CN103772699A - Novel chemical compound with anthracene ring antibiotic structure and preparation method and application thereof - Google Patents

Abstract

The invention discloses a novel chemical compound with an anthracene ring antibiotic structure and a preparation method and application thereof. The effect of the anthracene ring antibiotic coupling non-linear multi-block copolymer on treating choroid and retina diseases is better than that of medicines or drugs in other forms.

Description

The new compound that contains anthracycline antibiotics structure and preparation method and purposes

Technical field

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

Background technology

Anthracycline antibiotics is a kind of antimitotic and has Cytotoxic medicine.Anthracycline antibiotics can successfully suppress Several Kinds of Malignancy, comprises acute leukemia, lymphoma, soft tissue and osteosarcoma, children malignant tumors and becomes human solid tumor.Its mechanism of action is the penetrable cell that enters of medicine, is combined with karyomit(e).Thereby the planar rings in medicines structure is inserted between base pair and is combined with DNA and forms mixture, severe jamming DNA is synthetic, DNA dependent rna is synthetic and protein synthesis.But the drug level that produces tumor locus in the required Dx concentration ratio clinical treatment of anti-proliferative effect by this mechanism wants high.In addition anthracycline antibiotics is also relevant with redoxomorphism, can participate in reaction and obtain cytotoxic compound, as hydroxy and the hydrogen peroxide etc. of superoxide, activity.The site of action of anthracycline antibiotics may be at cytolemma, anthracycline antibiotics is not also reached common understanding in scientific circles for the pharmacodynamic action of various diseases at present, mechanism of action for every kind of disease does not all have general character, needs further scientific research to be explained.

Anthracycline antibiotics is widely used at present, but often needs to be become salt, for example Dx at Point of View of Clinical, common medicinal form is doxorubicin hydrochloride, needs to become hydrochloride, because of the solubleness of Dx own very low, do not transform the form of salify, cannot apply.And become after hydrochloride, the pharmacodynamics function of Dx declines to a great extent, and therefore this brings very large difficulty to clinical application.

Another shortcoming of anthracycline antibiotics is due to strong cytotoxicity itself, therefore the toxicity of itself is very large, after using through being everlasting, within about 10 days, there is obvious bone marrow depression, use after one week, can show very significantly gastrointestinal side effect and cardiac toxic, therefore must after accurate calculation, could apply when medication, and the transformation period of this medicine is very short, brings limitation equally to application.

Preparation method of the present invention, can make the new texture that contains anthracycline antibiotics of preparation dissolve in water, prolong half-life, and this preparation method has solved the application limitation of anthracene nucleus medicament in the lump.

Summary of the invention

Of the present invention theing contents are as follows:

As shown in the formula the non-linear segmented copolymer of the anthracycline antibiotics coupling shown in I, its structure is as follows:

Wherein R1, R2, R3, R4, R5 are independently H, OH, CH separately ₃, CH ₂oH or OCH ₃, R6 is H, OH, CH ₃, CH ₂oH, OCH ₃, COCH ₃or COCH ₂oH, R7, R8 are independently H, OH, CH separately ₃, OCH ₃, COCH ₃or OR9, the integer between n=1-200, preferably n=1-100; Wherein R9 is pyridyl, furyl, pyrryl, thienyl or pyranyl.Preferably wherein R1, R2, R3, R4 are independently H, OH or OCH separately ₃, R5 is OH, R6 is H, OH, COCH ₃or COCH ₂oH, R7, R8 are independently OH, CH separately ₃, OCH ₃, COCH ₃, the integer between n=1-20.More preferably wherein R1 is H or OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₃or COCH ₂oH, R7, R8 are independently OH, CH separately ₃or OR9, the integer (preferably n=1-100) between n=1-200; Wherein R9 is pyranyl.

Described polymkeric substance is respectively:

1) R1 is OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₂oH, R7 is CH ₃, R8 is OH, the integer (preferably n=1-100) between n=1-200;

2) R1 is OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₂oH, R7 is CH ₃, R8 is OR9, R9 is pyranyl, the integer (preferably n=1-100) between n=1-200;

3) R1 is OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₃, R7 is CH ₃, R8 is OH, the integer (preferably n=1-100) between n=1-200;

4) R1, R2 are H, and R3, R4 and R5 are OH, and R6 is COCH ₃, R7 is CH ₃, R8 is OH, the integer (preferably n=1-100) between n=1-200.

Select most structure as follows:

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

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

Compd A;

Polymer B;

Anthracycline antibiotic C.

Preferably, wherein anthracycline antibiotics C is selected from Dx, Zorubicin, pidorubicin, pirarubicin, daunorubicin, daunoblastin, idarubicin, aclarubicin, zhengdingmeisu, aclacinomycin or carminomycin.Wherein step 1) optionally use solvent, described solvent is selected from: one or more in benzene, toluene, pyridine, tetrahydrofuran (THF), chloroform, tetracol phenixin, methylene dichloride, dicyclohexylcarbodiimide; Step 3) optionally use solvent, described solvent is selected from one or more in methyl alcohol, ethanol, methylene dichloride, chloroform, tetracol phenixin, dimethyl sulfoxide (DMSO), DMF.

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 ℃, reaction times 20min to 2h with compd A; After question response mixture is cooling, washing, the dry polymer B that obtains;

4) by anthracycline antibiotics C and polymer B as for 0.5-24 hour in solvent, in subzero 30 ℃ of 0-, after ultrasonic reaction 2-20 minute, rotation volatilization obtains crude product, aftertreatment obtains finished product formula I.

Its reaction equation is as follows:

The purposes of polymkeric substance of the present invention, purposes preparation treatment choroid and retinal diseases.

The polymkeric substance that what the present invention obtained contain anthracycline antibiotics structure, is easy to dissolve in water, and its transformation period extend much than anthracycline antibiotics, such new compound has solved the application limitation of anthracene nucleus medicament.

Accompanying drawing explanation:

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

Fig. 2 embodiment 1, poly-sebacic acid ethylene glycol copolymer directly wrap up medicine accumulative releasing degree and the time chart of nanoparticle-associated doxorubicin and Dx ordinary preparation

Fig. 3 embodiment 1-4, directly wraps up solubleness comparison in the water of nanoparticle-associated doxorubicin

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 20g in 250ml diacetyl oxide refluxes, to form ethanoyl-sebacic acid;

2) methoxy poly (ethylene glycol) amine 6g, citric acid 70mg, dicyclohexylcarbodiimide 200mg and pyridine 10mg mix and add 30ml 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, at 150 ℃, reacts 1 hour; Treat that polymkeric substance is cooled to room temperature chloroform and dissolves, and also dry by petroleum ether;

4) by Dx and step 3) polymkeric substance of 800mg puts into the solution being mixed by 10ml methylene dichloride and 6ml dimethyl sulfoxide (DMSO); Then insert in baking oven 2 hours; In subzero 10-20 degree ultrasonic 10 minutes; Product is put into 5% cholic acid solution 600 and is turned and stir 2 hours; Freeze-drying after centrifugal collection, obtains end product.

Embodiment 2

1. methoxy poly (ethylene glycol) amine 3g, citric acid 45mg, dicyclohexylcarbodiimide 260mg 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;

Ethanoyl-sebacic acid 10g (being purchased) and the 1st step Product mix are put into flask by 2, reacts 1.5 hours at 120 ℃; Treat that polymkeric substance is cooled to room temperature chloroform and dissolves, and with petroleum ether the dry polymkeric substance that obtains; Pirarubicin and this polymkeric substance 1000mg are put into the solution being mixed by 3ml methyl alcohol and 18ml methylene dichloride by 3; Then insert in baking oven 14 hours; In subzero 10-20 degree, ultrasonic 20 minutes; Product is put into 3% cholic acid solution 400 and is turned and stir 4 hours; Freeze-drying after centrifugal collection, obtains end product.

Embodiment 3

The mixture of 1 sebacic acid 20g in diacetyl oxide 200ml refluxes, to form ethanoyl-sebacic acid;

2 methoxy poly (ethylene glycol) amine 2g, citric acid 28mg, dicyclohexylcarbodiimide 90mg and tetrahydrofuran (THF) mix and add 15ml chloroform, at room temperature stir and spend the night; Then wash with ether, and dry under vacuum, obtain polymkeric substance; The 1st step and the 2nd step Product mix are put into flask by 3, reacts 1 hour at 150 ℃; Treat that polymkeric substance is cooled to room temperature chloroform and dissolves, and also dry by petroleum ether;

The 900mg polymkeric substance of aclarubicin and step 3 is put into the solution being mixed by 12ml methylene dichloride and 8ml dimethyl sulfoxide (DMSO) by 4; Then insert in baking oven 20 hours; In subzero 20-30 degree, ultrasonic 2 minutes; Product is put into 5% cholic acid solution 700 and is turned and stir 2 hours; Freeze-drying after centrifugal collection, obtains end product.

Embodiment 4

The mixture of 1 sebacic acid 15g in diacetyl oxide 750ml refluxes, to form ethanoyl-sebacic acid;

2 methoxy poly (ethylene glycol) amine 6g, citric acid 80mg, dicyclohexylcarbodiimide 240mg mixes, and adds 40ml methylene dichloride, at room temperature stirs and spends the night; Then wash with ether, and dry under vacuum, obtain polymkeric substance;

The 1st step and the 2nd step Product mix are put into flask by 3, reacts 1 hour at 150 ℃; Treat that polymkeric substance is cooled to room temperature chloroform and dissolves, and also dry by petroleum ether;

The 1000mg polymkeric substance of daunorubicin and step 3 is put into the solution being mixed by 17ml methylene dichloride and 6mlDMF by 4; Then insert in baking oven 24 hours; Subzero 10 in 0-, ultrasonic 10 minutes; Product is put into 3% cholic acid solution 400 and is turned and stir 2 hours; Freeze-drying after centrifugal collection, obtains end product.

(there is not chemical coupling reaction in sample prepared by embodiment 1-4 and sebacic acid-glycol copolymer directly the nanoparticle medicine group of the Dx of parcel, anthracycline antibiotics does not have structural changes), anthracycline antibiotics ordinary preparation (powder injection) carries out respectively the drug action test of solubility test in the test of stability test, drug release in vitro, water and choroid cambium.

Stability test:

By the nanoparticle medicine group of the Dx of 1 group of sample of preparing of embodiment and sebacic acid-glycol copolymer parcel, Dx ordinary preparation is got with amount (in Dx) and is measured respectively absorbance and put into 20 degree incubator 3 months, take out subsequently and measure absorbance under 480 nanometers, the visible embodiment 1-4 group of result changes with the absorbance front and back nothing of the Dx of ordinary preparation group, and the nanoparticle group absorbancy of parcel declines 30%.

Drug release in vitro test:

By 1 group of embodiment, the nanoparticle medicine group of the Dx of sebacic acid-glycol copolymer parcel and Dx ordinary preparation component have another name called get equivalent medicine (in Dx, every group containing 10mg Dx), 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 480 nanometers, 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 1 discharges is more permanent, makes drug half-life longer.

Solubility test in water:

By embodiment 1-4 group, the nanoparticle medicine group of the Dx of sebacic acid-glycol copolymer parcel and Dx ordinary preparation component have another name called get equivalent medicine (in anthracycline antibiotics, every group containing 100mg anthracycline antibiotics), 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.

Table 1 solubleness comparison (see Fig. 3, from left to right, embodiment 1-4, directly wraps up nanoparticle-associated doxorubicin (layering))

The restraining effect of medicine to tela chorioidea's hyperplasia:

Get 56 of male rats, be divided at random 7 groups, i.e. the nanoparticle medicine group (chemical coupling reaction does not occur, and Dx does not have structural changes) of the Dx of control group, sebacic acid-glycol copolymer parcel, embodiment 1-4 group, Dx general formulation group.Each treated animal number is 8.Every rat is chosen a glance at random for experimental eye, and another eye is contrast eye.The equal intraocular injection 10 μ g medicines of each group or the medicine carrying prolonged action preparation containing 10 μ g medicines except control group, control group is given isopyknic PBS solution.

Use laser radiation rat eye, after light is solidifying, has Bubble formation or break up Bruch film with hyporrhea (sometimes with light sound) mark, be designated as available point.After laser photocoagulation, inject each group of medicine to rat right eye eyeball.14d after light is solidifying, tissues observed hyperplasia area also carries out histological examination.Result is as follows:

Table 2 retina and hyperplasia Area comparison (unit: mm of tela chorioidea ²)

With the comparison of control group group ^*p < 0.05, ^*p < 0.01, with the comparison of ordinary preparation group ^#p < 0.05, ^##p < 0.01

The result of upper table shows, control group retina and choroid generation hyperplasia area are larger, each treatment group respectively different limits dwindled hyperplasia area.Experiment to rat part tissue of eye hyperplasia area shows, each embodiment group can reduce pathology retina and choroidal hamartoplasia area simultaneously, but effect difference to some extent, and the embodiment 1-4 group effect that coupling wherein occurs is better.

The result of histological examination is that under light microscopic, control group light coagulates the visible outer retina in spot place and choroid structure disturbance, retinal pigment epithelium, choroid cambium hyperplasia, free companion's inflammatory cell infiltration.Medicine group compared with control group, the rarely found and less companion's limitation of cambium serous detachment of retina; Embodiment group compares that superpolymer simply wraps up Dx and that the nanoparticle medicine group effect of linked reaction does not occur is more remarkable, and the compound after coupling can significantly reduce tela chorioidea's hyperplasia.Histological examination shows, embodiment group is more thorough to the treatment of age related maculopathy than the nanoparticle medicine group of ordinary preparation group, and retina and choroid to pathology play a role simultaneously.

Claims (10)

1. the non-linear segmented copolymer of the anthracycline antibiotics coupling being shown below, its structure is as follows:

Wherein R1, R2, R3, R4, R5 are independently H, OH, CH separately ₃, CH ₂oH or OCH ₃, R6 is H, OH, CH ₃, CH ₂oH, OCH ₃, COCH ₃or COCH ₂oH, R7, R8 are independently H, OH, CH separately ₃, OCH ₃, COCH ₃or OR9, the integer between n=1-200; Wherein R9 is pyridyl, furyl, pyrryl, thienyl or pyranyl.

2. the multipolymer of claim 1, wherein R1, R2, R3, R4 are independently H, OH or OCH separately ₃, R5 is OH, R6 is H, OH, COCH ₃or COCH ₂oH, R7, R8 are independently OH, CH separately ₃, OCH ₃, COCH ₃, the integer between n=1-200; Preferably n=1-100.

3. the multipolymer of claim 1, wherein R1 is H or OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₃or COCH ₂oH, R7, R8 are independently OH, CH separately ₃or OR9, the integer (preferably n=1-100) between n=1-200; Wherein R9 is pyranyl.

4. the multipolymer of claim 1, it is respectively:

1) R1 is OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₂oH, R7 is CH ₃, R8 is OH, the integer (preferably n=1-100) between n=1-200;

2) R1 is OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₂oH, R7 is CH ₃, R8 is OR9, R9 is pyranyl, the integer (preferably n=1-100) between n=1-200;

3) R1 is OCH ₃, R2 is H, and R3, R4 and R5 are OH, and R6 is COCH ₃, R7 is CH ₃, R8 is OH, the integer (preferably n=1-100) between n=1-200;

4) R1, R2 are H, and R3, R4 and R5 are OH, and R6 is COCH ₃, R7 is CH ₃, R8 is OH, the integer (preferably n=1-100) between n=1-200.

5. the multipolymer of claim 1, its structure is as follows:

6. as the preparation method of any one multipolymer in claim 1-5, it is characterized in that:

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

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

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

Compd A;

Polymer B;

Anthracycline antibiotic C.

7. preparation method claimed in claim 6, wherein anthracycline antibiotics C is selected from Zorubicin, pidorubicin, pirarubicin, daunorubicin, daunoblastin, idarubicin, aclarubicin, zhengdingmeisu, aclacinomycin, Dx, or carminomycin.

8. the method for claim 6 or 7, wherein step 1) optionally use solvent, described solvent is selected from: one or more in benzene, toluene, pyridine, tetrahydrofuran (THF), chloroform, tetracol phenixin, methylene dichloride, dicyclohexylcarbodiimide; Step 3) optionally use solvent, described solvent is selected from one or more in methyl alcohol, ethanol, methylene dichloride, chloroform, tetracol phenixin, dimethyl sulfoxide (DMSO), DMF.

9. the purposes of claim 1-5 any one polymkeric substance, purposes is the medicine of preparation treatment choroid and retinal diseases.

10. the purposes of claim 9, wherein choroid and retinal diseases refer to choroid and retina age related disease.

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