CN103897041A - Novel compound containing vancomycin structure and preparation method and use thereof - Google Patents

Abstract

The invention discloses a novel compound containing a vancomycin structure and a preparation method and use thereof. Compared with other forms of drugs or agents, the drug prepared from the novel compound containing the vancomycin structure has a better effect on treatment of an endophthalmitis disease.

Description

The new compound that contains vancomycin structure and preparation method and purposes

Technical field

The invention discloses the new compound that contains vancomycin structure and preparation method and the purposes of this compound.

Background technology

Vancomycin (Vancomycin, INN) is a kind of glycopeptide antibiotics, is used for prevention and the infection that causes for the treatment of gram-positive microorganism.Traditionally, vancomycin is used as " last first-line drug ", is used for treating all invalid severe infections of all microbiotic.

Endophthalmitis is a kind of inflammation of involving eyeball internal layer, vitreum, sclera, and most endophthalmitis are due to bacterium or fungi infestation.Bacterium route of infection can be wound, and the eyeball wound that operation causes can be also haematogenous (bacterium menses spread and cast to intraocular).The symptom of endophthalmitis is usually heavier, mainly contains ophthalmodynia, hyperemia, serious photophobia and eyesight and sharply declines.The symptom of endophthalmitis is usually heavier, mainly contains ophthalmodynia, hyperemia, serious photophobia and eyesight and sharply declines.

The new compound that contains vancomycin of preparing at this patent directly wraps up the nanometer formulation that vancomycin makes and compares with polymkeric substance is simple when endophthalmitis in treatment, the medicine that this patent is invented medication effect in the time of the eye disease model for the treatment of endophthalmitis and severe infections is very good, and effect far surmounts the nanometer formulation of the simple directly parcel of polymkeric substance vancomycin.

Summary of the invention

Of the present invention theing contents are as follows:

The segmented copolymer that the invention discloses the vancomycin coupling shown in following formula I, its structure is as follows:

Wherein PEG refers to polyoxyethylene glycol, specifically finger-C ₂h ₄(OC ₂h ₄) W-, the wherein integer between W=1-500, molecular weight is about 100-21000, the preferred 1-300 of w, molecular weight is preferably about 100-13000; Integer between n=1-200; The preferably integer between n=1-100.

The preparation method of multipolymer of the present invention, is characterized in that:

1) obtain compd A with methoxy poly (ethylene glycol) amine and citric acid reactions;

2) reaction of the sebacic acid of compd A and ethanoyl obtains polymer B, the wherein integer of n=1-200, preferably 1-100;

3) polymer B is reacted and is obtained final product with vancomycin C;

compd A;

Polymer B;

Vancomycin C

Wherein vancomycin C is purchased.The required solvent of described chemical step of the final product of the synthetic new compound that contains vancomycin is selected from: one or more in benzene, toluene, pyridine, tetrahydrofuran (THF), chloroform, tetracol phenixin, methylene dichloride, methyl alcohol, ethanol, methylene dichloride, dimethyl sulfoxide (DMSO), DMF.

The compound of new system can be prepared into the nanometer formulation that is suitable for topical, microball preparation.Purposes is the medicine of preparation treatment endophthalmitis.

Preparation method of the present invention is specific as follows:

1) sebacic acid is refluxed in diacetyl oxide, form ethanoyl-sebacic acid;

2) methoxy poly (ethylene glycol) amine and citric acid are dissolved in solvent, and hybrid reaction is spent the night, the dry compd A that obtains;

3) ethanoyl-sebacic acid is mixed to reaction at 100-200 DEG C, reaction times 10min to 10h with compd A; After question response mixture is cooling, washing, the dry polymer B that obtains;

4) by vancomycin C and polymer B as for 1-96 hour in solvent, after ultrasonic reaction 1-30 minute, in baking oven, foster and obtain formula I polymkeric substance, then homogenizer high-speed stirring 1-10 minute in subzero 30 DEG C of 0-, rotation volatilization obtains crude product, and aftertreatment obtains the nanometer formulation of end product formula I polymkeric substance.

Its reaction equation is as follows:

The polymkeric substance that what the present invention obtained contain vancomycin structure, is easy to dissolve in water, and its transformation period extend much than vancomycin, such new compound effect in the time of the model for the treatment of endophthalmitis is splendid.

The nanometer formulation that the end product of preparing at this patent is made treatment eye disease relatively in, the medication effect of this patent new compound is very good, surmounts the nanometer formulation that is directly wrapped up vancomycin (not with vancomycin coupling) by polymkeric substance completely.

Brief description of the drawings:

The nuclear magnetic resonance map of the end product of Fig. 1 embodiment 1.

Nanoparticle, the sebacic acid-glycol copolymer that Fig. 2 embodiment 1 makes directly wraps up medicine accumulative releasing degree and the time chart of vancomycin nanoparticle and vancomycin ordinary preparation.

Embodiment

Specific embodiment is described in further detail the present invention below, but the present invention not only limits to following examples.Preparation Example is as follows:

Embodiment 1

1) mixture of sebacic acid 50g in 500ml diacetyl oxide refluxes, to form ethanoyl-sebacic acid;

2) methoxy poly (ethylene glycol) amine 6.5g, citric acid 73mg, dicyclohexylcarbodiimide 168mg and pyridine 8.2mg mix and add 32ml methylene dichloride, at room temperature stir and spend the night; Then wash with ether, and dry under vacuum, obtain polymkeric substance;

3) by the 1st) step and the 2nd) step Product mix puts into flask, and at 180 DEG C, decompression contains intermingle with reaction 1 hour; Treat that polymkeric substance is cooled to room temperature chloroform and dissolves, and also dry by petroleum ether;

4) the 800mg polymkeric substance of 150mg vancomycin and step 3 is put into the dichloromethane solution 48 hours of 8ml dimethyl sulfoxide (DMSO) and 12ml; Ultrasonic 5 minutes; Then insert in baking oven 1 hour; In subzero 10-20 degree, homogenizer ultra-high speed stirs 3 minutes, then puts into 2% polyvinyl alcohol solution 600 and turns and stir 3 hours; Freeze-drying after centrifugal collection, obtains the nanoparticle of end product.

Embodiment 2

1) methoxy poly (ethylene glycol) amine 3g, citric acid 45mg, dicyclohexylcarbodiimide 200mg and pyridine 6mg mix and add 20ml methylene dichloride, at room temperature stir and spend the night; Then wash with ether, and dry under vacuum;

2) ethanoyl-sebacic acid 20g (being purchased) and the 1st step Product mix are put into flask, at 170 DEG C, react 1.5 hours; Treat that polymkeric substance is cooled to room temperature chloroform and dissolves, and with petroleum ether the dry polymkeric substance (structure or with homemade slightly different, molecular weight or different because being purchased quality) that obtains;

3) 30mg vancomycin and this polymkeric substance 150mg are put into the solution being mixed by 1ml methyl alcohol and 1ml methylene dichloride; Ultrasonic 2 minutes; Then insert in baking oven 24 hours; In subzero 10-20 degree, homogenizer high-speed stirring 2 minutes, product is put into 3% cholic acid solution 600 and is turned and stir 4 hours; Freeze-drying after centrifugal collection, obtains the microballoon of new compound.

Effect experiment is as follows:

Sample prepared by embodiment 1-2 and and sebacic acid-glycol copolymer directly the nanoparticle medicine group of the vancomycin of parcel (there is not chemical coupling reaction, vancomycin does not have structural changes), vancomycin ordinary preparation (powder injection) carries out respectively the drug action test of stability test, drug release in vitro test and endophthalmitis.

Stability test:

By the nanoparticle medicine group of the vancomycin of 1 group of sample of preparing of embodiment and sebacic acid-glycol copolymer parcel, vancomycin ordinary preparation is got with amount (in vancomycin) and is measured respectively absorbance.Then put into 20 degree incubator 3 months for three groups, take out subsequently and measure 280 nanometer absorbances, the absorbance front and back of the vancomycin of the visible embodiment 1-2 group of result and ordinary preparation group are without changing, and the nanoparticle group absorbancy of parcel decline 26%.Drug release in vitro test:

By 1 group of embodiment, sebacic acid-glycol copolymer directly the nanoparticle medicine group of the vancomycin of parcel and vancomycin ordinary preparation component have another name called get equivalent medicine (in vancomycin, every group containing 10mg vancomycin), then by each group of medicine as in test tube, after soaking with PBS damping fluid, 37 degrees Celsius of lower joltings in shaking table, after timing sampling, under ultraviolet spectrophotometer, measure the content of medicine, and the medicament contg per-cent that after record, calculating discharges, do releasing curve diagram, X-coordinate be the time (my god), ordinate zou is the per-cent discharging.See Fig. 2, the medicine that visible embodiment discharges is more permanent, makes drug half-life longer.

Solubility test in water:

By embodiment 1-2 group, sebacic acid-glycol copolymer directly the nanoparticle medicine group of the vancomycin of parcel and vancomycin ordinary preparation component have another name called get equivalent medicine (in vancomycin, every group containing 100mg vancomycin), put into respectively test tube, with the dissolving of 10mlPBS damping fluid jolting, situation is dissolved in static rear observation.The dissolved state that records 3 minutes and 20 minutes, result is as follows.

The comparison of table 1 solubleness

The therapeutic action of medicine to eye inflammation

1 laboratory animal

Healthy Wistar rat, male and female are not limit, and body weight is at 220g, observes intraocular in good condition and carry out this test without other eye illness person under slit lamp.

2 experimental techniques

2.1 modeling method

Carry out eyeground trauma infection contamination model, rat sucks etherization, on simple fixing and mouse plate, uses ophthalmology micro-surgical instrument on right eye temporo centered by 10 positions after elongation limbus of sclera 2 millimeters, does the arc incision parallel with corneoscleral junction, is about 3.0mm.Cut wall of eyeball holostrome, deeply reach vitreous space.Incision causes pigmented film incarceration, and pigmented film exposed parts is parallel with cutting edge, and otch will not be sewed up, and allows it produce and infects.

2.2 clustering method

Modeling success laboratory animal is divided into model group, embodiment 1-2 group, vancomycin ordinary preparation group at random, polymkeric substance directly wraps up nanoparticle subgroup prepared by vancomycin, the equal intravitreal 50 μ l medicines of each group, model group gives isopyknic PBS solution.

3 inspection items and method

Conventional fundoscopy

Every daily slit lamp, indirect ophthalmoscope inspection.After intervening, each treated animal was carried out to the scoring of endophthalmitis clinical scale in 1,4,7 day, primary part observation anterior chamber scintillation, aqueous cell and vitreous opacity degree, its grade scale is that (inflammatory exudate enters aqueous humor in anterior chamber's scintillation classification, under the narrow light belt of slit lamp casts oblique rays on, visible light flash and ooze out particle float, 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 lens details); 3 grades, significantly aqueous flare (obvious milky white light beam, impalpable iris and lens details); 4 grades, serious anterior chamber's scintillation, aqueous humor becomes curdled appearance, with a large amount of fibrinous exudates.

4 statistical procedures

Data represent with x ± s, t inspection between relatively employing group between two groups.P < 0.05, for there being significant difference, has statistical significance.

5 results

Clinical inflammation scoring

24h after sudden and violent leakage wound, there is palpebral edema in all rats, and corneal edema is obvious, and conjunctiva is congested, oedema obviously, anterior chamber's scintillation (+), exudation of anterior chamber (+).After intervening, within 4,7 days, carry out clinical inflammation scoring, 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 and is tending towards absorbing, and vitreous opacity alleviates in various degree.Each treatment group and model group more all have significant difference (table 1).

The clinical inflammation scoring of the each group of the postoperative different time points of table 1 situation (n > 10x ± s)

With relatively * p < 0.05 of model group, * * p < 0.01.

Claims (9)

1. in the structure being shown below, contain the segmented copolymer of vancomycin, its structure is as follows:

。

2. the multipolymer of claim 1, wherein PEG refers to polyoxyethylene glycol, specifically finger-C ₂h ₄(OC ₂h ₄) W-, the wherein integer between W=1-500, molecular weight is 100-21000, the preferred 1-300 of w, the integer between n=1-200; The preferably integer between n=1-100.

3. the preparation method of multipolymer as claimed in claim 1, is characterized in that:

1) obtain compd A with methoxy poly (ethylene glycol) amine and citric acid reactions;

2) reaction of the sebacic acid of compd A and ethanoyl obtains polymer B, the wherein integer of n=1-200, preferably 1-100;

3) polymer B is reacted and is obtained final product with vancomycin C;

compd A;

Polymer B;

Vancomycin C.

4. the method for claim 3, wherein said chemical step 1-3 selects solvent to be selected from: one or more in benzene, toluene, pyridine, tetrahydrofuran (THF), chloroform, tetracol phenixin, methylene dichloride, methyl alcohol, ethanol, methylene dichloride, dimethyl sulfoxide (DMSO), DMF.

5. the compound of claim 1 can be prepared into the nanometer formulation that is suitable for topical, microball preparation.

6. described in claim 5, part is intravitreal injection administration, dosing eyes.

7. the purposes of the compound of claim 1, purposes is the medicine of preparation treatment eye disease.

8. the purposes of the compound of claim 1, purposes is the medicine of preparation treatment ocular infection.

9. the purposes of claim 7, wherein eye disease refers to endophthalmitis.

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