WO2021077242A1

WO2021077242A1 - Caspofungin synthesis method and intermediate thereof

Info

Publication number: WO2021077242A1
Application number: PCT/CN2019/112067
Authority: WO
Inventors: 陈迎会; 朱兵峰
Original assignee: 鲁南贝特制药有限公司
Priority date: 2019-10-20
Filing date: 2019-10-20
Publication date: 2021-04-29
Also published as: CN113286807B; CN113286807A

Abstract

A caspofungin synthesis method and an intermediate compound thereof, the synthesis method using pneumocandin B0 as the starting material and acetone fork protection to obtain a caspofungin intermediate III compound, reacting same with ethylenediamine to obtain caspofungin intermediate IV, and then further reducing same to obtain caspofungin.

Description

A kind of synthetic method of caspofungin and its intermediate

Technical field

The invention belongs to the technical field of medicine, and specifically relates to a method for synthesizing caspofungin and its intermediates.

Background technique

Caspofungin is a echinocandin antifungal agent with a new mechanism of action developed by Merck, and it is also the first echinocandin drug to be marketed (February 2001). Its chemical name is: 1-[(4R,5S)-5-[(2-aminoacetic acid)amino]-N ₂ -(10,12-dimethyl-1-carbonyltetradecyl)4-hydroxy- L-ornithine]-5-[(3R)-3-hydroxy-L-ornithine] echinocandin B ₀ . The molecular formula is C ₅₂ H ₈₈ N ₁₀ O ₁₅ , and the structural formula is as follows:

.

Caspofungin is an echinocandin antifungal drug approved by the FDA, and has always attracted much attention. There have been many reports on its synthesis and related literature. Its synthesis is based on the biological fermentation product neumocandine B ₀ as the starting material. . The synthetic route can be divided into two categories according to the reduction of amide: one is reduction by hydrogen after dehydration of amide to cyano group; the other is direct reduction with borane. There have been many reports on the synthesis of caspofungin in the prior art.

William R. Leonard (William R. Leonard, Jr., the Journal of Organic Chemistry, 2007, 7, vol72, 2335-2343) uses neumoconidine B ₀ and thiophenol to synthesize caspofungin, thiophenol As a stereoselective agent, it can carry out selective mono-substitution, but the highly toxic, malodorous, and pungent odor of thiophenol is not suitable for industrial production, especially mass production of drugs.

CN101648994A reported the use of mercapto aromatic compound R-SH as a stereoselective agent to synthesize caspofungin, where R is phenyl, 4-methoxyphenyl, methylimidazolyl, benzimidazolyl and the like. CN102367267A describes the use of hydroxyl, benzyloxy, phenoxy, substituted phenoxy, and substituted benzyloxy substituted aromatic ring mercapto compounds R'-SH as a strong leaving group to synthesize caspofungin, preferably p-hydroxy Thiophenol, but it is difficult to avoid the shortcomings of its highly toxic and malodorous. CN102219833A reported the use of stereostructure selector 2-mercaptobenzothiazole or 1-phenyl-5-mercapto-tetrazolium to synthesize caspofungin, but the actual yield was not high.

CN102112487A reports the use of a nitrogen-containing nucleophile RX-NH to replace a thiol or thiophenol compound as a strong leaving group, and X is an auxiliary group selected from the group consisting of i) including at least one N-atom as a subgroup Five- and six-membered heterocyclic aromatic rings and derivatives thereof that are part of an amine group, wherein the N-atom constitutes the link to the cyclic hexapeptide; and ii) tetrazole and its derivatives, wherein one nitrogen atom constitutes the ring Connection of hexapeptides. Although the use of foul-smelling and highly toxic thiols or thiophenol compounds is avoided, pyridine and other highly toxic and pungent odor solvents are introduced.

EP620232 and WO94/21677 use the starting material Numocantine B _{0 to} react with alkyl mercaptan or aryl mercaptan to obtain the sulfone intermediate after oxidation, and then react with amine compound in anhydrous aprotic solvent, and then pass chromatography The method is separated.

Previously reported or published methods for synthesizing caspofungin using thiol or thiophenol compounds as stereoselective agents or strong leaving groups, most of which are neumocandine B ₀ or caspofungin intermediates dissolved in The thiol or thiophenol compound in the acid is mixed. The selected acid is mainly strong acids such as methanesulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid, trifluoromethanesulfonic acid or trifluoroacetic acid, and the mixing temperature is demanding. Generally in -50~-15ºC.

For selective reduction of amide bond in Numocantine B ₀ , CN96191834.9 and CN201010144485.3 use borane complex (borane and dimethyl sulfide, diphenyl sulfide and dibenzyl sulfide), boron chloride Caspofungin is obtained by reduction of alkane complex, etc. CN97195496.8 uses borane complex (borane complexed with dimethyl sulfide, diphenyl sulfide, THF, etc.) or metal boride (borane chromium chloride or borane Titanium chloride) is reduced to obtain caspofungin. CN201010538951.6 uses borane and chloroborane for reduction.

The published method for synthesizing caspofungin is not suitable for large-scale industrial production in terms of yield, stability, purity and three wastes. Most methods require the use of highly toxic, foul-smelling, and pungent thiol or thiophenol compounds, which seriously damage the health of operators and seriously pollute the environment. In some of the existing synthesis methods, the introduction of cyano groups inevitably leads to the formation of isomers, the yield and stereoselectivity are not high, and the use of expensive metals as catalysts increases the industrial cost; most synthesis methods use borane and When analogues are reduced, borane and analogues are volatile and toxic; multiple use of preparative chromatography increases industrialization costs and produces a large amount of waste. In view of this, there is an urgent need to further study a method for synthesizing caspofungin suitable for industrial production.

technical problem

Aiming at the deficiencies in the prior art, the present invention provides a method for synthesizing caspofungin and its intermediates. It can effectively solve the problems of three wastes, low yield and low purity.

Technical solutions

The specific technical scheme of the present invention is as follows:

A caspofungin intermediate III, its structural formula is as follows:

The present invention also provides a method for preparing caspofungin intermediate III. Taking the compound of formula II numocandine B0 as a raw material, it is heated and refluxed with acetone under acid conditions to generate caspofungin intermediate III, the reaction formula is as follows:

Preferably, the mass ratio of the added volume of acetone to numocantine B0 is 10-30:1, mL/g, more preferably 20:1, mL/g.

Preferably, the acid is an inorganic acid or an organic acid, preferably p-toluenesulfonic acid, benzenesulfonic acid, 2,4,5-trimethylbenzenesulfonic acid, acetic acid, concentrated sulfuric acid or concentrated nitric acid, more preferably p-toluenesulfonic acid, benzenesulfonic acid, 2,4,5-trimethylbenzenesulfonic acid, concentrated sulfuric acid or concentrated nitric acid. Toluenesulfonic acid or 2,4,5-trimethylbenzenesulfonic acid.

Preferably, the molar ratio of the acid to numocantine B0 is 0.03 to 0.07:1, preferably 0.05:1.

The following content further details the preparation method of caspofungin intermediate III, including the following steps: adding acetone, molecular sieve, and acid to the reaction system, stirring and dissolving, adding neumocandine B0, heating and refluxing to the end, adding water and acetic acid After the ethyl ester is fully stirred and dissolved, it is allowed to stand for liquid separation. The organic layer is collected, the aqueous phase is extracted with ethyl acetate, the organic phases are combined, washed with water to neutral, dried with magnesium sulfate, filtered, and concentrated. The concentrated solution is crystallized with n-hexane to obtain carbopol Fungin Intermediate III.

Preferably, the molecular sieve is a 3A, 4A or 5A molecular sieve, and the mass ratio of the molecular sieve to Numocantine B0 is 0.1 to 0.3:1.

Further preferably, the molecular sieve is a 4A type molecular sieve, and the mass ratio of the molecular sieve to numocantine B0 is 0.2:1.

Preferably, after the reaction is completed, water and ethyl acetate are added for post-treatment, and the volume-to-mass ratio of water to neumoconidine B0 is 2-10:1, mL/g, preferably 5:1, mL/g; water and ethyl acetate The volume ratio of ester is 1:2-8, preferably 1:4.

The third aspect of the present invention provides a method for preparing caspofungin using caspofungin intermediate III.

A method for preparing caspofungin. Caspofungin intermediate III is reacted with ethylenediamine in the presence of a weak acid resin to form intermediate IV, and intermediate IV is reduced by sodium borohydride and organic acid to obtain caspofungin Fungin, the reaction formula is as follows:

Preferably, the reaction solvent in step b is one or more of methanol-water, ethanol-water, isopropanol-water, and acetonitrile-water, more preferably a methanol-water mixed solvent.

Preferably, the volume ratio of methanol, ethanol, isopropanol or acetonitrile to water is 7-12:1, more preferably 9:1.

Preferably, the weak acid resin is a resin containing a carboxylic acid group, a phosphoric acid group or a phenol group. More preferably, the weak acid resin is D110, D113, D151 or D152, and more preferably D113 resin.

Preferably, the mass ratio of the caspofungin intermediate III to the weakly acidic resin is 1:1.5-6, preferably 1:3.

Preferably, the volume mass ratio of ethylenediamine to caspofungin intermediate III in step b is 5.0-7.5:1, mL/g, and more preferably 6.25:1, mL/g.

Preferably, the reducing agent in step c is sodium borohydride and an organic acid, and the organic acid is selected from one of formic acid, acetic acid, propionic acid, butyric acid, valeric acid, benzoic acid or o-hydroxybenzoic acid, further preferably It is acetic acid.

Preferably, the molar ratio of sodium borohydride to organic acid in step c is 1:1 to 2, preferably 1:1.

Preferably, the molar ratio of caspofungin intermediate IV to sodium borohydride in step c is 1:1 to 2, more preferably 1:1.5.

Preferably, the reaction solvent in step c is one or more of 1,4-dioxane, dimethylsulfoxide, and dimethylimide, preferably 1,4-dioxane.

The following content further details the synthesis method of caspofungin, including the following steps:

(B) Under the protection of nitrogen, dissolve caspofungin intermediate III in a reaction solvent, add weak acid resin, and then add ethylenediamine, and react to completion at room temperature. After filtration, the filtrate was concentrated under reduced pressure, methanol and ethyl acetate were added, the solid was separated by filtration, and dried under vacuum to obtain Caspofungin Intermediate IV;

(C) Add sodium borohydride, caspofungin intermediate IV and reaction solvent to the reaction system, stir under the protection of nitrogen at room temperature, add organic acid and reaction solvent to another reaction flask, mix and stir evenly, add dropwise to the above reaction In the system, the reaction was stirred at room temperature. After the reaction was completed, the reaction was quenched by acetic acid, the organic solvent was removed by distillation under reduced pressure, and the liquid phase was prepared and purified to obtain caspofungin.

It should be noted that the liquid phase preparation and purification process in step c can be carried out according to the prior art or conventional technical methods known in the art, for example, the preparative chromatography in CN106478781A or CN107778360A can be used for purification.

Beneficial effect

(1) Avoid using mercaptan or thiophenol compounds that are highly toxic, malodorous, and pungent, which seriously damage the health of operators and seriously pollute the environment.

(2) Avoid using volatile and poisonous borane as a reducing agent, and use sodium borohydride and organic acid to form acyloxyborohydride for selective reduction to obtain the target product. The reaction conditions are mild and easy to scale up. .

(3) The synthetic caspofungin of the present invention has high purity, high yield, low impurity content, simple process steps, safety and environmental protection, strong operability, and is beneficial to large-scale production.

Embodiments of the present invention

The following examples are used to further illustrate the present invention. These examples are only for illustrative purposes and do not limit the scope of the present invention in any way. The processes and methods not described in detail in the following embodiments are conventional techniques or methods known in the art. The raw materials and solvents used can be obtained through commercial channels unless otherwise specified.

Example 1

Add 200mL acetone, 2.0g 4A molecular sieve and p-toluenesulfonic acid hydrate (89.30mg, 0.47mmol) to the reaction system, stir and wait for p-toluenesulfonic acid to dissolve, add numocidine B0 (10.0g, 9.39mmol), under nitrogen protection , The reaction was heated to reflux, and the reaction process was tracked and detected by HPLC. After the reaction was completed, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 50mL of water and 200mL of ethyl acetate, stir well to dissolve and then stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 50 mL of n-hexane to the concentrated solution, reflux for 0.5 h, and then naturally cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 96.67%, and the yield is 92.34%.

Example 2

Add 300mL acetone, 3.0g 3A molecular sieves, concentrated sulfuric acid (0.05mL, 0.66mmol) to the reaction system, stir and mix well, add neumoconidine B0 (10.0g, 9.39mmol), under nitrogen protection, heat reflux reaction, HPLC tracking The progress of the reaction was detected, after the reaction was completed, filtered, and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 20 mL of water and 100 mL of ethyl acetate, stir well to dissolve and then stand to separate the layers, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100 mL), combine the organic phases, wash with water until neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 80 mL of n-hexane to the concentrated solution, reflux for 1 hour and slowly cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 93.23%, and the yield is 86.11%.

Example 3

Add 100mL acetone, 1.0g 5A molecular sieve, benzenesulfonic acid (44.3mg, 0.28mmol) to the reaction system, stir to dissolve the benzenesulfonic acid, then add neumoconidine B0 (10.0g, 9.39mmol), under the protection of nitrogen, heat to reflux During the reaction, the reaction process was tracked and detected by HPLC. After the reaction was completed, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 100mL of water and 800mL of ethyl acetate, stir well to dissolve and then stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 120 mL of n-hexane to the concentrated solution, reflux for 2 hours and slowly cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 94.28%, and the yield is 88.07%.

Example 4

Add 200mL acetone, 2.0g 4A molecular sieve, 2,4,5-trimethylbenzenesulfonic acid (112.2mg, 0.56mmol) to the reaction system, stir and add after the 2,4,5-trimethylbenzenesulfonic acid is dissolved Numocandine B0 (10.0g, 9.39mmol), under the protection of nitrogen, the reaction was heated and refluxed, and the reaction process was tracked by HPLC. After the reaction, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 60mL of water and 300mL of ethyl acetate, stir well to dissolve and then let stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 100mL n-hexane to the concentrated solution, reflux for 0.5h and slowly cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 95.92%, and the yield is 91.19%.

Example 5

Add 200mL acetone, 2.0g 4A molecular sieve, concentrated nitric acid (3.0mg, 0.47mmol) to the reaction system, stir and mix well, add neumocondine B0 (10.0g, 9.39mmol), under nitrogen protection, heat reflux reaction, HPLC tracking The progress of the reaction was detected, and after the reaction was completed, it was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 50mL of water and 200mL of ethyl acetate, stir well to dissolve and then stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 50 mL of n-hexane to the concentrated solution, reflux for 0.5 h, and then naturally cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 92.45%, and the yield is 89.66%.

Example 6

Add 200mL acetone, 2.0g 4A molecular sieve, acetic acid (2.8mg, 0.47mmol) to the reaction system, stir and mix well, add neumoconidine B0 (10.0g, 9.39mmol), under nitrogen protection, heat reflux reaction, HPLC tracking detection During the reaction process, after the reaction is completed, filter and concentrate the reaction solution to obtain the crude caspofungin intermediate III. Add 50mL of water and 200mL of ethyl acetate, stir well to dissolve and then stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 50 mL of n-hexane to the concentrated solution, reflux for 0.5 h, and then naturally cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 92.03%, and the yield is 84.85%.

Example 7

Add 200mL acetone, 5.0g 4A molecular sieve, p-toluenesulfonic acid hydrate (178.8mg, 0.94mmol, 0.1eq) to the reaction system, stir and wait until p-toluenesulfonic acid is dissolved, then add numocantine B0 (10.0g, 9.39mmol) Under the protection of nitrogen, the reaction was heated and refluxed, and the reaction process was tracked and detected by HPLC. After the reaction was completed, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 50mL of water and 200mL of ethyl acetate, stir well to dissolve and then stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 50 mL of n-hexane to the concentrated solution, reflux for 0.5 h and then naturally cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 88.52%, and the yield is 41.93%.

Example 8

Add 200mL acetone, 2.0g 4A molecular sieve, 2,4,5-trimethylbenzenesulfonic acid (38.0mg, 0.19mmol, 0.02eq) to the reaction system, and stir for 2,4,5-trimethylbenzenesulfonic acid After dissolving, add Numocantine B0 (10.0g, 9.39mmol), under the protection of nitrogen, the reaction was heated to reflux, and the reaction process was tracked by HPLC. After the reaction, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 60mL of water and 300mL of ethyl acetate, stir well to dissolve and then let stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 100 mL of n-hexane to the concentrated solution, reflux for 0.5 h and slowly cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 90.66%, and the yield is 73.56%.

Example 9

Add 50mL acetone, 2.0g 4A molecular sieve, p-toluenesulfonic acid hydrate (17.88mg, 0.094mmol, 0.01eq) to the reaction system, stir and add numocantine B0 (10.0g, 9.39mmol) after p-toluenesulfonic acid is dissolved Under the protection of nitrogen, the reaction was heated and refluxed, and the reaction process was tracked and detected by HPLC. After the reaction was completed, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 60mL of water and 300mL of ethyl acetate, stir well to dissolve and then let stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 100 mL of n-hexane to the concentrated solution, reflux for 0.5 h and slowly cool down and crystallize to obtain Caspofungin Intermediate III with an HPLC purity of 84.85% and a yield of 57.34%.

Example 10

Add 500mL acetone, 5.0g 4A molecular sieve, 2,4,5-trimethylbenzenesulfonic acid (188.2mg, 0.94mmol, 0.1eq) to the reaction system, and stir until 2,4,5-trimethylbenzenesulfonic acid After dissolving, add Numocantine B0 (10.0g, 9.39mmol), under the protection of nitrogen, the reaction was heated to reflux, and the reaction process was tracked by HPLC. After the reaction, the reaction solution was filtered and the reaction solution was concentrated to obtain the crude caspofungin intermediate III. Add 50mL of water and 200mL of ethyl acetate, stir well to dissolve and then stand for liquid separation, collect the organic layer, extract the aqueous phase with ethyl acetate (3×100mL), combine the organic phases, wash with water to neutral, then use anhydrous magnesium sulfate Dry, filter, and concentrate; add 50 mL of n-hexane to the concentrated solution, reflux for 1 hour and slowly cool down and crystallize to obtain Caspofungin Intermediate III. The HPLC purity is 78.73%, and the yield is 43.85%.

Example 11

Under the protection of nitrogen, the Caspofungin Intermediate III (8.0g, 7.24mmol) was dissolved in a mixed solvent of 50mL methanol and water (methanol-water volume ratio 9:1), then 24.0g D113 resin was added, and then 50mL ethyl acetate was added. Diamine was added dropwise to the reaction system. After the addition, the reaction was stirred at room temperature. HPLC detected that the caspofungin intermediate Ⅲ was almost exhausted to stop the reaction; filtered, concentrated under reduced pressure, and added 100 mL methanol and 200 mL ethyl acetate The ester was stirred and separated by filtration to obtain solid crystals. The solid crystals were vacuum dried to obtain the compound of formula IV with a purity of 96.85% and a yield of 95.46%.

Example 12

Under the protection of nitrogen, dissolve Caspofungin Intermediate III (8.0g, 7.24mmol) in 50mL of isopropanol and water mixed solvent (isopropanol-water volume ratio 12:1), add 48.0g D110 resin, and then 40mL of ethylenediamine was added dropwise to the reaction system. After the addition was completed, the reaction was stirred at room temperature. HPLC detected that the caspofungin intermediate III was almost exhausted to stop the reaction; filtered, concentrated under reduced pressure, and added 100mL of methanol and 200 mL of ethyl acetate was stirred and separated by filtration to obtain solid crystals. The solid crystals were vacuum dried to obtain the compound of formula IV with a purity of 94.51% and a yield of 91.63%.

Example 13

Under the protection of nitrogen, the Caspofungin Intermediate III (8.0g, 7.24mmol) was dissolved in a mixed solvent of 60mL ethanol and water (volume ratio of ethanol to water 7:1), 40.0g D151 resin was added, and then 60mL ethyl acetate was added. The diamine was added dropwise to the reaction system. After the addition was completed, the reaction was stirred at room temperature. The reaction was stopped when the caspofungin intermediate Ⅲ was almost exhausted by HPLC; filtered, concentrated under reduced pressure, and added 150mL methanol and 200mL ethyl acetate The ester was separated by filtration to obtain solid crystals, and the solid crystals were vacuum-dried to obtain the compound of formula IV with a purity of 93.93% and a yield of 92.74%.

Example 14

Under the protection of nitrogen, dissolve caspofungin intermediate III (8.0g, 7.24mmol) in a mixed solvent of 60mL of acetonitrile and water (the volume ratio of acetonitrile to water is 10:1), add 30.0g of D152 resin, and then add 50mL of acetonitrile to water. Diamine was added dropwise to the reaction system. After the addition, the reaction was stirred at room temperature. HPLC detected that the caspofungin intermediate Ⅲ was almost exhausted to stop the reaction; filtered, concentrated under reduced pressure, and added 100 mL methanol and 200 mL ethyl acetate The ester was separated by filtration to obtain solid crystals, and the solid crystals were vacuum dried to obtain the compound of formula IV with a purity of 95.67% and a yield of 94.56%.

Example 15

Under the protection of nitrogen, the caspofungin intermediate III (8.0g, 7.24mmol) was dissolved in a mixed solvent of 60mL methanol and water (methanol-water volume ratio 7:1), 40.0g D151 resin was added, and then 60mL ethyl acetate was added. The diamine was added dropwise to the reaction system. After the addition was completed, the reaction was stirred at room temperature. HPLC detected that Caspofungin Intermediate III was almost exhausted to stop the reaction; filtered, concentrated under reduced pressure, and added 150mL methanol and 250mL acetic acid The ethyl ester was stirred and separated by filtration to obtain solid crystals. The solid crystals were vacuum dried to obtain the compound of formula IV with a purity of 93.46% and a yield of 93.35%.

Example 16

Under the protection of nitrogen, the Caspofungin Intermediate III (8.0g, 7.24mmol) was dissolved in a mixed solvent of 50mL methanol and water (methanol-water volume ratio 3:1), 80.0g D113 resin was added, and 30mL ethyl acetate was added. Diamine was added dropwise to the reaction system. After the addition, the reaction was stirred at room temperature. HPLC detected that the caspofungin intermediate Ⅲ was almost exhausted to stop the reaction; filtered, concentrated under reduced pressure, and added 100 mL methanol and 200 mL ethyl acetate The ester was stirred and separated by filtration to obtain solid crystals. The solid crystals were vacuum dried to obtain the compound of formula IV with a purity of 92.67% and a yield of 81.68%.

Example 17

Under the protection of nitrogen, the Caspofungin Intermediate III (8.0g, 7.24mmol) was dissolved in a mixed solvent of 50mL methanol and water (methanol-water volume ratio 9:1), 48.0g D751 resin was added, and then 50mL ethyl acetate was added. Diamine was added dropwise to the reaction system. After the addition, the reaction was stirred at room temperature. HPLC detected that the caspofungin intermediate Ⅲ was almost exhausted to stop the reaction; filtered, concentrated under reduced pressure, and added 100 mL methanol and 200 mL ethyl acetate The ester was stirred and separated by filtration to obtain solid crystals. The solid crystals were vacuum dried to obtain the compound of formula IV with a purity of 88.91% and a yield of 63.45%.

Example 18

Add sodium borohydride (256.5mg, 6.78mmol), compound of formula IV (5.0g, 4.52mmol) and 100mL 1,4-dioxane into the reaction flask, and stir for 0.5h at room temperature under nitrogen protection for use; Add acetic acid (0.41g, 6.78mmol) and 100mL 1,4-dioxane into the reaction flask. After stirring, replace with nitrogen three times, and add the mixed solution dropwise to the above reaction system. After the addition is complete, Continue to stir the reaction. HPLC detects the end of the reaction. After the reaction is completed, 5.0 mL of acetic acid is added dropwise to the reaction system. After the addition is complete, the reaction is continued for 1 hour. The solvent is distilled off under reduced pressure. The concentrate is diluted with methanol and loaded onto the preparative column. Using 0.2% formic acid water as mobile phase A and methanol as mobile phase B for gradient elution, the effluent containing caspofungin was collected and lyophilized to obtain caspofungin with a purity of 99.89% and a yield of 85.96%.

Example 19

Add sodium borohydride (171.0mg, 4.52mmol), compound of formula IV (5.0g, 4.52mmol) and 120mL dimethyl sulfoxide into the reaction flask, and stir for 0.5h under the protection of nitrogen at room temperature for use; put into another reaction flask Add o-hydroxybenzoic acid (1.25g, 9.04mmol) and 150mL dimethyl sulfoxide, stir well, replace with nitrogen three times, and add the mixed solution dropwise to the above reaction system. After the addition is complete, continue to stir and react HPLC detects the end of the reaction. After the reaction is completed, 8.0 mL of acetic acid is added dropwise to the reaction system. After the addition is complete, the reaction is continued for 1 hour. The solvent is distilled off under reduced pressure. The concentrated solution is diluted with methanol and loaded onto the preparative chromatography column. % Formic acid water was used as mobile phase A, methanol was used as mobile phase B, gradient elution was performed, the effluent containing caspofungin was collected, and caspofungin was lyophilized to obtain caspofungin with a purity of 99.81% and a yield of 84.39%.

Example 20

Add sodium borohydride (342.0mg, 9.04mmol), compound of formula IV (5.0g, 4.52mmol) and 150mL dimethylimide into the reaction flask, and stir for 0.5h under the protection of nitrogen at room temperature for later use; put into another reaction flask Add benzoic acid (1.10g, 9.04mmol) and 150mL dimethylimide. After stirring, replace with nitrogen three times, and add the mixed solution dropwise to the above reaction system. After the addition is complete, continue to stir and react, HPLC The end point of the reaction was detected. After the reaction was completed, 10.0 mL of acetic acid was added dropwise to the reaction system. After the addition, the reaction was continued for 1 hour. The solvent was distilled off under reduced pressure. The concentrated solution was diluted with ethanol and loaded on the preparation column with 0.8% formic acid. Water was used as mobile phase A, and acetonitrile was used as mobile phase B for gradient elution. The effluent containing caspofungin was collected and lyophilized to obtain caspofungin with a purity of 99.78% and a yield of 86.03%.

Example 21

Add sodium borohydride (256.5mg, 6.78mmol), compound of formula IV (5.0g, 4.52mmol) and 100mL 1,4-dioxane into the reaction flask, and stir for 0.5h at room temperature under nitrogen protection for use; Add formic acid (0.42g, 9.04mmol) and 100mL to the reaction flask 1,4-Dioxane, after stirring uniformly, nitrogen replacement three times, and the mixed solution was added dropwise to the above reaction system, after the dropwise addition, continue to stir the reaction, HPLC to detect the end of the reaction, after the completion of the reaction, Add 5.0 mL of acetic acid dropwise to the reaction system. After the addition is complete, continue the reaction for 1 hour. The solvent is distilled off under reduced pressure. The concentrate is diluted with methanol and loaded onto the preparative chromatographic column with 0.5% formic acid water as mobile phase A and methanol as the mobile phase. In mobile phase B, gradient elution was performed, and the effluent containing caspofungin was collected, and caspofungin was lyophilized to obtain caspofungin with a purity of 99.71% and a yield of 85.89%.

Example 22

Add sodium borohydride (171.0mg, 4.52mmol), compound of formula IV (5.0g, 4.52mmol) and 100mL dimethyl sulfoxide into the reaction flask, stir for 0.5h under the protection of nitrogen at room temperature for use; put into another reaction flask Add propionic acid (0.34g, 4.52mmol) and 100mL dimethyl sulfoxide, stir well, replace with nitrogen three times, and add the mixed solution dropwise to the above reaction system. After the addition is complete, continue to stir and react, HPLC The end point of the reaction was detected. After the reaction was completed, 5.0 mL of acetic acid was added dropwise to the reaction system. After the addition was completed, the reaction was continued for 1 hour. The solvent was distilled off under reduced pressure. The concentrated solution was diluted with methanol and loaded onto the preparative column with 0.2% formic acid Water was used as mobile phase A and methanol was used as mobile phase B for gradient elution. The effluent rich in caspofungin was collected and lyophilized to obtain caspofungin with a purity of 99.79% and a yield of 84.57%.

Example 23

Add sodium borohydride (342.0mg, 9.04mmol), compound of formula IV (5.0g, 4.52mmol) and 100mL dimethylimide into the reaction flask, and stir for 0.5h under nitrogen protection at room temperature for later use; put into another reaction flask Add valeric acid (1.85g, 18.08mmol) and 100mL dimethylimide, stir well, replace with nitrogen three times, and add the mixed solution dropwise to the above reaction system. After the addition is complete, continue to stir and react, HPLC The end point of the reaction was detected. After the reaction was completed, 5.0 mL of acetic acid was added dropwise to the reaction system. After the addition was completed, the reaction was continued for 1 hour. The solvent was removed by distillation under reduced pressure. Water was used as mobile phase A and methanol was used as mobile phase B for gradient elution. The effluent rich in caspofungin was collected and lyophilized to obtain caspofungin with a purity of 99.59% and a yield of 88.17%.

Example 24

Add sodium borohydride (513.0mg, 13.56mmol)), compound of formula IV (5.0g, 4.52mmol) and 100mL 1,4-dioxane to the reaction flask, and stir for 0.5h at room temperature under nitrogen protection for later use; Add butyric acid (0.81g, 13.56mmol) and 100mL 1,4-dioxane to a reaction flask. After stirring, replace with nitrogen three times, and add the mixed solution dropwise to the above reaction system. The addition is complete After that, the reaction was continued to stir, and the end of the reaction was detected by HPLC. After the reaction was completed, 5.0 mL of acetic acid was added dropwise to the reaction system. After the addition, the reaction was continued for 1 hour. The solvent was distilled off under reduced pressure. Chromatographic column, with 0.2% formic acid water as mobile phase A and methanol as mobile phase B for gradient elution, collect the effluent rich in caspofungin, lyophilize to obtain caspofungin, purity 97.54%, yield 89.09% .

Example 25

Add sodium borohydride (119.7mg, 3.16mmol), compound of formula IV (5.0g, 4.52mmol) and 100mL of dichloromethane to the reaction flask, stir for 0.5h under the protection of nitrogen at room temperature for later use; add three to another reaction flask Fluoroacetic acid (0.36g, 3.16mmol) and 100mL of dichloromethane, after stirring uniformly, nitrogen replacement three times, and the mixed solution was added dropwise to the above reaction system, after the addition is complete, continue to stir the reaction, HPLC detects the end of the reaction After the reaction is complete, add 5.0 mL of acetic acid dropwise to the reaction system, continue the reaction for 1 hour after the addition is complete, and distill under reduced pressure to remove the solvent. The concentrated solution is diluted with methanol and loaded onto the preparative chromatography column with 0.2% formic acid water as the flow In phase A, methanol was used as mobile phase B for gradient elution. The effluent rich in caspofungin was collected and lyophilized to obtain caspofungin with a purity of 97.14% and a yield of 68.51%.

Caspofungin intermediate III was analyzed by ¹ H-NMR and MS mass spectrometry:

¹ HNMR (600.13MHz, CD ₃ OD): 7.13 (m, 1H), 7.03 (t, 2H), 6.91 (d, 1H), 6.71 (m, 2H), 6.67 (s, 1H), 4.56 (dd, 1H), 4.42-4.50 (m, 3H), 4.43 (dd, 1H), 4.23 (m, 3H), 4.14 (dd, 1H), 3.89 (d, 1H), 3.69-3.72 (m, 3H), 3.54 (d, 1H), 3.25 (d, 1H), 2.44 (m, 2H), 2.37 (dd, 1H), 2.11-2.15 (m, 3H), 1.86-1.93 (m, 5H), 1.75 (s, 2H) ),1.45(m,6H)1.20-1.35(m,15H),0.96(m,2H),0.86(m,1H),0.79(t,3H),0.75-0.78(m,6H);

MS (ESI): 1105.6027 (M+H ⁺ ).

formula

The compound was analyzed by ¹ H-NMR and MS mass spectrometry:

¹ HNMR (600.13MHZ, CD ₃ OD): 7.13 (m, 2H), 7.03 (t, 2H), 6.71 (m, 2H), 4.89 (d, 1H), 4.85 (d, 1H), 4.60 (d, 1H), 4.56 (dd, 1H), 4.42-4.50 (m, 2H), 4.43 (dd, 1H), 4.23 (m, 3H), 4.14 (dd, 1H), 4.11 (d, 1H), 3.89 (d ,1H), 3.69-3.72 (m, 3H), 3.54 (d, 1H), 3.25 (d, 1H), 2.98 (t, 2H), 2.72-2.95 (m, 2H), 2.44 (m, 2H), 2.37(dd,1H),2.11-2.15(m,3H),1.86-1.93(m,3H),1.75(s,6H),1.20-1.35(m,15H),0.96(m,2H),0.86( m,1H),0.79(t,3H),0.75-0.78(m,6H);

MS (ESI): 1108.3215 (M+H ⁺ ).

Caspofungin is analyzed by ¹ H-NMR, ¹³ C-NMR nuclear magnetic resonance spectroscopy, IR infrared spectroscopy and MS mass spectrometry:

¹ HNMR (600.13MHz, CD ₃ OD): 7.13 (m, 2H), 6.71 (m, 2H), 4.89 (d, 1H), 4.85 (d, 1H), 4.60 (d, 1H), 4.56 (dd, 1H), 4.42-4.50 (m, 2H), 4.43 (dd, 1H), 4.23 (m, 3H), 4.14 (dd, 1H), 4.11 (d, 1H), 3.89 (d, 1H), 3.69-3.72 (m, 3H), 3.54 (d, 1H), 3.25 (d, 1H), 2.98 (t, 2H), 2.72-2.95 (m, 4H), 2.37 (dd, 1H), 2.11-2.15 (m, 3H) ),1.86-1.93(m,5H),1.75(s,6H),1.46(m,2H),1.20-1.35(m,15H),0.96(m,2H),0.86(m,1H),0.79( t,3H),0.75-0.78(m,6H);

¹³ C-NMR(150.9MHZ): 176.0, 172.8, 172.3, 172.1, 171.4, 169.0, 167.5, 157.2, 131.6, 128.2, 114.9, 76.0, 74.2, 73.7, 70.7, 67.0, 68.7, 68.0, 66.8, 63.0, 61.4 ,57.0,55.8,54.8,54.5,49.8,48.0,47.9,47.8,47.6,47.5,47.3,47.2,45.7,44.5,42.4,38.3,37.6,37.1,36.0,35.2,34.2,33.2,31.5,29.8,29.4 ,29.3,29.2,29.0,26.7,25.7,19.3,18.8,10.4;

IR(KBr): 3775-2348,3345,2920,1634,1545,1461,1231,1093cm ^-1

MS (ESI): 1093.6423 (M+H ⁺ ).

Claims

A caspofungin intermediate III compound, its structural formula is as follows:
.
A method for preparing caspofungin intermediate III, which is characterized in that the compound of formula II numocandine B0 is used as a raw material, and it is heated and refluxed with acetone under acid conditions to generate caspofungin intermediate III. The reaction formula is as follows:

.
The preparation method according to claim 2, characterized in that the mass ratio of the added volume of acetone to the mass ratio of Numocantine B0 is 10-30:1, mL/g; more preferably 20:1, mL/g.
The preparation method according to claim 2, wherein the acid is an inorganic acid or an organic acid, preferably p-toluenesulfonic acid, benzenesulfonic acid, 2,4,5-trimethylbenzenesulfonic acid, acetic acid , Concentrated sulfuric acid or concentrated nitric acid, more preferably p-toluenesulfonic acid or 2,4,5-trimethylbenzenesulfonic acid.
The preparation method according to claim 2, characterized in that the molar ratio of the acid to numocantine B0 is 0.03 to 0.07:1, preferably 0.05:1.
A method for synthesizing caspofungin, characterized in that caspofungin intermediate III reacts with ethylenediamine in the presence of a weak acid resin to form intermediate IV, and intermediate IV is reduced by sodium borohydride and organic acid To obtain caspofungin, the reaction formula is as follows:

.
The synthesis method according to claim 6, wherein the reaction solvent in step b is one or more of methanol-water, ethanol-water, isopropanol-water, and acetonitrile-water, more preferably methanol-water的mixed solvents.
The synthesis method according to claim 6, wherein the weakly acidic resin is a carboxylic acid, phosphoric acid or phenol-based resin, preferably, the weakly acidic resin is D110, D113, D151 or D152, and further Preferably it is D113 resin.
The synthesis method according to claim 6, wherein in step c, the reducing agent is sodium borohydride and an organic acid, and the organic acid is selected from the group consisting of formic acid, acetic acid, propionic acid, butyric acid, valeric acid, and benzoic acid. Or one of ortho-hydroxybenzoic acid, more preferably acetic acid.
The synthesis method according to claim 6, wherein the molar ratio of sodium borohydride to organic acid in step c is 1:1 to 2, preferably 1:1; in step c, the intermediate IV of caspofungin and The molar ratio of sodium borohydride is 1:1 to 2, more preferably 1:1.5.