhodopsin and regeneration period, the effect of vitamin A and pigment epithelium cell) .Exp.Eye.Res.22,435-455.
[0606] F.J., Law, W.C.and Rando, R.R. (1990) .Substratespecificities and mechanism in the enzymatic processing of vitamin Ainto 11-cis-retinol (substrate specificity and enzymatic processing vitamin A is 11-cis-retinol mechanism) .Biochemistry29,9690-9697.
[0607] Chen, C.A.and Manning D.R. (2001) .Regulation of Gproteins by covalent modification (regulating G albumen) .Oncogene20.1643-1652. by covalent modification
[0608] Deignet, P.S., Law, W.C., F.J., and Rando, R.R. (1989) .Membranes as the energy source in the endergonic transformationof vitamin A to 11-cis-retinol (is to make the energy with film in 11-cis-retinol in the vitamin A endothermic disintergration) .Science244,968-971.
[0609] Dunphy, J.T.and Linder, M.E. (1998) .Signaling functionsof protein palmitoylation (semiotic function of albumen palmitoylation) .Biochim BiophysActa.1436,245-261.
[0610] E1-Husseini; A.E.and Bredt D.S. (2002) ProteinPalmitoylation:A regulator of neuronal developmental and function (albumen palmitoylation: the .Nat.ReV.3 regulator of neuronal development and function), 791-802.
[0611] Gollapalli, D.R., Rando R.R. (2003) .All-trans-Retinylesters are the Substrates for Isomerization in the Vertebrate VisualCycle (alltrans-retinyl ester is an isomerized substrate in the vertebrates visual cycle) .Biochemistry42,5809-18.
[0612] Gollapalli, D.R., Maiti, P., Rando, R.R. (2003) RPE65operates in the vertebrate visual cycle by stereospecifically binding all-trans-retinyl esters (RPE65 works in conjunction with alltrans-retinyl ester by Stereoselective in the vertebrates visual cycle) .Biochemistry 42,11824-30.
[0613] Gonzalez-Fernandez, F. (2002) .Evolution of the visualcycle:the role of retinoid-binding proteins (evolution of visual cycle: the protein-bonded effect of retinoid) .J.Endocrinol.175,75-88.
[0614] Hamel, C.P., Tsilou, E., Pfeffer, B.A., Hooks, J.J., Detrick, B., and Redmond, T.M. (1993) .Molecular cloning and expression ofRPE65, a novel retinal pigment epithelium-specific microsomal proteinthat is post-transcriptionally regulated in vitro.J.Biol.Chem (molecular cloning and expression RPE65, the retinal pigment epithelium specific particulate body protein of regulating behind a kind of new in vitro transcription) .268,15751-15757.
[0615] Hanna-Rose, W.and Han, M. (2002) The Caenorhabditiselegans EGL-26 protein mediates vulval cell morphogenesis (Caenorhabditis elegans EGL-26 protein mediated pudendum cellular morphology take place) .Dev.Biol.241,247-258.
[0616] Houslay; M.D; (1996) Regulation of cellular signaling byfatty acid acylation and prenylation of signal transduction proteins (regulating cell signal) .Cell.Signal.8 by fatty acid acidylate and signal transducer isoprenylation, 403-412.
[0617] Jahng; W.J.; David, C., Nesnas; N.; Nakanishi, K., andRando; R.R. (2003) .A cleavable afifnity biotinylating agent reveals aretinoid binding role for RPE65 (can decompose the retinoid combination that the biological acylating agent of affinity shows RPE65) .Biochemistry42,6159-6168.
[0618] Jahng; W.J.; Xue; L.; Rando; R.R. (2003) .Lecithin retinolacyltransferase is a founder member of a novel family of enzymes (the lecithin retinol acyltransferase is that rising of new enzyme family makes the member) .Biochemistry42,12805-12.
[0619] Kammer; B.; Schmidt; M.F.G.; Veit; M. (2003) Functionalcharacterization of palmitoylation and nonacylated SNAP-25 purifiedfrom insect cells infected with recombinant baculovirus (SNAP-25 at the insect cell matter purification that infects recombinant baculovirus of functional sign palmitoylation and on-acylated) .Mol.Cell.Neuroscience.23,333-340.
[0620] Linder, M.E.and Deschenes, R.J. (2003) .New insights intothe mechanisms of protein palmitoylation (to the new visual field of Protein Palmitoylation mechanism) .Biochemistry42,4311-4320.
[0621] Lowry, O.H., Rosebrough, N.J., Farr, A.L.and Randall, R.J. (1951) .Protein measurement with the Folin phenol reagent (measuring albumen) .J.Biol.Chem.193 with forint-phenol reagent, 265-275.
[0622] Ma, J., Zhang, J., Othersen, K.L., Moiseyev, G., Ablonczy, Z., Redmond, T.M., Chen, Y., Crouch, R.K. (2001) Expression, purification, and MALDI analysis of RPE65 (expression, purification and maldi analysis RPE65) .Invest Ophthalmol Vis Sci.42,1429-35.
[0623] Mata, N.L., Moghrabi, W.N., Lee, J.S., Bui, T.V., Radu, R.A., Horwitz, J., Travis, G.H. (2004) .Rpe65is a retinyl-ester bindingprotein that presents insoluble substrate to the isomerase in retinalpigment epithelial cells (Rpe65 be representative conjugated protein to the retinyl ester of the insoluble substrate of isomerase in retinal pigment epithelium) .J Biol.Chem.279,635-43.
[0624] Milligan; G.Parenti; M.and Magee, A.I. (1995) .TheDynamic role of palmitoylation in signal transduction (kinetics function of palmitoylation in signal transduction) .Trends Biochem.Sci.20,181-187.
[0625] Morello JP; Bouvier M. (1996) .Palmitoylation:a post-translational modification that regulates signaling from G-protein coupledreceptors (palmitoylation: regulate the post translational modification of the signal of g protein coupled receptor) .Biochem.Cell Biol.74,449-457.
[0626] Mumby, S.M., (1997) .Reversible palmitoylation ofsignaling proteins (making signal protein reversibility palmitoylation) .Curr.Opin.Cell Biol.9,148-154.
[0627] Patterson; P.L. (2002) .Posttranslational protein S-palmitoylation and the compartmentalization of signaling molecules inneurons (signaling molecule in posttranslational protein S-palmitoylation and the compartmentation neuron) .BiolRes.35,139-50.
[0628] Qanbar; R; Bouvier; M. (2003) .Role ofpalmitoylation/depalmitoylation reactions in G-protein-coupled receptorfunction is (in G-albumen-coupled receptor function; the effect of palmitoylation/take off palmitoylation reaction) .Pharmacol.Ther.97,1-33.
[0629] Rando, R.R.and Bangerter, F.W. (1982) The Rapidintermembraneou-s transfer of retinoid (intermembranous retinoid transfer) .Biochem.Biophys.Res.Commun.104,430-436.
[0630] Rando, R.R. (1990) The chemistry of vitamin A and thevisual process (chemistry of vitamin A and vision process) .Angew.Chemie (Int.Ed.Engl.) 29,461-480.
[0631] Rando, R.R. (1991) Membrane phospholipids as an energysource in the operation of the visual cycle (in service in visual cycle, as to make the energy) .Biochemistry30,595-602. with membrane phospholipid
[0632] Rando, R.R. (2001) The biochemistry of the visual cycle (biochemistry of visual cycle) .Chem.Rev101,1881-1896.
[0633]Redmond，T.M.，Yu，S.，Lee，E.，Bok，D.，Hamasalci，D.，Chen，N.，Goletz，P.，Ma，J.X.，Crouch，R.K.，and?Pfeifer，K.(1998)Nat.Genet.20，344-351.
[0634] Resh; M.D. (1999) .Fatty acylation of proteins:new insightsinto mernbrane targeting of myristoylated and palmitoylated proteins (fatty acylated protein: the new visual field of the proteic film target of Semen Myristicae acidylate and palmitoylation) .Biochi m.Biophys.Acta.1451,1-16.
[0635] Saari, J.C. (2000) .Biochemistry of visual pigmentregeneration:the Friedenwald lecture (biochemistry of visual pigment regeneration: the Friedenwald lecture) .Invest.Ophth.Vis.Sci.41,337-348.
[0636] Saari, J.C., Bredberg, L., and Farrell, D.F. (1993) Retinolesterification in bovine retinal pigment epithelium; Reversibility ofLecithin:retinol acyltransferase (carries out the retinol esterification: the reversibility of lecithin: .Biochem.J.291 retinol acyltransferase), 697-700. in the bovine retina pigment epithelium cell
[0637]Segal，I.H.(1993).Enzyme?Kinetics.(New?York；Wiley-Interscience).
[0638] Shi, Y.-Q., Furuyoshi; S., Hubacek, T.; and Rando, R.R. (1993) .Affinity labeling of lecithin retinol acyltransferase (the affinity labelling of lecithin retinol acyltransferase) .Biochemistry32,3077-3080.
[0639] Stacie, R.L., Yuechueng L. (1997) .Characterization of thePalmitoylation Domain of SNAP-25 (characterizing SNAP-25 palmitoylation domain) .J.Neurochem.69,1864-1869.
[0640] Stryer L. (1986) Cyclic GMP cascade of vision (the cyclo GMP cascade of vision) .Annu.Rev.Neurosci.9,87-119.
[0641] Thompson, D.A.and Gal A. (2003) .Genetic defects invitamin A metabolism of the retinal pigment epithelium (genetic defect in the retinal pigment epithelium in the Vitamin A Metabolism) .Dev.Ophthalmol.37,141-54.
[0642] Trehan, A., Canada, F.J.and Rando, R.R. (1990) .Inhibitorsof retinyl ester formation also prevent the biosynthesis of 11-cis-retinol (retinyl ester inhibitor form also stop biosynthesis 11-cis-retinol) .Biochemistry29,309-312
[0643] von Lintig, J., Vogt, K. (2000) .Filling the gap in vitamin Aresearch.Molecular identification of an enzyme cleaving beta-carotene toretinal (remedies the blank in the vitamin A research.It is retinal that the Molecular Identification enzyme decomposes beta-carotene) .J.Biol.Chem.275,11915-11920.
[0644]Waid，G.The?biochemistry?of?vision(1953)Annu.Rev.Biochem.22，497-526.
[0645] West, K.A., Yan, L., Shadrach, K., Sun, J., Hasan, A., Miyagi, M., Crabb, J.S., Hollyfield, J.G., Marmorstein, A.D., Crabb, J.W. (2003) .Protein database, human retinal pigment epithelium (albumen database, human retina pigment epithelium) .Mol.Cell.Proteomics.2,37-49.
[0646] Winston, A.and Rando, R.R. (1998) .Regulation ofIsomerohydrolase Activity in the Visual Cycle (regulating isomery hydrolytic enzyme activities in the visual cycle) .Biochemistry37,2044-2050.
[0647] Wolf, G. (1996) .The regulation of retinoic acid formation (regulating tretinoin forms) .Nutr.Rev.54,182-l84.
[0648] Embodiment 2: the short circuit medicine is in vivo to the effect of visual cycle
[0649] presses 50mg/kg dosage, give and inject (i.p.) in the mouse peritoneum at 25 μ l DMSO The compound of enumerating of middle preparation. Press 50mg/kg dosage, give positive control injected in mice 13-25 μ l DMSO solution of cis-retinoic acid (ACCUTANE ). Give the negative control mouse Inject 25 μ l DMSO.
[0650] after the administration, makes in the given time mouse be exposed to the substantial light photograph, caused Full bleaching (bleaching) visual cycle. Then, in Mingguang City or half-light, carry out retinal current Figure (Electroretinograms) (ERG) measures the b-wave amplitude. The b-wave amplitude present with visual purple again The generation ratio, thereby relevant with the visual cycle function (be that the b-wave amplitude is more high, the visual cycle function More strong).
[0651] A.4-butyl-aniline and 3-benzocaine
[0652] the DMSO solution of preparation 4-butyl-aniline and 3-benzocaine. Give 7 monthly age wild type (wt; C57BL/6J X 129/SV; Rpe65 Leu450Leu) mouse i.p. Inject 25 each compound of μ l (50mg/kg). Use respectively ACCUTANE (13-cis-look Huang Acid, 25 μ l, 50mg/kg) and DMSO (be labeled as wt; 25 μ l) animal of injection do positive and Negative control. Two mouse of every group of injection. Carrying out ERG measures.
[0653] Figure 14 A represents the effect of compound injection after 1 hour. The wild type feminine gender is right Return to 50% baseline b-wave amplitude (with recovering to compare fully) according to showing, and positive control and survey The examination compound shows that the visual cycle infringement increases the weight of.
[0654] Figure 14 B represents the effect after 1 week (7 days) behind the compound injection. Testization Compound has continuous action, and positive control returns fully recovery.
[0655] Figure 14 C represents the effect in sky of rear 2 weeks (14) of injection. Test compounds is still right Visual cycle has effect.
[0656] B ethyl-(3-N-methyl) Aminobenzoate, N-methyl-4-butylaniline
[0657] Figure 15 A-B, 16A-B and 17A-B represent that respectively these compounds are at half-light (A) and three experiments in Mingguang City (B).
[0658] C. ethyl-(2-N-methyl) Aminobenzoate, N-methyl-4-butylaniline
[0659] Figure 18 A-B represents with these compounds in half-light (A) and Mingguang City (B) The experiment of carrying out.
[0660] Embodiment 3: enzyme inhibitor and RPE65 antagonist are in vivo to visual cycle Effect
[0661] other compound is repeated the experiment described in the embodiment 2.
[0662] A. retinyl palmityl ketone and retinyl decyl ketone
[0663] Figure 19 represents the experiment carried out with these compounds.
[0664] B. alltrans-retinyl palmityl ketone, alltrans-retinyl palmityl Base ether
[0665] Figure 20 A-B and 21A-B represent respectively with these compounds half-light (A) and Two experiments in Mingguang City (B).
[0666] C. octyl group method acid amides (famestimide), palmityl method acid amides
[0667] Figure 22 A represents to give behind these compounds soon, the experiment knot in half-light Really. Figure 22 B represents the result after one week of administration.
[0668] E. farnesyl-octyl group ketone, farnesyl-decyl ketone
[0669] Figure 23 A-C represents the experiment carried out with these compounds in half-light. Administration Obtain the data shown in Figure 23 B after 3 days. Administration obtains the data among Figure 23 C after 8 days.
[0670] F. farnesyl-palmityl ketone, farnesyl-decyl ketone
[0671] Figure 24 represented to inject these compounds after 1 hour, carried out in half-light The result of experiment.
[0672] embodiment 4: form A in the presence of arylamine2E。
[0673] 100mg (355 μ mol) alltrans-retinene is dissolved among the 3mL, subsequently Add 9.5 μ L glacial acetic acids, 9.5 μ L monoethanolamines (155 μ mol). From this solution, take out 300 μ L Sample fraction. Respectively at 0,2,3.75,14,16.25,18.0833 19.917 23.75 and During 48h, benzoic acid 3-amino ethyl ester (15.5 μ mol) is added sample. Control sample does not comprise Arylamine. Solution is at room temperature vibrated 48h. Then, at-80 ℃ of lower solution of preserving, with 15 μ L Be diluted to 250 μ L (methyl alcohol). Then, at the anti-phase (post of C18-5 μ m-4.6mm * 150mm) On, in the linear gradient 85%-96% methanol/water, analyze with the UV detector at the 430nm place Solution (15 μ L) is measured A2The amount that E forms. With A2A when E forms % and do not have no arylamine 48h2The amount contrast that E forms.
[0674] table 1:A2E forms
Time (hour) % % forms (48 hours=100) TG-AUC
 0    3.98     422.5
 2    8.67     920
 3.75    7.71     818.5
 14    19.73     2094
 16.25    19.25     2043
 18.0833    23.70     2515
 19.91667    27.65     2934
 23.75    22.44     2382
 48    100.00     10613
[0675] lists table 1 data among Figure 25, show that arylamine can not form A in the standard vitro reactions2E; In addition, they stop A2E forms.
[0676] embodiment 5: A in the body2E forms
[0677] research farnesyl-decyl ketone and N-palmityl method acid amides are to rejecting A in the mouse at abcr2The impact that E accumulates. Press 50mg/kg dosage, weekly or give little twice 25 μ l DMSO solution of mouse injectable drug or only inject DMSO for control group. 2-2.5 month After, put to death mouse, results A2E and different-A2E, for every kind of medicine or contrast, the A of quantitative 4 eyes2E and different-A2E. The results are shown in Table 2.
[0678] table 2: the A in processing or the abcr mouse of being untreated2E accumulates
(pmol/ eyes)
A 2E Different-A2E Amount to
No medicine 11.01  2.57  13.58
Farnesyl-decyl ketone 2.00  0.66  2.66
N-palmityl method acid amides 5.38  1.70  7.09
[0679] the bright farnesyl-decyl of these tables of data ketone reduces by 80% above A2E, about 74% different-A2E accumulates, and amounts to about 80%. N-palmityl method acid amides reduces by 50% above A2E, about 33% different-A2E accumulates, and amounts to about 47%.
[0680] these embodiment should be considered as nonrestrictive. The list of references of all references (list of references that comprises the document of quoting among the application, the patent Shen of patent granted, announcement Content please) is clear and definite incorporated herein by reference.
[0681] only use normal experiment, those skilled in the art can understand maybe can understand this The multiple replacement scheme that is equal to of the specific embodiments described in the literary composition.

Claims (253)

1. the purposes of medicine in the medicament of preparation treatment or prevention eye disease, wherein said medicine inhibition, antagonism or shortening occur in the visual cycle circuit on the outer visual cycle ladder of dish of shaft-like photoreceptor cell.
2. the purposes of claim 1, wherein said eye disease comprises degeneration of macula.
3. the purposes of claim 1 or claim 2, wherein said eye disease comprises the Shi Tajiate disease.
4. the purposes of each aforementioned claim, wherein said eye disease comprise that lipofuscin accumulates.
5. the purposes of each aforementioned claim, wherein said medicine improve 11-cis-retinal isomery and turn to the speed of alltrans-retinal.
6. the purposes of each aforementioned claim, the inhibition in retinal pigment epithelium of wherein said medicine, antagonism or shorten the visual cycle circuit.
7. the purposes of each aforementioned claim, wherein said medicine suppresses in retinal pigment epithelium or the antagonism visual cycle.
8. the purposes of claim 7, wherein said medicine suppress combining of at least a lecithin retinol acyltransferase, isomery hydrolytic enzyme and 11-cis-retinol dehydrogenase or inhibition and RPE65.
9. the purposes of each aforementioned claim, wherein said medicine shortens the visual cycle circuit in retinal pigment epithelium.
10. each purposes among the claim 1-9, wherein said medicament is given for a long time.
11. each purposes among the claim 1-9, wherein said medicament is given once.
12. each purposes among the claim 1-9, wherein said medicament gives once weekly.
13. each purposes among the claim 1-9, wherein said medicament gives weekly twice.
14. each purposes among the claim 1-13, the second kind of medicine that wherein is different from first kind of medicine is used to prepare described medicament.
15. the purposes of claim 14, wherein second kind of medicine inhibition, antagonism or shortening occur in the visual cycle circuit on the outer visual cycle ladder of dish of shaft-like photoreceptor cell.
16. the purposes of claim 14, wherein first kind of medicine and second kind of medicine all suppress or antagonism occurs in the outer visual cycle ladder of dish of shaft-like photoreceptor cell.
17. each purposes among the claim 14-16, wherein said first kind of medicine and second kind of medicine suppress or the antagonism different molecular.
18. each purposes among the claim 14-17, wherein said first kind of medicine suppress with second kind of medicine or the antagonism visual cycle in different ladders.
19. the purposes of claim 18, wherein said first kind of medicine and second kind of medicine suppress or the antagonism visual cycle in sequential ladder.
20. each purposes among the claim 14-17, wherein said first kind of medicine suppress with second kind of medicine or the antagonism visual cycle in identical ladder.
21. each purposes among the claim 14-20, wherein said first kind of medicine suppresses RPE65.
22. the purposes of claim 21, wherein said second kind of medicine suppresses lecithin retinol acyltransferase, isomery hydrolytic enzyme and/or 11-cis-retinol dehydrogenase.
23. each purposes among the claim 14-21, wherein said first kind of medicine inhibition or antagonism occur in the outer visual cycle ladder of dish of shaft-like photoreceptor cell, and second kind of medicine shortens the visual cycle circuit on the visual cycle ladder outside the dish that occurs in shaft-like photoreceptor cell.
24. treatment or prevention patient's the method for eye disease, described method comprises: the medicine that gives the visual cycle circuit on the outer visual cycle ladder of dish that described patient's inhibition, antagonism or shortening occur in shaft-like photoreceptor cell.
25. method for the treatment of or preventing degeneration of macula, described method comprises: need patient's medicine of treatment or prevention degeneration of macula, wherein said medicine inhibition, antagonism or shortening occur in the visual cycle circuit on the outer visual cycle ladder of dish of shaft-like photoreceptor cell.
26. each method among the claim 24-55, described method also comprises the degeneration of macula of diagnosing described patient.
27. each method among the claim 24-26, described method also comprises the degeneration of macula of monitoring described patient.
28. each method among the claim 24-27, wherein said eye disease comprises degeneration of macula.
29. each method among the claim 24-28, wherein said eye disease comprises the Shi Tajiate disease.
30. each method among the claim 24-29, wherein said eye disease comprise that lipofuscin accumulates.
31. each method among the claim 24-30, wherein said medicine improve 11-cis-retinal isomery and turn to the speed of alltrans-retinal.
32. each method among the claim 24-31, the inhibition in retinal pigment epithelium of wherein said medicine, antagonism or shortening visual cycle circuit.
33. each method among the claim 24-32, wherein said medicine suppress in retinal pigment epithelium or the antagonism visual cycle.
34. the purposes of claim 33, wherein said medicine suppress combining of at least a lecithin retinol acyltransferase, isomery hydrolytic enzyme and 11-cis-retinol dehydrogenase or inhibition and RPE65.
35. each method among the claim 24-34, wherein said medicine shorten the visual cycle circuit in retinal pigment epithelium.
36. each method among the claim 24-35, wherein said medicament is given for a long time.
37. each method among the claim 24-35, wherein said medicament is given once.
38. each method among the claim 24-35, wherein said medicament gives once weekly.
39. each method among the claim 24-35, wherein said medicament gives weekly twice.
40. each method among the claim 24-39, the second kind of medicine that wherein is different from first kind of medicine is used to prepare described medicament.
41. the method for claim 40, wherein second kind of medicine inhibition, antagonism or shortening occur in the visual cycle circuit on the outer visual cycle ladder of dish of shaft-like photoreceptor cell.
42. the method for claim 40, wherein first kind of medicine and second kind of medicine suppress or antagonism occurs in the outer visual cycle ladder of dish of shaft-like photoreceptor cell.
43. the method for claim 41 or 42, wherein first kind of medicine suppresses or the different molecule of antagonism with second kind of medicine.
44. each method among the claim 40-43, wherein first kind of medicine suppress with second kind of medicine or the antagonism visual cycle in different ladders.
45. the method for claim 44, wherein first kind of medicine and second kind of medicine suppress or the antagonism visual cycle in sequential ladder.
46. each method among the claim 40-43, wherein first kind of medicine suppress with second kind of medicine or the antagonism visual cycle in identical ladder.
47. each method among the claim 40-46, wherein first kind of medicine suppresses RPE65.
48. the method for claim 47, wherein second kind of medicine suppresses lecithin retinol acyltransferase, isomery hydrolytic enzyme and/or 11-cis-retinol dehydrogenase.
49. each method among the claim 40-47, wherein first kind of medicine suppresses or antagonism occurs in the ladder of the outer visual cycle of dish of shaft-like photoreceptor cell, and the visual cycle circuit on the outer visual cycle ladder of dish that second kind of medicine shortens to occur in shaft-like photoreceptor cell.
50. each method among each purposes or the claim 24-49 among the claim 1-23, wherein said medicine is included in the chemical compound of each definition among the claim 51-257.
51. formula I chemical compound:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X is-O-,-N (R a)-,-C (R b) p-or-S-;
Z be alkyl, haloalkyl ,-(CH 2CH 2O) pR bOr-C (=O) R b
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl or haloalkyl; With
Expression singly-bound, cis-double bonds or trans double bond.
52. formula Ia, Ib, Ic or Id chemical compound:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
R 4Do not exist or hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
Y is-C (R b) 2-or-C (=O)-;
X is-O-,-N (R a)-,-C (R b) 2-or-S-;
Z be alkyl, haloalkyl or-C (=O) R b
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl or haloalkyl; With
Figure A2005800115130007C1
Expression singly-bound, cis-double bonds or trans double bond.
53. the chemical compound of claim 52, wherein R 1It is methyl.
54. the chemical compound of claim 52 or claim 53, wherein n is 0.
55. the chemical compound of claim 52 or claim 53, wherein n is 1.
56. each chemical compound among the claim 52-55, wherein Y is-CH 2-.
57. each chemical compound among the claim 52-56, wherein X is-O-.
58. each chemical compound among the claim 52-56, wherein X be-N (H)-.
59. each chemical compound among the claim 52-58, wherein Z is-C (=O) R b
60. each chemical compound among the claim 52-58, wherein Z is-C (=O) R bAnd R bIt is haloalkyl.
61. each chemical compound among the claim 52-58, wherein Z is an alkyl.
62. each chemical compound among the claim 52-58, wherein Z is a haloalkyl.
63. each chemical compound, wherein R among the claim 52-62 3Be hydrogen.
64. each chemical compound, wherein R among the claim 52-63 4Be hydrogen, methyl or do not exist.
65. the chemical compound of formula Ie, If, Ig or Ih:
Figure A2005800115130008C1
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
X is-O-,-N (R a)-,-C (R b) 2-or-S-;
Z be alkyl, haloalkyl or-C (=O) R b
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bBe hydrogen, alkyl or haloalkyl.
66. the chemical compound of claim 65, wherein n is 0.
67. the chemical compound of claim 65, wherein n is 1.
68. each chemical compound among the claim 65-67, wherein X is-O-.
69. each chemical compound among the claim 65-67, wherein X be-N (H)-.
70. each chemical compound among the claim 65-69, wherein Z is-C (=O) R b
71. each chemical compound among the claim 65-69, wherein Z is-C (=O) R bAnd R bIt is haloalkyl.
72. each chemical compound among the claim 65-69, wherein Z is an alkyl.
73. each chemical compound among the claim 65-69, wherein Z is a haloalkyl.
74. each chemical compound, wherein R among the claim 65-73 3Be hydrogen.
75. the chemical compound of each or claim 74 among the claim 65-67, wherein X is-O-; With Z be alkyl.
76. the chemical compound of each or claim 74 among the claim 65-67, wherein X is-O-; With Z be haloalkyl.
77. the chemical compound of each or claim 74 among the claim 65-67, wherein X be-N (H)-; With Z be alkyl.
78. the chemical compound of each or claim 74 among the claim 65-67, wherein X be-N (H)-; With Z be haloalkyl.
79. a chemical compound, described chemical compound have by
The structure of representative.
80. formula II chemical compound:
Wherein n is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) p-;
Z does not exist or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-(CH 2CH 2O) pR b,
-C(＝O)R b，-C(＝O)CH 2F，-C(＝O)CHF 2，-C(＝O)CF 3，-C(＝O)CHN 2，-C(＝O)OR b，
-C (=O) CH 2OC (=O) R b,-C (=O) C (=C (R b) 2) R b,
Figure A2005800115130009C3
Or
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
Expression singly-bound, cis-double bonds or trans double bond.
81. the chemical compound of formula IIa, IIb, IIc or Iid:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
R 4Do not exist or hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
Y is-C (=O)-or-C (R b) 2-;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) 2-;
Z does not exist or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-C (=O) R b,-C (=O) CH 2F ,-C (=O) CHF 2,-C (=O) CF 3,-C (=O) CHN 2,-C (=O) CH 2OC (=O) R b,-C (=O) OR b,-C (=O) C (=C (R b) 2) R b,
Or
Figure A2005800115130011C2
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
Figure A2005800115130011C3
Represent singly-bound, cis-double bonds or trans double bond.
82. the chemical compound of claim 81, wherein n is 0.
83. the chemical compound of claim 81, wherein n is 1.
84. each chemical compound, wherein R among the claim 81-83 1Be hydrogen or methyl.
85. each chemical compound, wherein R among the claim 81-84 3Be hydrogen.
86. each chemical compound, wherein R among the claim 81-85 4Be hydrogen or methyl.
87. each chemical compound among the claim 81-86, wherein Y is-CH 2-.
88. each chemical compound among the claim 81-87, wherein X is-O-.
89. each chemical compound among the claim 81-87, wherein X is-NH-.
90. each chemical compound among the claim 81-87, wherein X is-C (R b) 2-.
91. each chemical compound among the claim 81-87, wherein X be-C (=O)-.
92. each chemical compound among the claim 81-91, wherein Z is an alkyl.
93. each chemical compound among the claim 81-91, wherein Z is a haloalkyl.
94. formula IIe, IIf, IIg or IIh chemical compound:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
R 3Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
X be hydrogen ,-O-,-S-,-N (R a)-,-N (R a)-N (R a)-,-C (=O)-,-C (=NR a)-,-C (=NOH)-,-C (=S)-or-C (R b) 2-;
Z does not exist or hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-CN ,-OR b,-C (=O) R b,-C (=O) CH 2F ,-C (=O) CHF 2,-C (=O) CF 3,-C (=O) CHN 2,-C (=O) CH 2OC (=O) R b,
-C (=O) OR b,-C (=O) C (=C (R b) 2) R b,
Figure A2005800115130012C2
Or
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl.
95. the chemical compound of claim 94, wherein n is 0.
96. the chemical compound of claim 94, wherein n is 1.
97. each chemical compound, wherein R among the claim 94-96 1Be hydrogen or methyl.
98. each chemical compound, wherein R among the claim 94-97 3Be hydrogen.
99. each chemical compound, wherein R among the claim 94-98 4Be hydrogen or methyl.
100. each chemical compound among the claim 94-99, wherein X is-O-.
101. each chemical compound among the claim 94-99, wherein X is-NH-.
102. each chemical compound among the claim 94-99, wherein X is-CH 2-.
103. each chemical compound among the claim 94-99, wherein X be-C (=O)-.
104. each chemical compound among the claim 94-103, wherein Z is an alkyl.
105. each chemical compound among the claim 94-103, wherein Z is a haloalkyl.
106. each chemical compound among the claim 94-103, wherein Z is-C (=O) R b
107. each chemical compound among the claim 94-98, wherein X is-O-; With Z-C (=O) R b
108. each chemical compound among the claim 94-98, wherein X is-CH 2-; With Z-C (=O) R b
109. each chemical compound among the claim 94-98, wherein X is-NH-; With Z-C (=O) R b
110. a chemical compound, described chemical compound have the structure by the following formula representative:
Figure A2005800115130013C1
111. formula III chemical compound:
Figure A2005800115130013C2
Wherein
N is 0-10;
R 1It is hydrogen or alkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
Y is-CR b(OR b)-,-CR b(N (R a) 2)-,-C (R b) p-,-C (=O)-or-C (R b) pC (=O)-;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) p-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-(CH 2CH 2O) pR b,-C (=O) R b,-C (=NR a) R b,-C (=NOR b) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
P is 0-20;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
Expression singly-bound or trans double bond.
112. formula III a, IIIb, IIIc or IIId chemical compound:
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
Y is-C (=O)-,-CR b(OR b)-,-CR b(N (R a) 2)-or-C (R b) 2-;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) 2-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-C (=O) R b,-C (=NR a) R b,-C (=NOH) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl;
P is 0-10; With
Expression singly-bound or trans double bond.
113. the chemical compound of claim 112, wherein n is 0.
114. the chemical compound of claim 112, wherein n is 1.
115. each chemical compound, wherein R among the claim 112-114 1Be hydrogen or methyl.
116. each chemical compound, wherein R among the claim 112-115 3Be hydrogen.
117. each chemical compound, wherein R among the claim 112-116 4Be hydrogen or methyl.
118. each chemical compound among the claim 112-117, wherein X is-O-.
119. each chemical compound among the claim 112-117, wherein X is-NH-.
120. each chemical compound among the claim 112-117, wherein X is-C (R b) 2-.
121. each chemical compound among the claim 112-117, wherein X be-C (=O)-.
122. each chemical compound among the claim 112-121, wherein Z is an alkyl.
123. each chemical compound among the claim 112-121, wherein Z is a haloalkyl.
124. formula III e, IIIf, IIIg or IIIh chemical compound:
Figure A2005800115130015C1
Wherein, the implication of following group when at every turn occurring is independently,
N is 0-4;
R 1It is hydrogen or alkyl;
X is-O-,-S-,-N (R a)-,-C (=O)-or-C (R b) 2-;
Z be hydrogen, alkyl, haloalkyl, aryl, aralkyl ,-OR b,-N (R b) 2,-C (=O) R b,-C (=NR a) R b,-C (=NOH) R b,-C (OR b) (R b) 2,-C (N (R a) 2) (R b) 2Or-(CH 2CH 2O) pR b
R aBe hydrogen, alkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
P is 0-10.
125. the chemical compound of claim 124, wherein n is 0.
126. the chemical compound of claim 124, wherein n is 1.
127. each chemical compound, wherein R among the claim 124-126 1Be hydrogen or methyl.
128. each chemical compound among the claim 124-127, wherein Y be-C (=O)-.
129. each chemical compound among the claim 124-127, wherein Y is-CH 2-.
130. each chemical compound among the claim 124-129, wherein Z is-C (=O) R b
131. each chemical compound among the claim 124-129, wherein Z is-CH (OH) R b-.
132. each chemical compound among the claim 124-129, wherein Z is-CH (NH) R b
133. each chemical compound among the claim 124-129, wherein Z is an alkyl.
134. each chemical compound among the claim 124-129, wherein Z is a haloalkyl.
135. a chemical compound, described chemical compound have the structure by the following formula representative:
136. formula IV chemical compound:
Figure A2005800115130016C2
Wherein, the implication of following group when at every turn occurring is independently,
N is 1,2,3 or 4;
Y is-C (R b) 2-or-C (=O)-;
X is-O-,-NR a-,-C (R b) 2-or-C (=O)-;
Z is-C (=O) R b,-OR b,-N (R b) 2, alkyl or haloalkyl;
R aBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl.
137. the chemical compound of claim 136, wherein Y is-CH 2-.
138. the chemical compound of claim 136 or claim 137, wherein X is-O-.
139. each chemical compound among the claim 136-138, wherein Z is-C (=O) R bAnd R bIt is alkyl.
140. each chemical compound among the claim 136-138, wherein Z is an alkyl.
141. the chemical compound of claim 136, wherein Y is-CH 2-; X is-O-; Z is-C (=O) R bAnd R bIt is alkyl.
142. the chemical compound of claim 136, wherein Y is-CH 2-; X is-O-; With Z be alkyl.
143. the chemical compound of claim 136, wherein Y is-CH 2-; X is-C (=O)-; With Z be alkyl.
144. the chemical compound of claim 136, wherein Y is-CH 2-; X is-C (=O)-; Z is-N (R b) 2And R bIt is alkyl.
145. a chemical compound, described chemical compound have the structure by the following formula representative.
146. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130017C2
147. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130017C3
148. a chemical compound, described chemical compound have the structure by the following formula representative.
149. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130017C5
150. a chemical compound, described chemical compound have the structure by the following formula representative.
151. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130018C1
152. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130018C2
153. a chemical compound, described chemical compound have the structure by the following formula representative.
154. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130018C4
155. a chemical compound, described chemical compound have the structure by the following formula representative.
156. a chemical compound, described chemical compound have the structure by the following formula representative.
157. a chemical compound, described chemical compound have the structure by the following formula representative.
158. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130018C8
159. formula V chemical compound:
Figure A2005800115130018C9
Wherein, the implication of following group when at every turn occurring is independently,
N is 1,2 or 3;
Y is-C (R b) 2-,-C (=O)-or-CH (OH)-;
X is-O-,-NR a-or-C (R b) 2-;
Z is-C (=O) R b, hydrogen ,-(CH 2CH 2O) pR b, alkyl or haloalkyl;
R aBe hydrogen, alkyl, haloalkyl, aryl or aralkyl;
R bBe hydrogen, alkyl, haloalkyl, aryl or aralkyl; With
P is 1-10.
160. the chemical compound of claim 159, wherein Y is-CH 2-.
161. the chemical compound of claim 159, wherein Y be-C (=O)-.
162. the chemical compound of claim 159, wherein Y be-CH (OH)-.
163. each chemical compound among the claim 159-162, wherein X is-O-.
164. each chemical compound among the claim 159-162, wherein X is-NR a-.
165. each chemical compound among the claim 159-162, wherein X is-C (R b)-.
166. each chemical compound among the claim 159-165, wherein Z is an alkyl.
167. each chemical compound among the claim 159-165, wherein Z is-C (=O) R bAnd R bIt is alkyl.
168. each chemical compound among the claim 159-165, wherein Z is-(CH 2CH 2O) pR bAnd R bIt is alkyl.
169. a chemical compound, described chemical compound have the structure by the following formula representative.
170. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130019C2
171. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130020C1
172. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130020C2
173. a chemical compound, described chemical compound have the structure by the following formula representative.
174. a chemical compound, described chemical compound have the structure by the following formula representative.
175. a chemical compound, described chemical compound have the structure by the following formula representative.
176. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130020C6
177. a chemical compound, described chemical compound have the structure by the following formula representative.
178. a chemical compound, described chemical compound have the structure by the following formula representative.
179. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130021C1
180. a chemical compound, described chemical compound have the structure by the following formula representative.
181. a chemical compound, described chemical compound have the structure by the following formula representative.
182. a chemical compound, described chemical compound have the structure by the following formula representative.
183. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130021C5
184. a chemical compound, described chemical compound have the structure by the following formula representative.
185. a chemical compound, described chemical compound have the structure by the following formula representative.
186. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130021C8
187. a chemical compound, described chemical compound have the structure by the following formula representative.
188. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130022C2
189. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130022C3
190. a chemical compound, described chemical compound have the structure by the following formula representative.
191. a chemical compound, described chemical compound have the structure by the following formula representative.
192. a chemical compound, described chemical compound have the structure by the following formula representative.
193. a chemical compound, described chemical compound have the structure by the following formula representative.
194. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130022C8
195. formula VI chemical compound:
Figure A2005800115130023C1
Wherein, the implication of following group when at every turn occurring is independently,
R 1Be hydrogen, alkyl, aryl or aralkyl;
X be alkyl, alkenyl ,-C (R b) 2-,-C (=O)-,-C (=NR a)-,-C (OH) R bOr-C (N (R a) 2) R b-;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bIt is hydrogen or alkyl.
196. the chemical compound of claim 195, wherein R 1Be hydrogen.
197. the chemical compound of claim 195 or claim 196, wherein X is-C (R b) 2-.
198. the chemical compound of claim 195 or claim 196, wherein X be-C (=O)-.
199. formula VIa or VIb chemical compound:
Wherein, the implication of following group when at every turn occurring is independently,
R 1Be hydrogen, alkyl, aryl or aralkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R 3It is hydrogen or alkyl;
R aBe hydrogen, alkyl, aryl or aralkyl;
R bIt is hydrogen or alkyl; With
Expression singly-bound, cis-double bonds or trans double bond.
200. the chemical compound of claim 199, wherein R 1Be hydrogen.
201. the chemical compound of claim 199 or claim 200, wherein R 2It is alkyl.
202. each chemical compound, wherein R among the claim 199-201 3Be hydrogen or methyl.
203. formula VIc, VId or VIe chemical compound:
Wherein, the implication of following group when at every turn occurring is independently,
N is 1-5;
M is 0-30;
R 1Be hydrogen, alkyl, aryl or aralkyl;
R 2Be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl group, alkynyl, aryl or aralkyl;
R 3It is hydrogen or alkyl;
R 4Be hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, arylalkenyl, sweet-smelling alkynyl, heteroarylalkyl, impure aromatic ene base, hetaryne base, cyano group, nitro, sulfydryl, hydroxyl, sulfonyl, amino, acyl amino, acylamino-, alkylthio group, carboxyl, carbamoyl, alkoxyl, sulfonate radical, sulfate radical, sulfonamido, sulfamoyl, sulfonyl and sulfoxide group;
R aBe hydrogen, alkyl, aryl or aralkyl; With
R bIt is hydrogen or alkyl.
204. the chemical compound of claim 203, wherein R 1Be hydrogen.
205. the chemical compound of claim 203, wherein R 4Be hydrogen.
206. the chemical compound of claim 203, wherein R 1Be hydrogen; And R 4Be hydrogen.
207. each chemical compound among the claim 203-206, wherein n is 1,2 or 3.
208. each chemical compound, wherein R among the claim 203-207 3It is methyl.
209. each chemical compound, wherein R among claim 203 or the 207-208 1Be hydrogen.
210. each chemical compound among the claim 203-206, wherein n is 1,2 or 3; R 3It is methyl.
211. the chemical compound of claim 203, wherein n is 1,2 or 3; R 3It is methyl; R 1Be hydrogen.
212. each chemical compound among the claim 203-211, wherein m is 1-10.
213. each chemical compound among the claim 203-211, wherein m is 11-20.
214. the chemical compound of claim 203, wherein m is 11-20; R 1Be hydrogen.
215. formula VII chemical compound:
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or carbonyl;
L is a hydrophobic part, or any two adjacent L are joined together to form condensed aromatic ring or hetero-aromatic ring.
216. the chemical compound of claim 215, wherein L is alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, carbonyl, ether or multi-ring base.
217. the chemical compound of claim 215, wherein L has formula VIIa:
Figure A2005800115130025C2
Wherein, the implication of following group when at every turn occurring is independently:
R ' and X are hydrogen, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, carbonyl, alkoxyl, hydroxyl, sulfydryl, alkylthio group or amino; With m be 1-6.
218. the chemical compound of formula VIIb:
Figure A2005800115130026C1
Wherein n is 1-8.
219. the chemical compound of formula VIIc:
Wherein, the implication of following group when at every turn occurring is independently,
R is H, alkyl or acyl group; With
R ' is alkyl or ether.
220. the chemical compound of claim 219, wherein R all is H when occurring for twice.
221. the chemical compound of claim 219, wherein at least one R is an alkyl.
222. the chemical compound of claim 219, wherein at least one R is a methyl.
223. formula VIId chemical compound:
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl or acyl group; With
R ' is alkyl or ether.
224. the chemical compound of claim 223, wherein R equal H when occurring for twice.
225. the chemical compound of claim 223, wherein at least one R is an alkyl.
226. the chemical compound of claim 223, wherein at least one R is a methyl.
227. formula VIIe chemical compound:
Figure A2005800115130027C1
Wherein, the implication of following group when at every turn occurring is independently:
X be hydrogen or-C (=O) OR ';
R is H, alkyl or acyl group; With
R ' is an alkyl.
228. the chemical compound of claim 227, wherein R is H.
229. the chemical compound of claim 227, wherein at least one R is an alkyl.
230. the chemical compound of claim 227, wherein R is a methyl.
231. formula VIIf chemical compound:
Wherein, the implication of following group when at every turn occurring is independently
R is H, alkyl or acyl group; With
R ' is an alkyl.
232. the chemical compound of claim 231, wherein R is H.
233. the chemical compound of claim 231, wherein at least one R is an alkyl.
234. the chemical compound of claim 231, wherein R is a methyl.
235. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130027C3
236. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130027C4
237. a chemical compound, described chemical compound have the structure by the following formula representative.
238. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130028C2
239. a chemical compound, described chemical compound have the structure by the following formula representative.
240. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130028C4
241. a chemical compound, described chemical compound have the structure by the following formula representative.
242. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130028C6
243. a chemical compound, described chemical compound have the structure by the following formula representative.
Figure A2005800115130028C7
244. formula VIII chemical compound:
Wherein R ' is hydrogen, alkyl or ether; Or any two adjacent L are joined together to form condensed aromatic ring or hetero-aromatic ring.
255. formula IX chemical compound:
ANR 2
IX
Wherein, the implication of following group when at every turn occurring is independently:
R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroarylalkyl or carbonyl; With
A is optional aryl or the heteroaryl that replaces.
256. formula X chemical compound:
AC(＝O)NHNH 2
X
The implication of wherein following group when at every turn occurring is independently:
R ' is hydrogen, alkyl or ether; With
A is optional aryl or the heteroaryl that replaces.
257. a preparation, described preparation comprise the chemical compound of each definition among the claim 51-257 and are different from second kind of chemical compound as each definition among the claim 51-257 of first kind of chemical compound.
258. a method of identifying treatment or prevention eye disease, described method comprises: suffer from or have patient's candidate medicine that the eye disease risk takes place; With accumulating of retinal toxicity chemical compound in measurement patient's retinal pigment epithelium (RPE); Wherein in the presence of drug candidate, compare with there not being drug candidate, the retinal toxicity chemical compound in RPE is accumulated and is shown that the candidate medicine is the medicine of treatment or prevention eye disease.
259. a method of identifying treatment or prevention eye disease, described method comprises: one or more cells of the external visual cycle model of representative are contacted with drug candidate; With accumulating of measurement visual cycle midbody product at least one cell;
Wherein in the presence of the candidate medicine, compare with there not being the candidate medicine, accumulating of midbody product shows that the candidate medicine is the medicine of treatment or prevention eye disease.
260. the method for claim 259 or claim 259, wherein said eye disease comprises degeneration of macula.
261. each method among the claim 259-260, wherein said eye disease comprises the Shi Tajiate disease.
262. each method among the claim 259-161, wherein said eye disease comprise that lipofuscin accumulates.
263. each method among claim 259 or the 261-262, wherein said retinal toxicity chemical compound are N-Asia retinyl-N-retinyl ethanolamine.
264. each method among the claim 260-262, wherein said intermediate is alltrans-retinal.
CN 200580011513 2004-02-17 2005-02-17 Management of ophthalmologic disorders, including macular degeneration Pending CN1997359A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109689033A (en) * 2016-04-28 2019-04-26 星火治疗有限公司 The relative effectivenes of coding of the viral vectors isomery hydrolysis alcohol measure
CN114907246A (en) * 2021-02-08 2022-08-16 复旦大学 Total synthesis method of vitamin A, derivative thereof and deuterated compound thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109689033A (en) * 2016-04-28 2019-04-26 星火治疗有限公司 The relative effectivenes of coding of the viral vectors isomery hydrolysis alcohol measure
CN114907246A (en) * 2021-02-08 2022-08-16 复旦大学 Total synthesis method of vitamin A, derivative thereof and deuterated compound thereof
CN114907246B (en) * 2021-02-08 2024-02-06 复旦大学 Total synthesis method of vitamin A, derivative thereof and deuterated compound thereof

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