CN103169717A - Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation - Google Patents
Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation Download PDFInfo
- Publication number
- CN103169717A CN103169717A CN2011104312738A CN201110431273A CN103169717A CN 103169717 A CN103169717 A CN 103169717A CN 2011104312738 A CN2011104312738 A CN 2011104312738A CN 201110431273 A CN201110431273 A CN 201110431273A CN 103169717 A CN103169717 A CN 103169717A
- Authority
- CN
- China
- Prior art keywords
- eye
- inflammation
- anthracycline antibiotics
- anthracene nucleus
- purposes according
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The invention discloses an application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation. The invention found that the anthracene nucleus antibiotic and its pharmaceutical salt have good effect on treatment of exogenous ocular inflammation and endogenous ocular inflammation, and especially the anthracene nucleus antibiotics can be prepared to prolonged action preparations such as a nano preparation, a microballoon preparation, liposome and hydrogel, the curative effect can be directly performed on ocular surface and eye ground, the toxicity of anthracene nucleus antibiotic is reduced, administration dosage is simultaneously enlarged through ophthalmic administration by prolonged action preparations, the duration time of drug effect is long, the medicament curative effect is stronger than that of the common preparations of the anthracene nucleus antibiotic, when the ophthalmic administration is carried out, the anthracene nucleus antibiotic has good treatment effect for exogenous and endogenous ocular surface, and eye ground inflammation, and especially has good treatment effect for the ocular surface and eye ground diseases caused by external wound, so that the problems of certain eyes inflammation with long treatment and more severe eye diseases due to eyes inflammation can be solved.
Description
Technical field
The present invention relates to a kind of medicine for the treatment of eye inflammation, being specifically related to a series of traditional antitumor antibiotics is anthracycline antibiotics and the new purposes of officinal salt in preparation treatment eye inflammation thereof, especially, the preparation of described anthracycline antibiotics and officinal salt thereof becomes collyrium or the long-acting preparations such as nanometer formulation, microball preparation, liposome and hydrogel.
Background technology
Eye inflammation mainly refers to endophthalmitis, keratitis, conjunctivitis, iritis, retinitis, choroiditis, cyclitis, panuveitis, scleritis, optic neuritis, hyalitis, phakitis etc., these inflammation can be produced by exogenous and endogenic reason, and the endogenous reasons such as exogenous reasons such as mechanical injury, chemical injury, operation wound and some traumatic secondary infection or autoimmune systemic disease, self virus, self-blood, liver and gall, urinary disease produce.
Can roughly be divided into the exogenous eye inflammation of eye table, eye according to fall ill source and site of pathological change and show four kinds of endogenous eye inflammation, the exogenous eye inflammation in optical fundus and optical fundus endogenous eye inflammation.
The common drug for the treatment of clinically eye inflammation is 1. chloromycetin of following various medicine, as chloromycetin; 2. the general class of amino sugar is as streptomycin, gentamycin, tobramycin, kanamycin, amikacin and Yi Nuomi magnitude; 3. vinegar class in macro ring is as erythromycin eye ointment; 4. promise ketone is as ciprofloxacin, ofloxacin, levofloxacin and lomefloxacin etc.At present, antibiotic ophthalmic liquid medicine is take promise ketone eye drop and amino sugar two class eye drops as main.Antifungal antibiotic has amphotericin B and fluorine health etc.Antiviral medicine commonly used has acyclovir and ribavirin etc.The viral conjunctivitis and the keratitis that are caused by multiple virus such as adenovirus, enterovirus, simple hcrpesviruses, chickenpox zona shingles virus etc., usually exist simultaneously with bacillary oculopathy, but clinical existing medicine is single antibacterial action or antivirus action, is unfavorable for bringing into play good therapeutic effect.The indication of above medicine is comparatively single repetition all, and effect is not good enough.
The pathogenic factor of eye inflammation is many and complicated, and the eye inflammation disease progression is very fast, delays slightly treatment, can produce to have a strong impact on, and secondary goes out multiple serious ophthalmic, even causes visual loss.And the medication of present eye inflammation unsatisfactory curative effect not only, and often indication is single, clinically, often best treatment time is incured loss through delay in diagnosis or the reagent meeting of the state of an illness.and often a lot of eye inflammation are understood mutual secondary or concurrent, the Cornea and conjunctiva of eye table, the retina on optical fundus and choroid, ectogenic and endogenic a lot of inflammation are secondary or concurrent mutually all, at present a lot of medicine indications are all comparatively single, can't satisfy the medication requirement, need to develop at present a kind of for most of eye inflammation, no matter fall ill in eye table or ophthalmic, no matter come from ectogenic or endogenic, extraordinary therapeutic effect is arranged, will save Diagnostic Time, improve therapeutical effect, delay treating time not, farthest reduce various eye inflammation to the infringement of eye.
In sum, if search out a kind of new medicine of effectively treating eye inflammation, if this medicine can solve the drug resistance problem, there is no again toxicity, can be used for bacillary oculopathy, can be used for fungoid or viral oculopathy again, namely can be used for exogenous eye inflammation, comprise the inflammation and the complication that produce after corneal transplantation or operation wound, can be used for again endogenic eye inflammation, comprise a series of common ophthalmic and eye table inflammation, bring huge clinical meaning will for the treatment ophthalmic diseases.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of anthracycline antibiotics and the new purposes of officinal salt in preparation treatment eye inflammation medicine thereof, especially the preparation of described anthracycline antibiotics and officinal salt thereof is become the long-acting preparations such as eye drop or nanometer formulation, microball preparation, liposome, hydrogel, can obtain better therapeutic effect.
The concrete technical scheme that the present invention adopts is:
Anthracycline antibiotics of the present invention is the following compound of structure and stereoisomer thereof:
Wherein, R1 is H, OH, CH
3, CH
2OH or OCH
3, R2 is COR1 or CR1R1, R3 is H, OH, CH
3, CH
2OH, OCH
3, pyridine radicals, furyl, pyrrole radicals, thienyl or pyranose.
More preferably, described anthracycline antibiotics is doxorubicin, table doxorubicin, pyrans doxorubicin, mitoxantrone, daunorubicin, daunoblastin, idarubicin, aclarubicin, rubidomycin, aklavine or carminomycin.
Above-claimed cpd can by in prior art disclosed method prepare, some compounds can be bought acquisition by commercial sources.
The present invention is prepared into anthracycline antibiotics and officinal salt thereof and pharmaceutically acceptable auxiliary material combination the disease that various preparations commonly used are used for the treatment of eye inflammation, and these preparations comprise: common flour injection, eye drop, long-acting sustained-release injection.Preferably, described long-acting sustained-release injection can be nano particle preparations, microball preparation, aqueogel or Liposomal formulation.Preferably, described aqueogel is the temperature-sensitive hydrogel preparation.Described anthracycline antibiotics is 0.5-2000 μ g/kg for the eye drops dosage of human body, preferred 1-1500 μ g/kg, more preferably 3-1000 μ g/kg.
When the described nanoparticle of preparation, microsphere, aqueogel, the pharmaceutically acceptable adjuvant that adopts comprises macromolecular material, described macromolecular material is selected from: one or more in poly-anhydride, polyoxyalkylene, polyamide, polyester, polyacrylic resin, polyethers or polysaccharide, or be selected from copolymer between the different monomers of described several macromolecular materials.preferably, described macromolecular material is selected from poly-anhydride, polyvinyl alcohol, Polyethylene Glycol, poly-anhydride-ethylene glycol copolymer, NIPA-acrylic copolymer, polybutylcyanoacrylate, polylactic acid, octadecane diacid acid anhydride-glycol copolymer, PLGA, polylactic-co-glycolic acid-ethylene glycol copolymer, PGA, Acetic acid, hydroxy-, bimol. cyclic ester lactide-ethylene glycol-Acetic acid, hydroxy-, bimol. cyclic ester lactide triblock copolymer, polyether sulfone, gelatin, poly(hydrobutyl ester), poloxamer, collagen protein, Fibrinogen, albumin, cellulose, glucosan, alginate, dextran, hyaluronic acid, one or more in polypeptide or chitosan etc.
When the described Liposomal formulation of preparation, the pharmaceutically acceptable adjuvant of employing comprises natural or synthetic phospholipid, lipoid or its combination.
The said medicine preparation can prepare by disclosed method in prior art, also can adopt the method for putting down in writing in the embodiment of the present invention to prepare.
The pathogenic factor of eye inflammation is many and complicated, and eye inflammation that sb.'s illness took a turn for the worse is very fast, delay slightly treatment, can produce serious disease, secondary goes out multiple serious ophthalmic, even causes visual loss.And the medication of present eye inflammation unsatisfactory curative effect not only, and often indication is single.And often a lot of eye inflammation are understood mutual secondary or concurrent, the Cornea and conjunctiva of eye table, the retina on optical fundus and choroid, ectogenic and endogenic a lot of inflammation are secondary or concurrent mutually all, and at present a lot of medicine indications are all comparatively single, can't satisfy the medication requirement, need at present a kind of for most of eye inflammation, no matter fall ill in eye table or ophthalmic, no matter come from ectogenicly or endogenic, the ophthalmic administration of extraordinary therapeutic effect is arranged.
At present, anthracycline antibiotics is usually used in treating leukemia and antitumor, has less people to use it for the treatment proliferative retinopathy.the inventor has carried out the research of new indication to anthracycline antibiotics, and delightedly find it with the eye dropping of eye drops agent or with nanometer, liposome, after the form eye drops of the long-acting preparation such as microsphere, anthracycline antibiotics is to treating exogenous eye inflammation and the endogenous eye inflammation all has very good effect, wherein exogenous eye inflammation refers to be caused by exopathogenic factor, especially refer to the various secondary ocular disease that caused by exterior trauma, for example comprise surgical operation, the inflammation that is caused by surgery operating wound after corneal transplantation or the inflammation that is caused by chemical injury, endogenic eye inflammation many by self inside with inflammation or virus, thereby the inflammation that might have each position of eyeball that produces, prove by experiment, anthracycline antibiotics can be to comprising the exogenous eye inflammation of eye table, eye table endogenous eye inflammation, the exogenous eye inflammation in optical fundus and optical fundus endogenous eye inflammation all have extraordinary therapeutical effect, and can not produce the multiple side effect that anthracycline antibiotics very easily produces eye.Research finds that simultaneously the anthracycline long-acting slow-release preparation not only has therapeutical effect to eye inflammation, have more targeting, it has long action time, toxic and side effects is little, can obviously improve the advantages such as bioavailability, make it to be used for bacillary oculopathy, can be used for again fungoid or viral oculopathy, namely can be used for traumatic endophthalmitis and keratitis, comprise the inflammation and the complication that produce after corneal transplantation, can be used for again non-traumatic endophthalmitis and keratitis, have huge clinical meaning.
The specific embodiment
Following specific embodiment is described in further detail the present invention, but the present invention not only limits to following examples.In following embodiment, method therefor is conventional method if no special instructions, and test material used if no special instructions, is the purchase of routine biochemistry reagent suppliers and obtains.
Preparation Example is as follows:
Embodiment 1
1. getting 300mg PLGA (PLGA) joins in the 1ml dichloromethane and is prepared into solution;
2. in the solution that makes in joining the 50mg aclarubicin 1., the high speed homogenization device stirs;
3. the solution in will be 2. slowly is injected in the Oleum Gossypii semen that contains 0.05% egg yolk ovum Oletum Trogopterori with syringe, after 1000r/min stirs 20min, reduces rotating speed and continues to stir 4h to 200r/min;
4. add the 20ml petroleum ether, the centrifugal 5min of 8000r/min after 30min to step in 3.;
5. collect microsphere, petroleum ether and volatilizing.
Embodiment 2
1. getting 200mg PLGA (PLGA) joins in the 1ml tetrahydrofuran solution and is prepared into solution;
2. the 50mg mitoxantrone is added in 0.2ml glycerol, join after mixing in the solution that 1. makes, homogenizer obtains colostrum after stirring 1min;
3. it is joined in 2% polyvinyl alcohol (PVA) solution of 12ml, 1000r/min adds the 0.5%PVA solution of 120ml to get emulsion after stirring 5min again;
4. reduce rotating speed to 200r/min continuation stirring 4h, the centrifugal 5min of 8000r/min;
5. collect microsphere, the distilled water wash postlyophilization.
Embodiment 3
1. getting the poly-anhydride-ethylene glycol copolymer (PSA-PEG) of 100mg joins in the 1ml dichloromethane solution and is prepared into solution;
2. in the solution that makes in joining the 50mg doxorubicin 1., ultrasonic mixing;
The solution that 3. will 2. make joins in 15ml 1%PVA solution;
4. reduce rotating speed to 200r/min continuation stirring 4h, the centrifugal 5min of 8000r/min;
5. collect microsphere, collect after distilled water wash.
Embodiment 4
1. getting the poly-anhydride-ethylene glycol copolymer (PSA-PEG) of 160mg joins in 2ml dimethyl sulfoxide and dichloromethane (8: 2) and is prepared into organic facies;
2. in joining the 40mg doxorubicin 1., after mixing, be positioned in 1% polyvinyl alcohol ultrasonic;
3. in will be 2., solution be placed in poly-vinyl alcohol solution and stirs;
4. organic solvent in removing 3., centrifugal collection obtains nanoparticle solution.
Embodiment 5
1. with daunorubicin and polylactic-co-glycolic acid-ethylene glycol copolymer (PLGA-PEG) 800mg 3ml dichloromethane and 1mlN, dinethylformamide acts in 30 ℃ of water-baths and makes its dissolving and high-speed stirred;
2. contain 1% polyvinyl alcohol to 100ml joining in 1., fully mixing;
3. stirring at room volatilization 2h, get nanoparticle solution;
4. at 4 ℃, collect the gained nanoparticle after centrifugal 20min;
5. 4. middle gains are drug-carrying nanometer particle three times with the distilled water washing.
Embodiment 6
1. getting the poly-anhydride (PSA) of 300mg joins in the 3ml dichloromethane and is prepared into organic facies;
2. in the organic facies that makes in joining the 40mg idarubicin 1.;
3. get propylene glycol block polyether and Tween 80 (1: 1) and make that to contain emulsifying agent be 4% aqueous solution, and to add wherein macrodex, regulator solution PH be 8;
4. use the nanometer sedimentation organic facies that makes to be dropped to the aqueous phase that 3. makes;
5. stir and remove organic solvent, obtain nanoparticle solution, centrifugal collection.
Embodiment 7
1. use 0.01molL
-1Sodium hydroxide solution as solvent;
2. get 1. that solvent 10ml adds 100 μ l hydroxypropyl emthylcelluloses, stir and make it even;
3. add 50 μ l carbomer 934s in the solution that 2. makes to step, stir and make its mix homogeneously;
4. add 40mg to gather anhydride-ethylene glycol copolymer (PSA-PEG) in the solution that 3. makes to step, stir and make its mix homogeneously;
5. add the 1mg carminomycin in mentioned solution;
6. take the 1.8g poloxamer188 and join 4. in prepared solution, under magnetic agitation, it is uniformly dispersed, 4 ℃ of conditions were placed more than 24 hours, made the abundant swelling of gel, and being uniformly dispersed obtains clear and bright solution, both the carminomycin hydrogel.
Embodiment 8
1. use 0.01molL
-1Sodium hydroxide solution as solvent;
2. get 1. that solvent 10ml adds 80 μ l hydroxypropyl emthylcelluloses, stir and make it even;
3. add 50 μ l sodium alginates in the solution that 2. makes to step, stir and make its mix homogeneously;
4. add 40mg polylactic-co-glycolic acid-ethylene glycol copolymer (PLGA-PEG) in the solution that 3. makes to step, stir and make its mix homogeneously;
5. add the 1mg epirubicin in mentioned solution;
6. take the 1.8g poloxamer188 and join 4. in prepared solution, under magnetic agitation, it is uniformly dispersed, 4 ℃ of conditions were placed more than 24 hours, made the abundant swelling of gel, and being uniformly dispersed obtains clear and bright solution, both the epirubicin hydrogel.
Embodiment 9
1. take 0.9g phospholipid, 0.3g cholesterol in the 50ml small beaker, add dehydrated alcohol 1-2ml, be placed in 65-70 ℃ of water-bath, be stirred to dissolve, rotate ethanol film forming on wall of cup that this small beaker makes phospholipid, with rubber pipette bulb featheriness wind, ethanol is flung to;
2. separately get pirarubicin solution 30ml in small beaker, with being placed in 65-70 ℃ of water-bath, be incubated, stand-by;
3. get the pirarubicin solution 30ml of preheating, add in the small beaker that contains phospholipid and cholesterol ester plasma membrane, 65-70 ℃ of stirred in water bath aquation 10min.Subsequently small beaker is placed on magnetic stirring apparatus, stirring at room 30-60min, mixing namely get Pirarubicin liposome.
Embodiment 10
1. take boric acid 0.8g, Borax 0.3g, sodium chloride 0.3g, disodiumedetate 0.01g, sodium sulfite 0.05g is dissolved in the 100ml distilled water;
2. 1. the solution of gained is the filtering with microporous membrane sterilization of 0.22 μ m by diameter;
3. in the solution of processing in the 500mg doxorubicin being joined 2.;
4. again filter, namely get stand-by collyrium by the demand packing.
Embodiment 11
1. take boric acid 0.8g, Borax 0.5g, PEG400 25g, disodiumedetate 0.02, sodium sulfite 0.05g, bromo geramine 0.02g is dissolved in the 100ml distilled water;
2. in the solution of processing in the 500mg aclarubicin being joined 1.;
3. again filter, namely get stand-by collyrium by the demand packing.
Embodiment 12
1. take thimerosal 0.05g, sodium sulfite 0.05g, sodium chloride 0.3g bromo geramine 0.02g is dissolved in the 100ml distilled water;
2. in the solution of processing in the 500mg mitoxantrone being joined 1.;
3. again filter, namely get stand-by collyrium by the demand packing.
The effect of drugs experiment:
The therapeutical effect of 1 medicine to endogenous eye inflammation (eye table)
1.1 laboratory animal
200 of healthy adult rabbit, male and female are not limit, and body weight is at 2.0-2.5kg, observes ophthalmic under slit lamp in good condition and without selected this test of other ophthalmic person.
1.2 experimental technique
1.2.1 modeling method
Every rabbit right eye is experimental eye, and left eye is the contrast eye.Weigh before experiment, drip with 0.75% bupivacaine hydrochloride inj eye drip 2-3, massage for a moment, is 3 * 10 with 1mL syringe extraction concentration after absent corneal reflex gently
8CFU/mL Pseudomonas aeruginosa 0.04mL, nearly central authorities inject in the cornea essential layer from cornea, cause the white bacterial plaque of diameter 4-6mm, after put single cage and raise.
1.2.2 group technology
Infect for the first time after charrin disease and occurred the keratitis symptom in rear 1 day: photophobia, shed tears, eyelid is red and swollen, secretions increase, cornea be dispersed in or diffusivity muddy, bulbar conjunctiva is congested and red and swollen.After the success of observation animal model, it being divided into 15 groups at random is model group, 12 groups of embodiment, anthracycline antibiotics ordinary preparation group, amphotericin collyrium, the carrying medicine that each group all adopts injected into anterior chambers 100 μ l medicines or contains 100 μ l medicines except model group and collyrium group, model group gives isopyknic PBS solution, and collyrium directly splashes in rabbit eyes.Treatment time is 10d.
1.3 inspection item and method
1.3.1 bacterial keratitis judgment criteria
Observe the pathological changes situation every day, emphasis to administration after the 1st, 4,7, the scoring of 10d adds up, take simple eye as the object of observation, comprehensive multiple symptom is marked, according to classification, the scoring when observing is commented to the arithmetic point one, is averaged timesharing and gets 2 significant digits.
The symptom classification of table 1 bacterial keratitis animal model and standards of grading
Symptom score is cumulative, and 0-4 is divided into without the Corneal inflammation shape, and 5-7 divides slight keratitis, and 8-12 divides moderate keratitis, 13-16 severe keratitis
1.3.2 cornea pathologic finding
After respectively organizing the rabbit completed treatment, get 5 rabbit for every group and extract eyeball, eyeball is fixed in 48h in 10% neutral formalin, through the conventional dehydration of ethanol, the transparent rear waxdip embedding of dimethylbenzene is made 4 μ m sections and is used for HE dyeing.
1.4 statistical procedures
Data represent with x ± s, t check between relatively employing group between two groups.P＜0.05 for significant difference is arranged, has statistical significance.
1.5 result
1.5.1 the bacterial keratitis efficacy result is estimated
After administration, the 2nd day keratitis symptom begins under control and mitigation, and secretions reduced in the 3rd day.Shed tears in the 4th day, the photophobia phenomenon is controlled, the symptom of the 5th day Cornea and conjunctiva obtains the effective treatment.The 6th day basic the healing.Continue to be administered into recovery from illness in 10 days.Model group infects for the first time and occurred the keratitis symptom in rear 1 day: photophobia, shed tears, eyelid is red and swollen, secretions increase, cornea is dispersed in or the diffusivity muddiness, bulbar conjunctiva is congested and red and swollen, and various keratitis symptoms increased the weight of in the 2nd day, and cornea is seriously muddy, eyelid is obviously red and swollen, cornea was seriously muddy in the 3rd day, and secretions makes whole eye district's humidity or adhesion, and part infects the serious pseudomonas aeruginosa corneal ulcer that is transformed into.There were 7 to change pseudomonas aeruginosa corneal ulcer into by the 5th day in 10 rabbit: blepharospasm swelling, the bulbar conjunctiva Combination is congested and treating serious edema caused, anterior corneal surface presents the canescence muddiness, anterior chamber structure can't be got a glimpse of, anterior chamber and interlayer empyema have a large amount of corneal epithelial cells downright bad, and tissue comes off, a large amount of yellow green purulent secretions are arranged, and eyes are blind.Concrete scoring situation sees Table 2.
Table 2 charrin's disease Rabbit keratitis is respectively organized Symptoms index (n＞10x ± s)
| Group | 1d | 4d | 7d | 10d |
| Model group | 11.25±0.87 | 11.29±0.93 | 11.54±0.89 | 11.69±0.98 |
| The doxorubicin ordinary preparation | 10.75±1.31 | 8.07±1.27 | 6.27±1.00 ** | 5.65±1.09 ** |
| The administration group | ||||
| Common collyrium | 10.72±1.18 | 9.24±1.31 | 8.43±1.15 ** | 7.71±1.01 ** |
| Embodiment 3 | 10.64±1.03 | 6.78±1.09 | 4.02±1.31 ** | 3.10±1.03 ** |
| Embodiment 4 | 10.65±1.30 | 6.06±1.08 * | 3.69±1.11 ** | 2.34±0.96 ** |
| Embodiment 6 | 10.22±0.99 | 6.13±0.98 * | 3.87±1.09 ** | 2.73±0.99 ** |
| Embodiment 7 | 10.15±1.12 | 6.04±1.02 * | 3.54±1.13 ** | 2.97±1.00 ** |
| Embodiment 9 | 10.28±1.05 | 6.57±1.02 | 4.01±1.08 ** | 3.17±1.05 ** |
| Embodiment 12 | 10.21±1.13 | 6.78±1.34 | 4.15±1.07 ** | 3.21±1.01 ** |
Compare with model group
*P＜0.05,
*P＜0.01
1.5.2 cornea pathological examination results
Observe as seen under light microscopic, the swelling of (1) model group corneal height, exfoliation of corneal epithelium, visible a large amount of neutrophilic granulocytes, lymphocyte, fibroblast and slough in substrate, upper subcutaneous wound ulcer does not heal, partially sliced visible perforation of cornea.(2) common drug treatment group corneal height swelling, but the neutrophilic granulocyte in substrate, lymphocyte, fibroblast and slough lack than model group, and rare corneal ulcer and perforation, as seen this dosage can be alleviated cell infiltration.(3) the slight swelling of durative action preparation treatment group cornea, rare neutrophilic granulocyte, lymphocyte, fibroblast and slough in substrate, rare corneal ulcer and perforation.This shows, medicine can effectively reduce the quantity of inflammatory cell, for bacterial keratitis, certain curative effect is arranged, and durative action preparation is better than ordinary preparation.
The therapeutical effect of 2 medicine exogenous eye inflammation (the eye off-balancesheet is hindered secondary infection)
2.1 laboratory animal
90 of healthy adult rabbit, male and female are regardless of, and body weight is at 2.0-2.5kg, observes ophthalmic under slit lamp in good condition and without selected this test of other ophthalmic person.
2.2 experimental technique
2.2.1 modeling method
Rabbit is fixed with fixed case, and the tinsel of about cut-off footpath 0.5mm is adopted the circular rings that an end is twisted into the 4mm size, circular rings burn on alcohol burner red after, iron rapidly to scald on cornea and cause annular lesions and form traumatic keratitis.
2.2.2 group technology
Determine that after animal model successfully, it being divided into nine groups at random is model group, drug-loaded liposome, medicine carrying microballoons, medicine carrying hydrogel, drug-carrying nanometer particle 1, drug-carrying nanometer particle 2, medicine carrying collyrium, anthracycline antibiotics ordinary preparation, common collyrium, the carrying medicine that each group all adopts injected into anterior chambers 100 μ l medicines or contains 100 μ l medicines except model group and collyrium group, model group gives isopyknic PBS solution, and collyrium directly splashes in rabbit eyes.Treatment time is 10d.
2.3 traumatic keratitis treatment effectiveness evaluation
Observe the pathological changes situation every day, emphasis to administration after the 1st, 4,7, the scoring of 10d adds up, mark as the object of observation take simple eye.According to classification, the scoring when observing is commented to the arithmetic point one, is averaged timesharing and gets 2 significant digits.
Therapeutic effect is estimated by following stage division:
0 grade of cornea ring-type damage disappears, corneal transparency;
1 grade of cornea ring-type damaged portion disappears, the cornea partially transparent;
2 grades of cornea ring-type damages are slight muddy, and conjunctiva is without redness and secretions;
3 grades of cornea ring-type damages are obvious and muddy, the conjunctiva mild hyperaemia;
The damage of 4 grades of cornea ring-types is obvious and muddy, conjunctival congestion swelling and white secretions is in a small amount arranged.
2.4 statistical procedures
Data represent with x ± s, t check between relatively employing group between two groups.P＜0.05 for significant difference is arranged, has statistical significance.
2.5 the traumatic keratitis efficacy result is estimated
Experimental result (seeing Table 3) shows, after corneal injury, owing to being upset, 1-2 days, every group had inflammatory reaction, it is muddy that corneal injury is obvious ring-type, conjunctiva, scleral injection, swelling has a small amount of from color secretions, other were respectively organized conjunctival congestion, swelling and secretions and all disappear except model group in the 3rd day.The durative action preparation group, its ring-type damage just part disappearance at the 5th day, corneal injury place partially transparent, each durative action preparation group the 6th day or corneal injury in the 7th day all disappear, cornea is fully transparent, and common treatment group and normal saline model group lasted till the 7th day, and the damage of cornea ring-type is still high-visible, observed the 10th day basic disappearance of common treatment group cornea ring-type damage.This presentation of results medicine has good therapeutic effect, if it is better to be made into the durative action preparation effect.
Table 3 rabbit traumatic keratitis is respectively organized Symptoms index (n＞10x ± s)
Compare with model group
*P＜0.05,
*P＜0.01
The therapeutical effect of 3 medicines to endogenous eye inflammation (optical fundus)
3.1 laboratory animal
80 of healthy adult rabbit, male and female are not limit, and body weight is at 2.0-2.5kg, observes ophthalmic under slit lamp in good condition and carry out this test without other ophthalmic person.
3.2 experimental technique
3.2.1 modeling method
Every rabbit right eye is experimental eye, and left eye is the contrast eye.Before modeling, the loose large pupil of 30min compound recipe holder pyrrole amide, then use chloromycetin flush operation eye.Compound ketamine injection (50mg/kg) intramuscular anesthesia, the Oxybuprocaine hydrochloride eye drops topical anesthesia.Conventional ophthalmology sterilization, drape, in right eye temporo top approximately after 10 limbus of corneae 3mm sentence aseptic 1ml syringe inserting needle with the 25G injection needle, first extract approximately 0.1ml of vitreous body out, inject subsequently staphylococcus aureus bacterial suspension 0.1ml to keep the intraocular pressure balance.
3.2.2 group technology
After intravitreal injection antibacterial 24h, all first extract the 0.1ml vitreous body out after all animals operate by aforementioned anesthesia and sterilization method and do the antibacterial cultivation, subsequently laboratory animal being divided at random eight groups is model group, drug-loaded liposome, medicine carrying microballoons, medicine carrying hydrogel, drug-carrying nanometer particle 1, drug-carrying nanometer particle 2, anthracycline antibiotics ordinary preparation, collyrium, respectively organize equal intravitreal 100 μ l medicines or contain the carrying medicine of 100 μ l medicines except model group and collyrium group, model group gives isopyknic PBS solution, and collyrium directly splashes in rabbit eyes.
3.3 inspection item and method
3.3.1 conventional examination of ocular fundus
Every daily slit lamp, indirect ophthalmoscope inspection.Respectively at after intervening 1,4,7,14d carries out the scoring of endophthalmitis clinical scale to each treated animal, primary part observation anterior chamber scintillation, aqueous cell and vitreous opacity degree, its grade scale is as follows:
Anterior chamber's scintillation classification (inflammatory exudate enters aqueous humor, under the narrow light belt of slit lamp casts oblique rays on, and visible light flash and ooze out granule and floating, this phenomenon is called aqueous flare): 0 grade, without aqueous flare (light beam is transparent shinny); 1 grade, slight aqueous flare (the faint light beam that turns white); 2 grades, moderate aqueous flare (the milky white light beam of moderate can be distinguished iris and crystalline lens details); 3 grades, remarkable aqueous flare (obvious milky white light beam, impalpable iris and crystalline lens details); 4 grades, serious anterior chamber's scintillation, aqueous humor becomes curdled appearance, with a large amount of fibrinous exudates.Aqueous cell classification (in the darkroom, be 45 °～60 ° with the angle adjustment of light source and slit lamp microscope, light beam is 1mm * 0.5mm, and light beam is calculated cell number in all light beams by lesser ring of Merkel): 0 grade, acellular; 1 grade, 5～10 cells; 2 grades, 11～20 cells; 3 grades, 21～50 cells; 4 grades,＞50 cells.The vitreous opacity classification: 0 grade, vitreous body is clear without muddy; 1 grade, slight muddy, optic nerve, retinal vessel and nerve fiber can be offered an explanation; 2 grades, slight muddy, optic nerve, retinal vessel can be recognized, but the fuzzy difficulty of nerve fiber is debated; 3 grades, moderate is muddy, and optic nerve, retinal vessel can recognize, but very fuzzy; 4 grades, severe is muddy, and rear utmost point section structure can not be seen.
3.3.2 the bacteriology cultivates
After intervening 5d, all animals carry out the vitreous chamber puncture under aseptic condition, get vitreous humor 0.1ml, are inoculated on Colombia's flat board, and antibacterial culturing 24h under 37 ℃ of conditions observes the Detection of pathogenic bacteria situation.
3.3.3B super the inspection
After bacterial injection, rear all animals of 14d of 24h and intervention carry out ultrasound diagnosis, sight glass body and detachment of retina situation.
3.3.4 histological examination
Each is got 5 rabbit for every group and extracts eyeballs after organizing rabbit intervention 14d, the same 1.3.2 of preparation dicing method.
3.4 statistical procedures
Data represent with x ± s, t check between relatively employing group between two groups.Between each group, positive rate of bacteria, detachment of retina incidence rate relatively adopt X 2 test.P＜0.05 for significant difference is arranged, has statistical significance.
3.5 result
3.5.1 clinical inflammation scoring
24h after the injection antibacterial, all laboratory animal blepharoedemas, conjunctiva is congested, edema obviously, and corneal edema is obvious, anterior chamber's scintillation (+), exudation of anterior chamber (+).After intervention, 4,7,14d carries out the scoring of clinical inflammation, the model group inflammatory symptom increases the weight of gradually, and treatment group laboratory animal cornea is all transparent gradually, and anterior chamber's scintillation alleviates gradually, oozes out to be tending towards absorbing, and vitreous opacity alleviates in various degree.Each treatment group and model group more all have significant difference (table 4).
Table 4 postoperative different time points is respectively organized clinical inflammation scoring situation (n＞10x ± s)
Compare with model group
*P＜0.05,
*P＜0.01
3.5.2 bacteriology's cultivation results
5d Detection of pathogenic bacteria situation after intervening: use X 2 test respectively to organize medicine carrying hydrogel group positive bacterial culture recall rate and compare with model group statistical significance (table 5) is all arranged, other each groups also have the trend that reduces positive rate.Can be found out by result, medicine is made durative action preparation can accurately arrive inflammation part, bacteria growing inhibiting, and then play the purpose that inflammation is alleviated.
Table 5 positive bacterial culture recall rate relatively
Compare with model group
*P＜0.05,
3.5.3B super check result
After injecting antibacterial 24h, ultrasound diagnosis shows that all experiments were animal vitreous chamber all has empyema to form, and choroid thickens.Intervene rear 14d capable ultrasound diagnosis again, each is organized all has empyema to exist in vitreous chamber, but treatment group has the trend that obviously alleviates, especially the durative action preparation group.Through X 2 test, each is organized the detachment of retina situation and compares with model group the trend that alleviates is all arranged, and wherein medicine carrying hydrogel group effect is best, with it, significant difference (table 6) is arranged relatively.
Table 6 detachment of retina rate relatively
Compare with model group
*P＜0.05,
3.5.4 histological examination result
After intervening 14d, the visible model group moderate of gross specimen is to severe corneal clouding, edema, vitreous chamber is filled by empyema fully, the eyeball distortion, common treatment group cornea is moderate or Mild edema, long-acting treatment group cornea is Mild edema, and wherein the aqueogel group levels off to normal condition, and each treatment group vitreous chamber all has local empyema.
Under light microscopic, the model group corneal edema, the extensive hypertrophy of squamous epithelial cancer, holostrome all has inflammatory cell infiltration; Be full of inflammatory cell in the anterior chamber, iris congestion and edema, massive inflammatory cells infiltrated, granulation tissue hyperplasia; Lenticular opacity, pus cell and degeneration vitreous body filling glass body cavity; Normal retinal structure disappears fully, and most of nethike embrane breaks away from fracture, retina cell extensive necrosis.Common treatment group cornea hyperplasia of squamous epithelium, inflammatory cell infiltration, the inter-lamination of cornea visible vessels is grown into; The visible obviously cell infiltration in place, angle, room, iris edema, local proliferation, part-blood enlargement of pipe, acute and chronic cell infiltration; Vitreous chamber massive inflammatory cells infiltrated, sphacelus ooze out and the vitreous body of degeneration; The retina level still can be differentiated, and structure is substantially complete, and local holostrome retina cell is downright bad, red dying, and durative action preparation group cornea hyperplasia of squamous epithelium is not obvious, rare inflammatory cell infiltration; Room Jiao Chu has no cell infiltration, iris edema, local proliferation, part-blood enlargement of pipe, acute and chronic cell infiltration; The rare inflammatory cell infiltration of vitreous chamber, sphacelus oozes out and the vitreous body of degeneration; The retina level is clearly more demarcated, the structure normal, and inside and outside granular layer distributes normal.The description of test medicine has therapeutical effect to bacterial endophthalmitis, after it is prepared into durative action preparation, plays a role more obvious, and longer duration, action effect is good.
The therapeutical effect of 4 medicine exogenous eye inflammation (infection of optical fundus traumatic secondary)
4.1 laboratory animal
80 of healthy Wistar rats, male and female are not limit, and body weight is at 220-260g, observes ophthalmic under slit lamp in good condition and carry out this test without other ophthalmic person.
4.2 experimental technique
4.2.1 modeling method
Suck etherization, on simple fixing and Mus plate, use ophthalmology micro-surgical instrument 2.0mm after the elongation limbus of sclera centered by 10 positions on the right eye temporo, do the curved incision parallel with corneoscleral junction, be about 3.0mm.Cut the wall of eyeball holostrome, deeply reach vitreous chamber.Incision causes the pigmented film incarceration, and the pigmented film exposed parts is parallel with cutting edge.Otch will not be sewed up.
4.2.2 group technology
It is model group, 12 groups of embodiment, doxorubicin ordinary preparation group that modeling success laboratory animal is divided into 14 groups at random, respectively organize equal intravitreal 100 μ l medicines or contain the carrying medicine of 100 μ l medicines except model group and collyrium group, model group gives isopyknic PBS solution, and collyrium directly splashes in rabbit eyes.
4.3 inspection item and method
4.3.1 conventional examination of ocular fundus
Every daily slit lamp, indirect ophthalmoscope inspection.Respectively at after intervening 1,4,7,14d carries out the scoring of endophthalmitis clinical scale to each treated animal, primary part observation anterior chamber scintillation, aqueous cell and vitreous opacity degree, its grade scale is described with 3.3.1.
4.3.4 histological examination
After each organizes rat intervention 14d, get 5 for every group and extract eyeballs, frozen section is made 4 μ m sections and is used for HE dyeing.
4.4 statistical procedures
Data represent with x ± s, t check between relatively employing group between two groups.P＜0.05 for significant difference is arranged, has statistical significance.
4.5 result
4.5.1 clinical inflammation scoring
24h after the injection antibacterial, blepharoedema appears in all rats, and corneal edema is obvious, and conjunctiva is congested, edema obviously, anterior chamber's scintillation (+), exudation of anterior chamber (+).After intervention, 4,7,14d carries out the scoring of clinical inflammation, the model group inflammatory symptom increases the weight of day by day, and treatment group laboratory animal cornea is all transparent gradually, and anterior chamber's scintillation alleviates gradually, oozes out to be tending towards absorbing, and vitreous opacity alleviates in various degree.Each treatment group and model group more all have significant difference (table 7).
Table 7 postoperative different time points is respectively organized clinical inflammation scoring situation (n＞10x ± s)
Compare with model group
*P＜0.05,
*P＜0.01
4.5.2 histological examination result
Under light microscopic, as seen, (1) model group: be all cellular infiltration in ball, take neutral grain as main, retina, choroid disappear, and the lymphocytic infiltration of inequality occurs.Choroid is take a large amount of lymphocytic infiltrations as main, the obvious hypertrophy of choroid.Retina, choroid structure disappear.The retina dissolving is downright bad, (draw materials and see ball wall out-of-shape, eyeball obviously dwindles the serious atrophy of choroid.In retina, external granular layer is covered with pus cell, the serious atrophy of the granular layer unclear retina choroid of identification.(2) common treatment group: retina, choroid anthorisma, the core off normal is without hypertrophy; A little oozes out interior external granular layer, has edema hemorrhage between granular layer.The obvious plump hypertrophy of choroid has hemorrhage.Each layer of retina edema, boundary are unclear, and take lymphocytic infiltration as main, granular layer has and oozes out, a large amount of lymphoaccumulations.The obvious plump hypertrophy of choroid, each stratum boundary limit of retina is unclear, a large amount of lymphocytic infiltrations.Retina, choroidal atrophy, optic atrophy.Choroid neutrophilic granulocyte, lymphocytic infiltration.The corpus ciliare choroideae cell infiltration, the angle, room is stopped up.(3) long-acting treatment group: rare neutrophil infiltration scleral walls in ball.Can distinguish retina, choroid, each layer has no atrophy.Choroid is without obviously hypertrophy, necrosis.The cellular atrophy of part ciliary epithelium.Optic atrophy is not obvious.Explanation thus, medicine Trauma endophthalmitis has certain therapeutical effect, and especially after being prepared into durative action preparation, the curative effect of performance is more obvious, directly arrives affected area and works.
In sum, by each group of anthracycline antibiotics embodiment, the therapeutic effect of the exogenous eye inflammation of eye table, eye table endogenous eye inflammation, the exogenous eye inflammation in optical fundus and optical fundus endogenous eye inflammation is analyzed, each group of embodiment shows extraordinary therapeutical effect effect, and also the therapeutic effect than anthracycline ordinary preparation is good for each group of embodiment.Each group of anthracycline antibiotics embodiment has the effect of extraordinary treatment eye inflammation.
Obviously, the above embodiment of the present invention is only for example of the present invention clearly is described, and is not to be restriction to embodiments of the present invention.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here can't give all embodiments exhaustive.Everyly belong to the row that apparent variation that technical scheme of the present invention extends out or change still are in protection scope of the present invention.
Claims (10)
1. anthracycline antibiotics and officinal salt thereof the purposes in preparation treatment eye inflammation medicine, is characterized in that, described eye inflammation is exogenous eye inflammation and/or endogenous eye inflammation.
2. purposes according to claim 1, is characterized in that, described anthracycline antibiotics is compound and the stereoisomer thereof of following structure:
Wherein, R1 is H, OH, CH
3, CH
2OH or OCH
3, R2 is COR1 or CR1R1, R3 is H, OH, CH
3, CH
2OH, OCH
3, pyridine radicals, furyl, pyrrole radicals, thienyl or pyranose.
3. purposes according to claim 1, is characterized in that, described anthracycline antibiotics is doxorubicin, table doxorubicin, pyrans doxorubicin, mitoxantrone, daunorubicin, idarubicin, aclarubicin or carminomycin.
4. according to claim 1-3 arbitrary described purposes, is characterized in that, described medicine is by described anthracycline antibiotics and officinal salt and the preparation of pharmaceutically acceptable adjuvant.
5. purposes according to claim 4, is characterized in that, described medicine is common flour injection, eye drop or long-acting sustained-release injection, and wherein, described long-acting sustained-release injection is nano particle preparations, microball preparation, aqueogel or Liposomal formulation.
6. purposes according to claim 5, it is characterized in that, the adjuvant of described nano particle preparations, microball preparation, employing when aqueogel prepares comprises macromolecular material, described macromolecular material is selected from: one or more in poly-anhydride, polyoxyalkylene, polyamide, polyester, polyacrylic resin, polyethers, polyphosphazene, starch, collagen, polypeptide or polysaccharide, or be selected from copolymer between the different monomers of described macromolecular material; The adjuvant that adopts when described Liposomal formulation prepares comprises natural or synthetic phospholipid, lipoid or its combination.
7. purposes according to claim 6, it is characterized in that, described macromolecular material is selected from: poly-anhydride, polyvinyl alcohol, Polyethylene Glycol, poly-anhydride-ethylene glycol copolymer, NIPA-acrylic copolymer, polybutylcyanoacrylate, polylactic acid, octadecane diacid acid anhydride-ethylene glycol block copolymer, PLGA, polylactic-co-glycolic acid-ethylene glycol copolymer, PGA, Acetic acid, hydroxy-, bimol. cyclic ester lactide-ethylene glycol-Acetic acid, hydroxy-, bimol. cyclic ester lactide triblock copolymer, polyether sulfone, gelatin, poly(hydrobutyl ester), poloxamer, collagen, Fibrinogen, albumin, cellulose, glucosan, alginate, dextran, hyaluronic acid, one or more in polypeptide or chitosan.
8. purposes according to claim 1, is characterized in that, described anthracycline antibiotics is 0.5-2000 μ g/kg for the eye drops dosage of human body.
9. purposes according to claim 8, is characterized in that, described anthracycline antibiotics is 1-1500 μ g/kg for the eye drops dosage of human body.
10. purposes according to claim 8, is characterized in that, described anthracycline antibiotics is 3-1000 μ g/kg for the eye drops dosage of human body.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104312738A CN103169717A (en) | 2011-12-21 | 2011-12-21 | Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104312738A CN103169717A (en) | 2011-12-21 | 2011-12-21 | Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103169717A true CN103169717A (en) | 2013-06-26 |
Family
ID=48630207
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011104312738A Pending CN103169717A (en) | 2011-12-21 | 2011-12-21 | Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103169717A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104910373A (en) * | 2013-08-25 | 2015-09-16 | 张雅珍 | New polymer, preparation and application thereof |
| CN105037710A (en) * | 2013-09-01 | 2015-11-11 | 韩冰 | Preparation method of new compound and new application of the compound in medical treatment |
-
2011
- 2011-12-21 CN CN2011104312738A patent/CN103169717A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| A.COHEN等: "Antivial effect of daunomycin", 《NATURE》 * |
| 边藏丽等: "检测六种抗肿瘤抗生素对五种耐药细菌的杀灭效果", 《药学服务与研究》 * |
| 陈舒等: "柔红霉素对兔晶状体上皮细胞及眼前节组织的影响", 《眼视光学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104910373A (en) * | 2013-08-25 | 2015-09-16 | 张雅珍 | New polymer, preparation and application thereof |
| CN105037710A (en) * | 2013-09-01 | 2015-11-11 | 韩冰 | Preparation method of new compound and new application of the compound in medical treatment |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8901131B2 (en) | Gatifloxacin-containing ophthalmic gel and preparation method thereof | |
| CN102085203B (en) | Ophthalmic preparation of levofloxacin and prednisolone acetate and preparation method thereof | |
| Gerhart et al. | Depletion of Myo/Nog cells in the lens mitigates posterior capsule opacification in rabbits | |
| CN101125149B (en) | Ophthalmic operating special-purpose colored perfusate | |
| US20210290432A1 (en) | Kit for producing keratoconjunctival cover sheet and method for producing keratoconjunctival cover sheet | |
| Pathengay et al. | Acute postoperative endophthalmitis following cataract surgery: a review | |
| Mietz et al. | Histopathology of an avascular filtering bleb after trabeculectomy with mitomycin-C | |
| JP3603129B2 (en) | Therapeutic agent for diabetic keratosis | |
| CN106619573A (en) | Timolol maleate cubic liquid crystal nanoparticle eye drops and preparation method thereof | |
| CN105142670B (en) | Eye drop composition for treating ocular inflammatory disease and preparation method thereof | |
| CN103169717A (en) | Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation | |
| CN114129889B (en) | Annular microneedle for ophthalmology | |
| CN103006706B (en) | Amniotic membrane liposome for ocular surface reconstruction and preparation method and application thereof | |
| Xiao et al. | Staining the anterior capsule with indocyanine green or trypan blue for capsulorhexis in eyes with white cataract | |
| CN103169718B (en) | Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating neovascular glaucoma | |
| CN112891326B (en) | Natamycin-loaded alginic acid gel medicine film and preparation method thereof | |
| US8349005B2 (en) | Method for burying implant to choroid | |
| CN102349901B (en) | Application of pirfenidone in preparation of medicaments for controlling proliferative diseases after ophthalmologic operation and eye drops thereof | |
| RU2284181C2 (en) | Pharmaceutical composition for prophylaxis of ophthalmological infections | |
| Carney et al. | Vitrectomy in endophthalmitis | |
| CN107638433A (en) | A kind of triperygium wilfordii extractive and preparation method thereof and the application in eye drops is prepared | |
| CN111358771B (en) | Atropine sulfate eye film agent and preparation method thereof | |
| US20210283174A1 (en) | Oxygenated emulsion for treatment of ocular injury | |
| CN116172885A (en) | Pharmaceutical preparation for resisting after-cataract and preparation method thereof | |
| JP3953322B2 (en) | Use of trypan blue to visualize the anterior lens capsule of the eyeball |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130626 |