CN106146536B

CN106146536B - A kind of preparation method of everolimus

Info

Publication number: CN106146536B
Application number: CN201510205424.6A
Authority: CN
Inventors: 张贵民; 白文钦; 黄传青; 徐勇; 王增日
Original assignee: Shandong New Time Pharmaceutical Co Ltd
Current assignee: Shandong New Time Pharmaceutical Co Ltd
Priority date: 2015-04-25
Filing date: 2015-04-25
Publication date: 2019-07-26
Anticipated expiration: 2035-04-25
Also published as: CN106146536A

Abstract

The invention discloses a kind of preparation methods of everolimus；For this method the following steps are included: rapamycin is reacted with trim,ethylchlorosilane by after the double protections of 31,40 hydroxyls, acidic hydrolysis obtains the intermediate 1 of 31 hydroxyl protections；Intermediate 1 and ethylene oxide carry out condensation reaction, obtain intermediate 2,2 acidic hydrolysis of intermediate obtains everolimus crude product, and crude product is isolated and purified with preparation liquid phase, obtains everolimus.

Description

A kind of preparation method of everolimus

Technical field

The invention belongs to field of medicaments, and in particular to a kind of preparation method of everolimus.

Background technique

Everolimus (everolimus；Trade name: Afinitor) chemical name [40-O- (2- ethoxy)-rapamycin], It is macrolides rapamycin derivative drug of new generation, structural formula is as follows:

Everolimus is the tool researched and developed by Novartis Co., Ltd, Switzerland centainly water-soluble rapamycin derivative, can take orally to Medicine is clinically mainly used to prevent the rejection after kidney transplant and heart transplant operation.Its mechanism of action mainly includes being immunized Inhibiting effect, antitumor action, antivirus action, vascular protection effect, often combining with other immunosuppressor such as cyclosporines makes To reduce toxicity.Additionally for treating advanced renal cell cancer.Also it is carrying out to neuroendocrine tumor, lymthoma, other cancers The research of disease and tuberous sclerosis can be used as single formulation or share with existing cancer treatment method.

WO9409010 reports the method for preparing everolimus as Material synthesis using rapamycin earliest；Document J.labelled Compd Radiopharm.2000,43,113-120 also report the everolimus similar with the patent document Synthesis preparation method.The above-mentioned method for preparing everolimus as Material synthesis using rapamycin includes two-step reaction: first two It is reaction with rapamycin and 2- (tert-butyl diphenyl silicon substrate) oxygen ethyl triflate under wopropyl ethyl amine alkaline condition Object, reaction prepares intermediate 1 in certain organic solvent；Intermediate 1 sloughs protecting group getting the product everolimus, above-mentioned Preparation process low yield, at high cost, the yield for preparing intermediate is only 6%；The product yield of second step reaction is also only 21%.

CN102127092A discloses a kind of preparation method of everolimus, which has continued to use above-mentioned with thunder substantially Pa mycin is the process route that Material synthesis prepares everolimus, only implements recycling to unreacted rapamycin raw material.

Since rapamycin reaction site is more, property is unstable, above-mentioned to prepare by Material synthesis of rapamycin according to dimension In the preparation method that do not take charge of, reaction needs to control to be carried out under the conditions of relatively mild, and typical temperature is controlled at 50-60 DEG C, temperature Spend low, reaction cannot be carried out sufficiently；Temperature is excessively high, and rapamycin and its intermediate are easy degradation or to generate other unknown miscellaneous Matter, reaction temperature is too low or the excessively high total recovery that will lead to product is lower.Existing literature report scheme reaction temperature is effectively controlled At 50-60 DEG C, can still have more than 50% rapamycin cannot effectively be able to participate in reaction system.

Summary of the invention

For deficiency existing for everolimus synthetic method in the prior art, the present invention provides a kind of everolimus preparation side Method.Under alkaline condition by rapamycin, and after trim,ethylchlorosilane reaction double hydroxyl protection, in acid condition with epoxy Ethane carries out condensation reaction, then hydrolyzed under acidic conditions obtains everolimus, and this method is easy to operate, rapamycin is easy to react, Reaction yield is high, in order to achieve the above-mentioned object of the invention.

The present invention provides following technical scheme, the program includes tetra- steps of A, B, C, D:

Step A: rapamycin under alkaline condition, is reacted with trim,ethylchlorosilane by after 31,40 hydroxyl protections, acid Hydrolysis, obtains the intermediate 1 of 31 hydroxyl protections；

Step B: in acid condition, intermediate 1 and ethylene oxide carry out condensation reaction, obtain intermediate 2,

Step C: intermediate 2 dissolves acidic hydrolysis in a solvent, obtains everolimus crude product,

D step: the crude product is isolated and purified with preparation liquid phase, obtains everolimus.

Specific reaction route is as follows:

Step A:

Step B

Step C:

D step:

Step A of the present invention: rapamycin under alkaline condition, is reacted with trim,ethylchlorosilane by 31,40 hydroxyl protections Afterwards, acidic hydrolysis, obtain the intermediate 1 of 31 hydroxyl protections: the alkaline condition of this step is provided by organic base, organic base It include: triethylamine, pyridine, diisopropyl ethyl ammonia, 2,6- lutidines, imidazoles etc.；As long as reaction dissolvent can dissolve reaction Object, and do not have influential solvent to be ok reaction, such as ethyl acetate, methylene chloride, chloroform tetrahydrofuran.Reaction temperature with The variation of reagent or solvent and change, usually -40 DEG C~30 DEG C, the reaction time is also the variation with reagent or solvent And change, usually 1-8 hours；TLC detection, no rapamycin spot, then double protection reactions are completed；Then acidic hydrolysis obtains To the intermediate 1 of 31 hydroxyl protections；Acid is mainly various inorganic acids, and sulfuric acid, hydrochloric acid, hydrobromic acid etc., reaction temperature is with reagent Or solvent variation and change, usually -20 DEG C~30 DEG C, the reaction time is also the variation with reagent or solvent and becomes Change, usually 3-28 hours；TLC detection, unparalleled protection rapamycin spot, then hydrolysis is completed；After the reaction was completed, with normal The post processing mode processing of rule can be obtained by intermediate 1.

Step B of the present invention: intermediate 1 in a solvent, in acid condition, carries out condensation reaction with ethylene oxide, obtains Intermediate 2；Solvent is the one or several kinds such as anhydrous ether, tetrahydrofuran, chloroform, methylene chloride, ethyl acetate, toluene；Make Acid for catalyst is mainly micro or a small amount of various inorganic acids, such as sulfuric acid, hydrochloric acid, hydrobromic acid etc.；Reaction temperature is with examination Agent or the variation of solvent and change, usually -20 DEG C~40 DEG C, the reaction time be also the variation with reagent or solvent and Variation, usually 2-12 hours；TLC detection, no 1 spot of intermediate, then condensation reaction is completed；After the reaction was completed, with routine Post processing mode processing can be obtained by intermediate 2.

Step C of the present invention: intermediate 2 dissolves acidic hydrolysis in a solvent, obtains everolimus crude product: as long as reaction dissolvent Reactant can be dissolved, and do not have influential solvent to be ok reaction, as ethyl acetate, methylene chloride, chloroform, tetrahydrofuran, Methanol etc.；Acid mainly various inorganic acids, sulfuric acid, hydrochloric acid, hydrobromic acid etc., reaction temperature become with the variation of reagent or solvent Change, usually -10 DEG C~20 DEG C, the reaction time is also the variation with reagent or solvent and changes, usually 3-10 hours； TLC detection, no 2 spot of intermediate, then hydrolysis is completed；After the reaction was completed, it is handled with conventional post processing mode, then mistake Silicagel column processing can be obtained by everolimus crude product.

D step of the present invention: crude product is isolated and purified with preparation liquid phase, obtains everolimus；Crude product acetonitrile and purified water Mixed liquor dissolution, with 500ml/min flow velocity sample introduction, collects the part of the main peak containing everolimus after the filtration of 0.45 micron membrane filter It is handled, obtains everolimus.

Specific embodiment

Beneficial effects of the present invention are now further described by following embodiment, it is thus understood that these embodiments are only used for The purpose of illustration, does not limit the scope of the invention, at the same those of ordinary skill in the art done according to the present invention it is apparent Change and modification be also contained within the scope of the invention.

Embodiment 1: the preparation of everolimus intermediate 1:

By 9g flower bud pa mycin, 4.5g2,6- lutidines, 100ml tetrahydrofuran is added in the there-necked flask of 250ml, ice Brine bath cools down, stirring and dissolving, starts (the CH that 7ml is added dropwise after 0.5h₃)₃SiCl, insulation reaction, after reaction 3 hours, TLC detection Reaction process, no rapamycin spot, then double protection reactions are completed, and 5mlHCl is then added dropwise, after reaction 5 hours, TLC detection, Unparalleled protection rapamycin spot, then hydrolysis is completed.Reaction solution 200ml ethyl acetate is extracted, ethyl acetate layer is used Water 50ml, NaHCO₃Saturated solution 50ml washing, then washed with 50ml salt, aqueous layer with ethyl acetate extracts 2 times, merges organic Layer, anhydrous Na₂SO₄It is dry；It removes ethyl acetate under reduced pressure, obtains single protection 9.6g.

Embodiment 2: the preparation of everolimus intermediate 2

At 0~10 DEG C, 500ml toluene is added in 1L three neck round bottom flask, 4.4g ethylene oxide, cooling is then added Afterwards, 9.6g rapamycin intermediate 1 is added in reaction solution, while a small amount of hydrochloric acid is added, the reaction was continued 6 hours, TLC monitoring To fully reacting, with water 250ml, NaHCO₃Saturated solution 250ml washing, then washed with 250ml salt, water layer extracts 2 with toluene It is secondary, merge organic layer, anhydrous Na₂SO₄It is dry；It removes toluene under reduced pressure, obtains 2 sterling 6.8g of everolimus intermediate.

Embodiment 3: the preparation of everolimus crude product

6.8g coupled product is dissolved in a 1L single port bottle with 500ml methanol, ice-water bath cooling is added dropwise after half an hour 10ml HCl after being added dropwise to complete, reacts 4 hours, TLC tracing detection, after completion of the reaction, then evaporated under reduced pressure uses 500ml second Acetoacetic ester dissolves residue, is washed with saturated brine, and ethyl acetate extracts water layer 3 times, merges organic layer, and anhydrous sodium sulfate is dry Dry, evaporated under reduced pressure obtains yellow oil, and column separation obtains crude product 5.6g.

Embodiment 4: the preparation of everolimus

By everolimus crude product 5.6g, is dissolved with 200ml acetonitrile and 120ml purified water, sample solution is configured to, through 0.45 After micron membrane filter filtration, to sample introduction；Start sampling pump and gradient elution journey is then turned on 500ml/min flow velocity sample introduction 1.4min Sequence starts to elute, until adjusting switching valve when target peak occurs, collects target peak, sampling is detected by HPLC, target peak other parts It is collected to rejected product container including isomers peak, is then switched to waste collection bucket；It repeats the above process, until this batch Sample is finished completely；Collection liquid is distilled with Rotary Evaporators low-temperature reduced-pressure, obtains everolimus pulverulent solids 3.8g.

Everolimus sample structure confirmation:

The element percentage composition of everolimus sample

The IR of everolimus sample composes each absorption peak ownership

Everolimus sample is in DMSO-d6¹H-NMR data

Claims

1. a kind of preparation method of everolimus, this method comprises the following steps:

Step A: rapamycin under alkaline condition, reacts after carrying out the double protections of hydroxyl, containing acid item with trim,ethylchlorosilane Part hydrolysis, obtains the intermediate 1 of hydroxyl protection

,

Step B: in acid condition, intermediate 1 and ethylene oxide carry out condensation reaction, obtain intermediate 2

,

Step C: intermediate 2 dissolves acidic hydrolysis in a solvent, obtains everolimus crude product

,

Step D: preparation liquid phase isolates and purifies after the dissolution of everolimus crude product solvent, obtains everolimus product

,

Wherein, the alkali under alkaline condition described in step A is selected from triethylamine, pyridine, diisopropyl ethyl ammonia, 2,6- dimethyl One of pyridine and imidazoles are a variety of；Acid under the acid condition is one of sulfuric acid, hydrochloric acid or hydrobromic acid；Step Solvent used in condensation reaction is one of anhydrous ether, tetrahydrofuran, chloroform, methylene chloride, ethyl acetate and toluene in B Or it is several；Acid under the acid condition is one of sulfuric acid, hydrochloric acid or hydrobromic acid.

2. the method as described in claim 1, it is characterised in that the bis- protection reaction dissolvents of step A be ethyl acetate, methylene chloride, One of chloroform and tetrahydrofuran are a variety of.

3. the method as described in claim 1, it is characterised in that the bis- protection reaction temperatures of step A are -40 DEG C -30 DEG C, when reaction Between be 1-8 hours.

4. the method as described in claim 1, it is characterised in that step B reaction temperature is -20 DEG C -40 DEG C, reaction time 2- 12 hours.

5. the method as described in claim 1, it is characterised in that step C reaction dissolvent be ethyl acetate, methylene chloride, chloroform, One of tetrahydrofuran and methanol are a variety of；The acid of step C hydrolysis is one of sulfuric acid, hydrochloric acid or hydrobromic acid.

6. the method as described in claim 1, it is characterised in that step C reaction temperature is -10 DEG C -20 DEG C, reaction time 3- 10 hours；Step D crude product acetonitrile and the dissolution of the mixed liquor of purified water.