CN1390124A - Medicine for preventing or treating myopia - Google Patents

Abstract

The invention relates to the use of NO synthase substrate and/or NO donor, or inducible NO synthase induce inhibitor and/or inducible NO synthase activity inhibitor for the preparation of drugs for treating or preventing myopia; the pharmaceutical compositions containing one or more of the compound(s) mentioned above and pharmaceutically acceptable excipients; new arginine derivatives, the method for treating or preventing myopia, and the commercial kits.

Description

The prevention of myopia, curative

The prevention of myopia, curative technical field

The present invention relates to the anti-of myopia, curative specifically, it is related to NO synthase (lower abbreviation N0S) substrate, NO donors, purposes of immune N0S (lower abbreviation iNOS) the induction inhibitor and iNOS activities inhibitor in the medicine for treating or preventing myopia is prepared and the Yao Wu Group compounds containing it, the method for treating or preventing myopia, and a kind of new arginine derivative.

Background technology

Although the teiology that myopia is the common eyesight patient's condition or disease myopia can pass away in many factors, it is faint including heredity, environment and light, but it is attributable to common factor, i.e. the axis of eyeball is elongated, and the reason for unsuitable refracting power of eyes or both is combined is caused.（Chen, W.C, etc. Myopia Updates II. Lin L. etc., Ed. pp.39-42, Springer, New York (1998)).A nearly a conduit made of long bamboo to it is serious when can cause retinopathy, the macular degeneration relevant with the age（) and the such as cataract AMD

It is due to initially to see near objects to cause one of near-sighted factor most often mentioned（Such as reading, eyesight display terminal（VDT), closely see TV, worked on fine assembly line）And make the long-term excessive contraction of ciliary muscle.One of theory of occurrence and development of refractive myopic eye thinks, operationally too near from obtaining, and becomes tetanic due to ciliary muscle excess shrinkage, as a result causes crystalline lens to produce the refracting power of interval.The ciliary muscle of myopia can not suitably relaxing so that on crystalline lens focusing image a long way off（Tokoro, T. Nippon Ganka Gakkai Zasshl. 102:796-812 (1998)), the contraction of ciliary muscle by a few types acceptor（Including muscarinic receptor, prostaglandin receptor and endothelin receptor）Mediate (Erickson-Larry, K. etc., Invest. Ophthalmol. Vis. Sci 32:492-495(1991) ).Ciliary muscle relaxation is adjusted by 2 independent mechanism：The mechanism relied on cAMP-, it includes the relexation that beta-adrenergic and prostaglandin receptor are adjusted, and the mechanism with cAMP- non-dependent（Goh, Y., etc. Invest. Ophthalmol. Vis. Sci. 36:1188-1192(1995) ) .Ciliary muscle relaxation is as the pharmacological intervention of myopia in humans and animals model with anticholinergic agent such as atropine (Shih, Y ,-F. etc., J. Ocular Pharmacol. Ther. 15:85-90 (1999)) and send logical sequence Xiping (Cottriall, C Shang and McBrien, N. A., Invest. Ophthalmol. Vis. Sci. 37:1368-1379 (1996)), cycloplegic Such as cyclopentolate and Tropicamide, and parasympathomimetic agent such as neo-synephrine and adrenaline carry out overtesting (Kamikavatoko, S. In Myopia Updates, Tokoro, T., ed., Proc. 6th Int. Conf. Myopia, Tokyo, pp. 147- 154 (1998)).However, side effect of the upper magic potion thing to autonomic nerve makes them unsuitable for prolonged application.In near-sighted possible study medication, the mechanism acted on the ciliary muscle relaxation of the non-plant of cAMP- non-dependent is recently by sizable concern.

Another reason for causing myopia is due to that the sclera lesion such as inflammation of eyeground portion causes meronecrosis, causes scleromalacia or hardness to reduce, as a result because the relation of intraocular pressure, makes sclera deformation as protruded backward, so that axis oculi is elongated and causes myopia.For such myopia, the medicine for the treatment of or prevention there is no.

Pharmacologically finding nitrogen oxide（NO effect) is nearer thing（From 1979）Generally, NO is by L-arginine as substrate and the NO synthase as synzyme（N0S) interact and produce.Isoform N0S includes the neuronal NOS (nNOS) in central nervous system and peripheral neverous system or the endothelial cell N0S (eNOS or ecN0S) in brain NOS (bNOS) and neuron and non-neuronal cell.NNOS and eNOS is in the short time generation NO after calcium/calmodulin activation, to produce different biological respinses.Immune N0S (iNOS) is by lipopolysaccharides and some cell factors（Such as interleukin -1, interleukin -6, TNF and interferon gamma) the 3rd isoform N0S of induction.They produce substantial amounts of NO to continue for some time, and cause Pathological Physiology to be reacted（Szabo, C. etc., Nitric Oxide, Biochemistry, Molecular Biology, and Therapeutic Implication. Ignarro, L. and Mur d, F., pp. 113- 153, Academic Press, New York (1995)).

Under normal operation, a small amount of NO of picomole level are produced by L-arginine as substrate and 2 type N0S (nNOS or eNOS) interactions.Then NO makes guanylate enzyme activation produce cGMP, in turn, and cGMP induces many biological respinses, including vasodilation, increase eye blood flow, reduction intraocular pressure again（I0P), prevention platelet aggregation, reduction polymorphonuclear cell（PMN the signal transduction of chemical toxicant), central nervous system and peripheral neverous system, suppresses tumour cell etc.（Ignarro, L. J. Nitric Oxide, Principles and Actions. Lancaster J., Ed. pp. 111-137 Academic Press, New York (1996)).

But in the case of improper, produced with the substantial amounts of NO of nanomolar range by L-arginine by iNOS, NO will be further oxidized to N0₂-, nitrite, peroxide it is sub- Nitrate（0N00_) and free radical, and with the interaction such as mercaptan, the iron-sulphur center, cytochrome oxidase, glycolytic enzyme, liver cell, macrophage, calmodulin of various enzymes, as a result the biological function of cell is changed, DNA has been damaged, and has caused the blunt nerve degeneration of Apoptosis, neurotoxicity, B and many ophthalmology diseases.INOS is only under pathophysiological conditions（Such as inflammation）Induced by lipopolysaccharides, endotoxin and cell factor.The rate-limiting step that NO is generated by iNOS is L-arginic availability, and L-arginine is recycled by argininosuccinate hydrochlorate synzyme, and the latter when iNOS is induced while being induced.In addition, iNOS induction has also triggered L-arginic membrane transport system to strengthen utilization rate of the L-arginine to iNOS, as a result therefore, it is a main policies for eliminating the reaction of NQ Pathological Physiology to suppress iNOS to a large amount of generation N0（Dawson, V. L. etc., Nitric Oxide, Biochemistry, Molecular Biology, and Therapeutic Implication. Ignarro, and Murad, F., Ed. pp. 323-349 L., Academic Press, New York (1995)).

It has been found that NO is relevant with ophthalmology disease such as glaucoma（Nathanson, J. A. J. Pharmacol. Exp. Ther. 956-965 (1992)), but the relation of NO and myopia then there is no data to show.The present inventor is by carefully studying, and NO can be occurred in knitting by Yan Group supply falls short of demand and excess supply by having been surprisingly found that, this causes the dysfunction of above-mentioned eye and causes disease, the generation of such as myopia respectively.Therefore, the dysfunction of eye and disease , Te Do are that the prevention or treatment of myopia can be not enough by making up NO, or reduce NO excess supplies by suppressing iNOS activity or suppressing induction.As a result, may reform the basic concepts of the treatment of myopia, and the NO levels being finally related in regulation ocular tissue.

In addition, in existing treatment or prevention myopia method, in addition to medication, also having attempted physical therapy such as low-frequency therapeutic, ultrasonic therapy etc., no one kind can reach gratifying effect.Therefore, in the urgent need to developing a kind of effective prevention or treating the medicine of myopia.

To solve the above problems, prevention or the curative of a kind of effective myopia are provided, the present inventor is by concentrating on studies, success finds that N0S substrates and NO donors can be by increasing NO concentration too low in eye, so that ciliary muscle relaxation with the mechanism that the ciliary muscle relaxation of the non-plant of cAMP- non-dependent is acted on；INOS induces inhibitor and iNOS activities inhibitor can reduce the concentration of too high NO in eye, suppresses the protrusion of central sclera；Two kinds of effects of upper fan may finally all suppress the elongation of axis oculi, reach the effect of prevention or treatment myopia, So as to complete the present invention.

Summary of the invention

The first aspect of the present invention is related to the application of N0S substrates and/or NO donors or iNOS induction inhibitor and/or iNOS activities inhibitor in the medicine of preparation or pre- Anti-myopic eye.

Another aspect of the present invention is related to the Yao Wu Group compounds for containing N0S substrates and/or NO donors or iNOS induction inhibitor and/or iNOS activities inhibitor as active ingredient.

Further aspect of the present invention is related to a kind of new arginine ester derivative.

Another aspect of the present invention is related to a kind of prevention and treatment method of myopia, and it includes giving patient by therapeutically effective amount above-claimed cpd or composition.

Finally, the present invention relates to a kind of commercial kit, it includes the above-claimed cpd or composition of therapeutically effective amount, and records medical compounds Huo Group compounds can be used in the treatment or prevention of myopia or the data of the medical compounds or composition should be used.

That is, the present invention is as described below.

(1) application of N0S substrates and/or NO donors or iNOS induction inhibitor and/or iNOS activities inhibitor in the medicine for treating or preventing myopia is prepared.

The wherein preferred 4- phenyl -3- N- Yangization oxadiazole formonitrile HCN of N0S substrates, arginine, canavanine, the arginine of Ν alpha-substituted and its officinal salt or ester；More preferably 4-stupid base-3- Ν-Yangization oxadiazoles formonitrile HCN, L-arginine, L- canavanine, Not-substituted L-arginine and its officinal salt or ester；L-arginine, the L-arginine and its officinal salt or ester of Net-hydrocarbon carbonyl oxygen substitution of the further excellent-N- Yangization oxadiazole formonitrile HCN of ease 4- phenyl -3, L-arginine, L- canavanine, Not-acyl group substitution；Particularly preferred 4- phenyl -3- N- Yangization oxadiazoles formonitrile HCN, L-arginine, L-canavanine, Ν α-benzoyls-L-arginine ethyl ester, two -0- (Not-benzoyl-L-arginine）Glycol ester, three -0- (No-stupid formoxyl-L-arginine）Glyceride；Most preferably L-arginine, L- canavanine, Noc-Ben Jia Ugly bases-L-arginine ethyl ester.

The preferred minoxidil of NO donors, hydrolazine, nitroglycerin, isobide and its ester, nitroprusside, nitrite, glutathione derivative and N- nitropyrazole derivatives；More preferably minoxidil, hydrolazine, nitroglycerin, ISDN, nitroprusside, natrium nitrosum, glutathione, GSNO, 3,5-dimethyl-N- nitropyrazoles, N- nitropyrazoles or 3- methyl-N- nitropyrazoles；Most preferably hydrazine bends paint. The preferred polyamine of 1N0S induction inhibitor, calcium antagonist, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody（TNFab), interleukin, kinases inhibitor, Tyrokine kinase inhibitors；More preferably spermine aldehyde, dihydropyridine, w- (N, N ,-diethylamino）- n- alkyl trimethoxybenzoic acid salt（TMB), platelet-derived growth factor, platelet activating factor antagonist, TNFab, interleukin -4, interleukin -8, interleukin -10, interleukin -13, interleukin-lra, kinases inhibitor, cloricromen；Most preferably spermine aldehyde, cloricromen.

The preferred L-arginine analog of 1N0S activities inhibitor, methylene blue, aminoguanidine, S- methyl isothioureas；More preferably N^G- methyl-L-arginine, N^G- amino-blood sperm propylhomoserin, N^G- amino-L-arginine, N^G- nitro-L-arginine, diethylarginine, methylene blue, aminoguanidine, most preferably S- methyl isothioureas, aminoguanidine.

(2) pharmaceutical composition, containing selected from N0S substrates and/or NO donors or iNOS induction inhibitor and/or the one or more kinds of compound of iNOS activities inhibitor and pharmaceutical acceptable carrier or excipient.

The wherein stupid bases -3 of the preferred 4- of N0S substrates ~ N-Yangization oxadiazoles formonitrile HCN, arginine, canavanine, Net-substituted arginine and its officinal salt or ester；More preferably 4-phenyl-3- N- Yangization oxadiazoles formonitrile HCN, L-arginine, L- canavanine, Not-substituted L-arginine and its officinal salt or ester；L-arginine, the L-arginine and its officinal salt or ester of Ν α-hydrocarbon carbonyl oxygen substitution of the stupid-N-Yangization oxadiazole formonitrile HCNs of base -3 of further preferred 4-, L-arginine, L- canavanine, Net-barefoot base substitution；The L-arginine and its officinal salt or ester of acyl group substitution；Stupid base -3- Ν-Yangization oxadiazole the formonitrile HCNs of particularly preferred 4-, L-arginine, L- canavanine, Not-stupid formoxyl-L-arginine ethyl ester, two -0- (Ν α-benzoyl-L-arginine）Glycol ester, three -0- (Noc-benzoyl-L-arginine）Glyceride；Most preferably L-arginine, L- canavanine, Ν α-benzoyl-L-arginine ethyl esters.

The preferred minoxidil of NO donors, hydrolazine, nitroglycerin, isobide and its ester, nitroprusside, nitrite, glutathione derivative and N- nitropyrazole derivatives；More preferably minoxidil, hydrolazine, nitroglycerin, ISDN, nitroprusside, natrium nitrosum, glutathione, GSNO, 3,5-dimethyl-N- nitropyrazoles, N- nitropyrazoles or 3- methyl-N-nitropyrazole；Most preferably hydrolazine.

INOS induction inhibitor preferably is selected from polyamine, calcium antagonist, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody（TNFab), Bai Jie Element, kinases inhibitor, Tyrokine kinase inhibitors；More preferably spermine aldehyde, dihydropyridine, w- (N, N ,-diethylamino）- n- alkyl trimethoxybenzoic acid salt（TMB), platelet-derived growth factor, platelet activating factor antagonist, TNFab, interleukin -4, interleukin -8, interleukin -10, interleukin -13, interleukin-lra, kinases inhibitor, chlorine gram year Meng；Most preferably spermine aldehyde, cloricromen.

The preferred L-arginine analog of iNOS activities inhibitor, methylene blue, aminoguanidine, S- methyl isothioureas；More preferably N^G- methyl-L-arginine, N^G- amino-blood sperm propylhomoserin, N^G- amino-L-arginine, N^G- nitro-L-arginine, diethylarginine, methylene blue, aminoguanidine, S- methyl isothioureas；Most preferably aminoguanidine.

Aforementioned pharmaceutical compositions, can also contain anticholinergic agent, cycloplegic and/or parasympathomimetic agent.

The wherein preferred atropine of anticholinergic agent and pirenzepine, the preferred cyclopentolate of cycloplegic and Tropicamide, the preferred phenylephrine of parasympathomimetic agent is He Kidney upper parathyrines；More preferably atropine.

(3) compound, it is two -0- (Ν α-benzoyl-L-arginine）Glycol ester, three -0- (Net-benzoyl-L arginine）Glyceride.

(4) prevention of myopia or treatment method, including by therapeutically effective amount（2) composition described in or（3) compound gives patient.

5. a kind of commercial kit, including therapeutically effective amount are above-mentioned（2) Group compounds and（3) compound, and record the Group compounds or compound can be used in the treatment or prevention of myopia or the data of said composition or compound should be used.

The brief description of accompanying drawing

Fig. 1 is the suppression for illustrating to induce iNOS by the interleukin -1P iNOS induced and spermine aldehyde.

Fig. 2 is the relaxation effect for illustrating L- canavanine to muscle

Fig. 3 is the relaxation effect for illustrating hydrolazine to muscle

Detailed description of the invention

It is used as the specific example of the myopia of the present invention, curvature myopia, malignant myopia, progressive myopia, preceding drive myopia, simple myopia, index myopia can for example be enumerated, and the high myopia divided from myopic degree, low degree short sightedness, the true myopia divided from the stage of myopia, pseudo-myopia etc..In addition, also including relevant disease, such as retinopathy, the macular degeneration relevant with the age caused by myopia（) and cataract etc. AMD. If not otherwise stated, following term used in the entire disclosure of the present invention is interpreted as with following implication：

" acyl group " refers to the group that RC0- is represented; wherein R represents alkyl or aryl; the wherein preferred CI- C6 alkyl of alkyl; refer to the straight or branched alkyl with 6 carbon atoms of 1-; methyl, ethyl, propyl group, isopropyl, butyl, isobutyl group, sec-butyl, the tert-butyl group, amyl group, isopentyl, neopentyl, tertiary pentyl, 1- ethyl propyls, hexyl, isohesyl, 1 can be enumerated; 1_ dimethylbutyls, 2; 2- dimethylbutyls, 3,3- dimethylbutyls, 2- ethyl-butyls etc.；Aryl refers to benzene, naphthalene etc..Wherein, the preferred ethyls of R, phenyl.Above-mentioned group R can also be replaced with following groups, for example, low-grade alkenyl（C2- C6 alkenyl etc. such as vinyl, cyclobutenyl, acrylic）, cycloalkyl（C3- C7 cycloalkyl etc. such as cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl）, aryl（The C6- C10 aryl such as stupid base, naphthyl）, fragrant heterocyclic radical（Such as furyl, thienyl, pyrrole radicals, oxazolyl, isoxazolyls, thiazolyl, isothiazolyl, imidazole radicals, pyrazolyl）, amino, one or two elementary alkyl amido（One or two CI- C6 alkyl aminos etc. such as methylamino, ethylamino, dimethylamino）, three low alkyl group ammoniums（Three-CI- C6 alkyl ammonium groups etc. such as front three ammonium, three second ammoniums, 3 third ammoniums）, the substituent number, excellent ease for 1- 3, in the occasion for having all substituent, they can be with identical, can also be different.

" hydrocarbon carbonyl oxygen " is the group that formula R0C0- is represented, wherein R represents alkyl, special bags of willow branches, framed by bamboo, for building dykes base or aryl；Wherein alkyl can enumerate CI- C6 alkyl, refer to the straight or branched alkyl with 6 carbon atoms of 1-, methyl, ethyl, propyl group, isopropyl, butyl, isobutyl group, sec-butyl, the tert-butyl group, amyl group, isopentyl, neopentyl, tertiary pentyl, 1- ethyl propyls, hexyl, isohesyl, 1 can be enumerated, 1- dimethylbutyls, 2,2- dimethylbutyls, 3,3- dimethylbutyls, 2- ethyl-butyls etc.；For example, the C2- such as vinyl, cyclobutenyl, acrylic C6 alkenyl for alkenyl；Aryl refers to benzene, naphthalene etc..Wherein, the preferred methyl of R, ethyl, the tert-butyl group or phenyl.Above-mentioned group R can also be replaced with following groups, for example, low-grade alkenyl（C2- C6 alkenyl etc. such as vinyl, cyclobutenyl, acrylic）, cycloalkyl（C3- C7 cycloalkyl etc. such as cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl）, aryl（The C6- C10 aryl such as phenyl, naphthyl）, fragrant heterocyclic radical（Such as furyl, thienyl, pyrrole radicals, Evil oxazolyl, isoxazolyls, thiazolyl, isothiazolyl, imidazole radicals, pyrazolyl）, amino, one or two elementary alkyl amido（One or two C1 such as methylamino, ethylamino, dimethylamino - C6 alkyl aminos etc.）, three low alkyl group ammoniums（Three-CI-C6 alkyl ammonium groups etc. such as front three ammonium, three second ammoniums, 3 third ammoniums）, the substituent number, preferably 1 ~ 3, in the occasion for having several substituents, they can be with identical, can also be different.

In this manual, " patient " includes people and other mammals.

In this manual, term " high myopia " refer to more than 500 degree myopia, preferably more than 600 degree of myopia, " low degree short sightedness " refers to less than 500 degree of myopia.

In this manual, N0S substrates are that can be changed into NO material in the presence of NO synzyme.- N-Yangization oxadiazole the formonitrile HCN of 4-phenyl-3, canavanine, and the compound with basic arginine structure can be enumerated.So-called " having basic arginine structure " refers to arginine compounds, has different substituted arginine derivatives on arginic nitrogen, or be used as other compounds such as Ν of arginine metabolism thing_α- hydroxyarginine derivative and specific Ν^ω- hydroxyarginine and its various Ν_αThe ester-formin of-protection.Also include that arginic " prodrug " compound can be hydrolyzed to.Above-mentioned canavanine, and the compound with basic arginine structure can be the forms such as its ester or salt or amidate.

In the present invention, the excellent ease of canavanine has L _ isomer configuration.Preferably the compound with basic arginine structure has L-isomer configuration, preferably L-arginine and Ν_αL-arginine derivative of-substitution.Suitable N_tt- substituent includes above-mentioned acyl group, excellent ease benzoyl or acetyl group；And above-mentioned hydrocarbon carbonyl oxygen, preferably tert-butoxycarbonyl (Boc) and benzyloxycarbonyl (Cbz).

Wherein, some N0S substrates contain hydroxyl; suitable ester containing hydroxy compounds is; such as acetic acid esters, citrate, lactate, tartrate, malonate, oxalate, salicylic acid ester, propionic ester, succinate, fumarate, maleate, methylene-two-β oxynaphthoic acid ester, the stupid formic acid esters of 2,5- dihydroxy, isethionic acid ester, two-to the stupid formoxyl tartrate of first, methanesulfonates, esilate, benzene sulfonate, p-methyl benzenesulfonic acid ester, cyclohexylsulfamates and quinate.

Upper fan N0S substrates with carboxyl, can form base addition salts, ester or amidate.The alkali of officinal salt can be formed when being mixed with free acid by preferably including those available for the alkali for preparing base addition salts, the officinal salt refers to its cation salt nontoxic to patient under the dosage of salt, so that the beneficial inhibitory action of free alkali will not be disturbed by the side effect caused by cation.Officinal salt in the scope of the invention, including from salt derived from alkali and alkaline earth metal ions salt, including from salt made from following alkali：Sodium hydride, Sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminium hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, ammonia, ethylenediamine, N- methyl-glucamines, lysine, arginine, ornithine, choline, N, N ,-dibenzyl-ethylenediamin, chloroprocanine, diethanol amine, procaine, N- benzyl-1-phenylethylamines, diethylamine, piperazine, three（Methylol）Aminomethane, TMAH etc..The carboxylate formed can be nontoxic to patient ester under dosage, suitable esters can enumerate methyl esters, ethyl ester, isopropyl ester, butyl ester, isobutyl ester, the tert-butyl ester, pentyl ester and isopentyl ester, glycol ester, glyceride and benzyl ester, it is preferred that ethyl ester, glycol ester and glyceride, more preferably ethyl ester.The amidate formed can be that amidate nontoxic to patient under dosage, suitable can enumerate formamide, formyl ethamine, formyl propylamine, formyl butylamine, formailide etc..

Some above-mentioned N0S substrates are alkaline, and these compounds can be used in the form of the free alkali or in the form of its pharmaceutically acceptable acid addition salts.The acid of officinal salt can be formed when being mixed with free alkali by including those available for the sour excellent ease for preparing acid-addition salts, the officinal salt refers to its anion salt nontoxic to patient under the dosage of salt, so that the beneficial inhibitory action of free alkali will not be disturbed by the side effect caused by anion.Officinal salt in the scope of the invention is included from salt made from inorganic acid and organic acid, therefore including hydrogen from hydrochlorate, such as hydrochloride and hydrobromate, sulfate, phosphate, nitrate, sulfamate, acetate, citrate, lactate, tartrate, malonate, oxalates, salicylate, propionate, succinate, fumarate, maleate, methylene-two-β oxynaphthoic acid salt, 2, 5- dihydroxy-benzoic acid salt, isethionate, two-to toluoyl tartaric acid salt, mesylate, esilate, benzene sulfonate, tosilate, cyclohexyl-n-sulfonate and quinate.It is preferred that hydrochloride.

In this manual, term " NO donors " has no other limitations, for example vasodilator nitroglycerin plays its pharmacotoxicological effect by discharging nitrogen oxide as long as releasable NO plays the compound of its pharmacological action.Minoxidil, hydrolazine, nitroglycerin, isobide and its ester, nitroprusside, nitrite, glutathione derivative and N-nitropyrazole derivative can be enumerated." ester " therein and " salt " is same as described above.Glutathione derivative can enumerate glutathione, S-GSNO for that can carry the glutathione of substituent." N-nitropyrazole derivative " can for example enumerate 3,5-dimethyl-N-nitropyrazole, N-nitropyrazole and 3-methyl-N-nitropyrazole for that can carry N-nitropyrazole of substituent.More excellent ease minoxidil, hydrazine bend paint, nitroglycerin, ISDN, nitroprusside, natrium nitrosum, glutathione, S-nitroso paddy The sweet peptide of Guang, 3,5- dimethyl-N- nitropyrazoles, N- nitropyrazoles and 3- methyl-N

- fluorine base pyrazoles；Most preferably hydrolazine.

As long as iNOS induces induction of the inhibitor to iNOS to have the material of inhibitory action, other limitations are had no.It preferably is selected from polyamine, calcium antagonist, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody（TNFab), interleukin, kinases inhibitor, Tyrokine kinase inhibitors.Wherein " polyamine " can be enumerated：Spermine aldehyde." calcium antagonist " can enumerate dihydropyridine, W- (N, N ,-diethylamino）- n- alkyl trimethoxybenzoic acid salt（TMB)." interleukin " can enumerate interleukin _ 4, interleukin -8, interleukin -10, interleukin -13, interleukin _ lra.

" Tyrokine kinase inhibitors " can enumerate cloricromen.

More preferably spermine aldehyde, dihydropyridine, w- (N, N ,-diethylamino）- n- alkyl trimethoxybenzoic acid salt（TMB), platelet-derived growth factor, platelet activating factor antagonist, TNFab, interleukin -4, interleukin -8, interleukin -10, interleukin -13, interleukin-lra, kinases inhibitor, cloricromen；Most preferably spermine aldehyde, cloricromen.

INOS activities inhibitor are the material for having inhibitory action to iNOS activity, have no other limitations.It is preferred that the different thin urea of L-arginine analog, methylene blue, aminoguanidine, S- methyl.Wherein " L-arginine analog " can be enumerated： N^G- methyl-L-arginine, N^G- amino-blood sperm propylhomoserin, Ν⁶- amino-L-arginine, r-nitro-L-arginine, diethylarginine.

More excellent ease-methyl-L-arginine, N^G- amino-blood sperm propylhomoserin, N^G- amino-L-arginine, N^G- nitro-L-arginine, diethylarginine, methylene blue, aminoguanidine, most preferably S- methyl isothioureas, aminoguanidine.

Above-mentioned N0S substrates and NO donors make ciliary muscle relaxation by improving NO concentration too low in eye；Above-mentioned iNOS induction inhibitor and iNOS activities inhibitor suppress the protrusion of central sclera by reducing NO concentration too high in eye；Two kinds of effects of upper fan all can shorten the axis of eyeball, reach the purpose for treating or preventing myopia.Wherein, above-mentioned iNOS induction inhibitor and iNOS activities inhibitor are especially effective to high myopia.

In the present invention, two -0- (Ν α-stupid formoxyl-L-arginine）Glycol ester, three -0- (Ν α-benzoyl-L-arginine）Glyceride is noval chemical compound.Other compounds are all known compound, and wherein 4- phenyl -3- Ν-Yangization oxadiazole formonitrile HCN is disclosed in W099/30716, and remaining can be obtained by commercially available.Wherein, minoxidil, hydrazine It is antihypertensive to bend piperazine, nitroprusside；Nitroglycerin, isobide and its ester are antianginal drug；Calcium antagonist is anti-hypertension and vasodilator agent；Platelet-derived growth factor is hemostatic；Platelet activating factor antagonist is anticoagulation；TNFab, interleukin, kinases inhibitor, Tyrokine kinase inhibitors are immunomodulator；Methylene blue is antidote；Aminoguanidine, S-methyl isothiourea are common agents.

The compound of the present invention has effective pharmacological activity, therefore can be incorporated into Yao Wu Group compounds, present invention additionally comprises containing at least one the compounds of this invention, pharmaceutical acceptable carrier or excipient, and/or other drugs etc. Yao Wu Group compounds.

Other drugs refer to other medicines for being used to treat or prevent myopia, such as anticholinergic agent, cycloplegic and/or parasympathomimetic agent.

The wherein preferred atropine of anticholinergic agent and pirenzepine, the preferred cyclopentolate of cycloplegic and Tropicamide, the preferred phenylephrine of parasympathomimetic agent and adrenaline.Most preferably atropine.

Above-mentioned Hua compound Huo Group compounds can be administered by suitable method.In practice, it can be generally administered by parenteral, part, rectum, oral, suction or embedment.Such as treat posterior retina degeneration damage also needs Formulations for systemic administration in addition to locally administration.

According to administering mode, composition can be solid such as tablet, pill, powder, granule, glue Nang agent, sugar-coat agent etc., semi-solid such as ointment, gel, liquid dosage form such as injection, solution, suspension liquor, syrup, eye drops etc., the unit dosage form of the suitable single-dose of gas formulation such as inhalant etc., preferably exact dose.In view of the influence to other circulatory systems and its effect, the form of the local administration of eye is preferably used in, particularly preferred eye drops, ophthalmic ointment, gel, ocular administration construct, it is referred to as blind pipe to be suitable for insertion into（With the region similar to pocket of conjunctiva formation）Ocular formulation（The solid carrier for being soaked with medicine such as inserted in eye）.

The preparation of above-mentioned form of medication, necessary additive on the preparations such as common carrier, excipient, bonding agent, stabilizer can be added in this compound, preparation is conventionally made.For example, the carrier that will allow on the compounds of this invention and preparation（Excipient, bonding agent, disintegrant, flavouring, flavoring agent, emulsifying agent, diluent, isotonic agent, dissolving adjuvant etc.）Mixing, is made tablet, pill, powder, granule, glue Nang agent, sugar-coat agent, ointment, gel, injection, solution, suspension liquor, syrup, suppository, inhalant, cutaneous permeable agent, eye drops or ocular administration construct etc. and is suitable for the form of medication such as mouth month, parenteral, part, rectum, suction or embedment. When solid pharmaceutical preparation is made, additive can be used such as sucrose, lactose, cellulose sugar, PEARLITOL 25C, maltitol, glucan, starch, agar, arginine ester, chitin, chitosan, pectin, bassora gum, Arabic gum, gelatin, collagen, casein, albumin, calcium phosphate, sorbierite, glycine, carboxymethyl cellulose, polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, glycerine, polyethylene glycol, sodium acid carbonate, magnesium stearate, talcum.Moreover, the tablet for generally having coating, such as sugar coated tablet, enteric coatings piece, film coating ply or two-ply, multilayer tablet can be made in tablet when necessary.

When semisolid preparation is made, vegetable and animals oils can be used（Olive oil, corn oil, castor oil etc.）, mineral oil（Vaseline, White petrolatum, solid paraffin etc.）, wax class（Jojoba oil, Brazil wax, beeswax etc.）, polyethylene kind mineral oil gel, it is partially synthetic or all synthesis fatty acid glyceride（Laurate, myristic acid, palmitic acid etc.）.

When preparation is ointment, it can suitably escape and select ointment bases（Vaseline, lanolin, plastic substrate）, preservative（Benzalkonium chloride, parabens, methaform etc.）Deng being prepared.

When liquid preparation is made, additive is such as sodium chloride, glucose, sorbierite, glycerine, olive oil, propane diols, ethanol.

Liquid is implemented such as injection, eye drops.

When injection is made, aseptic aqueous solution can be used（Such as physiological saline）, isotonic solution, oiliness liquid（Such as sesame oil, soybean oil）.In addition, if necessary can also be simultaneously using appropriate suspended agent, such as plain sodium of Suo Jia Ji Yu dimensions, nonionic surfactant, dissolving adjuvant（Such as benzyl benzoate, phenmethylol）Deng.

When eye drops is made, waterborne liquid or the aqueous solution, particularly sterile aqueous solution for injection can be used.Slow electuaries, stabilizer, wetting agent, emulsifying agent, suspended agent, surfactant, isotonic agent, preservative, tackifier class various additives can also be properly added in eye drops.

Slow electuaries can be used such as upright stone tablet hydrochlorate buffer, boric acid Slow electuaries, citrate Slow electuaries, tartaric acid Slow electuaries, acetate Slow electuaries, amino acid.

Stabilizer can be used such as sodium ethylene diamine tetracetate, citric acid.

Wetting agent is such as glycerine.

Emulsifying agent is such as polyvinylpyrrolidone.

Suspended agent is such as hydroxypropyl methyl cellulose, methylcellulose. Surfactant is such as Tween 80, Crodaret.

Isotonic agent can use the carbohydrate such as sorbierite, glucose, mannitol, the polyalcohol such as glycerine, propane diols, the salt such as sodium chloride.

Preservative can use the quaternary ammonium salt such as benzalkonium chloride, benzethonium chloride, to the parabenses such as the stupid methyl formate of hydroxyl, ethyl-para-hydroxybenzoate, phenmethylol, benzyl carbinol, sorbic acid and its salt, thimerosal, methaform etc..

PH adjusting agent such as sodium hydroxide, hydrochloric acid or sulfuric acid etc..

Tackifier can be used such as hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose and its salt.

If using suspension, then its granular size should be less than Ι Ο μ ι η, so as to which the stimulation to eye is minimized.As eye drops in use, pH is usually adjusted to about 3-9. 5, be preferably adjusted to about 6-8. 5, most preferably can close to tears Ρ Η values, i.e. Ρ Η 7. 4 or so.

If using micro- fat body preparation, by the way that above-claimed cpd or composition are mixed together with suitable lipids or phospholipid substance, be added into various micro- fat bodies.Such a preparation, is particularly useful form for N0S substrates and NO donors.Some arginine class compounds（Such as three-（ Ν_α- benzoyl-L-arginyl-）- glycerine）Nang bubbles itself can be spontaneously formed under suitable conditions, as a result there is provided the form with the compound for changing release and/or metabolic rate.

If using ocular administration construct, various polymer may be used as preparing opthalmological carrier.Document, which is chatted, below has been confused such polymer：（Saettone, M. F., etc. J. Pharm. Pharmocol (1984) 36：229, and Park, K. etc., in Recent Advances in Drug Del ivery Systems. Anderson etc., eds., Plenum Press (1984) 163-183), full text is introduced into the application and referred to.The general bio-digestion through dissolution or polymer of insoluble drug release, infiltration or its process merged.The preparation of construct should make the rate of release of medicine not significantly interfere with tears tension force.

More specifically, several matrix type induction systems can be used for the present invention.These systems have in the following references chats fan in detail.（Ocular Pharm. of Drug Release Devices, in Controlled Drug Del ivery. Bruck, ed., vol. II, Chap 4, CRC Press Inc (1983)), its full text is introduced herein as reference.The system includes the hydrophily soft contact ophthalmic len with medicine needed for being soaked with or being marked with, and biodegradable or soluble construct, and this composition thing need not be removed again after inserting in eye.The soluble eye insert can be by any degradable material composition, it Can be resistant to by eyes and be compatible with the medicine given.Be confused material includes but is not limited to gather（Vinyl alcohol）, polymer and copolymer, the ethyl acrylate and vinyl pyrrolidone of polyacrylamide, and the polypeptide or polysaccharide being crosslinked, such as chitin.

Glue Nang type induction systems can be used for the present invention.These system application polymer films control the release of the medicine.The construct is used especially for conveying hydrophilic medicament.Hydrophobic drug can be given through silicon rubber construct.

The amount of above-claimed cpd or composition in myopia prevention or curative is preparation

The weight % of 0. 01- 100, suitably 0. 1-10 weight %.Dosage, administration number of times according to symptom, administration number of times, the age, body weight, administering mode and it is different, generally for adult, for example when being used as eye drops, it can be thrown to the preparation containing its 0. 001-10w/v%, preferably 0. 01-1. 0$ ^% for N0S substrates；For NO donors, it can throw to the preparation containing its 0. 001-10w/v%, the preferably 3w/v% of 0. 03- 0.；Inhibitor or iNOS activities inhibitor are induced for iNOS；It can throw to containing its 0. 001- 10w/v%, preferably (the ^ % preparations of 0. 1- 1.；For several times daily, preferably 1-6 times, preferably few drops every time, 1-3 drops.As injection in use, administration range can be 0. 1-20mg/kg, preferably 8-15mg/kg, most preferably 10mg/kg or so, for several times daily, preferably 1-6 times.Mode is released as implantation Slow, the effective dose of myopia is controlled generally in 0. 1- 20mg/kg/ days scopes.The system can be implanted into surgical site infections, so as to pre- myopia prevention.In order to treat myopia, the new implant of implantation in every 3-10 days, until the 60th day.The new implant of excellent ease implantation in every 5-7 days, until the 30th day.

Amount of the cholinolytic class medicine in myopia prevention or curative is 0. 03-0. 25 weight % of preparation, and concentration is too low, does not have corresponding effect；If but excessive concentration, the side effect to autonomic nerve occurs.

Present invention additionally comprises the prevention of myopia or treatment method, this method includes giving patient by the above-claimed cpd or composition of therapeutically effective amount.

In the present invention, two-0-(N ot-stupid formoxyl-L-arginine）Ethylene glycol vinegar, three-0-(N oc-benzoyl-L-arginine）Glyceride can be by can be by the way that N et-benzene first barefoot base-L-arginine be made with ethylene glycol or glycerine.

The acid-addition salts of the present invention can be by the way that free alkali be prepared with suitable acid using known method reaction.For example, free alkali can be by being dissolved in the water containing suitable acid or aqueous alcohol solutions or other suitable solvents and preparing solution evaporation separation salt by the acid-addition salts of the compounds of this invention, can also be by anti-in organic solvent by free alkali and acid It should prepare, in such a case it is possible to be directly separated out salt or salt can be obtained by concentrate solution.

The base addition salts of the present invention can be by the way that free acid be prepared with suitable alkali using known method reaction.For example, free acid can be by being dissolved in the water containing suitable alkali or aqueous alcohol solutions or other suitable solvents and preparing solution evaporation separation salt by the base addition salts of the compounds of this invention, it can also be prepared by the way that free bronsted lowry acids and bases bronsted lowry is reacted in organic solvent, in such a case it is possible to be directly separated out salt or salt can be obtained by concentrate solution.

The ester of the present invention can be obtained the compound of free acid or hydroxyl by known ester preparation method.

The amidate of the present invention can be by the way that free acid be obtained with amine using known preparation method

The present invention will be described in detail by embodiment and test example below, but the present invention is not limited by the embodiment and test example.

Embodiment

The eye drops 1 of embodiment 1

The compound of embodiment 1 is dissolved in distilled water for injection, pH to 7 is adjusted with sodium hydroxide, eye drops is prepared according to following formulas.

The 5g of L-arginine 0.

The lg of 2 hydrate of sodium dihydrogen phosphate 0.

The 9g of sodium chloride 0.

Appropriate distilled water for injection

The eye drops 2 of full dose 100ml embodiments 2

According to method same as Example 1, prepare with 1. 0_g/ 100ml concentration contains the eye drops of L-canavanine.

The eye drops 3 of embodiment 3

According to method same as Example 1, the eye drops for containing N-benzoyl-L-arginine ethyl ester with 0. lg/100ml concentration is prepared.

The eye drops 4 of embodiment 4

According to embodiment 1, the eye drops for containing hydrolazine with the m 1 of 0. 1 g 8 00 concentration is prepared. Blunt dose 5 of the drop of embodiment 5 H

According to embodiment 1, the eye drops for containing aminoguanidine with 0. 1 g/ 100ml concentration is prepared.

The eye drops 6 of embodiment 6

According to method same as Example 1, the eye drops for containing spermine aldehyde with 0. 7g/100ml concentration is prepared.

The eye drops 7 of embodiment 7

According to method same as Example 1, the eye drops for containing cloricromen with 1. Og/lOOml concentration is prepared.

The tablet of embodiment 8

The compound of mix embodiment 1, lactose, cornstarch and avicel cellulose, are integrated with PVP K30 paste liquid, are granulated by 20 mesh sieves.After being dried 2 hours under 50 °C, by 24 mesh sieves, the tablet that every is 120mg is made with a diameter of 7mm punch press for mixing talcum and magnesium stearate.

N ot-the 00mg of benzoyl-L-arginine ethyl ester 10.

The 04mg of atropine 0.

The OOrng of lactose 50.

The OOrng of cornstarch 20.

The 66mg of avicel cellulose 28.

The OOrng of PVP K30 5.

The OOrng of talcum 5.

The 30mg of magnesium stearate 0.

120. OOrng

The capsule of embodiment 8

The compound of mix embodiment 1, lactose and cornstarch, are integrated with PVP K30 paste liquid, are granulated by 20 mesh sieves.After being dried 2 hours under 50 °C, by 24 mesh sieves, mixing talcum and magnesium stearate are filled into ebonite Nang shells（No. 4）In, the capsule that every is 120mg is made. Ν α-the OOmg of benzoyl-L-arginine ethyl ester 5.

The OOmg of glutathione 3.

The OOmg of lactose 72.

The OOmg of cornstarch 35.

The OOmg of PVP K30 2.

Talcum 2.70mg

Magnesium stearate 0.30mg

120. OOmg test examples

This embodiment of test example 1 illustrates that NO donors and N0S substrates can effectively relax and causes the contraction of ciliary muscle by carbachol and Endothelin -1

Method

Material

Carbachol（Formyl chloride choline U.S. P) it is purchased from City Chemical Corp (New York-NY) Endothelin -1 (ET- 1, people and pig Chong Group bodies）, hydralazine hydrochloride, and natrium nitrosum are purchased from Sigma Chemical Co (St. Louis. MO).Hydrochloric acid L- arginine, sulfuric acid L- canavanine, and hydrochloric acid N- benzoyls-L-arginine ethyl ester (BAEE) are purchased from Pfales ＆Bauer companies (Water bury CT).Improved Krebs liquid is prepared by SILVER REAGENT pure material（Group is into mM:NaCl, 142.0;NaHC0₃, 18.0;Glucose, 11.0; KC1,5.4; CaCl₂, 2.0; MgCl₂, 1.2) 。

Group knits preparation

Fresh buphthalmos ball is obtained from local slaughterhouse, is immediately placed on and carries on ice to laboratory.After extraction ciliary body is isolated in 3 hours from eyeball.Under surgical operation microscope, from ciliary body, carefully solution cuts longitudinal ciliary muscle, and is cut into length about 3- 6mm flesh bar.With two ends of the silk suture colligation flesh bars of 5- 0, and flesh bar is placed in immediately in the organ bath equipped with the improved Krebs Slow fliud flushings of 10ml in 34 °C.

Organ bath and tape deck

Ciliary muscle flesh bar is vertically mounted in 10ml double glazing organ bath with suture, one suture is connected to the positioning pestle in bath, and another suture is connected on the isometric tension converters of Grass FT- 03 on adjustable gripper.It is full of in bath Acyclic Krebs Slow fliud flushings, and with 95 % 0₂ r 5 % 0)₂Mixed gas constantly effervescent.Often the pH of monitoring Krebs liquid, makes pH be maintained at 7. 4 ± 0. 4 by adding IN NaOH or IN HC1.Bath and Krebs liquid is set to be maintained at 34 °C with recirculated water.Tape deck by Grass 7D1 low level D. C preamplifiers, Grass 74DA amplifiers, with Grass 79D tetra- lead platform-type paper-ink Ji Lu Yi Group into.The record Shou Shrink power that offsets up of pen increases, and offsets downward record Shou Suo Li Minus small.Tape deck is calibrated before record every time.

Drug-treated scheme

By randomised way, add and in vitro test is carried out to ciliary muscle flesh bar for 5 testing compounds of this research.Before medicine is applied, first by muscle specimen in organ bath, at least balanced 1 hour with 300mg resting tensions, closely monitoring resting tension baseline, and be adjusted by vertical micro- manipulation transforms device.Once medicine is added in bath, then no longer manual adjustment tension force.In order to determine the effect that testing compound shrinks to carbachol excitant, to adding carbachol in the bath of some muscle specimens, reach 1x10-⁶M ultimate density, and have recorded contractile response.By 1x10-⁶M is defined as that the carbachol least concentration of the longitudinal ciliary muscle maximum collapse effect of ox can be caused（Data are not shown）.After maximum collapse about 5 minutes, testing compound is added to bath, and determine tension change.The flesh bar that each testing compound is handled is handled with from a sequence concentration of 0. 01%-3% weight.After the addition before continuous concentration, reach tension change and be spaced between metastable state, usual two concentration about 5-10 minutes.The flesh bar for causing control with carbachol shrinks, and is allowed to remain retracted state during testing, or is at least kept for 60 minutes.Ru Guo Group, which are knitted, not to react carbachol, then fills this sample.

In order to determine the effect that testing compound shrinks to ET-1 excitants, according to the identical test step received to carbachol inductivity, carbachol is replaced with ET-1, its final bath concentration be 1x10-⁷M.By 1x10-⁷M is defined as that ET -1 least concentrations of the longitudinal ciliary muscle maximum collapse effect of ox can be caused（Data are not shown）.Cause control flesh bar to shrink with ET -1, be allowed to remain retracted state during testing, or at least kept for 60 minutes.Ru Guo Group, which are knitted, not to react ET -1, then discards this sample.

Statistical analysis

All data are provided as the native standard error of average（ SE ) .Student's t- measure is merged, to analyze the conspicuousness of difference between two averages.In P<Think that difference has conspicuousness when 0. 05. As a result

The effect of carbachol and the in vitro ciliary muscle of 1 couple of ET-.

For all samples, 1x10-⁶M carbachols generate 552 ± 43mg average maximum collapse tension force from 300mg baseline resting tension above baseline.For all samples, lxl (T⁷M ET- 1 generate 408 ± 35mg average maximum collapse tension force from 300mg baseline resting tension above baseline.For the tissue of drug-treated, allow muscle to shrink about 5- 10 minutes in maximum after carbachol is added, then add medicine to be measured.For control tissue, it is allowed to contraction of muscle 60 minutes, and by the data point of the last tension force effect maximum collapse percentage after 60 minutes.Compare flesh bar for carbachol, it is 560 ± 90mg tension force above baseline that average last after 60 minutes, which shrinks, or 96.0 ± 2.5% maximum collapse.Flesh bar is compareed for ET- 1, it is 340 ± 21mg tension force above baseline that average last after 60 minutes, which shrinks, or 71.7 ± 5.4% maximum collapses.The effect of N0S substrates

For the muscle of carbachol processing, it is compared with a control, 0.3% weight and 1% weight L-canavanine and 0.1%-1% weight BAEE generate significant relaxation effect（Table 1).It is low up to 0.1% low concentration L- canavanine and the BAEE of the low low concentration up to 0.03% weight, the relexation of aobvious chopsticks is less produced compared with the control.For the muscle of carbachol processing, it is compared with a control L-arginine and produces relexation (table 1) in 3% weight concentration.

For the muscle of the processing of ET- 1, it is compared with a control, the L- canavanine of all concentration all produces significant relexation（Table 2).It is compared with a control, BAEE is only in 0.01%-0. iy.The low concentration of weight generates significant relexation (table 2).It is compared with a control, L-arginine produces the relexation of aobvious chopsticks under 3% weight concentration（Table 2).

The effect of the various concentration of comparative drug, it is found that result is different between the muscle that the muscle and ET- 1 of carbachol processing are handled（Tables 1 and 2).For the muscle of the processing of ET- 1, compared with the muscle handled carbachol, L- canavanine is all to produce aobvious chopsticks compared with unique N0S substrates that prednisone relaxation is acted in all measured concentration（Fig. 2).For the muscle of the processing of ET- 1, BAEE generates aobvious chopsticks in the weight low concentration of 0.01% weight -0.1% and acted on compared with prednisone relaxation（P≤0.05), still, for the muscle of carbachol processing, aobvious chopsticks is actually generated under the higher concentration of 0.3% weight and 1% weight and is acted on compared with prednisone relaxation.L-arginine is in the concentration determined, to the muscle of carbachol processing with the effect of the muscle to the processing of ET- 1, being shown in 3% weight There is significant difference（Tables 1 and 2).

Table 1

N0S substrates are to 1 χ 10_⁶The effect of the ciliary muscle of Μ carbachols processing

The percentage of maximum collapse sharp kappa ^

0.01% 0.03% 0.1% 0.3% 1% 3% degree of medicine/dense

The .0 of 101.+2.7 100.8 ± 3.4 98. d3.3 of L " ^ propylhomoserin 102.342.3 102.942.3 90.

103.3+2.6 100.8+2.6 95.3±6.0 89.6+3.8* 60.5±14.4* -

All data of mo.0* of 101.2 ± 3.4 94.6+4.0 35.3 ± 3.6* of 56.3+4.2* 28.5+6.5* of BAEE 21. are expressed as the average scholar SE of N=6,

* represent that there is significant difference, P≤0.05 with 100%

Table 2

NOS substrates to 1x10-⁷The effect of the ciliary muscle of the processing of M Endothelins -1

The percentage for the maximum collapse that Endothelin -1 is stimulated

Medicine/0.01% 0.03% 0.1% 0.3% 1% 3%

The ￡ L 8 96.2 ± 3.6 87.3 ± 5.2 of L " ^ cities 98.6^b89.7 ± 6.2 85. .4 70.6+13.5* L-r of 88.3 ± 3.4* of knife, 75. 32.9 ± 5.6* of .3* 59.7+4.3* 8.

Data all 96.3 2.2 27.7+8.4* of β .6* 74.6db6.6* 77., 96. .1 of BAEE 87.4 are expressed as the average soil SE of Ν=6,（Except^aN= 5, ^bN= 8)

* represent that there is aobvious chopsticks sex differernce, P≤0.05 with 100%

The effect of nitric oxide donor

For the muscle of carbachol processing, it is compared with a control, the natrium nitrosum of 0.1% weight hydrolazine and all concentration generates significant relexation（Table 3).For the muscle of the processing of ET- 1, it is compared with a control, the natrium nitrosum of the hydrolazine of all concentration and all concentration all generates significant relexation（Table 4).

Compared with the muscle of carbachol processing, for the muscle of the processing of ET- 1, it has been suggested that hydrolazine can be produced significantly compared with the effect of prednisone relaxation under the concentration of 0.01% weight and 0.03% weight（Fig. 3).The relexation of muscle of the 0.1% weight hydrolazine to the muscle handled of ET- 1 and to carbachol processing, close to 100% relaxation.Natrium nitrosum is produced significantly compared with the effect of prednisone relaxation in the muscle that the concentration of 0.03% weight and 0.1% weight is handled ET- 1（P≤0.05), still, the muscle handled in the concentration of 0.3% weight carbachol produces aobvious chopsticks and acted on compared with prednisone relaxation（P≤0.05) .It is respectively 0.01% in natrium nitrosum The effect of the muscle handled under the least concentration and maximum concentration of weight and 1 % weight for the muscle and carbachol of ET -1 processing, does not show chopsticks sex differernce（Table 3 and table 4).

Table 3

NO donors to 1x10-⁶The effect of the ciliary muscle of M carbachols processing

The percentage of maximum collapse sharp kappa M ' j

o

The 1+1. 8* N/A N/A of ± 1. 9 97. 8+2. of hydrolazine 101. 8 7-1. of medicine/concentration 0. 01% 0. 03% 0. 1% 0. 3% 1%

The sour 8+4. 9 of sodium 80.^a* 75. 7+6. 2^a* 63. 4+5. 9^a* 22. 1±3. 5^a* -15. 6+3. * o

All data are expressed as the o averages soil SE of N=6, (except^aN = 5 )

* represent that there is significant difference, P≤0. 05 with 100 %

Table 4

NO donors to 1x10-⁷The effect of the ciliary muscle of the processing of M Endothelins -1

The percentage for the maximum collapse that Endothelin -1 is stimulated

The 8+4. 5* N/A N/A of 57. 5+4. 0* of hydrolazine, 18. 9 ± 4. 1* 2. of medicine/concentration 0. 3% 1%

Data all the 2* -8. 5i3. 6* of 47. 3+3. 9* of Righteousness 75. 3+3. 1*, 58. 5+2. 1* 38. 2 are expressed as the average soil SE of N=6,

* represent that there is aobvious chopsticks sex differernce, P≤0. 05 with 100 %

(result）

The N0S substrates and NO donors of the present invention can produce significant ciliary muscle relaxation effect.The cGMP of test example 2 measure

(method）

In order to determine after administration in RPE cells（Purchased from ATCC companies）The generation of middle intracellular cGMP, we will use cyclo GMP enzyme immunoassay kit（Purchased from Cayman Chemical companies）.This detection method is that the striving property unexpectedly between the cGMP tracers based on free cGMP and acetylcholinesterase connection is combined.CGMP tracers yellowing and there is strong absorbance in 412nm when adding Ellman's reagents.Giving NO donors or N0S substrates respectively, and after the control sample containing only carrier, will crack cell according to the specification of kit, and preparing lysate, cGMP is being measured.

(result）

As a result show, compared with the carrier of control, using the N0 donors or N0S substrates of the present invention, cGMP amount shows chopsticks and is higher than control sample. The application Western blot of test example 3 detects iNOS

(method）

By retinal pigment epithelium (RPE cells, purchased from American Tissue and Cell Collection companies) and Interleukin -1β (100ng/ml, Sigma Chemical companies) or carrier or the β of interleukin -1 (100ng/ml, Sigma Chemical) and iNOS induction inhibitor（LOOng/ml after) incubating 12 hours altogether, culture medium is taken out, the lysis buffer that 0.5ml boils is added to each hole pond（ImM Tris pH7.4,1 % lauryl sodium sulfate（ SDS) ) .For half bore pond therein, to prevent NO volatilization, the generation of nitrogen oxide in its culture medium is determined immediately（See test example 2).Net each Kong Chi inclusion is taken, is transferred in 1.5ml centrifuge tube.Make the cell inclusion of cracking together with Slow fliud flushings, by No. 26 syringe needles 6 times, to split off the inclusion of core.Can be by storage of samples in -70.C.With BCA detection methods（Pierce companies）, the standard curve for the bovine serum albumin(BSA) prepared in 0.1% SDS is applied, the protein concentration of each sample is determined.This measure is to apply Packard Spectra Cont spectrophotometers, is carried out in the microtest plate in 96 hole ponds.In order to reduce SDS to BCA detection reagents（Purchased from Pierce companies）Interference, it is necessary to water by sample 1:10 dilutions, make SDS concentration be reduced to 0.1%.And determine each sample and contain 40 μ₈The volume of total protein.

By sample and 10 °/dyeing Slow fliud flushings and 3% reduction Slow fliud flushings（Purchased from Novex companies）Mixing, then in company with prestained standard protein gradient（Gibco companies）(Novex companies are purchased to the small gels of NuPAGE Bis-Tris together）Sample-adding.In MOPS- SDS electrophoresis Slow fliud flushings（Purchased from Novex companies）In, electrophoresis is carried out to this gel with about 200V constant voltage.Bio- Rad Trans- Blot transfer devices are used after gel electrophoresis, by Protein transfer to Immobilon-P PVDF transfer membranes（Micropore）On.After transfer is completed, by with rushing salt solution in the Tris-Slow containing 0.1% Tween-20（TBS-T) middle 5% confining liquid prepared（Purchased from Amersham companies）Incubate altogether at room temperature, to membrane closure 60 minutes.After washing away confining liquid in TBS- T, by with exempting from anti-iNOS polyclonal antiserums（Mouse area phagocyte from cytokine induction, Cayman Chemical), measured enrichment stage is incubated 60 minutes altogether at room temperature, the iNOS albumen on film is detected.After washing away first antibody solution, by the goat-anti-rabbit secondary antibody of this film and peroxide binding marker（Purchased from Bio- Rad companies）, measured enrichment stage is incubated 60 minutes altogether at room temperature.Then (it is purchased from using ECL Western Blot Detection kits (ECL Western Blot Detection Kit) Amersham companies）, by chemoluminescence method, and this film is exposed X-radiographic film, the protein belt on observation film.

To the statistical analysis of data

Will be with the native standard deviation of average value（SE form) provides all data.Using the conspicuousness between two averages of Stndent's t- check analyses of merging.

(result）

As a result show, only add the sample of carrier, do not there is iNOS protein belt；The β of interleukin -1 sample is added, there is the generation of iNOS protein belt；The sample of the β of interleukin -1 and spermine aldehyde is added, the protein belt that iNOS is not shown is produced.INOS induction inhibitor, particularly the spermine aldehyde of the present invention shows the significant blocking effect induced for IL -1 β iNOS, and the generation of iNOS albumen is suppressed by spermine aldehyde（Accompanying drawing 1).

Measure of the test example 4 to nitrogen oxide

(method）

By RPE cells (being purchased from American Tissue and Cel l Col lection companies) and the β of interleukin -1 (100ng/ml, S igma Chemical companies) or carrier or interleukin-11 (100ng/ml, Sigma Chemical companies）Inhibitor is induced with iNOS（After 100ng/ml) incubating 12 hours altogether, culture medium is taken out, nitrate/nitrite colorimetric detection kit is used（Purchased from Cayman Chemical), by the content that nitrogen oxide in culture medium is determined the step of its specification.

To the statistical analysis of data

With test example 1

(result）

As a result show, compared with the β of interleukin -1 of control, inhibitor, particularly spermine aldehyde are induced using iNOS, NO amount is suitable with the NO amounts produced using only carrier considerably less than the NO amounts that the β of interleukin -1 is produced are used alone（Table 5). In RPE cells, the suppression of the spermine aldehyde NO generation beta induced to il-1

* aobvious chopsticks increases P compared with control vector<0. 05.

* is compared with being used alone the β of interleukin -1, while significantly inhibiting P to the generations of Ν 0 using spermine aldehyde<0. 05.

N0 donors or N0S substrates can make the amount for the cGMP that N0 produces to guanylate enzyme activation show chopsticks increase as can be seen from the above results, make ciliary muscle relaxation, have good result for the too low myopia of N0.And iNOS induction inhibitor has a blocking effect of the obvious induction to iNOS, this blocking effect can long lasting for a large amount of N0 of elimination generation.INOS activities inhibitor can make iNOS activity reductions, significantly reduce N0 amount.So, both because NO produces excessive caused myopia, particularly high myopia for having good result.Also, they are for retinopathy caused by myopia, the macular degeneration relevant with the age（AMD) and cataract etc. effectively.

Industry application possibility

The N0S substrates and N0 donors of the present invention can be by increasing N0 concentration too low in eye, so that ciliary muscle relaxation with the mechanism that the ciliary muscle relaxation of the non-plant of cAMP- non-dependent is acted on；INOS induces inhibitor and iNOS activities inhibitor can reduce N0 too high in eye concentration, suppresses the protrusion of central sclera；Above two effect may finally all suppress the elongation of axis oculi, reach the effect of prevention or treatment myopia.

Prevention or curative that thus the compound or composition of the present invention can be effectively as various myopia or with near-sighted relevant disease, such as curvature myopia, malignant myopia, progressive myopia, preceding drive myopia, simple myopia, index myopia, and the high myopia divided from myopic degree, low degree short sightedness, the true myopia divided from the stage of myopia, pseudo-myopia etc..In addition, also include by myopia to it is serious when meeting caused by retinopathy, the macular degeneration relevant with the age（) and cataract etc. AMD.

Claims (34)

1. Claim

   1. the application of NO synthase substrates and/or NO donors or immune NO synthase induction inhibitor and/or immune NO synthase activities inhibitor in the medicine for treating or preventing myopia is prepared.

   2. the application of claim 1, wherein NO synthase substrates are selected from the arginine and its officinal salt or ester of the stupid base -3- N-Yangization oxadiazole formonitrile HCNs of 4-, arginine, canavanine, Ν α-acyl group substitution.

   3. the application of claim 2, wherein NO synthase substrates are selected from the L-arginine and its officinal salt or ester of 4- phenyl -3- N- Yangization oxadiazoles formonitrile HCN, L-arginine, L- canavanine, Net-acyl group substitution.

   4. the application of claim 3, wherein NO synthase substrates are selected from the stupid-N-Yangization oxadiazole formonitrile HCNs of base -3 of 4-, L-arginine, L- canavanine, Net-benzoyl-L-arginine ethyl ester, two-O- (Ν α-benzoyl-L-arginine）Glycol ester, three-Ο-(Ν α-stupid formoxyl-L-arginine）Glyceride.

   5. the application of claim 4, wherein NO synthase substrates are selected from L-arginine, L-canavanine, Ν α-stupid formoxyl-L-arginine ethyl ester.

   6. any one of claim 1- 5 application, wherein NO donors are selected from minoxidil, hydrolazine, nitroglycerin, isobide and its ester, nitroprusside, nitrite, glutathione derivative and N-nitropyrazole derivative.

   7. the application of claim 6, wherein NO donors are selected from minoxidil, hydrolazine, nitroglycerin, ISDN, nitroprusside, natrium nitrosum, glutathione, GSNO, N- nitropyrazoles, 3,5- dimethyl-N -s nitropyrazole, 3- methyl-N-nitropyrazole：■

   8. the application of claim 7, wherein NO donors are hydrolazines.

   9. any one of claim 1- 8 application, wherein immune NO synthase induction inhibitor is selected from polyamine, calcium antagonist, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody, interleukin, kinases inhibitor, Tyrokine kinase inhibitors.

   10. the application of claim 9, wherein immune NO synthase induction inhibitor is selected from spermine aldehyde, dihydropyridine, w- (N, N ,-diethylamino）The stupid formates of-n- alkyl trimethoxies, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody, Interleukin -4, interleukin -8, interleukin -10, interleukin -13, interleukin-lra, kinases inhibitor, cloricromen.

   11. the application of claim 10, wherein immune NO synthase induction inhibitor is spermine aldehyde or cloricromen.

   12. any one of claim 1- 11 application, wherein immune NO synthase activities inhibitor is selected from L-arginine analog, methylene blue, aminoguanidine, S- methyl isothioureas.

   13. the application of claim 12, wherein immune NO synthase activities inhibitor is selected from N^G- methylarginine, N^G- amino-blood sperm propylhomoserin, N^G- amino-L-arginine, N^G- nitro-L-arginine, diethylarginine, methylene blue, aminoguanidine, S- methyl isothioureas.

   14. the application of claim 13, wherein immune NO synthase activity inhibitor is aminoguanidine.

   15. Yao Wu Group compounds, containing selected from NO synthase substrates and/or NO donors or immune NO synthase induction inhibitor and/or exempting from the one or more kinds of compound of phlegm NO synthase activities inhibitor and pharmaceutical acceptable carrier or excipient.

   16. the composition of claim 15, wherein NO synthase substrates are aoxidized selected from-the N- of 4- phenyl -3.The arginine and its officinal salt or ester of-azoles formonitrile HCN, arginine, canavanine, Ν α-acyl group substitution.

   17. claim 16 Group compounds, wherein NO synthase substrates are selected from the L-arginine and its officinal salt or ester that 4- phenyl -3 aoxidizes ^ azoles formonitrile HCN, L-arginine, L- canavanine, Ν α-acyl group substitution.

   18. the composition of claim 17, wherein NO synthase substrates are selected from the-N- Yangization Evil azoles formonitrile HCN of 4- phenyl -3, L-arginine, L- canavanine, Ν α-benzoyls-L-arginine ethyl ester, two-Ο-(Ν α-benzoyl-L-arginine）Glycol ester, three -0- (Ν α-benzoyl-L-arginine）Glyceride.

   19. claim 18 Group compounds, wherein NO synthase substrates are selected from L-arginine, L- canavanine, Not-benzoyl-L-arginine ethyl ester.

   20. any one of claim 15- 19 Group compounds, wherein NO donors are selected from minoxidil, hydrolazine, nitroglycerin, isobide and its ester, nitroprusside, nitrite, glutathione derivative and N-nitropyrazole derivative.

   21. claim 20 Group compounds, wherein NO donors are selected from minoxidil, hydrolazine, nitroglycerin, ISDN, nitroprusside, natrium nitrosum, glutathione, S- GSNO, N- nitropyrazoles, 3,5- dimethyl-N -s nitropyrazole, 3- methyl-N- nitropyrazoles.

   22. claim 21 Group compounds, wherein NO donors are hydrolazines.

   23. any one of claim 15- 22 Group compounds, wherein immune NO synthase induction inhibitor is selected from polyamine, calcium antagonist, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody, interleukin, kinases inhibitor, Tyrokine kinase inhibitors.

   24. claim 23 Group compounds, wherein immune NO synthase induction inhibitor is selected from spermine aldehyde, dihydropyridine, w- (N, N ,-diethylamino）- n- alkyl trimethoxybenzoic acids salt, platelet-derived growth factor, platelet activating factor antagonist, tnf antibody, interleukin-4, interleukin-8, interleukin -10, interleukin -13, interleukin-lra, kinases inhibitor, cloricromen.

   25. claim 24 Group compounds, wherein immune NO synthase induction inhibitor is spermine aldehyde or cloricromen.

   26. any one of claim 15- 25 composition, wherein immune NO synthase activities inhibitor is selected from L-arginine analog, methylene blue, aminoguanidine, S- methyl isothioureas.

   27. the composition of claim 26, wherein immune NO synthase activities inhibitor is selected from N^G- methyl-L-arginine, N^G- amino-blood sperm propylhomoserin, N^G- amino-L-arginine, N^G- nitro-L-arginine, diethylarginine, methylene blue, aminoguanidine, S-methyl isothiourea.

   28. the composition of claim 27, wherein immune NO synthase activity inhibitor is aminoguanidine.

   29. any one of claim 15- 28 pharmaceutical composition, also containing anticholinergic agent, cycloplegic and/or parasympathomimetic agent.

   30. the pharmaceutical composition of claim 29, wherein anticholinergic agent are selected from atropine and pirenzepine, cycloplegic is selected from cyclopentolate and Tropicamide, and parasympathomimetic agent is selected from phenylephrine and adrenaline.

   31. the pharmaceutical composition of claim 29, wherein also containing atropine.

   32. compound, it is two -0- (Ν α-benzene first barefoot base-L-arginine）Glycol ester, three-Ο-(Ν α-benzoyl-L-arginine）Glyceride.

   33. prevention or the treatment method of myopia, including give patient by the Yao Wu Group compounds of any one of claim 15-31 or the compound of claim 32 of pharmacologically effective dose.

   34. a kind of commercial kit, Group compounds or compound are recorded including any one of claim 15-32, and records the Yao Wu Group compounds or compound can be used in the treatment or prevention of myopia or the data of the pharmaceutical composition or compound should be used.