CN103833997A - New compound with tozasertib structure as well as preparation method and application thereof - Google Patents

Abstract

The invention discloses a new compound with a tozasertib structure as well as a preparation method and application thereof. Compared with other forms of medicines or medicaments, the medicine prepared by the new compound with the tozasertib structure has better effects in treating glaucoma and protecting optic nerve.

Description

The new compound that contains MK-0457 structure and preparation method and purposes

Technical field

The invention discloses the new compound that contains MK-0457 structure and preparation method and the purposes of this compound.

Background technology

MK-0457 (English: Tozasertib; Chemical name: N-[4-[[4-(4-methyl isophthalic acid-piperazinyl)-6-[(5-methyl isophthalic acid H-pyrazole-3-yl) amino]-2-pyrimidyl] sulphur] phenyl] cyclopropane carboxamide; Another name: MK 0457 or VX 680 are a kind of antitumor drugs of discovered in recent years, after have its effect in neural and glaucoma of report.By the end of 2012, there are in the world 6,005 million peoples to suffer from glaucoma, wherein 4,005 million peoples are primary open angle glaucomas, 1,005 million peoples are primary angle-closure glaucomas.A glaucomatous main blinding factor is exactly that intraocular pressure (intraocular pressure, IOP) raises.At present, mainly treat glaucoma from two aspects, the one, reduce intraocular pressure, thereby its mechanism of action is to reduce intraocular pressure by promoting aqueous humor drainage, the generation of minimizing aqueous humor and height to ooze dehydration; The 2nd, protection optic nerve; Deterioration of Optic Nerve in Glaucoma finally causes visual function to be lost; optic nerve protection treatment measure comprises reduction intraocular pressure, uses calcium antagonist, antioxidant; immunotherapy; supplement and alternative medicine, potential cytothesis and gene therapy, be not that all optic nerve protection means all can be used for clinical; much, still in the experimental study stage, its effect is not good enough.MK-0457 coupling segmented copolymer of the present invention, can make the new texture that contains MK-0457 of preparation dissolve in water, well controls drug delivery amount, and then reaches treatment glaucoma disease or derivative optic nerve injury illness.

The new compound that contains MK-0457 of preparing at this patent in treatment when glaucoma and polymkeric substance simply wrap up the nanometer formulation that MK-0457 makes and compare, the effect of drugs that this patent is invented far surmounts polymkeric substance and simply wraps up the nanometer formulation of MK-0457, and medication effect is very good.

Summary of the invention

Of the present invention theing contents are as follows:

The segmented copolymer that the invention discloses the MK-0457 coupling shown in following formula I, its structure is as follows:

Wherein PEG refers to polyoxyethylene glycol, and molecular weight is 100-100000, the 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 the C that contains MK-0457;

compd A;

polymer B;

Containing the C of MK-0457

The compound that wherein contains MK-0457 C is prepared according to the disclosed preparation method of CN100484934C.Wherein the required solvent of described chemical step of synthetic final product 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 the preparation glaucomatous medicine for the treatment of and neuroprotective.

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 10min to 10h with compd A; After question response mixture is cooling, washing, the dry polymer B that obtains;

4) by the C that contains MK-0457 structure 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 ℃ 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 MK-0457 structure, is easy to dissolve in water, and its transformation period extend much than MK-0457, such new compound is splendid in treatment glaucoma kind effect, obvious in the effect of protection optic nerve.

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 MK-0457 (not with MK-0457 coupling) by polymkeric substance completely.

Accompanying drawing explanation:

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 MK-0457 nanoparticle and MK-0457 ordinary preparation.

Nanoparticle, the sebacic acid-glycol copolymer that Fig. 3 embodiment 1 makes directly wraps up nanoparticle that MK-0457 the makes comparison diagram in the provide protection of optic nerve

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 68mg, dicyclohexylcarbodiimide 160mg 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 ℃, 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 200mg MK-0457 and step 3 is put into the dichloromethane solution 48 hours of 10ml dimethyl sulfoxide (DMSO) and 10ml; 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 ℃, 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 MK-0457 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 MK-0457 of parcel (there is not chemical coupling reaction; MK-0457 does not have structural changes), MK-0457 ordinary preparation (powder injection) carries out respectively the drug action test of stability test, drug release in vitro test and glaucoma and neuroprotective.

Stability test:

By the nanoparticle medicine group of the MK-0457 of 1 group of sample of preparing of embodiment and sebacic acid-glycol copolymer parcel, MK-0457 ordinary preparation is got with amount (in MK-0457) and is measured respectively absorbance.Then put into 20 degree incubator 3 months for three groups, take out subsequently and measure absorbance, the absorbance front and back of the MK-0457 of the visible embodiment 1-2 group of result and ordinary preparation group are without changing, and the nanoparticle group absorbancy of parcel decline 18%.

Drug release in vitro test:

By 1 group of embodiment, sebacic acid-glycol copolymer directly the nanoparticle medicine group of the MK-0457 of parcel and MK-0457 ordinary preparation component have another name called get equivalent medicine (in MK-0457, every group containing 10mg MK-0457), 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 MK-0457 of parcel and MK-0457 ordinary preparation component have another name called get equivalent medicine (in MK-0457, every group containing 100mg MK-0457), 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

Treatment experiment to rat angle closure glaucoma:

1.1 laboratory animal and grouping

Experiment adopts Thirty male rats, body weight 220g left and right.Before modeling, by animal random packet: i.e. blank group; Model group; Embodiment 1-2 group, polymkeric substance directly wraps up MK-0457 group, and common MK-0457 preparation group is administered once every 5 days intraocular injections, dosage 0.1mg injection/kg; Positive drug group selection pilocarpine eye drop, be administered once every day, one time one.All administrations continue 15 days.

The foundation of 1.2 animal models

Abdominal injection rat 4% Chloral Hydrate (200mg.kg-1) before experiment, then uses 1% lignocaine toponarcosis cornea of rats.During high intraocular pressure, keep every half an hour 0.2 milliliter of abdominal injection 4% Chloral Hydrate make the state of animal continuous narcosis.Adopt pulley-suture system that rat right eye intraocular pressure is raise, left side eyeball contrasts simultaneously.Be respectively an identical heavy counterweight by surgical sutures two ends, hitch a circle in the middle of suture for around after rat canthus limbus of sclera 2 millimeters, the counterweight at two ends is exerted pressure to rat eyes continually and steadily by pulley.It is 20 grams that counterweight is exerted pressure, and continuous 6 hours causes rat angle closure glaucoma model.Raise and measured rathole pressure record to the 15th day.

1.3 experimental result

Experimental result is in table 1

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

With model group comparison ^*p < 0.05 ^*p < 0.01

Provide protection experiment to optic nerve:

2.1 laboratory animal and grouping

Experiment adopts Adult female rats, body weight 200g left and right.By animal random packet: i.e. blank group; Model group; Embodiment 1-2 group, it is thin that polymkeric substance directly wraps up MK-0457, and common MK-0457 preparation group is administered once every 5 days intraocular injections, and dosage 0.1mg injection/kg finishes after 20 days to observe.

The foundation of 2.2 animal models

Carry out abdominal injection rat 6% Chloral Hydrate (200mg.kg-1) anesthesia before the preparation experiment of nerve injury model according to the method for the people such as Chen Xiaorui report, the local cleaning-sterilizing of eye, go in ring and cut off outer canthus portion bulbar conjunctiva, open Tenons capsule, blunt separation Suspensory ligament, tractive eyeball forward, exposes optic nerve, eye scissors 2mm place after ball enters the vertical optic nerve that cuts off in socket of the eye and causes optic nerve injury, avoids damaging optical fundus blood vessel.Guarantee consistence, all optic nerve transections hinders all in identical position, and stitch ball conjunctiva is also received eyeball, painting Antibiotic Eye Ointment.Within postoperative 2 days, occur Marcus-gun pupil, eyeball is without obviously outstanding, and catacleisis is complete, and eyeground is successful model without obvious bleeder.False wound eye only separates and exposes optic nerve, not all right cross-section wound.Administration was won eyeball after 20 days.After disconnected neck is put to death, win whole eyeball (retaining ball optic nerve 2mm), make the capable electron microscopic observation of sample.The results are shown in Figure 3, the effect that the neural action effect of visible embodiment protection is better than polymkeric substance far away and directly wraps up MK-0457 neuroprotective.

Claims (9)

1. in the structure being shown below, contain the segmented copolymer of MK-0457, its structure is as follows:

。

2. the multipolymer of claim 1, wherein PEG refers to polyoxyethylene glycol, and molecular weight is 100-100000, 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 with react and obtain final product containing MK-0457 C;

compd A;

polymer B;

MK-0457 analogue 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 eye, head administration.

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 to prepare the medicine of neuroprotective.

9. the purposes of claim 6, wherein eye disease refers to glaucoma.

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