CN103834003B - Noval chemical compound and preparation method and purposes containing Crizotinib structure - Google Patents

Abstract

The invention discloses the preparation methods and purposes of the noval chemical compound containing Crizotinib structure and the compound.The drug that the noval chemical compound for containing Crizotinib structure is prepared into treatment glaucoma disease and protects the effect on optic nerve more preferable than the drug of other forms or medicament.

Description

Noval chemical compound and preparation method and purposes containing Crizotinib structure

Technical field

The invention discloses the preparation methods and purposes of the noval chemical compound containing Crizotinib structure and the compound.

Background technique

Crizotinib, Chinese name also known as gram in replace Buddhist nun, English name crizotinib, Food and Drug Adminstration of the US (FDA) advanced stage of approval (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (commodity English name Xalkori) treatment anaplastic lymphoma kinase abnormal gene expression (Locally Advanced or metastatic) non-small cell lung cancer.The clinical treatment of Xalkori be single drug, after have been reported that it in mind Through with the effect on neovascular glaucoma.By the end of 2012, there are 6,005 million peoples to suffer from glaucoma in the world, In 4,005 million peoples be primary open-angle glaucoma, 1,500 power people are primary angle-closure glaucomas.Glaucoma One main blinding factor is exactly that intraocular pressure (intraocular pressure, IOP) is increased.Currently, mainly in terms of two Glaucoma is treated, first is that reduce intraocular pressure, mechanism of action is generated and hypertonic de- by promoting aqueous humor drainage, reducing aqueous humor Water effect is to reduce intraocular pressure；Second is that protection optic nerve, Deterioration of Optic Nerve in Glaucoma eventually leads to visual function forfeiture, optic nerve Protecting remedy measures includes reducing intraocular pressure, using calcium antagonist, antioxidant, immunotherapy, supplement and alternative medicine, potentially Cell repair and gene therapy, and not all optic nerve protection means are used equally for clinic, much still in the experimental study stage, Its less effective.Crizotinib of the invention is coupled segmented copolymer, can make preparation containing the new of Crizotinib Structure dissolves in water, controls drug delivery amount well, and then reaches treatment glaucoma disease or derivative optic nerve injury disease Disease.

It is simply wrapped with polymer when treating glaucoma in the noval chemical compound containing Crizotinib of this patent preparation Nanometer formulation made of Crizotinib is compared, and the effect of drugs that this patent is invented far surmounts simply wrapped gram of polymer In azoles base of a fruit Buddhist nun nanometer formulation, medication effect is very good.And the tool of the ordinary preparation of this patent discovery Crizotinib There is drop intraocular pressure effect well.

Summary of the invention

The contents of the present invention are as follows:

The invention discloses the segmented copolymer of the coupling of Crizotinib shown in following formula I, structure is as follows:

Wherein PEG refers to polyethylene glycol, the integer between molecular weight 100-100000, n=1-200；It is preferred that n=1- Integer between 100.

The preparation method of present copolymer, it is characterised in that:

1) compound A is obtained with methoxy poly (ethylene glycol) amine and citric acid reactions；

2) compound A and acetylating decanedioic acid react to obtain polymer B, wherein the integer of n=1-200, preferably 1- 100；

3) polymer B reacts to obtain final product with the C containing Crizotinib；

Compound A；

Polymer B；

Crizotinib C

The chemical step optional solvents for wherein synthesizing final product are selected from: benzene, toluene, pyridine, tetrahydrofuran, chlorine One of imitative, carbon tetrachloride, methylene chloride, methanol, ethyl alcohol, methylene chloride, dimethyl sulfoxide, n,N-Dimethylformamide or It is a variety of.

The compound of brand-new can be prepared into the nanometer formulation suitable for local administration, microball preparation.Purposes is preparation treatment The drug of glaucoma and the drug of protection nerve.

Preparation method of the invention is specific as follows:

1) decanedioic acid is flowed back in acetic anhydride, forms acetyl group-decanedioic acid；

2) methoxy poly (ethylene glycol) amine and citric acid dissolve in a solvent, and hybrid reaction is overnight, are dried to obtain compound A:

3) acetyl group-decanedioic acid is mixed with compound A, is reacted at 100-200 DEG C, reaction time 10min to 10h； After reaction mixture is cooling, washing is dried to obtain polymer B；

4) by Crizotinib C and polymer B as after 1-96 hours in solvent, ultrasonic reaction 1-20 minutes, in baking oven In foster to obtain Formulas I polymer, then homogenizer high-speed stirred 1-10 minutes in subzero 30 DEG C of 0-, rotation volatilization is slightly produced Object, post-processing obtain the nanometer formulation of final product Formulas I polymer.

Its reaction equation is as follows:

The polymer containing Crizotinib structure that the present invention obtains is easy to dissolve in water, and its half-life period ratio Crizotinib extends very much, such noval chemical compound acts on obvious in treatment glaucoma kind excellent in protection optic nerve.

The nanometer formulation made of the final product of this patent preparation is in the comparison for the treatment of eye disease, the new chemical combination of this patent The medication effect of object is very good, surmount completely directly wrapped up by polymer Crizotinib (not with Crizotinib idol Connection) nanometer formulation.

Detailed description of the invention:

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

Nanoparticle made of Fig. 2 embodiment 1, decanedioic acid-glycol copolymer directly wrap up Crizotinib nanoparticle With the drug accumulative releasing degree and time chart of Crizotinib ordinary preparation.

Nanoparticle made of Fig. 3 embodiment 1, decanedioic acid-glycol copolymer directly wrap up made of Crizotinib Comparison figure of the nanoparticle in the protective effect of optic nerve

Specific embodiment

Invention is further described in detail for specific embodiment below, but the present invention not only limits to following embodiment.

It is as follows to prepare embodiment:

Embodiment 1

1) mixture reflux of the decanedioic acid 45g in 500ml acetic anhydride, to form acetyl group-decanedioic acid；

2) methoxy poly (ethylene glycol) amine 6g, citric acid 72mg, dicyclohexylcarbodiimide 185mg and pyridine 8mg mixing add Enter 32ml methylene chloride, is stirred at room temperature overnight；Then it is washed, and is dried under vacuum with ether, obtain polymer；

3) by the 1) step and the 2) step product mix be put into flask, decompression contains intermingle with reaction 1 hour at 180 DEG C；To poly- It closes object to be cooled to room temperature and is dissolved with chloroform, and with petroleum ether and drying；

4) by 200mg Crizotinib is stand-by and the 700mg polymer of step 3 is put into 8ml dimethyl sulfoxide and 10ml 24 hours in dichloromethane solution；Ultrasound 5 minutes；Then it is placed in baking oven 2 hours；Homogenizer ultrahigh speed is stirred in subzero 10 degree It mixes 2 minutes, is then put into 1.5% poly-vinyl alcohol solution and stirs 3 hours for 500 turns；Freeze-drying is after being collected by centrifugation to get final product Nanoparticle.

Embodiment 2

1) methoxy poly (ethylene glycol) amine 4g, citric acid 55mg, dicyclohexylcarbodiimide 260mg and pyridine 6mg mixing add Enter 20ml methylene chloride, is stirred at room temperature overnight；Then it is washed, and is dried under vacuum with ether；

2) acetyl group-decanedioic acid 20g (commercially available) and step 1 product mix are put into flask, react 1.5 at 170 DEG C Hour；Object to be polymerized is cooled to room temperature to be dissolved with chloroform, and with petroleum ether and be dried to obtain polymer (structure or with self-control It is slightly different, molecular weight or due to commercially available quality it is different)；

3) 30mg Crizotinib and polymer 150mg are put into mixed by 0.5ml methanol and 1ml methylene chloride it is molten In liquid；Ultrasound 2 minutes；Then it is placed in baking oven 24 hours；In minus 20 degrees, homogenizer high-speed stirred 3 minutes, product was put into It is stirred 2 hours for 600 turns in 5% cholic acid solution；The microballoon to get noval chemical compound is lyophilized after being collected by centrifugation.

Effect experiment is as follows:

By the embodiment 1-2 sample prepared and the Crizotinib directly wrapped up with decanedioic acid-glycol copolymer (chemical coupling reaction does not occur for nanoparticle pharmaceutical group, Crizotinib does not have structure change), Crizotinib ordinary preparation (powder Injection) stability test, drug release in vitro test and the drug action of letter out eye and neuroprotection test are carried out respectively.

Stability test:

By the sample of example 1 group preparation and the nanoparticle of decanedioic acid-glycol copolymer package Crizotinib Medicine group, Crizotinib ordinary preparation take same amount (in terms of Crizotinib) to measure absorbance value respectively.Then it is put into for three groups 3 months in 20 degree of incubators, then take out measurement absorbance value, as a result visible embodiment 1-2 group and ordinary preparation group gram in azoles Without change before and after the absorbance value of base of a fruit Buddhist nun, and the nanoparticle group absorbance decline 21% wrapped up.Drug release in vitro test:

The nanoparticle pharmaceutical group for the Crizotinib that example 1 group, decanedioic acid-glycol copolymer are directly wrapped up and Crizotinib ordinary preparation component also known as takes the drug (in terms of Crizotinib, every group of Crizotinib containing 10mg) of equivalent, Then each group drug after being impregnated with PBS buffer solution, is shaken under 37 degrees Celsius as in test tube in shaking table, after timing sampling, The medicament contg percentage that release is calculated in the content of ultraviolet specrophotometer measurement drug, and after recording, does releasing curve diagram, Abscissa is time (day), and ordinate is the percentage of release.See Fig. 2, it is seen that the drug of embodiment release is more permanent, makes drug Half-life period is longer.

Solubility test in water:

By embodiment 1-2 group, the nanoparticle pharmaceutical group of the Crizotinib that decanedioic acid-glycol copolymer directly wraps up with And Crizotinib ordinary preparation component also known as takes the drug of equivalent (in terms of Crizotinib, every group containing the azoles base of a fruit in 100mg grams Buddhist nun), it is respectively put into test tube, with 10mlPBS buffer solution and shakes, observe dissolution situation after static.Record 3 minutes and 20 The dissolved state of minute, it is as a result as follows.

1 solubility of table compares

To the Experiment on therapy of rat angle-closure glaucoma:

1.1 experimental animals and grouping

Experiment uses Thirty male rats, weight 220g or so.Before modeling, animal is grouped at random: i.e. blank Control group；Model group；Embodiment 1-2 group, polymer directly wrap up Crizotinib group, common Crizotinib preparation group every Intraocular injection is administered once within 5 days, dosage 0.1mg injection/kg；Positive drug group selection pilocarpinum eye drops, daily administration Once, one time one drop.All administrations continue 15 days.

The foundation of 1.2 animal models

Experiment before be injected intraperitoneally 4% chloraldurate of rat (200mg.kg-1), then with 1% lidocaine local anaesthesia Cornea of rats.During high intraocular pressure, keep making animal continuous narcosis every 0.2 milliliter of 4% chloraldurate of half an hour intraperitoneal injection State.Increase rat right eye intraocular pressure using pulley-suture system, while left side eyeball compares.By surgical sutures two Holding each is an identical heavy counterweight, a circle is hitched among suture for 2 millimeters after the rat canthus limbus of sclera, both ends Counterweight continually and steadily presses to rat eye by pulley.Counterweight pressure is 20 grams, continuous 6 hours, rat is caused to close angle Type glaucoma model.Raising is pressed and is recorded to the 15th day measurement rathole.

1.3 experimental result

Experimental result is shown in Table 1

The intraocular pressure result (mmHg, n=10) of glaucoma rat under 2 each group drug treatment of table

Compared with model group^*P < 0.05^**P < 0.01

Protective effect experiment to optic nerve:

2.1 experimental animals and grouping

Experiment uses Adult female rats, weight 200g or so.Animal is grouped at random: i.e. blank control group； Model group；Embodiment 1-2 group, polymer directly wrap up Crizotinib group, and common Crizotinib preparation group is intraocular every 5 days Drug administration by injection is primary, dosage 0.1mg injection/kg, terminates observation after 20 days.

The foundation of 2.2 animal models

6% water of rat is injected intraperitoneally before carrying out the preparation experiment of nerve injury model according to the method for Chen Xiaorui et al. report Chloral (200mg.kg-1) anesthesia is closed, eye part cleaning-sterilizing goes in ring and cuts off outer canthus portion bulbar conjunctiva, opens Tenons capsule, passivity Suspensorium is separated, pulls eyeball forward, exposure optic nerve, eye scissors are made 2mm at into vertically cutting off optic nerve in socket of the eye from after ball At optic nerve injury, optical fundus blood vessel is avoided damage to.Guarantee consistency, all optic nerve transection wounds are all in identical position, seam Bulbar conjunctiva is closed, also receives eyeball, applies Antibiotic Eye Ointment.There is within postoperative 2 days Marcus-gun pupil, eyeball is without obvious prominent, eyelid Completely, eyeground is successful model without obvious bleeder to closure.Vacation hurts eye and only separates exposure optic nerve, not all right cross-section wound.Administration 20 Eyeball is won after it.After disconnected neck is put to death, entire eyeball (retaining ball optic nerve 2mm) is won, the observation of sample row Electronic Speculum is made into.Knot Fruit sees Fig. 3, it is seen that the function and effect of the nerve of embodiment protection are much better than polymer and directly wrap up Crizotinib patron saint The effect of warp.

Claims (7)

1. structure is as follows containing the segmented copolymer of Crizotinib in the structure being shown below:

Wherein PEG is polyethylene glycol, the integer between molecular weight 100-100000, n=1-200.

2. the preparation method of copolymer as claimed in claim 1, it is characterised in that:

1) compound A is obtained with methoxy poly (ethylene glycol) amine and citric acid reactions；

2) compound A and acetylating decanedioic acid react to obtain polymer B, wherein the integer of n=1-200；

3) polymer B reacts to obtain final product with Crizotinib C；

3. the preparation method of claim 2, wherein the step 1) -3) select solvent to be selected from: benzene, toluene, pyridine, tetrahydro furan It mutters, one of chloroform, carbon tetrachloride, methylene chloride, methanol, ethyl alcohol, dimethyl sulfoxide, N,N-dimethylformamide or more Kind.

4. the purposes of the segmented copolymer of claim 1, purposes is the nanometer formulation being prepared into suitable for local administration, microballoon system The drug of agent.

5. the purposes of the segmented copolymer of claim 1, purposes is the drug of preparation treatment eye disease.

6. the purposes of claim 5, wherein the eye disease is nonneovascular glaucoma.

7. the purposes of the segmented copolymer of claim 1, purposes is the drug of preparation protection nerve.