CN118108806A

CN118108806A - Industrial refining and purifying method of Fuciclosporin precursor

Info

Publication number: CN118108806A
Application number: CN202211516092.XA
Authority: CN
Inventors: 张特; 朱丽君; 杨北; 陈群; 陈磊; 胡锦亮; 彭深振; 魏伟业; 吴小涛; 李战
Original assignee: Nanjing Gritpharma Co ltd
Current assignee: Nanjing Gritpharma Co ltd
Priority date: 2022-11-30
Filing date: 2022-11-30
Publication date: 2024-05-31

Abstract

The invention provides an industrialized refining and purifying method of a Fu-cyclosporine precursor isomer, which relates to the field of drug synthesis. The method simplifies the purification steps of the chromatographic column in the traditional process of the cyclosporine, has simple operation and low production cost, and is easy for industrialized popularization.

Description

Industrial refining and purifying method of Fuciclosporin precursor

Technical Field

The invention relates to the technical field of medicine synthesis, relates to medicine purification, and in particular relates to an industrialized refining and purifying method of a Fusarium oxysporum precursor.

Background

Lupkynis the active ingredient, VCS (vollosporin), is a derivative of CsA, developed by Isotechnika company (now renamed as AURINIA PHARMACEUTICALS company) and is mainly used for treating rejection after organ transplantation and autoimmune diseases such as lupus nephritis, psoriasis and noninfectious uveitis. It is a new generation calcineurin inhibitor (CNi), has stronger enzyme inhibition activity, lower nephrotoxicity compared to cyclosporin, and lower incidence of post-transplant new diabetes (NODAT) compared to tacrolimus.

Lupkynis on month 22 of 2021, FDA approved for marketing in the united states in the form of a cyclosporin capsule in combination with immunosuppressive therapy for adult active Lupus Nephritis (LN). Lupkynis is the first FDA approved oral therapy for lupus nephritis. The compound of the Fulvine is of an isomer structure, and the separation and purification preparation of the isomer is an important step in the synthesis process of the Fulvine. In the prior art, in the preparation method of the cyclosporine, a chromatographic column method is adopted to separate and match with repeated recrystallization to reduce isomers. In the industrialized production, chromatographic column separation is adopted, so that the requirements on equipment are high, the operation is complex, the production cost is high, and the industrialized production is not easy.

Disclosure of Invention

Aiming at the problem of complex removal and purification operations of the isomer of the Fulvine in the prior art, the invention provides an industrialized refining and purifying method of the precursor isomer of the Fulvine.

In the prior art, synthesis of procyanidins the synthesis of procyanidins is carried out by deacylation of the procyanidins as follows. The invention provides an industrialized purification and refining method of a precursor isomer of the following cyclosporine.

The aim of the invention is achieved by the following technical scheme:

a composition of a cyclosporine precursor, wherein the purity of the cyclosporine precursor in said composition is not less than 95% and the isomer ratio Z/E in the cyclosporine precursor is not more than 0.15/1;

preferably, the purity of the cyclosporine precursor in the composition is not less than 97%, and the isomer ratio Z/E in the cyclosporine precursor is not more than 0.15/1;

Preferably, the purity of the procyanidin is not less than 97%, and the isomer ratio Z/E in the procyanidin is not more than 0.09/1.

Further preferably, the purity of the procyanidin is not less than 99% and the isomer ratio Z/E in the procyanidin is not more than 0.09/1.

The composition is prepared by recrystallizing a crude product of the procyanidin for 2 times or more times.

Specifically, the first recrystallization solvent is at least one of organic solvent A methyl tertiary butyl ether, ethyl tertiary butyl ether, isopropyl ether and ethyl propyl ether; methyl tertiary butyl ether is preferred. And secondly, dissolving and crystallizing by using acetone and an antisolvent C to prepare the Fuciclosporin precursor with high purity and low Z-type isomer content, wherein the antisolvent C is water, ethanol and acetonitrile, and water is preferred.

A method for purifying a procyanidin isomer by purification, comprising the steps of:

(1) Adding the crude product of the Fuciclosporin precursor into the organic solvent A for dissolving and clarifying, continuously stirring until crystallization, filtering, washing and drying the Fuciclosporin precursor I;

(2) Adding the Fuciclosporin precursor I into the organic solvent B, stirring, dissolving and clarifying, then dripping the antisolvent C, crystallizing, filtering, washing and drying to obtain a Fuciclosporin precursor pure product;

At least one of methyl tertiary butyl ether, ethyl tertiary butyl ether, isopropyl ether and ethyl propyl ether is organic solvent A; methyl tertiary butyl ether is preferred.

The organic solvent B is acetone; the antisolvent C is water, ethanol, acetonitrile, preferably water.

And (3) repeating the operation for not less than 2 times in the step (2).

In some embodiments, the procyanidin I comprises organic solvent B and antisolvent C in a ratio of 1:5-10:3-6 (Kg/L/L).

In some preferred embodiments, the operation steps of step (2) are specifically:

(a1) Adding the Fuciclosporin precursor I into the organic solvent B, stirring, dissolving and clarifying, then dripping the anti-solvent C, crystallizing, filtering, washing and drying to obtain a Fuciclosporin precursor pure product II;

(a2) Adding the Fuciclosporin precursor II into the organic solvent B, dropwise adding the antisolvent C, crystallizing, filtering, washing and drying to obtain a Fuciclosporin precursor pure product;

Wherein the step (a 1) comprises the steps of preparing a cyclosporine precursor I, an organic solvent B and an antisolvent C in a ratio of 1:5:3 (Kg/L/L);

the step (a 2) is characterized in that the ratio of the Fusarium cyclosporine precursor II to the organic solvent B to the antisolvent C is 1:10:3-6 (Kg/L/L).

In some preferred embodiments, the step (a 2) is performed with a 1:10:6 (Kg/L/L) of the cyclosporine precursor II to the organic solvent B to the antisolvent C.

In some preferred embodiments, the number of repeated operations of step (a 2) is not less than 1.

In some embodiments, the crude product of the cyclosporine precursor in step (1) is 1:1 to 4, preferably 1:2, as organic solvent A.

In some embodiments, the washing in step (1) employs n-heptane, an n-hexane solvent, preferably n-heptane.

In some embodiments, the washing in step (2) employs a mixed solution of an organic solvent B and an anti-solvent C, preferably an organic solvent B: anti-solvent C of 1:1 to 5, more preferably an organic solvent B: anti-solvent C of 1:1.

In some embodiments, the stirring, dissolving, clarifying and specific stirring time in the step (1) is 10-60min, preferably 30min.

In some embodiments, the crystallization time of step (1) is 1 to 8 hours, preferably 4 hours.

In some embodiments, the step (1) is performed in an amount of 2 to 4 times, preferably 2 times, the mass of the crude product of the procyanidin.

3. Advantageous effects

Compared with the prior art, the invention has the advantages that:

(1) After preliminary purification by methyl tertiary butyl ether, the method further adopts a solvent system of acetone and water to carry out multiple recrystallization operation on the purified cyclosporine precursor product, and the purity of the finally obtained product reaches more than 99.90 percent, so that the impurity of the Fu Huan sporine precursor isomer is almost completely removed. Simple operation, low production cost and easy industrialized popularization.

(2) According to the embodiment 2 of the invention, the prepared cyclosporine precursor is further synthesized into the cyclosporine, and finally the obtained cyclosporine product has high purity and less Fu Huanbao isomer impurities, so that the prepared cyclosporine precursor is synthesized into the cyclosporine, the purity of the cyclosporine is high, the content of related substances is low, the content of the cyclosporine isomer is controlled within a standard range, the chromatographic purification step in the traditional process of synthesizing the cyclosporine is simplified, the problem that the chromatographic purification and amplification production is difficult is effectively solved, and the method is suitable for industrial mass production of the cyclosporine.

(3) The method of the invention does not contain high-toxicity, inflammable and explosive substances, is safe to operate, can recycle and reuse the solvent, and is environment-friendly and pollution-free.

Drawings

FIG. 1 is a nuclear magnetic resonance spectrum of crude cyclosporine precursor of example 1 of the present invention (the Z/E of the cyclosporine precursor isomer is 0.35/1);

FIG. 2 is a liquid chromatogram of the cyclosporine precursor I-1 of example 1 (at peak time RT38.355min for the cyclosporine precursor) of the invention;

FIG. 3 is a nuclear magnetic spectrum of the cyclosporine precursor I-1 of example 1 (the cyclosporine precursor isomer Z/E is 0.19/1) of the present invention;

FIG. 4 is a liquid chromatogram of a pure product of the cyclosporine precursor of example 1 (at peak time RT38.303min for the cyclosporine precursor);

FIG. 5 is a nuclear magnetic resonance spectrum of a pure product of a cyclosporine precursor of example 1 (the Z/E of the cyclosporine precursor isomer is 0.11/1);

FIG. 6 is a liquid chromatogram of a pure product of the cyclosporine precursor of example 2 (at a cyclosporine precursor off-time RT38.210 min);

FIG. 7 is a nuclear magnetic resonance spectrum of a pure product of a cyclosporine precursor of example 2 (the Z/E of the cyclosporine precursor isomer is 0.08/1);

FIG. 8 is a liquid chromatogram of the cyclosporine of example 2 (at peak time RT29.483min for cyclosporine);

FIG. 9 is a nuclear magnetic resonance spectrum of a cyclosporine of example 2 of the invention (cyclosporine isomer Z/E is 0.05/1);

FIG. 10 is a nuclear magnetic resonance spectrum of a pure product of a cyclosporine precursor of example 3 (the Z/E of the cyclosporine precursor isomer is 0.08/1);

FIG. 11 is a nuclear magnetic resonance spectrum of a pure product of a cyclosporine precursor of example 4 of the present invention (Z/E is 0.09/1 for the cyclosporine precursor isomer);

FIG. 12 is a nuclear magnetic resonance spectrum of a pure product of a cyclosporine precursor of example 5 of the present invention (Z/E of the cyclosporine precursor isomer is 0.08/1);

FIG. 13 is a nuclear magnetic resonance spectrum of a pure product of a cyclosporine precursor of example 6 of the present invention (Z/E of the cyclosporine precursor isomer is 0.06/1);

FIG. 14 is a nuclear magnetic spectrum of pure product I-11 of the cyclosporine precursor of example 11 of the invention (Z/E is 0.19/1 for the cyclosporine precursor isomer);

FIG. 15 is a nuclear magnetic resonance spectrum of a pure product of the cyclosporine precursor of example 11 of the present invention (Z/E of the cyclosporine precursor isomer is 0.17/1).

Detailed Description

The invention will now be described in detail with reference to the drawings and the accompanying specific examples.

Example 1

2Kg of crude product of the procyanidin with the purity of 70% (shown in figure 1, the isomer Z/E is 0.35/1) is added into 2L of methyl tertiary butyl ether, the mixture is stirred for 32min at normal temperature to dissolve and clarify, the mixture is continuously stirred until solid is separated out, the mixture is stirred for 2h at normal temperature, the reaction liquid is pumped and filtered, the filter cake is leached by 2L of n-heptane, and the leaching is carried out and then is dried for 4-6h at the temperature of 35-40 ℃ in vacuum, thus obtaining 1.2kg of white solid, namely the procyanidin precursor Ia. HPLC purity 98.395% of the Fulvin precursor I-1 (peak time RT38.35min) as shown in FIG. 2, and isomer residue as judged by nuclear magnetic resonance spectroscopy, as shown in FIG. 3, was 0.19/1 for the Fulvin precursor Ia isomer Z/E. Adding 6L of acetone into 1.2kg of the Fuciclosporin precursor I, stirring at normal temperature for 25min, dissolving and clarifying, dripping 3.6L of water, preserving heat, crystallizing and stirring for 2h after dripping, leaching the reaction solution, leaching the filter cake by 1.2L of acetone/water (volume ratio of 1:1), and drying for 4-6h at the temperature of 35-40 ℃ in vacuum after leaching to obtain 0.9kg of Fuciclosporin precursor pure product. The HPLC purity of the pure product of the procyanidin can be seen as 99.751% (peak time RT38.303 min) as shown in FIG. 4, and the Z/E of the pure product of the procyanidin is 0.11/1 as shown in FIG. 5.

Example 2

And (3) adding 5.4L of acetone into 0.9kg of the prepared cyclosporine precursor obtained in the example 1, stirring for 30min at normal temperature, dissolving and clarifying, dripping 2.7L of water, preserving heat, crystallizing and stirring for 2h after dripping is finished, leaching the reaction solution, leaching the filter cake by 1.8L of acetone/water (volume ratio of 1:1), and drying for 4-6h at the temperature of 35-40 ℃ in vacuum to obtain 0.72kg of pure cyclosporine precursor. As can be seen from FIG. 6, the HPLC purity of the pure product of the cyclosporine precursor is 99.978% (peak time RT38.210 min), and as shown in FIG. 7, the isomer Z/E of the pure product of the cyclosporine precursor prepared in this example is 0.08/1.

The pure product of the procyanidin prepared in this example was synthesized into procyanidin according to the method disclosed in example 15 of the original-research enterprise patent CN1798759a for procyanidin. The pure product of the cyclosporine precursor is dissolved in methanol, the aqueous solution of potassium carbonate is added for reaction, the methanol is evaporated, the residue is dissolved in ethyl acetate, and the aqueous phase is separated and discarded. The organic phase is washed by citric acid aqueous solution and half saturated sodium chloride aqueous solution, sodium sulfate is dried, filtered and concentrated to obtain the Fuciclosporin product.

As shown in FIG. 8, the HPLC purity of the Fulvine is 99.812% (peak time RT29.483 min), and as shown in FIG. 9, the isomer Z/E of the Fulvine is 0.05/1, the purity of the Fulvine is high, and the isomer impurity content is low.

In CN17987959A example 15, synthesis of Fulvine from an unpurified Fulvine precursor, product E-ISA247 (85% content) was synthesized and purification was performed by chromatographic HPLC. Thus, E-ISA247 synthesized directly from the unpurified cyclosporin precursor of example 15 is only 85% pure and requires subsequent purification by chromatographic HPLC. The application relates to an amplified production process of the cyclosporine, which adopts a chromatographic purification technology to industrially refine the cyclosporine in large batches, is not easy to operate, and has high production cost, long time consumption and low production efficiency.

Example 3

2Kg of crude product (Z/E is 0.30/1) of the Fuciclosporin precursor with the purity of 75 percent is added with 4L of methyl tertiary butyl ether, stirred at normal temperature for 32min for dissolving and clarifying, stirred continuously until solid is separated out, stirred continuously at normal temperature for 2h, the reaction liquid is filtered by suction, the filter cake is leached by 4L of n-heptane, and dried for 4-6h at the temperature of 35-40 ℃ in vacuum after leaching, thus obtaining 1.1Kg of white solid, namely, fuciclosporin precursor I-3 (purity 97.265 percent, Z/E is 0.18/1). Adding 3.3L of acetone into 1.1Kg of the Fuciclosporin precursor I, stirring at normal temperature for 28min, dissolving and clarifying, dripping 3.3L of water, preserving heat and crystallizing after dripping is finished, stirring for 3h, leaching the reaction solution by suction, leaching the filter cake by 2.2L of acetone/water (volume ratio of 1:1), and drying at 35-40 ℃ for 4-6h after leaching to obtain 0.803Kg (purity 98.954%, Z/E is 0.12/1) of Fuciclosporin precursor II. Adding acetone 4L into 0.803Kg of the Fuciclosporin precursor II-3, stirring at normal temperature for 25min, dissolving and clarifying, dripping water 2.4L, preserving heat and crystallizing after dripping is finished, stirring for 2h, leaching the reaction liquid, leaching the filter cake by 1.6L of acetone/water (volume ratio of 1:1), and drying at 35-40 ℃ for 4-6h after leaching to obtain 0.626Kg (purity 99.845%) of Fuciclosporin precursor pure product. As shown in FIG. 10, the pure isomer Z/E of the cyclosporine precursor prepared in this example was 0.08/1.

Example 4

2Kg of crude product of the procyanidin with the purity of 50 percent (Z/E is 0.40/1) is added with 4L of methyl tertiary butyl ether, stirred at normal temperature for 30min for dissolving and clarifying, stirred continuously until solid is separated out, stirred at normal temperature for 2h, the reaction liquid is filtered by suction, the filter cake is leached by 4L of n-heptane, and dried for 4-6h at the temperature of 35-40 ℃ in vacuum after leaching, thus obtaining 0.9Kg of white solid, namely the procyanidin precursor I-4 (the purity is 95.357 percent, and the Z/E is 0.23/1). Adding 4.5L of acetone into 0.9Kg of the Fuciclosporin precursor I, stirring at normal temperature for 28min, dissolving and clarifying, dripping 2.7L of water, preserving heat and crystallizing after dripping is finished, stirring for 2h, leaching the reaction solution by suction, leaching the filter cake by 2L of acetone/water (volume ratio of 1:1), and drying at 35-40 ℃ for 4-6h after leaching to obtain 0.65Kg (purity 97.265% and Z/E of 0.17/1) of Fuciclosporin precursor II. And (3) adding 6.5L of acetone into 0.65Kg of the Fuciclosporin precursor II, stirring at normal temperature for 25min, dissolving and clarifying, dripping 1.95L of water, preserving heat and crystallizing and stirring for 2h after dripping is finished, leaching the reaction solution, leaching the filter cake by 1.5L of acetone/water (volume ratio of 1:1), and drying at the temperature of 35-40 ℃ for 4-6h after leaching to obtain 0.525Kg of Fuciclosporin precursor pure product (the purity is 99.417%). As shown in FIG. 11, the pure isomer Z/E of the procyanidin in this example is 0.09/1.

Example 5

2Kg of crude product of the procyanidin with the purity of 60 percent (Z/E is 0.32/1) is added with 4L of methyl tertiary butyl ether, stirred at normal temperature for 35min for dissolving and clarifying, stirred continuously until solid is separated out, stirred at normal temperature for 2h, the reaction liquid is pumped and filtered, the filter cake is leached by 4L of n-heptane, and the leaching is placed at the temperature of between 35 and 40 ℃ for drying for 4to 6h after leaching, thus obtaining 1.04Kg of white solid, namely the procyanidin precursor I-5 (the purity is 97.297 percent, and the Z/E is 0.2/1). Adding 5.2L of acetone into 1.04Kg of Fuciclosporin precursor I, stirring at normal temperature for 30min, dissolving and clarifying, dripping 3.1L of water, preserving heat and crystallizing after dripping is finished, stirring for 2h, leaching the reaction solution by suction, leaching the filter cake by 2.0L of acetone/water (volume ratio of 1:1), and drying at 35-40 ℃ for 4-6h after leaching to obtain 0.759Kg (purity 97.895% and Z/E of 0.12/1) of Fuciclosporin precursor II. 7.6L of acetone is added into 0.759Kg of the Fulvine precursor II, the solution is stirred for 30min at normal temperature, the solution is dissolved and clarified, 4.55L of water is added dropwise, the solution is stirred for 2h after the dripping is finished by heat preservation crystallization, the reaction solution is filtered by suction, 1.5L of acetone/water (1:1) is used for leaching the filter cake, and the filter cake is dried for 4-6h at the temperature of 35-40 ℃ in vacuum after leaching, so that 0.569kg (the purity is 99.494%) of the Fulvine precursor is obtained. As shown in FIG. 12, the pure isomer Z/E of the procyanidin in this example is 0.08/1.

Example 6

2G of a crude product of the Fuciclosporin precursor with the purity of 75 percent (Z/E is 0.3/1) is added with 8mL of methyl tertiary butyl ether, stirred at normal temperature for 5min for dissolution and clarification, stirred continuously until solid is separated out, stirred at normal temperature for 2h, the reaction liquid is filtered, the filter cake is leached by 8mL of n-heptane, and dried for 4-6h at the temperature of 35-40 ℃ in vacuum after leaching, thus obtaining 1.2g of white solid, namely, fuciclosporin precursor I-6 (the purity 98.560 percent, and Z/E is 0.18/1). Adding 6mL of acetone into 6g of the Fuciclosporin precursor I-61.2g, stirring at normal temperature for 10min, dissolving and clarifying, dripping 3.6mL of water, preserving heat, crystallizing and stirring for 2h after dripping, leaching the reaction liquid, leaching the filter cake by 2.4mL of acetone/water (volume ratio of 1:1), and drying at 35-40 ℃ for 4-6h after leaching to obtain 60.96g of Fuciclosporin precursor II-60.96g (purity 99.432% and Z/E of 0.10/1). 9.6mL of acetone is added into 9-60.96 g of the Fuciclosporin precursor, the solution is stirred at normal temperature for 10min, the solution is dissolved and clarified, 5.8mL of water is added dropwise, the solution is stirred for 2h after the dropwise addition is finished by heat preservation crystallization, the reaction solution is filtered by suction, 1.9mL of acetone/water (volume ratio of 1:1) is used for leaching the filter cake, and the leaching is carried out and then the leaching is carried out for 4-6h at the temperature of 35-40 ℃ in vacuum, thus 0.806g (the purity of 99.965%) of Fuciclosporin precursor pure product is obtained. As shown in FIG. 13, the pure isomer Z/E of the procyanidin is 0.06/1.

Example 7

2G of the Fusarium oxysporum precursor Ia prepared in the method of example 1 is added into 10mL of acetone, the mixture is stirred at normal temperature for 30min to dissolve and clarify, 6mL of n-heptane is added dropwise, and the mixture is subjected to heat preservation crystallization and stirring for 2h after the dropwise addition is finished, so that no solid precipitation is observed.

Example 8

2G of the Fusarium oxysporum precursor Ia prepared in the method of example 1 is added into 10mL of acetone, and the mixture is stirred at normal temperature for 30min to dissolve and clarify, 6mL of n-hexane is added dropwise, and after the dropwise addition is finished, the mixture is subjected to heat preservation crystallization and stirring for 2h, and no solid precipitation is observed.

Example 9

2Kg of the Fusarium oxysporum precursor Ia prepared in the method of example 1 is added into 8mL of methanol, the mixture is stirred at normal temperature for 30min to dissolve and clarify, 4.8mL of water is added dropwise, the mixture is subjected to heat preservation crystallization and stirring for 2h after the dropwise addition is finished, and oily matters are separated out and are not obvious solid particles.

Example 10

Taking 2g of the crude product of the cyclosporine precursor in the same batch of example 1, adding 8mL of acetone, stirring at normal temperature for 30min, dissolving and clarifying, dripping 4.8mL of water, and preserving heat and crystallizing for 2h after dripping is finished, wherein no solid precipitation is found.

Example 11

2G of a crude product of the procyanidin with the purity of 70% is added with 8mL of methyl tertiary butyl ether, stirred at normal temperature for 15min for dissolution and clarification, stirred until solid is separated out, stirred at normal temperature for 2h, the reaction liquid is filtered, the filter cake is leached by 8mL of n-heptane, and dried for 4-6h at the temperature of 35-40 ℃ in vacuum after leaching, so as to obtain 1.1g of white solid, namely the procyanidin I-11. As shown in FIG. 14, the Z/E of pure I-11 isomer of the procyanidin is 0.19/1. And (3) adding 4.4mL of methyl tertiary butyl ether into 1.1g of the Fuciclosporin precursor I, stirring at normal temperature for 15min for dissolving and clarifying, separating out solids, stirring at normal temperature for 2h, filtering the reaction liquid, leaching a filter cake by 4.4mL of n-heptane, and drying at 35-40 ℃ for 4-6h after leaching to obtain 0.66g of white solid, namely Fuciclosporin precursor II-11. Adding 2.4mL of methyl tertiary butyl ether into 0.6g of pure product II of the Fuciclosporin, stirring for 15min at normal temperature, dissolving and clarifying, separating out solid, stirring for 2h at normal temperature, leaching the reaction liquid, leaching a filter cake by 2.4mL of n-heptane, placing the leached filter cake in vacuum at 35-40 ℃ for drying for 4-6h, and obtaining 0.4g of white solid, namely the pure product of the Fuciclosporin precursor. As shown in FIG. 15, the pure isomer Z/E of the procyanidin is 0.17/1.

The method is characterized in that the procyanidin precursor is subjected to 3-methyl tertiary butyl ether purification, the removal effect of the isomer Z/E is not obvious compared with that of an acetone/water system, and the yield is lower than that of the acetone/water system.

And after the crude product of the Fugu cyclosporine precursor is subjected to preliminary purification by adopting methyl tertiary butyl ether to obtain a Fugu cyclosporine precursor I, the Fugu cyclosporine precursor I is further recrystallized and purified by utilizing methyl tertiary butyl ether again, and the purity and the Z/E ratio of the Fugu cyclosporine precursor I purified by utilizing the methyl tertiary butyl ether are not further improved.

From the above examples 1 and 2, it can be seen that the preparation of the precursor of the cyclosporine by multiple recrystallization of the precursor of the cyclosporine, the preparation of the cyclosporine from the purified precursor of the cyclosporine has high purity and low isomer content, and the purification step of the chromatographic column is not required, so that the requirement on equipment is low, the production cost is low, and the industrial mass production is easy.

In combination with examples 3-6, the crude procyanidins of different purities were treated as above and the isomers Z/E were all satisfactory.

In comparative examples 7 to 10, the crude product of the procyanidin is purified and crystallized, the requirement on a solvent system is high, the crude product of the procyanidin is directly purified by adopting an acetone solvent system, no solid precipitation is found, and the crystallization cannot be purified. And acetone is adopted to be matched with other solvents for crystallization and purification, and no solid precipitation is found. Comparative example 11 it can be seen that the purification of the crude product of the precursor of the procyanidin by repeated recrystallization directly with methyl tert-butyl ether does not finally allow to obtain a product of the precursor of the procyanidin in high purity. According to the invention, after preliminary crystallization and purification are carried out by using methyl tertiary butyl ether, repeated recrystallization is carried out by using an acetone/water solvent system, so that the final obtained Fuciclosporin precursor has high purity, and the isomer impurities in the Fuciclosporin precursor are effectively removed.

Claims

1. A composition comprising a cyclosporine precursor, wherein the purity of the cyclosporine precursor in the composition is not less than 95% and the isomer ratio Z/E in the cyclosporine precursor is not more than 0.15/1;

2. The industrial refining and purifying method of the cyclosporine precursor is characterized by comprising the following steps of:

(1) Adding the crude product of the Fuciclosporin precursor into the organic solvent A for dissolving and clarifying, continuously stirring until crystallization, filtering, washing and drying to obtain the Fuciclosporin precursor I

3. The method for industrially purifying a cyclosporine precursor according to claim 2, wherein said step (2) is repeated not less than 2 times.

4. The method for industrially refining and purifying the cyclosporine precursor according to claim 2, wherein the ratio of the cyclosporine precursor I to the organic solvent B to the antisolvent C is 1:5-10:3-6 (Kg/L/L).

5. The method for industrially purifying a cyclosporine precursor according to claim 4, wherein the operation of said step (2) is specifically:

(a1) Adding the Fuciclosporin precursor I into the organic solvent B, stirring, dissolving and clarifying, then dripping the antisolvent C, crystallizing, filtering, washing and drying to obtain the Fuciclosporin precursor II;

The organic solvent B and the antisolvent C in the step (a 1) are 1:3-5:3 (Kg/L/L);

6. The method for industrially purifying a cyclosporine precursor according to claim 5, wherein said step (a 2) comprises the steps of (a) and (B) mixing said cyclosporine precursor II with an organic solvent (B) and mixing said mixture with an antisolvent (C) at a ratio of 1:10:6 (Kg/L/L).

7. The method for industrially purifying a cyclosporine precursor according to claim 5, wherein the number of repeating operations of said step (a 2) is not less than 1.

8. The method for industrially purifying and refining a procyanidin precursor according to claim 2, wherein the crude procyanidin precursor in step (1) is 1:1 to 4 (Kg/L), preferably 1:2 (Kg/L), of organic solvent a.

9. The method for industrially purifying a cyclosporine precursor according to claim 2, wherein the washing in step (1) is performed with n-heptane, an n-hexane solvent, preferably n-heptane.

10. The method for industrially purifying a cyclosporine precursor according to claim 2, wherein the washing in step (2) is performed using a mixed solution of an organic solvent B and an antisolvent C, preferably an organic solvent B: antisolvent C of 1:1 to 5, and more preferably an organic solvent B: antisolvent C of 1:1.