CN104496843B

CN104496843B - Method for synthesizing ubenimex

Info

Publication number: CN104496843B
Application number: CN201410742120.9A
Authority: CN
Inventors: 高肇林; 王彦厚; 史永强; 张兴柱; 魏正风
Original assignee: SHANDONG YIKANG PHARMACEUTICAL CO Ltd
Current assignee: SHANDONG YIKANG PHARMACEUTICAL CO Ltd
Priority date: 2014-12-08
Filing date: 2014-12-08
Publication date: 2017-02-22
Anticipated expiration: 2034-12-08
Also published as: CN104496843A

Abstract

The invention discloses a method for synthesizing ubenimex. The method comprises the following steps: by taking D-Boc-phenyl alaninal as a raw material, sequentially preparing hydroxynitrile, carrying out a nitrile hydrolysis reaction, performing amino protection, performing chiral resolution on (2RS,3R)-3-tert-butoxyacylamino-2-hydroxy-4-phenylbutyric acid, and synthesizing (S)-phenyl-2-((2S,3R)-3-(tert-butoxyacylamino)-2-hydroxy-4-phenylbutyramide)-4-mevalonate, thereby synthesizing the ubenimex. According to the synthetic method disclosed by the invention, the defect that the resolving agent used in the conventional synthetic method is high in toxicity (such as strychnine) or high in price (such as alpha-phenylethylamine) is overcome, the synthetic method is simple in steps and low in cost, and use of reagents such as strychnine is avoided; and moreover, a dextro-amino compound is taken as the chiral resolution agent, the price is low, the operation is simple, the stirring and recrystallization time is shortened, and the synthetic efficiency is greatly improved.

Description

A kind of synthetic method of ubenimex

Technical field

The invention belongs to the synthesis field of chemicals, it is related to a kind of synthetic method of antineoplastic and in particular to one Plant the synthetic method of ubenimex.

Background technology

Ubenimex (Ubenimex), its medicine name is bestatin；Another name：By Bi Nimei；Trade name：Hundred scholars are glad； Chemical name：N- [(2S, 3R) -3- amino -2- hydroxyl -4- benzene butyryl]-L-Leu.Ubenimex is 1976 by Japanese scholars Mei Ze shore husband separates, from the nutrient solution of the netted streptomycete of olive, the small molecule dipeptide compound obtaining, and can suppress multiple aminopeptidases Activity, such as APN, APB, APP, LAP and MetAP.Additionally, ubenimex can also strengthen host immune function, suppress tumor group Knit Angiogenesiss, thus suppressing invasion and attack and the transfer of tumour.1987, ubenimex was as cancer therapy drug on Japan is formal City.

Clinical studies show, ubenimex is the cytotoxic activity by antineoplastic known to raising, thus realizing it Curative effect in treatment of cancer.Chemotherapy, the auxiliary treatment of radiotherapy and senile immunologic function that this product is used for tumor patient lack The disease such as sunken.Indication includes leukaemia, Huppert's disease, RAEB, and other solid tumor.With The increase of global consumption, obviously can not meet the demand of drug market by fermenting and producing ubenimex.Therefore, pharmacy work Industry research team and scientific research and academic institution take great effort, to find the production practicable route of this medicine.

Ubenimex complex structure, it comprises a peptide bond, and this key is very sensitive to soda acid, and unstable at high temperature Fixed.Additionally, it comprises three asymmetric carbon atoms：C2, C3 and C5.Therefore, 7 stereoisomers in it and other coexist in theory. So properly forming 3 chiral centres, and keep the complete of its chirality so that the synthesis of ubenimex has greatly has Challenge.

1976, the reported first such as Suda ubenimex fully synthetic, its key technology is to improve chipal compounds Optical purity.Hereafter, in more than 30 year, the synthesis of ubenimex is always the focus of chemist and pharmacy corporation.Substantial amounts of The document report synthesis of ubenimex, from traditional Split Method to asymmetric syntheses, then arrives biological catalysis.However, because Asymmetric syntheses relatively costly, biological catalysis at present also in laboratory stage, industrial use now remain biography The Split Method of system.The Split Method of document report mainly has two classes：The first kind, by chromatographing post separation, this method is big due to adsorbance, Greatly reduce total recovery；Equations of The Second Kind, by chiral resolving agent separate, this method be at present industrial production chipal compounds Common method.However, the resolving agent of current document report or toxicity too big (as strychnia) or expensive (α-benzene second Amine).

Chinese patent CN 101891647 A discloses a kind of preparation method of ubenimex, and described preparation method includes Prepare highly purified key intermediate (2S, 3R) -3- ammonia with 1B, L-arginine or L-Histidine as resolution reagent Base -2- hydroxy-4-phenyl butyric acid and be used EDCI/HOAt to keep peptide chain to form the chirality of middle C-5 as condensing agent.The present invention Efficiently solve that existing resolving agent exists can not completely will (2S, 3R) -3- amino -2- hydroxy-4-phenyl butyric acid and (2R, 3S) the problem of the detached problem of -3- acetylaminohydroxyphenylarsonic acid 2- hydroxy-4-phenyl butyric acid and amide condensed middle racemization.The present invention exists Following defect：In the present invention, the synthesis step of ubenimex is loaded down with trivial details, and the reaction time is long, and intermediate product is many, high cost, thus efficiency Low；, as resolution reagent, stirring, recrystallization time are longer afterwards, lead to reaction efficiency for 1B, L-arginine or L-Histidine Low.

Content of the invention

The purpose of the present invention is exactly that this method has in order to solve the above problems, provide a kind of synthetic method of ubenimex Have the advantages that cheap, simple to operate.

To achieve these goals, the present invention adopts the following technical scheme that：

A kind of synthetic method of ubenimex, with D-Boc- phenylpropyl alcohol ammonium aldehyde ((R)-(+) -2- (tertbutyloxycarbonylamino) - 3- hydrocinnamicaldehyde) it is raw material, sequentially pass through the preparation of hydroxyl nitrile, the protection of itrile group hydrolysis and amino, (2RS, 3R) -3- uncle The chiral resolution of fourth oxanamide base -2- hydroxy-4-phenyl butyric acid, (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- Hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters synthesis, the synthesis of ubenimex；

The preparation of described hydroxyl nitrile is specially：D-Boc- phenylpropyl alcohol ammonium aldehyde is dissolved in ethyl acetate, adds water, NaHSO₃, instead Answer 8～15h (preferably 10h), phase of fetching water, add NaCN in aqueous phase, continue reaction 8～15h (preferably 10h), with ethyl acetate Extraction, washing, organic phase is dried, filters, be evaporated (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile, its Middle D-Boc- phenylpropyl alcohol ammonium aldehyde：Ethyl acetate：Water：NaCN：NaHSO₃=1:2～3:8～12:0.3～0.8:0.1～0.4, g: ml：ml：g：g；

The protection of itrile group hydrolysis and amino is specially：By (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxyl -4- benzene Base butyronitrile adds 2～4 times amount (i.e.：Dioxane concentrated hydrochloric acid mixed liquor volume ml be (2RS, 3R) -3- tert-butoxycarbonylamino - The multiple of 2- hydroxy-4-phenyl butyronitrile quality g) dioxane concentrated hydrochloric acid mixed liquor (v/v=1/0.8～2), backflow overnight, It is dissolved in water after being evaporated, is washed with ethyl acetate, 3N NaOH solution adjusts pH to 11～12, under ice bath, drip 4～6 times amount (i.e.：Drip Plus oxolane volume (ml) is 4-6 times of raw material D-Boc- phenylpropyl alcohol ammonium aldehyde weight (g)) containing dimethyl dicarbonate butyl ester (Boc)₂Tetrahydrofuran solution (the w/v=1g of O：3～6ml) after temperature reaction 10～15h (preferably 12h), THF is evaporated off, with oil Ether is washed, and 1N HCl adjusts pH to 2～3, and ethyl acetate extracts, and is dried, and filters, is evaporated, recrystallizes (2RS, 3R) -3- tertiary fourth oxygen acyl Amido -2- hydroxy-4-phenyl butyric acid；

The chiral resolution of (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid is specially：By (2RS, 3R)- 3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid is dissolved in absolute ethyl alcohol, adds (1S, 2S) -2- amino -1- (4- nitrobenzene Base) propane -1,3- glycol, let cool crystallization after 50～70 DEG C (preferably 60 DEG C) stirrings 1.5～2.5h (preferably 2h), filter, will tie Crystalline substance is adjusted after PH to 2～3 with 1N HCl with ethyl acetate extraction after being dissolved in water, is dried, filters, be evaporated, obtain the tertiary fourth of (2S, 3R) -3- Oxanamide base -2- hydroxy-4-phenyl butyric acid, wherein (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid：Anhydrous Ethanol：(1S, 2S) -2- amino -1- (4- nitrobenzophenone) propane -1,3- glycol=1:8～13:1～2, g:ml:g；

(S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters Synthesis be specially：By (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid be dissolved in 4- (dimercapto methylene) - 2- methyl -6- (to dimethylaminostyryl) -4H- pyrans (DCM) or anhydrous methylene chloride, (preferably DCM), add under ice bath 1- hydroxy benzo triazole (HoBt), 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride (EDCI), 20～ 40min (preferably 30min) adds L-Leu benzyl ester tosilate and triethylamine afterwards, and room temperature reaction overnight, is divided with quality Number is 10% citric acid, saturation NaHCO₃, saturated common salt is washed, and is dried, and filters, is evaporated (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters, wherein (S)-phenyl 2- ((2S, 3R) -3- (uncle Fourth oxanamide base) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters：DCM：HoBt：EDCI：L-Leu benzyl ester is to first Benzene sulfonate：Triethylamine=1:10～20:0.2～0.8:0.3～0.9:1～2:0.2～0.8, g:ml:g:g:g:ml；

The synthesis of ubenimex is specially：By (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- Fenbutyramidum base) -4- methylpent acid esters be dissolved in 4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) - 4H- pyrans (DCM), dropping trifluoroacetic acid (TFA), room temperature reaction 1.5～2.5h (preferably 2h), it is evaporated, adds water, with NaHCO₃Molten Liquid adjusts pH to 8.5～9.5 (preferably 9.0), and ethyl acetate extracts, and is dried, and filters, is evaporated to obtain colorless oil, by colorless oil Thing with methyl alcohol as solvent, H₂/ Pd-C environment is (in catalyst Pd-C and H₂In the environment of) under react 4.5～5.5 (preferably 5h), mistake Filter, concentrates, and standing filters, dry ubenimex, wherein (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl Base -4- fenbutyramidum base) -4- methylpent acid esters：DCM：TFA=1:10～20:2～5, g:ml:g.

Room temperature reaction is overnight:Refer to that this day unit terminates the time to working continuation in second day.

Beneficial effects of the present invention：

1st, synthetic method of the present invention solves used resolving agent in existing synthetic method or toxicity too greatly (as Strychnos nux-vomica Sub- alkali) or expensive (α-phenylethylamine) shortcoming, and synthetic method step is simple, low cost, avoids strychnia Use Deng reagent.

2nd, the present invention adopts dextrorotation amino substance as hand-type resolving agent, and cheap, simple to operate, stirs, ties again The brilliant time shortens, and substantially increases combined coefficient.

3rd, the present invention, with D-Boc- phenylpropyl alcohol ammonium aldehyde as initiation material, is synthesized ubenimex, synthetic route merely through 5 steps Modern design, synthesis step is short, greatlys save production time and production cost, and midbody product is few, decreases discarded The generation of thing and discharge, reduce the pollution to environment, more environmentally-friendly saving.

4th, while simplifying step, the synthesis yield of ubenimex can reach more than 87.3% to the present invention, and purity reaches To more than 98.5%.

Brief description

The HPLC-UV detection of the ubenimex that Fig. 1 is prepared for the present invention.

Specific embodiment

With reference to embodiment, the invention will be further described.

Embodiment 1

The preparation of (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile (1)：

36g (0.143mol) D-Boc- phenylpropyl alcohol ammonium aldehyde is dissolved in 100mL ethyl acetate, adds 400mL distilled water, 17.6g (0.171mol) sodium hydrogensulfite, thin-layer chromatography (TLC) monitors the reaction mechanism mechanism of reaction, and reaction after 10h terminates, point water intaking phase, uses 300mL Ethyl acetate is washed 2 times, adds 7.7g (0.157mol) Cymag to aqueous phase, and room temperature reaction overnight, is extracted with 300mL ethyl acetate After 3 times, with 300mL wash 3 times, organic phase with anhydrous sodium sulfate drying 24h, be evaporated (2RS, 3R) -3- tert-butoxycarbonylamino - 2- hydroxy-4-phenyl butyronitrile (1) 27g, yield 68.5%.

The preparation of (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (2)：

25g (0.09mol) (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile (1) is dissolved in 100mL Isosorbide-5-Nitrae-dioxane, instills 100mL concentrated hydrochloric acid under ice bath, back flow reaction overnight, is evaporated, plus 100mL water dissolves, 100mL acetic acid Ethyl ester is washed 3 times, with 3mol L^-1Sodium hydroxide solution controls pH 11.0～12.0, and ice bath is cooled to 0 DEG C, is slowly added to 100mL Containing 23.5g (0.108mol) dimethyl dicarbonate butyl ester (Boc)₂The anhydrous tetrahydro furan (THF) of O, uses 1mol L simultaneously^-1Hydrogen-oxygen Change sodium solution and control pH 8.0～9.0, room temperature reaction overnight, keeps whole process pH 8.0～9.0, concentrates after completion of the reaction Reactant liquor, is evaporated off THF, and 200mL petroleum ether washes 3 times, aqueous phase 1mol L^-1HCl solution adjusts pH 2.0～3.0, uses 200mL Ethyl acetate extracts three times, anhydrous sodium sulfate drying 24h, filters, and concentrates, recrystallized with ethyl acetate-hexane (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (2) 17.2g, yield 65.0%, (2S/2R=2:1).

The preparation of (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (3)：

17.0g (0.058mol) (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (2) is dissolved in 200mL absolute ethyl alcohol, adds 18.4g (0.087mol) (1S, 2S) -2- amino -1- (4- nitrobenzophenone) propane -1,3- glycol (dextrorotation amino substance), lets cool crystallization after 60 DEG C of stirring 2h, filters, crystallization is dissolved in 100mL water, adjust PH 2.0 with 1N HCl solution After～3.0 with 100mL ethyl acetate extract 3 times, be dried, filter, be evaporated, obtain (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxyl - 4-phenylbutyrate (3) 7.65g, yield 45.0%.

1.4 (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent The preparation of acid esters (4)：

6.0g (0.020mol) (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (3) is dissolved in 100mL 4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) -4H- pyrans (DCM), adds 3.0g under ice bath (0.022mol) 1- hydroxy benzo triazole (HoBt), 4.54g (0.022mol) 1- ethyl-(3- dimethylaminopropyl) carbon two Inferior amine salt hydrochlorate (EDCI), adds 8.65g (0.022mol) L-Leu benzyl ester tosilate, 3.0mL after half an hour (0.022mol) triethylamine, room temperature reaction overnight, wash 3 times for 10% citric acid solution, 50mL saturation by 50mL mass concentration NaHCO₃Solution is washed 3 times, and the 50mL saturated common salt aqueous solution is washed 1 time, adds anhydrous magnesium sulfate to be dried overnight, filters, concentrate, places Crystallize (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters (4) 8.0g, yield 80%.

The synthesis of ubenimex

By 8.0g (0.016mol) (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum Base) -4- methylpent acid esters (4) are dissolved in 80mL4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) -4H- Pyrans (DCM), dropping 20mL trifluoroacetic acid (TFA), room temperature reaction 2h, it is evaporated, plus 50mL water, with saturation NaHCO₃Adjust pH extremely 9.0,100mL ethyl acetate extract 3 times, are dried, and filter, are evaporated to obtain colorless oil, colorless oil is dissolved in absolute methanol, Add 0.8g Pd-C, H₂React 5h under environment, filter, concentrate, standing, filters, dry ubenimex 4.4g, yield 90.0%, purity 98.5%.

Embodiment 2

21.6g (0.0858mol) D-Boc- phenylpropyl alcohol ammonium aldehyde is dissolved in 43mL ethyl acetate, adds 173mL distilled water, 6.5g (0.103mol) sodium hydrogensulfite, thin-layer chromatography (TLC) monitors the reaction mechanism mechanism of reaction, and reaction after 10h terminates, point water intaking phase, uses 300mL Ethyl acetate is washed 2 times, adds 2.16g (0.094mol) Cymag to aqueous phase, and room temperature reaction overnight, is extracted with 300mL ethyl acetate After taking 3 times, washed 3 times with 300mL, organic phase, with anhydrous sodium sulfate drying 24h, is evaporated the tertiary fourth oxanamide of (2RS, 3R) -3- Base -2- hydroxy-4-phenyl butyronitrile (1) 15g, yield 63.4%.

10g (0.036mol) (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile (1) is dissolved in 20mL Isosorbide-5-Nitrae-dioxane, instills 16mL concentrated hydrochloric acid under ice bath, back flow reaction overnight, is evaporated, plus 100mL water dissolves, 40mL acetic acid second Ester is washed 3 times, with 3mol L^-1Sodium hydroxide solution controls pH 11.0～12.0, and ice bath is cooled to 0 DEG C, is slowly added to 40mL and contains 13g (0.059mol) dimethyl dicarbonate butyl ester (Boc)₂The anhydrous tetrahydro furan (THF) of O, uses 1mol L simultaneously^-1NaOH Solution controls pH 8.0～9.0, and room temperature reaction overnight, keeps whole process pH 8.0～9.0, concentrates reaction after completion of the reaction Liquid, is evaporated off THF, and 100mL petroleum ether washes 3 times, aqueous phase 1mol L^-1HCl solution adjusts pH 2.0～3.0, with 80mL acetic acid second Ester extracts three times, anhydrous sodium sulfate drying 24h, filters, and concentrates, recrystallizes (2RS, 3R) -3- uncle with ethyl acetate-hexane Fourth oxanamide base -2- hydroxy-4-phenyl butyric acid (2) 6.1g, yield 57.6%, (2S/2R=2:1).

20.0g (0.068mol) (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (2) is dissolved in 160mL absolute ethyl alcohol, adds 20g (0.102mol) (1S, 2S) -2- amino -1- (4- nitrobenzophenone) propane -1, and 3- glycol is (right Rotation amino substance, lets cool crystallization after 60 DEG C of stirring 2h, filters, crystallization is dissolved in 100mL water, with 1N HCl solution adjust PH 2.0～ Extracted 3 times with 100mL ethyl acetate after 3.0, be dried, filter, be evaporated, obtain (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxyl -4- Phenylbutyric acid (3) 7.7g, yield 38.5%.

10g (0.033mol) (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (3) is dissolved in 100mL 4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) -4H- pyrans (DCM), adds 2.0g under ice bath (0.015mol) 1- hydroxy benzo triazole (HoBt), 3g (0.0146mol) 1- ethyl-(3- dimethylaminopropyl) carbon two is sub- Amine hydrochlorate (EDCI), adds 10g (0.029mol) L-Leu benzyl ester tosilate, 2.0mL after half an hour (0.0146mol) triethylamine, room temperature reaction overnight, wash 3 times for 10% citric acid solution, 50mL saturation by 50mL mass concentration NaHCO₃Solution is washed 3 times, and the 50mL saturated common salt aqueous solution is washed 1 time, adds anhydrous magnesium sulfate to be dried overnight, filters, concentrate, places Crystallize (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters (4) 13.0g, yield 78%.

The synthesis of ubenimex：

By 10.0g (0.020mol) (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- benzene butyryl Amido) -4- methylpent acid esters (4) be dissolved in 100mL4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) - 4H- pyrans (DCM), dropping 20mL trifluoroacetic acid (TFA), room temperature reaction 2h, it is evaporated, plus 50mL water, with saturation NaHCO₃Adjust pH Extract 3 times to 9.0,100mL ethyl acetate, be dried, filter, be evaporated to obtain colorless oil, colorless oil is dissolved in no water beetle Alcohol, adds 0.8g Pd-C, H₂React 5h under environment, filter, concentrate, standing, filters, dry ubenimex 5.4g, yield 88.0%, purity 98.8%.

Embodiment 3

11g (0.043mol) D-Boc- phenylpropyl alcohol ammonium aldehyde is dissolved in 33mL ethyl acetate, adds 132mL distilled water, 8.8g (0.052mol) sodium hydrogensulfite, thin-layer chromatography (TLC) monitors the reaction mechanism mechanism of reaction, and reaction after 10h terminates, point water intaking phase, uses 300mL Ethyl acetate is washed 2 times, adds 4.4g (0.048mol) Cymag to aqueous phase, and room temperature reaction overnight, is extracted with 100mL ethyl acetate After 3 times, with 100mL wash 3 times, organic phase with anhydrous sodium sulfate drying 24h, be evaporated (2RS, 3R) -3- tert-butoxycarbonylamino - 2- hydroxy-4-phenyl butyronitrile (1) 8g, yield 66%.

17g (0.06mol) (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile (1) is dissolved in 68mL 1, 4- dioxane, instills 34mL concentrated hydrochloric acid under ice bath, back flow reaction overnight, is evaporated, plus 100mL water dissolves, 100mL ethyl acetate Wash 3 times, with 3mol L^-1Sodium hydroxide solution controls pH 11.0～12.0, and ice bath is cooled to 0 DEG C, is slowly added to 105mL and contains 17g (0.075mol) dimethyl dicarbonate butyl ester (Boc)₂The anhydrous tetrahydro furan (THF) of O, uses 1mol L simultaneously^-1NaOH Solution controls pH 8.0～9.0, and room temperature reaction overnight, keeps whole process pH 8.0～9.0, concentrates reaction after completion of the reaction Liquid, is evaporated off THF, and 100mL petroleum ether washes 3 times, aqueous phase 1mol L^-1HCl solution adjusts pH 2.0～3.0, uses 100mL acetic acid Ethyl ester extracts three times, anhydrous sodium sulfate drying 24h, filters, and concentrates, recrystallizes (2RS, 3R) -3- with ethyl acetate-hexane Tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (2) 13g, yield 72.2%, (2S/2R=2:1).

30g (0.102mol) (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (2) is dissolved in 400mL Absolute ethyl alcohol, adds 60g (0.142mol) (1S, 2S) -2- amino -1- (4- nitrobenzophenone) propane -1,3- glycol (dextrorotation amino Thing, 60 DEG C stirring 2h after let cool crystallization, filter, crystallization is dissolved in 100mL water, with 1N HCl solution adjust PH 2.0～3.0 after with 100mL ethyl acetate extracts 3 times, is dried, and filters, is evaporated, obtains (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl fourth Sour (3) 13g, yield 43.3%.

15g (0.050mol) (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid (3) is dissolved in 300mL 4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) -4H- pyrans (DCM), adds 12g under ice bath (0.088mol) 1- hydroxy benzo triazole (HoBt), 13.5g (0.022mol) 1- ethyl-(3- dimethylaminopropyl) carbon two Inferior amine salt hydrochlorate (EDCI), adds 30g (0.034mol) L-Leu benzyl ester tosilate, 12mL after half an hour (0.088mol) triethylamine, room temperature reaction overnight, wash 3 times for 10% citric acid solution, 50mL saturation by 50mL mass concentration NaHCO₃Solution is washed 3 times, and the 50mL saturated common salt aqueous solution is washed 1 time, adds anhydrous magnesium sulfate to be dried overnight, filters, concentrate, places Crystallize (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters (4) 20.0g, yield 80%.

The synthesis of ubenimex：

By 15.0g (0.030mol) (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- benzene butyryl Amido) -4- methylpent acid esters (4) be dissolved in 150mL4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) - 4H- pyrans (DCM), dropping 75mL trifluoroacetic acid (TFA), room temperature reaction 2h, it is evaporated, plus 50mL water, with saturation NaHCO₃Adjust pH Extract 3 times to 9.0,100mL ethyl acetate, be dried, filter, be evaporated to obtain colorless oil, colorless oil is dissolved in no water beetle Alcohol, adds 0.8g Pd-C, H₂React 5h under environment, filter, concentrate, standing, filters, dry ubenimex 8g, yield 87.3%, purity 98.8%.

Detection test

Chromatographic condition and system suitability

It is filler (250mm × 4.6mm) with octadecylsilane chemically bonded silica；With acetonitrile:0.6%NaH₂PO₄(17： 83) it is mobile phase A；With acetonitrile:0.6%NaH₂PO₄(2：1) it is Mobile phase B；Flow velocity is 1.0ml per minute；Gradient elution；Detection Wavelength is 210nm.Theoretical cam curve is calculated with ubenimex peak and should be not less than 5000, as seen from Figure 1, the inventive method institute The final material of preparation is ubenimex.

Time	Mobile phase A (%)	Mobile phase B (%)
			0	100	0
20	100	0
			50	0	100
70	0	100
			70.01	100	0
85	100	0

Although above-mentioned be described to the specific embodiment of the present invention in conjunction with the embodiments, not the present invention is protected The restriction of scope, one of ordinary skill in the art should be understood that on the basis of technical scheme, those skilled in the art Do not need to pay the various modifications that creative work can make or deformation still within protection scope of the present invention.

Claims

1. a kind of synthetic method of ubenimex is it is characterised in that comprise the following steps：

(1) preparation of hydroxyl nitrile：D-Boc- phenylpropyl alcohol ammonium aldehyde is dissolved in ethyl acetate, adds water, NaHSO₃, react 8～15h, take Aqueous phase, adds NaCN in aqueous phase, continues reaction 8～15h, with ethyl acetate extraction, washes, organic phase is dried, filter, be evaporated Obtain (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile, described D-Boc- phenylpropyl alcohol ammonium aldehyde：Ethyl acetate：Water： NaCN：NaHSO₃=1:2～3:8～12:0.3～0.8:0.1～0.4, g:ml：ml：g：g；

(2) protection of itrile group hydrolysis and amino：(2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile is added 2 The dioxane of～4 times amount and concentrated hydrochloric acid mixed liquor, described dioxane is 1 with the volume ratio of concentrated hydrochloric acid：0.8～2, backflow Overnight, be dissolved in water after being evaporated, washed with ethyl acetate, adjust pH to 11～12, under ice bath dropping 4～6 times amount containing carbonic acid two Tertiary butyl ester (Boc)₂The tetrahydrofuran solution of O, described dimethyl dicarbonate butyl ester (Boc)₂O：Oxolane=1：3～6, g：Ml, After room temperature reaction 10～15h, THF is evaporated off, is washed with petroleum ether, adjust pH to 2～3, ethyl acetate extracts, be dried, filter, be evaporated, Recrystallize (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid；Described 2-4 times amount is：Dioxane with dense Volume ml of hydrochloric acid mixed solution is 2-4 times of (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyronitrile quality g；Institute Stating 4-6 times amount is：Dropping oxolane volume is 4-6 times of raw material D-Boc- phenylpropyl alcohol ammonium aldehyde weight g for raw material ml；

(3) chiral resolution of (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid：By tertiary for (2RS, 3R) -3- fourth Oxanamide base -2- hydroxy-4-phenyl butyric acid is dissolved in absolute ethyl alcohol, add (1S, 2S) -2- amino -1- (4- nitrobenzophenone) propane - 1,3- glycol, 50～70 DEG C stirring 1.5～2.5h after let cool crystallization, filter, by crystallization be dissolved in after water adjust pH to 2～3 after with Ethyl acetate extracts, and is dried, and filters, is evaporated, obtains (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid, described (2RS, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid：Absolute ethyl alcohol：(1S, 2S) -2- amino -1- (4- nitrobenzene Base) propane -1,3- glycol=1:8～13:1～2, g:ml:g；

(4) (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters Synthesis：By (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid be dissolved in 4- (dimercapto methylene) -2- methyl - 6- (to dimethylaminostyryl) -4H- pyrans or anhydrous methylene chloride, add 1- hydroxy benzo triazole, 1- second under ice bath Base-(3- dimethylaminopropyl) carbodiimide hydrochloride, adds L-Leu benzyl ester tosilate after 20～40min And triethylamine, room temperature reaction overnight, with mass fraction for 10% citric acid, saturation NaHCO₃, saturated common salt is washed, and is dried, mistake Filter, is evaporated (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- fenbutyramidum base) -4- methylvaleric acid Ester, described (2S, 3R) -3- tert-butoxycarbonylamino -2- hydroxy-4-phenyl butyric acid：4- (dimercapto methylene) -2- methyl -6- is (right Dimethylaminostyryl) -4H- pyrans：1- hydroxy benzo triazole：1- ethyl-(3- dimethylaminopropyl) carbodiimide Hydrochloride：L-Leu benzyl ester tosilate：Triethylamine=1:10～20:0.2～0.8:0.3～0.9:1～2:0.2～ 0.8, g:ml:g:g:g:ml；

(5) synthesis of ubenimex：By (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- hydroxyl -4- benzene butyryl Amido) -4- methylpent acid esters are dissolved in 4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) -4H- pyrans, Dropping trifluoroacetic acid room temperature reaction 1.5～2.5h, is evaporated, adds water, and adjusts pH to 8.5～9.5, and ethyl acetate extracts, and is dried, mistake Filter, is evaporated to obtain colorless oil, by colorless oil with methyl alcohol as solvent, H₂4.5～5.5h, mistake is reacted under/Pd-C environment Filter, concentrates, and standing filters, dry ubenimex, described (S)-phenyl 2- ((2S, 3R) -3- (tert-butoxycarbonylamino) -2- Hydroxyl -4- fenbutyramidum base) -4- methylpent acid esters：4- (dimercapto methylene) -2- methyl -6- is (to dimethylamino styrene Base) -4H- pyrans：Trifluoroacetic acid=1:10～20:2～5, g:ml:g.

2. synthetic method as claimed in claim 1 is it is characterised in that D-Boc- phenylpropyl alcohol ammonium aldehyde is dissolved in second by described step (1) Acetoacetic ester, adds water, NaHSO₃, 10h, phase of fetching water, add NaCN in aqueous phase, continue reaction 10h.

3. synthetic method as claimed in claim 1 is it is characterised in that use 3N NaOH solution to adjust pH extremely in described step (2) 11～12.

4. synthetic method as claimed in claim 1 is it is characterised in that after room temperature reaction 12h in described step (2), be evaporated off THF, is washed with petroleum ether, and 1N HCl adjusts pH to 2～3.

5. synthetic method as claimed in claim 1 is it is characterised in that let cool crystallization after 60 DEG C of stirring 2h in described step (3).

6. synthetic method as claimed in claim 1 uses 1N it is characterised in that crystallization is dissolved in after water in described step (3) HCl adjusts PH to 2～3.

7. the synthetic method as described in claim 1 is it is characterised in that by tertiary for (2S, 3R) -3- fourth oxygen in described step (4) Amide groups -2- hydroxy-4-phenyl butyric acid be dissolved in 4- (dimercapto methylene) -2- methyl -6- (to dimethylaminostyryl) - 4H- pyrans, addition 1- hydroxy benzo triazole under ice bath, 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride, L-Leu benzyl ester tosilate and triethylamine is added after 30min.

8. synthetic method as claimed in claim 1 is it is characterised in that drip trifluoroacetic acid room temperature reaction in described step (5) 2h.

9. synthetic method as claimed in claim 1 is it is characterised in that with NaHCO in described step (5)₃Solution adjusts pH extremely 9.0.

10. synthetic method as claimed in claim 1 is it is characterised in that H in described step (5)₂5h is reacted under/Pd-C environment.