CN101759741A - Compound and application thereof in preparation of medicine for treating angiogenesis - Google Patents

Abstract

The invention relates to a compound and application thereof in preparation of medicine for treating angiogenesis, and the compound is shown in formula (1), wherein R2 is a hydrocarbon group which contains 1-2 heteroatom(s) and has less than or equal to ten carbon atoms. The compound is prepared by carrying out acylation reaction 21 on compound in formula (1.1). The compound shown in the formula (1) and solvate thereof which can be acceptable in physiology can be used for preparing medicine for treating age-related macular degeneration and the medicine for treating diseases caused by angiogenesis of human being or animals.

Description

A kind of compound and the application in the medicine of preparation treatment angiogenesis thereof

Technical field:

The present invention relates to a kind of medical compounds and synthetic method thereof and the application in the medicine of preparation treatment age-related macular degeneration.

Background technology:

Angiogenesis comprises two notions: brephic blood vessel take place (vasculogenesis, VG) and postnatal vasculogenesis (anglogenesis, AG).VG refers to not to be had under the situation of vascular system, by endothelial progenitor cell (endothelialprogenitor, cells, EPCs) or angioblast (angloblasts) be divided into endotheliocyte, and form vasoganglion.AG refers at the adult blood vessel, breaks through the migration of tube wall matrix by the mature endothelial cell that has existed, and propagation and reconstruct make the blood vessel branch continue to prolong in the germination mode.Angiogenesis among the present invention be meant postnatal vasculogenesis (anglogenesls, AG).Angiogenesis (anglogenesls) process that (as growth, wound healing) institute must have during still normal physiological changes, scientists finds that also the development of numerous diseases such as it and tumour, age-related macular degeneration, malignant hematologic disease has confidential relation in recent years.Suppress pathologic angiogenesis, can treat or slow down numerous diseases such as tumour, age-related macular degeneration, malignant hematologic disease

Age-related macular degeneration (age-related macular degeneration, AMD), be a kind of being mainly in more than 45 years old, the disease that morbidity increases with age growth, it is the major reason of current middle-aged and old blindings, choroidal artery is invaded retina and is constituted choroidal neovascularization down, and macular area retinal pigment epithelium serosity or hemorrhagic disciform detachment down or under the neuro epithelium take place, and the macular degeneration that causes is considered to important pathogenesis.

Having introduced anecortave acetate among the CN03818826 is that a kind of exploitation is used to suppress the vasoinhibitor that the eyes neovascularity generates.Preparation and method that this invention relates to and is used to prevent AMD dependency visual loss, keeps AMD patient's eyesight and suppresses AMD dependency infringement development.Said preparation and method relate to the sclera side dressing with the anecortave acetate of 3-30mg or its corresponding alcohol so that trans-scleral drug release to be provided.

International Application No. WO 95/26974 discloses the compound as shown in the formula (a), and R1 is the alkyl of 1～23 carbon atom, and this compound has the angiogenesis restraining effect, can be used as the therapeutical agent of malignant tumour, diabetic retinopathy, rheumatosis etc.

Summary of the invention:

In the research to formula (a) compound, we have found that the acceptable solvent compound has than the stronger effect of formula (a) compound on formula (1) compound and the physiology thereof aspect angiogenesis inhibiting, especially on by mucosal absorption.

R wherein ₂For containing 1-2 heteroatomic ten carbon with interior hydrocarbon group, heteroatoms is N, O, and one or both among the S, preferred but be not limited only to R ₂A kind of in furyl, thienyl, pyrryl, imidazolyl, pyrazolyl, thiazolyl, pyranyl, pyridyl, miaow pyridine base, pyrazinyl, indyl, purine radicals, the benzopyranyl, more preferably R ₂A kind of in furyl, thienyl, pyranyl, pyridyl, pyrazinyl, the indyl, preferred especially R ₂A kind of in furyl, pyrryl, thienyl, the pyridyl, most preferably R ₂Be the α-Fu Nan base.

The invention also discloses a kind of synthetic method of formula (1) compound:

Formula (1.1) compound dissolution in inert solvent, in the time of 0～25 ℃, is added excessive alkaline reagents, in the time of 0～25 ℃, slowly add excessive R then ₂COCl stirs and to carry out acylation reaction, and reaction finishes the excessive tertiary amine of post neutralization, with the separation method of routine as obtain formula (1) compound with the isolating method of silica gel column chromatography.The weightmeasurement ratio of described formula (1.1) compound and inert solvent is 1: 5～1: 20, described inert solvent includes but are not limited to one or more in methylene dichloride, trichloromethane, ethylene dichloride, tetracol phenixin, acetonitrile, pyridine, the tetrahydrofuran (THF), in preferred especially methylene dichloride and the trichloromethane one or both, the preferred triethylamine of described alkaline reagents, the mol ratio of described alkaline reagents and formula (1.1) compound is 3～6 times, described R ₂The mol ratio of COCl and formula (1.1) compound is 2.5～4 times, also can adopt a large amount of basic solvents such as pyridine to replace alkaline reagents when reaction.

Formula (1.1) compound can obtain according to disclosed method in the International Application No. WO 95/26974

The invention discloses the application of acceptable solvent thing in the medicine of preparation treatment people or mammalian diseases on formula (1) compound and the physiology thereof, the invention also discloses the application in the medicine of the disease that preparation treatment people or Mammals blood vessel new life cause of acceptable solvent thing on formula (1) compound and the physiology thereof, the disease that described angiogenesis causes includes but are not limited to tumour, malignant hematologic disease, diabetic retinopathy, psoriatic, rheumatoid arthritis, age-related macular degeneration.

Described tumour includes but are not limited to various noumenal tumours such as tumor stomach, lung tumors, liver's tumour, various body of gland tumour, nose tumour, ocular tumor, pharyngeal tumour, laryngeal neoplasm etc.

Described malignant hematologic disease is meant the malignant tumour of blood system, mainly comprises leukemia, lymphoma, multiple myeloma and malignant histocytosis etc.

Preferred this compound is applied to prepare the medicine of the new natural disposition disease of treatment ocular angiogenesis, and preferred especially this compound is applied to prepare the medicine for the treatment of age-related macular degeneration.

The eye new vessel generates the disease cause and relates to cornea, iris, choroid and retina etc., and pathological changes such as it causes oozes out, hemorrhage and hyperplasia are the major reasons that causes visual disorder to the infringement of eye structure and function.All there are pathologic processes such as inflammation, ischemic, anoxic in most of ophthalmic; Therefore, illness in eye and eye new vessel are formed with confidential relation.Wherein the eye new vessel generates the disease cause and comprises:

Cornea rebirth blood vessel illness in eye: in the eye disease relevant with cornea rebirth blood vessel, modal is cornea rebirth blood vessel disease due to the wearing of contact lens, other cause that the eye disease of cornea rebirth blood vessel has the cornea wound, alkali and other chemical substances burn, operation on cornea, various infection comprise infectation of bacteria, choamydiae infection, virus infection (herpes simplex virus and varicella zoster virus etc.), protozoan infection (leishmania) etc.

Iris neovascular illness in eye: common have neovascular glaucoma, and retinal detachment, wound, diabetic retinopathy, tumour (retinoblastoma), central vein of retina embolism etc. are its common inducements.

Retinal neovascularization illness in eye: diabetes, tumour, retinal detachment, central retinal vein occlusion, periphlebitis of retina, systemic lupus erythematosus, Eales disease, Coat disease, Takavas disease etc. all can cause.

Choroidal neovascularization illness in eye: age-related macular degeneration (age-related macular degeneration, AMD), high myopia, the exudative retinochoroiditis of centrality, wound, tumour, ocular histoplasmosis's syndrome, serpiginous choroidopathy etc. all can cause this kind illness in eye.

The application of acceptable solvent thing in the medicine of preparation treatment age-related macular degeneration on special preferred formula (1) compound and the physiology thereof.

" treatment " of the present invention can extend to the prevention and the diagnosis of disease.

The dosage of formula disclosed by the invention (I) compound in treatment people or mammalian diseases is 0.001mg-5mg/kg/ days, preferred 0.005mg-2mg/kg/ days.Formula (I) compound is 0.001mg-5mg/kg/ days at the dosage of treatment people or mammal neovascularization disease medicine, preferred 0.005mg-2mg/kg/ days.During as treatment people's disease, people's body weight is generally calculated according to 50kg, so the dosage of human described in the present invention can calculate according to ABW, also can calculate according to above-mentioned dosage * 50.

The invention also discloses a kind of pharmaceutical composition, it is characterized in that containing described any one or a few compound and one or more the pharmaceutically useful auxiliary materials of formula (1) as activeconstituents.

Described pharmaceutical composition dosage when treatment people or mammalian diseases is counted 0.001mg-5mg/kg/ days with formula (1) compound, preferred 0.005mg-2mg/kg/ days.The dosage of described pharmaceutical composition when treatment people or Mammals blood vessel new life disease is counted 0.001mg-5mg/kg/ days with formula (1) compound, preferred 0.005mg-2mg/kg/ days, described pharmaceutical composition adopts every day 1 time or more administering mode in use, preferred every day 1～3 time, also can adopt medication in many days administering mode once, dosage is the stack of Dan Tian.

Described pharmaceutically useful auxiliary material includes but are not limited to pH regulator agent, solubility promoter, osmotic pressure regulator, viscosity modifier, antioxidant, antibacterial sanitas, buffer reagent, suspending agent, local anesthetic, tensio-active agent, solubilizing agent, wetting agent, emulsifying agent, stablizer, weighting agent, protective material, solvent.

Described medicinal compositions can be mixed with liquid preparation, sterilization preparation and sterile preparation, solid preparation, semi-solid preparation, aerosol, above-mentioned preparation type can be according to pharmaceutics (the 5th edition, People's Health Publisher, Cui Fude chief editor) related definition in is understood.

Concrete, the present invention is when being used to prepare the medicine of treatment eye neovascular diseases, can make ophthalmic preparation, said preparation contains acceptable salt or solvate on formula (1) compound or its physiology, and one or more are as the pharmaceutical excipient of carrier.This ophthalmic preparation can be made eye drops, injection, implants, preferably makes the form of eyes such as Eye ointments, eye drop, capsula bulbi injection, eyeground injection with therapeutical agent.

Under the situation of making the eye preparation, under the situation that guarantees the preparation conformability, can add pharmaceutically useful other activeconstituentss, include but are not limited to anti-inflammatory agents such as epsilon-amino caproic acid, Rizinsan K2 A2, diclofenac sodium, Y-8004; Vasoconstrictors such as hydrochloric acid naphthane azoles beautiful jade, meta-synephrine hydrochloride, hydrochloric acid naphthalene first azoles beautiful jade, hydrochloric acid naphthane azoles beautiful jade; Anti-allergic agent such as Sodium Cromoglicate, ketotifen; Antihistaminic agent such as Toldrin, diphenhydramine; Vitaminss such as vitamin B6, lactofiavine phosphate, cyanocobalamin, pantothenic acid alcohol, tocopherol acetate, Flavin Adenin Dinucleotide Sodium; Amino acidses such as sodium chondroitin sulfate, L-potassium aspartate, taurine;

What can also add pharmaceutically useful non-active ingredient includes but are not limited to antibacterial sanitass such as benzalkonium chloride, benzethonium chloride, potassium sorbate, p-Hydroxybenzoate, Sorbic Acid, butylene-chlorohydrin; Tween-80, polyoxyethylene hydrogenated castor oil 60, polyoxyethylene glycol-stearate, Macrogol 4000, Yelkin TTS, sucrose ester, Voranol EP 2001, polyoxy stearate, tensio-active agents such as polyoxyethylene one polyoxypropylene diols and analogue thereof; Glycerine, propylene glycol, sodium-chlor, Repone K, Sorbitol Powder, N.F,USP MANNITOL isosmoticity conditioning agent; Viscosity modifiers such as Xylo-Mucine, Vltra tears, hydroxypropylcellulose, polyvinyl alcohol, carboxy vinyl polymer, Polyvinylpyrolidone (PVP).

When described medicinal compositions was Eye ointments, the matrix in the described auxiliary material can be but be not limited only in lanolin, Vaseline, the Liquid Paraffin one or more.

Described Eye ointments auxiliary material can also include but are not limited to fungistat, stablizer etc., and all can be used for the auxiliary material of Eye ointments.

When described medicinal compositions was made injection, described solvent can include but are not limited to water for injection, oil for injection, propylene glycol, polyoxyethylene glycol, N,N-DIMETHYLACETAMIDE, ethanol, glycerine, phenylcarbinol.

Concrete grammar can be prepared according to the formulation method preparation of anecortave and ester thereof or according to the method for the related preparations in the pharmaceutics (the 5th edition, People's Health Publisher, Cui Fude chief editor).The position of injecting when making eye with injection is identical with the position of anecortave and ester injection thereof.

Embodiment:

Column chromatography separation method among the present invention:

The minimum 70cm of the length of chromatographic column, inner filling 254-silica gel, and will need isolating organism to be dissolved in minimum chloroform entirely: methyl alcohol=in 1: 1, this solution absorption is placed on the top of silica gel in the chromatography column with minimum 254-silica gel, use the moving phase wash-out, chromatography column connects the solution that obtains through column chromatography with several 10ml test tubes down, the control flow velocity is 10ml/3min, the solution of each test tube is analyzed with HPLC, the test tube solution that retention time is identical merges, the compound of getting principal point carries out recrystallization, obtains corresponding product.Wash-out moving phase adopts chloroform: the mixed solvent of methyl alcohol=1: 1 (volume ratio).

Determine the method for principal point: will need isolating organism to analyze with HPLC, except that the point of the peak area maximum of raw material point is defined as principal point, its retention time is the retention time of principal point.

Equipment: HP 1084B liquid chromatograph, HP 79850 BLC terminals and UV detector

Column material: Hypersll C18,5um, 125 * 4.6mm

Detect wavelength: 242nm

Moving phase: methyl alcohol: water=5.5: 4.5

Column temperature: 45 ℃

Flow velocity: about 1.2ml/ branch

Embodiment 1:

Pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3, the preparation of 20-diketone-17-hydroxyl-17-furoate (compound 1-a)

Acidylate

Modus ponens (1.1) compound 3g is dissolved in the 30ml methylene dichloride, the solution of gained is cooled to 0～5 ℃, and under this temperature, add the 6ml triethylamine, then under temperature is 5-10 ℃ condition, slowly add the 2.5ml furoyl chloride, under 5-10 ℃ condition, stir resultant mixture then, reaction 3h afterreaction liquid washing three times, each 20ml water, with concentrated hydrochloric acid water layer pH is transferred to 1-2 during washing, and with the dichloromethane extraction water of 10ml * 3 time, organic phase merges, with the washing of 200ml water and with the 8N sodium hydroxide solution pH is transferred to 5-6, organic phase is concentrated, when condensing crystal is directly closely done, pour 10ml methyl alcohol, slowly cool to-5～5 ℃ of filtrations and obtain pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-furoate crude product 2.83g.

Refining

To obtain pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-furoate crude product is dissolved in methyl alcohol: the mixed solvent of chloroform=1: 1 (volume ratio), adopt the isolating method of aforementioned column chromatography to obtain pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-furoate elaboration 2.25g.

Ultimate analysis calculated value: C ₂₇H ₃₃FO ₅C 71.03% H 7.29% F 4.16% O 17.52%

Ultimate analysis measured value: C 71.08% H 7.31% F 4.12% O 17.49%

¹³C-NMR(CDCl ₃)：

The position of C	??1	??2	??3	??4	??5	??6	??7	??8
									?? 13C-NMR	??26.1	??34.3	??199.2	??123.4	??170.0	??23.6	??28.9	??42.7
The position of C	??9	??10	??11	??12	??13	??14	??15	??16
									?? 13C-NMR	??104.6	??47.8	??25.0	??23.9	??43.8	??45.5	??24.3	??22.4
The position of C	??17	??18	??19	??20	??21	??22	??23	??24
									?? 13C-NMR	??104.8	??13.4	??21.5	??210.0	??31.8	??158.8	??144.9	??118.0
The position of C	??25	??26	??27	??28	??29	??30	??31	??32
									?? 13C-NMR	??112.2	??145.9	??20.0

Embodiment 2:

Pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3, the preparation of 20-diketone-17-hydroxyl-17-(2-thiophenic acid)-ester (compound 1-b)

Acidylate

Modus ponens (1.1) compound 3g is dissolved in the 30ml methylene dichloride, gained suspension is cooled to 0～5 ℃, and under this temperature, add the 6ml triethylamine, then under temperature is 5-10 ℃ condition, slowly add 2.5ml 2-thiophene chloride, under 5-10 ℃ condition, stir resultant mixture then, reaction 3h afterreaction liquid washing three times, each 20ml water, with concentrated hydrochloric acid water layer pH is transferred to 1-2 during washing, and with the dichloromethane extraction water of 10ml * 3 time, organic phase merges, with the washing of 200ml water and with the 8N sodium hydroxide solution pH is transferred to 5-6, organic phase is concentrated, when condensing crystal is directly closely done, pour 10ml methyl alcohol, slowly cool to-5～5 ℃ of filtrations and obtain pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-(2-thiophenic acid)-ester crude product 2.76g.

Refining

To obtain pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-(2-thiophenic acid)-ester crude product is dissolved in methyl alcohol: the mixed solvent of chloroform=1: 1 (volume ratio), adopt the isolating method of aforementioned column chromatography to obtain pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-(2-thiophenic acid)-ester elaboration 2.12g.

Ultimate analysis calculated value: C ₂₇H ₃₃FSO ₄C 68.62% H 7.04% F 4.02% O 13.5% S 6.78%

Ultimate analysis measured value: C 68.67% H 7.01% F 3.99% O 13.48% S 6.85%

¹³C-NMR(CDCl ₃)：

The position of C	??1	??2	??3	??4	??5	??6	??7	??8
									?? 13C-NMR	??26.1	??34.3	??199.2	??123.4	??170.0	??23.6	??28.9	??42.7
The position of C	??9	??10	??11	??12	??13	??14	??15	??16
									?? 13C-NMR	??104.6	??47.8	??25.0	??23.9	??43.8	??45.5	??24.3	??23.0
The position of C	??17	??18	??19	??20	??21	??22	??23	??24

The position of C	??1	??2	??3	??4	??5	??6	??7	??8
									?? 13C-NMR	??106.87	??13.4	??21.5	??210.5	??31.8	??160.8	??134.7	??134.6
The position of C	??25	??26	??27	??28	??29	??30	??31	??32
									?? 13C-NMR	??128.9	??134.2	??20.0

The preparation of embodiment 3 capsules is to prepare 1000 capsules

Compound 1-a 0.1g

Starch 20g

With pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3,20-diketone-17-hydroxyl-17-furoate and starch mixing pour into Capsules No. 4, obtain capsule.Every capsules contains compound 1-a 0.1mg, starch 20mg

The preparation of embodiment 4 eye drops

Compound 1-a 0.05g (particle diameter 5～20 μ m)

Sodium-chlor 0.08g

SODIUM PHOSPHATE, MONOBASIC 0.65g

Sodium phosphate dibasic 0.5g

Benzalkonium chloride 0.01g

Vltra tears 0.2g

Tween-80 0.08g

Water for injection is to 100ml

Getting benzalkonium chloride is dissolved in recipe quantity 50% water for injection, be heated to 40-50 ℃, adding SODIUM PHOSPHATE, MONOBASIC, Sodium phosphate dibasic, tween-80 make dissolving, No. 4 sintered filter funnels filter conduct＜1〉liquid is stand-by, in addition Vltra tears is dissolved in the distilled water of recipe quantity 30, filter with the B that is lined with 200 order nylon cloths, be heated to 60-70 ℃, add compound 1-a and stir evenly, insulation 30min is cooled to 40-50 ℃, again with＜1〉the liquid merging, add the injection water to capacity, 200 order nylon mesh filter twice, packing, sterilize, seal.

5 preparations of embodiment with injection

Tween-80 0.4g

Glycerol 2.6g

Water for injection is to 100ml

Compound 1-a 2g (particle diameter≤5 μ)

Tween-80, glycerol, water for injection are filtered as＜1 with No. 4 sintered filter funnels as the mixed back of matrix〉liquid is stand-by, compound 1-a micro mist is scattered in＜1〉liquid in the B filtration that is lined with 200 order nylon cloths, filtration sterilization packing.

Pharmacology embodiment 1 experiment in vitro

Adopt the influence of chick chorioallantoic membrane (CAM) blood vessel hyperplasia model observation type (I) compound of growth factor-induced to blood vessel hyperplasia, understanding formula (I) compound is to the effect of vasculogenesis.

Experiment material:

The instar chicken embryo on the 3rd of being fertilized

Glass fiber filter paper

Vascular endothelial growth factor (VEGF) Chemlcon company product

Will with disclosed pregnant steroid-9 α-fluoro-6 α methyl-4-alkene-3 among the made compound 1-a of embodiment 1-2 and compound 1-b and International Application No. WO 95/26974 embodiment, 20-diketone-17-hydroxyl-17-acetic ester (as compound 1-c) dissolves with DMSO earlier respectively, (phosphate buffer soln pH=7.4) is diluted to needed concentration (DMSO concentration＜0.1%) to use PBS again

Experimental technique

1.CAM the foundation of model:

75% ethanol cleans chicken ovigerm shell and dries up in super clean bench, behind the horizontal positioned 5min, carefully chorion is broken into pieces at 100mm culture dish edge, and the ovum content places culture dish (culture dish is added with 10ml DMEM substratum in advance).This culture dish is put into the big culture dish of 150mm (big culture dish adds little water), cover the ware lid, place 37 ℃, 5%CO ₂Cell culture incubator in cultivate.

2. to the influence of CAM blood vessel hyperplasia

With tapping and plugging machine glass fiber filter paper is made the sequin of diameter 3mm, moist heat sterilization, each 10 μ l drips on glass fiber filter paper with reagent, makes the medicine film, dries up standby.Behind the chicken embryo culture 3d, be divided into 5 groups at random, every group 10, compound 1-a, compound 1-b, compound 1-c (open among International Application No. WO 95/26974 embodiment) are made as 3 administration groups (1g/L), give somatomedin (2 μ g/L) simultaneously, and independent stimulating group (positive controls) of somatomedin (VEGF) and PBS (phosphate buffered saline buffer, PH 7.4) stimulating group (negative control group).The medicine film is affixed on CAM and yolk cyst membrane (yolk sacmembrane, YSM) the outer less position of 2/3 place's blood vessel.Behind the dosing 48h, microscopically is observed, middle or small blood vessel number in the 5mm around the counting medicine film.

Table 1 pair CAM blood vessel hyperplasia influence table (

N=10)

Group name	Dosage μ g/egg	Medium vessels	Little blood vessel	P (each group is to the VEGF group in the little blood vessel)
					??PBS		??2.50±0.67	??8.20±0.35	??＜0.01
??VEGF		??2.50±0.33	??17.60±0.87
					Compound 1-a	??0.5	??2.50±0.34	??6.80±1.08	??＜0.01
Compound 1-b	??0.5	??2.50±1.05	??7.70±0.93	??＜0.01
					Compound 1-c	??1	??2.50±0.78	??7.90±0.96	??＜0.01

Annotate: the statistical method data with

Expression. relatively use the t check between group

3. result

1 formula (I) compound is to VEGF inductive CAM

Blood vessel hyperplasia influence table 1 as a result, it is obvious that the good .VEGF of PBS group angiogenic growth organizes little blood vessel hyperplasia, each experimental group blood vessel hyperplasia obviously is suppressed. little blood vessel significantly reduces, and compares with the VEGF group, and there is notable difference (P＜0.01) in little blood vessel number.

CAM is classical vasculogenesis evaluation model, has easy, the easy observation of method, advantage such as inexpensive. be at present the most frequently used in the phantom type.VEGF is important short angiogenesis factor, and the hyperplasia and the migration of energy stimulating endothelial cell promote vasculogenesis, and find overexpression in the kinds of tumors tissue.Originally studies show that: formula (I) compound suppresses VEGF inductive CAM blood vessel hyperplasia, shows that formula (I) compound has the short angiogenic action that suppresses VEGF.And little blood vessel is being suppressed under the suitable situation of effect, the required dosage of compound 1-a that makes in the embodiment of the invention and compound 1-b only is 50% of disclosed compound 1-c among reference International Application No. WO 95/26974 embodiment.

Pharmacology embodiment 2 experimentation on animalies

Formula (1) compound is used laser induced rat choroidal artery new life again, and (Chorodaial Neovascularization CNV) carries out subconjunctival injection in the model, record the angiogenesis restraining effect of gained formula of the present invention (1) compound.

(1) making of rat CNV model

Get 70 of BN (Brown Norway) rats, male and female are not limit, body weight 180-210g, be divided into 3 groups every group 10 at random, every rat eyes are according to Cai Chunmei (BN rat choroidal neovascularization Animal Model Making, 2005 the 26th volume third phases of Mudanjiang Medical College's journal, 1-3) disclosed method argon laser (520 years nanometers of wavelength, 100 microns of spot diameters, energy 150 milliwatts, 0.1 second time shutter) Photocoagulation carries out modeling, and every rat eyes all carry out modeling, every is coagulated 8 points around optic disk light between blood vessel, coagulates with light and carries out fundus fluorescein angiography inspection (FFA) after back 14 days discoid fluorescein seepage to occur serve as generation CNV.

(2) administration

After the modeling, inject the eye injection for preparing according to embodiment 5 methods with the micro-syringe that has No. 30 syringe needles with the dosage of 50 μ l/ eyes to the top of eyeball conjunctiva, according to different the be divided into matrix group (control group 1) of administration kind with dosage, compound 1-c 1000 μ g/ ocular administration groups (control group 2), anecortave 1000 μ g/ ocular administration groups (control group 3), compound 1-a1000 μ g/ ocular administration group (test group 1), compound 1-a 3000 μ g/ ocular administration groups (test group 2), compound 1-b 1000 μ g/ ocular administration groups (test group 3), compound 1-b 3000 μ g/ ocular administration group (test group 4) test-results are carried out the t check.

Each situation of organizing the CNV incidence sees the following form

N=10

Group	CNV incidence %	The P value
			?? *Control group 1	??75.00±8.33
?? **Control group 2	??58.75±6.04
			?? ***Control group 3	??48.75±7.10
Test group 1	??37.50±5.89	?? *P＜0.01， **P＜0.05， ***P＜0.05，
			Test group 2	??22.50±5.27	?? *P＜0.01， **P＜0.01， ***P＜0.05，

Group	CNV incidence %	The P value
			Test group 3	??45.75±7.10	?? *P＜0.01， **P＜0.05， ***P＜0.05，
Test group 4	??30.75±7.10	?? *P＜0.01， **P＜0.01， ***P＜0.05，

^*P represents the P value compared with control group 1, ^*P represents the P value compared with control group 1, ^{* *}P represents the P value compared with control group 1.

From the experimental result of last table as can be seen, the CNV incidence of comparing test group 1-4 with the control group 1 that uses matrix all has significance (P＜0.01).And also all have significance with the CNV incidence of using the control group 2 that has medicine now to compare test group 1-4 with control group 3, wherein, the dosage of test group 1,3 and control group 2,3 is 1000 μ g/ eyes, explanation is under same dose, adopt the test group 1,3 of gained compound of the present invention to compare and produced significant reduction, produced better result of treatment with the CNV incidence of control group 2,3.And adopt the test group 2,4 of 3000 μ g/ ocular administration dosage to compare better efficacy with the test group 1,3 that adopts respective compound 1000 μ g/ ocular administration dosage, the formula compound that gained of the present invention is described when treatment CNV except having the better effect than existing medicine, also has dosage correlation, use the test group 1,2 of compound 1-a especially, compare to show with test group 3,4 CNV is better suppressed effect.

Pharmacology embodiment 3 retinal functions influence test

After the mensuration of pharmacology embodiment 1 finishes, respectively organize of the influence of used medicine to retinal function with electroretinogram (ERG) mensuration.

Measuring method: the rat of the mensuration of the pharmacology that will be through with embodiment 2 was raised in the darkroom more than 1 hour, implemented dark adatpation.Following operating in the darkroom carried out under red light.With rat body anesthesia, the method identical with pharmacology embodiment 2 carried out the mydriasis operation.Rat under anesthesia splashes into 1 0.4% Oxybuprocaine hydrochloride eye drops (trade(brand)name: times promise happiness, Japan Santen Pharmaceutical Co. Ltd. system, import drugs registration certificate H20020245) after, be fixed on and measure on the platform, ground-electrode (pin electrode) and indifferent electrode (pin electrode) are installed respectively at tail and the nose of rat.After on the eye of rat the LED electrode being installed, extinguish ruddiness.After confirming recording status on the oscilloscope, record ERG (15 milliseconds of light stimulus times, luminosity 3000cd/m ²(500 μ w)).

Measure a ripple and b wave amplitude from the waveform that obtains with ERG determinator (NEC SYNAx ERllOO, NEC system), calculate the b wave amplitude of ERG with respect to the ratio of a wave amplitude (b/a than) and carry out the t check.Test-results sees the following form

Group	ERG b/a ratio
		?? *Control group 1	??2.02±0.23
?? **Control group 2	??2.03±0.34
		?? ***Control group 3	??2.07±0.26
Test group 1	??2.05±0.31
		Test group 2	??2.09±0.35

Group	ERG b/a ratio
		Test group 3	??2.08±0.29
Test group 4	??2.06±0.25

Can draw from last table, use each test group of gained formula of the present invention (1) compound, compare with the control group 1 that adopts matrix, the b/a value does not have significance, (P＞0.05) illustrates the retinal function not influence of formula (1) compound of gained of the present invention to laboratory animal.

Claims (10)

1. compound as shown in the formula (1)

R ₂For containing 1-2 heteroatomic ten carbon with interior hydrocarbon group.

2. the synthetic method reaction process of compound as claimed in claim 1 is as follows:

Formula (1.1) compound dissolution in inert solvent, in the time of 0～25 ℃, is added excessive alkaline matter, in the time of 0～25 ℃, slowly add excessive R then ₂COCl stirs and to carry out acylation reaction, and reaction finishes the excessive alkaline matter of post neutralization, with the separation method of routine as obtain formula (1) compound with the isolating method of silica gel column chromatography.

3. the application of acceptable solvent thing in the medicine of preparation treatment people or mammalian diseases on formula (1) compound as claimed in claim 1 and the physiology thereof.

4. the application of acceptable solvent thing in the human age-related macular degeneration of preparation treatment on formula (1) compound as claimed in claim 1 and the physiology thereof.

5. as the arbitrary described formula of claim 3 to 4 (1) application of compound, it is characterized in that the dosage that described formula (1) compound uses is 0.001mg-5mg/kg/ days when treatment people or Mammals.

6. pharmaceutical composition is characterized in that containing described any one or a few compound and one or more the pharmaceutically useful auxiliary materials of formula (1) as activeconstituents.

7. pharmaceutical composition as claimed in claim 6, the dosage when treatment people or mammalian diseases is counted 0.001mg-5mg/kg/ days with formula (1) compound.

8. as arbitrary described pharmaceutical composition in the claim 6 to 7, described pharmaceutically useful auxiliary material comprises one or more in pH regulator agent, solubility promoter, osmotic pressure regulator, viscosity modifier, antioxidant, antibacterial sanitas, buffer reagent, suspending agent, local anesthetic, tensio-active agent, solubilizing agent, wetting agent, emulsifying agent, stablizer, weighting agent, protective material, the solvent.

9. as arbitrary described pharmaceutical composition in the claim 6 to 8, when it is characterized in that described pharmaceutical composition is used to prepare the medicine of treatment treatment eye neovascular diseases, can make in eye drops, injection, implants, Eye ointments, eye drop, capsula bulbi injection, the eyeground injection one or more.

10. the pharmaceutical composition of treatment treatment eye neovascular diseases as claimed in claim 9 is characterized in that described pharmaceutical composition can also comprise anti-inflammatory agents such as adding epsilon-amino caproic acid, Rizinsan K2 A2, diclofenac sodium, Y-8004; Vasoconstrictors such as hydrochloric acid naphthane azoles beautiful jade, meta-synephrine hydrochloride, hydrochloric acid naphthalene first azoles beautiful jade, hydrochloric acid naphthane azoles beautiful jade; Anti-allergic agent such as Sodium Cromoglicate, ketotifen; Antihistaminic agent such as Toldrin, diphenhydramine; Vitaminss such as vitamin B6, lactofiavine phosphate, cyanocobalamin, pantothenic acid alcohol, tocopherol acetate, Flavin Adenin Dinucleotide Sodium; In the amino acidses such as sodium chondroitin sulfate, L-potassium aspartate, taurine one or more.