CN113929840A

CN113929840A - Hollow porous medium for separating and enriching taxane, preparation and application thereof

Info

Publication number: CN113929840A
Application number: CN202111182956.4A
Authority: CN
Inventors: 张华�; 余龙江; 付春华; 赵春芳; 徐航
Original assignee: Huazhong University of Science and Technology
Current assignee: Huazhong University of Science and Technology
Priority date: 2021-10-11
Filing date: 2021-10-11
Publication date: 2022-01-14
Anticipated expiration: 2041-10-11
Also published as: CN113929840B

Abstract

The invention belongs to the technical field of molecular imprinting polymers, and particularly relates to a hollow porous medium for separating and enriching taxane, and preparation and application thereof. The method comprises the steps of grafting a silanization reagent with double bonds on the surface of silica gel particles, forming a molecular imprinting polymerization layer with a certain thickness on the surface of the silica gel particles by using a surface molecular imprinting technology, and finally corroding amorphous silica gel and a cage-shaped silsesquioxane reagent in the molecular imprinting polymerization layer by using hydrofluoric acid, so that the hollow porous medium with high specific surface area and porosity and high selective adsorption performance on a taxane structure is obtained. The hollow porous medium obtained by the process can simultaneously have higher adsorption selectivity on various taxane substances, can realize high-efficiency synchronous separation and purification to obtain various taxanes, and has process application value.

Description

Hollow porous medium for separating and enriching taxane, preparation and application thereof

Technical Field

The invention belongs to the technical field of molecular imprinting polymers, and particularly relates to a hollow porous medium for separating and enriching taxane, and preparation and application thereof.

Background

Paclitaxel (Paclitaxel) is a diterpenoid compound, is used as a natural anticancer drug with high efficiency, broad spectrum, strong activity and unique action mechanism, especially for ovarian cancer, breast cancer, non-small cell lung cancer and the like, and is gradually popularized to more cancer treatments. However, because the content of paclitaxel in the branches and leaves of taxus chinensis is very low, which accounts for about 0.005-0.06% of the dry weight of the plant, and a large amount of taxane compounds with similar structures to the paclitaxel exist, in the traditional separation and purification process, the high-purity paclitaxel can be obtained only by combining multiple continuous normal phase silica gel column chromatographies with a reverse preparation chromatographic system, but various high value-added taxane products cannot be synchronously obtained. The paclitaxel has low production efficiency and high cost, and the current situation of short supply and short demand still exists in clinical application. Therefore, the technology for further developing the paclitaxel and the precursor substance thereof has good application prospect.

The molecular imprinting technique is a novel technique for preparing a high-molecular polymer material having strong affinity and high selectivity for a target compound (template molecule). The polymers produced are referred to as Molecularly Imprinted Polymers (MIPs). Since MIPs are synthesized using a specific target compound as a template, they are highly matched with template molecules in terms of spatial structure and functional base position, thereby having the ability to perform memory recognition on the template molecules and their structural analogs. This memory recognition is highly selective and specific. The molecular imprinting polymer is widely applied to various fields such as separation, analysis, catalysis, sensors and the like at present due to the unique property, and has very wide development prospect.

Patent document CN201611010222.7 discloses a preparation method of amino-modified mesoporous silica for separating and purifying paclitaxel, which has the application effect of increasing the content of paclitaxel in crude extract (0.4%) to about 10%, and is mainly used for crude separation, and is not used in the subsequent fine separation stage, and the separation efficiency in the subsequent fine separation stage still needs to be increased.

CN201510098761.X discloses a preparation method of a molecularly imprinted polymer capable of enriching paclitaxel, which comprises the steps of cross-linking and polymerizing paclitaxel serving as a template molecule and an acetone solvent serving as a pore-forming agent to obtain a block polymer, and washing away the template molecule by using methanol and acetic acid to obtain the molecularly imprinted polymer for enriching paclitaxel. Only in the examples, the treated samples filled with 200mg of filler demonstrated the enrichment effect on paclitaxel, but the specific adsorption enrichment efficiency was not quantitatively described. In addition, the MIPs obtained by the molecular imprinting technology are often seriously embedded with binding sites, a large number of binding sites cannot be utilized, and the adsorption capacity of the MIPs on template molecules is seriously influenced.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a hollow porous medium for efficiently and specifically separating and enriching taxane, a preparation method and application thereof, and the hollow porous medium for efficiently and specifically separating taxane has the following characteristics: the surface molecularly imprinted polymer layer with a certain thickness and a large number of binding sites which are partially or completely the same as the spatial structure of the paclitaxel are provided, so that the technical problems that the binding sites of molecularly imprinted polymer materials for separating and enriching the taxane in the prior art are seriously embedded, a large number of binding sites cannot be utilized, and the separation and enrichment efficiency of the taxane is low are solved.

In order to achieve the above object, the present invention provides a method for preparing a hollow porous medium for separating and enriching taxane, comprising the steps of:

(1) mixing the silica gel particles activated by the acid solution with a silanization reagent, modifying the silica gel particles, and washing and drying to obtain modified silica gel particles;

(2) mixing modified silica gel particles with a pore-foaming agent, template molecules, a functional monomer, a cross-linking agent, a cage-like silsesquioxane reagent and an initiator, degassing, reacting under the conditions of heating and stirring, and filtering, washing and drying after the reaction is finished to obtain a surface molecularly imprinted polymeric material;

(3) and (3) adding a hydrofluoric acid solution into the surface molecular imprinting polymer material obtained in the step (2), stirring, filtering, washing with water to be neutral, and drying to obtain the hollow porous medium for separating the taxane.

Preferably, the silylation reagent in the step (1) is one or more of vinyldimethylethoxysilane, vinyltrimethoxysilane, vinyltriethoxysilane, methacryloxypropyltrimethoxysilane and vinyltris (2-methoxyethoxy) silane, and the mass ratio of the silylation reagent to the acid-activated silica gel particles is 1: 30-1: 10, and more preferably 1: 15-1: 10.

Preferably, the porogen in step (2) is one or more of dichloromethane, chloroform and toluene, and further preferably dichloromethane.

Preferably, the weight parts of the template molecule in the step (2) are 0.16-0.64 part, the weight parts of the functional monomer are 0.12-0.48 part, the weight parts of the cross-linking agent are 0.8-2 parts, the weight parts of the cage-like silsesquioxane reagent are 0.16-0.8 part, and the weight parts of the initiator are 0.04-0.2 part.

Preferably, the template molecule of step (2) is one or more of 10-deacetylbaccatin III, baccatin III, paclitaxel, cephalomannine and 7-epi-paclitaxel.

Preferably, the functional monomer in step (2) is one or more of methacrylic acid, 2-vinylpyridine, 4-vinylpyridine and acrylamide, and is further preferably methacrylic acid.

Preferably, the crosslinking agent in the step (2) is one or more of ethylene glycol dimethacrylate, maleated rosin acrylate, trimethoxypropane trimethacrylate and divinylbenzene, and further preferably ethylene glycol dimethacrylate; the mass ratio of the modified silica gel particles to the cross-linking agent is 1: 1-1: 4, and the preferred mass ratio is 1: 1.5-1: 2.5.

Preferably, the caged silsesquioxane reagent in the step (2) is one or more of octavinyl-POSS, phenyl-POSS and amino-POSS, and further preferably octavinyl-POSS.

Preferably, the concentration of the hydrofluoric acid in the step (3) is 0.5-20 wt%, and more preferably 5-12 wt%.

According to another aspect of the invention, the hollow porous medium prepared by the preparation method is provided.

According to another aspect of the present invention, there is provided a use of the hollow porous medium for column chromatography for separation and enrichment of taxane; preferably used for the separation and enrichment of the taxane through medium and low pressure column chromatography, and further preferably used for the separation and enrichment of the taxane through normal phase medium and low pressure column chromatography.

Preferably, during the normal phase medium and low pressure column chromatography separation process, the mobile phase is one or more of dichloromethane, methanol, ethyl acetate and chloroform.

Generally, compared with the prior art, the above technical solution conceived by the present invention has the following beneficial effects:

(1) the invention prepares the hollow porous medium material for separating taxane, introduces a cage-shaped silsesquioxane reagent into a cross-linking polymerization system, and the reagent is polymerized with a template molecule under the action of a functional monomer and a cross-linking agent in the process of simultaneously participating in the preparation of a molecularly imprinted polymer, and then a large amount of pores can be obtained by means of the corrosion effect of hydrofluoric acid on silicon oxide and silicon-oxygen bonds in the cage-shaped silsesquioxane reagent, so that the utilization rate of binding sites generated by molecularly imprinted polymeric filler is improved, and the imprinting effect and the adsorption performance are improved. The method can prepare molecularly imprinted polymers with different templates.

(2) The preparation method of the hollow porous medium has simple process and easy implementation, and can better recover expensive template molecules, thereby greatly reducing the production cost. Meanwhile, under the treatment of hydrofluoric acid, a large number of pores are obtained, the specific surface area is increased, and the prepared molecularly imprinted filler with a shell-core structure can be used in the separation stage of taxus crude extracts, can synchronously obtain various high-purity taxane substances, and has industrial production value.

(3) The hollow porous medium prepared by the invention comprises the following structure: the surface molecularly imprinted polymer layer has binding sites with similar spatial structure with the taxane; the inside of the molecularly imprinted polymeric layer is of a hollow structure. The method comprises the steps of grafting a silanization reagent with double bonds on the surface of amorphous silica gel particles, forming a molecular imprinting polymerization layer with a certain thickness on the surface of the silica gel particles by using a surface molecular imprinting technology, and finally corroding the amorphous silica gel and a cage-shaped silsesquioxane reagent in the molecular imprinting polymerization layer by using hydrofluoric acid, so that a hollow porous medium with high specific surface area and porosity and high selective adsorption performance on a taxane structure is obtained. The invention has the advantages that: the hollow porous medium obtained by the process can simultaneously have higher adsorption selectivity on various taxane substances, can realize high-efficiency synchronous separation and purification to obtain various taxanes, and has process application value.

Drawings

FIG. 1 is a schematic flow chart of a method for preparing the hollow porous medium according to the present invention;

FIG. 2 is a morphology analysis diagram of the hollow porous medium porous core-shell structured filler prepared in example 1, wherein the content (a) is a scanning electron microscope diagram, and the content (b) is a transmission electron microscope diagram;

FIG. 3 is a morphology analysis diagram of the hollow porous medium porous core-shell structured filler prepared in example 2, wherein the content (a) is a scanning electron microscope diagram, and the content (b) is a transmission electron microscope diagram;

FIG. 4 is a morphology analysis diagram of the hollow porous medium porous core-shell structured filler prepared in example 3, wherein the content (a) is a scanning electron microscope diagram, and the content (b) is a transmission electron microscope diagram;

FIG. 5 is a morphology analysis diagram of the hollow porous medium porous core-shell structured filler prepared in example 4, wherein the content (a) is a scanning electron microscope diagram, and the content (b) is a transmission electron microscope diagram;

FIG. 6 shows FT-IR results for hollow porous media porous core-shell structured fillers prepared in examples 1, 2, 3 and 4.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The invention provides a preparation method of a hollow porous medium for separating taxane, which comprises the following steps as shown in figure 1:

(2) mixing modified silica gel particles with a pore-foaming agent, adding template molecules, a functional monomer, a cross-linking agent, cage-like silsesquioxane and an initiator, mixing, degassing, carrying out polymerization reaction under the conditions of heating and stirring, and filtering, washing and drying after the reaction is finished to obtain a surface molecularly imprinted polymeric material;

(3) and (3) adding a hydrofluoric acid solution into the surface molecular imprinting polymer material obtained in the step (2), stirring, filtering, washing with pure water to be neutral, and drying to obtain the hollow porous medium for separating the taxane.

The invention forms a molecular imprinting polymerization layer on the surface of amorphous silica gel particles by a surface molecular imprinting technology, and obtains a hollow porous medium with binding sites similar to a taxane spatial structure by an erosion technology.

In some embodiments, the silica gel activation and modification treatment in step (1) specifically comprises: placing the amorphous silica gel particles in a dilute nitric acid solution with the concentration of 20-50 wt% for stirring, filtering, washing with pure water to be neutral, and drying to obtain activated amorphous silica gel particles A; and (3) adding the A into toluene, adding a silanization reagent for modification, washing with methanol and pure water, and drying to obtain modified silica gel particles B.

In order to form a stable molecularly imprinted polymeric layer on the surface of silica gel, a silanization reagent with double bond groups is grafted. In some embodiments, the silylation agent is one or more of vinyldimethylethoxysilane, vinyltrimethoxysilane, vinyltriethoxysilane, methacryloxypropyltrimethoxysilane and vinyltris (2-methoxyethoxy) silane, and the mass ratio of the silylation agent to the acid-activated silica gel is 1:30 to 1:10, preferably 1:15 to 1: 10.

In some embodiments, the amorphous silica gel particles have a particle size of 20-100 μm, such as 20 μm, 25 μm, 32 μm, 46 μm, 51 μm, 71 μm, 98 μm, 100 μm, and the like.

In some embodiments, the surface molecular imprinting treatment in step (2) is specifically: adding B into a pore-foaming agent, and adding a certain amount of template molecules, functional monomers, a cross-linking agent, cage-like silsesquioxane and an initiator, wherein the pore-foaming agent comprises the following components in parts by weight: 0.16-0.64% of template molecules, 0.12-0.48% of functional monomers, 0.8-2% of cross-linking agents, 0.04-0.2% of initiators, 0.05-0.5% of cage-like silsesquioxane reagents and the balance of pore-forming agents to 100%, ultrasonically mixing and degassing for 20-50min, treating for 5-24 h under the conditions of heating and stirring at 30-60 ℃, and filtering, washing and drying after the reaction is finished to obtain the surface molecularly imprinted polymeric material C.

In the process of bulk molecular imprinting polymerization, a polymerization monomer and a free radical initiator are added into a reaction solution to initiate double bond polymerization; the double bond polymerization can occur between the polymerized monomer groups grafted on the silica gel and between the polymerized monomer groups and the functional monomer or the cage-shaped silsesquioxane, so that the template molecules can be better embedded into the imprinting polymerization layer, and then the utilization rate of the imprinting binding sites is improved by utilizing the corrosion technology.

In some embodiments, the porogen is one or more of dichloromethane, chloroform, and toluene, preferably dichloromethane.

In some embodiments, the template molecule is one or more of 10-deacetylbaccatin III, baccatin III, paclitaxel, cephalomannine, and 7-epi-paclitaxel.

In some embodiments, the functional monomer is one or more of methacrylic acid, 2-vinylpyridine, 4-vinylpyridine, and acrylamide, preferably methacrylic acid.

In some embodiments, the crosslinking agent is one or more of ethylene glycol dimethacrylate, maleated rosin acrylate, trimethoxypropane trimethacrylate, and divinylbenzene, preferably ethylene glycol dimethacrylate; the mass ratio of the modified silica gel particles to the cross-linking agent is 1: 1-1: 4, and preferably 1: 1.5-1: 2.5. The initiator may be AIBN (azobisisobutyronitrile) or other commonly used initiators.

In some embodiments, the caged silsesquioxane reagent is one or more of octavinyl-POSS, phenyl-POSS, amino-POSS, preferably octavinyl-POSS.

In some embodiments, after the polymerization reaction is finished, the template molecules are eluted and recovered by using a mixed solution of methanol and acetic acid (V: V ═ 9:1) by using a combined mode of ultrasound and soxhlet extraction, and residual acetic acid is washed by using methanol to obtain the surface molecularly imprinted polymeric material with molecularly imprinted binding sites.

In some embodiments, the step (3) of etching to obtain the hollow porous structure specifically comprises: and adding a hydrofluoric acid solution with a certain volume into the C, stirring, filtering, washing with pure water to be neutral, and drying to obtain the porous shell-core structure material D.

The solvent used in the corrosion technology is hydrofluoric acid, in some embodiments, the concentration of the hydrofluoric acid is 0.5-20 wt%, preferably 5-12 wt%, and the solution is stirred and corroded for 1-6 hours.

The hollow porous medium prepared by the preparation method provided by the invention has the same particle size as the initial silica gel, and can be used for column chromatography and a stationary phase for column chromatography separation. Preferably used for medium and low pressure column chromatography separation, and further preferably used for normal phase medium and low pressure column chromatography separation; in a preferred embodiment, during the normal phase medium-low pressure column chromatography separation process, the mobile phase is one or a mixture of dichloromethane, methanol, ethyl acetate and chloroform.

The molecular imprinting technology is a novel technology for preparing a high-molecular polymer material with strong affinity and high selectivity to a target compound (template molecule). The polymers produced are referred to as Molecularly Imprinted Polymers (MIPs). Since MIPs are synthesized using a specific target compound as a template, they are highly matched with template molecules in terms of spatial structure and functional base position, thereby having the ability to perform memory recognition on the template molecules and their structural analogs. MIPs materials obtained by molecular imprinting technology are often seriously embedded with binding sites, a large number of binding sites cannot be utilized, and the adsorption capacity of the MIPs on template molecules is seriously influenced. The caged silsesquioxane (POSS) has an inorganic core consisting of a silica skeleton, is mostly polyhedral in shape, is named polyhedral silsesquioxane, has three-dimensional size of 1nm to 3nm, can be polymerized with paclitaxel molecules under the action of functional monomers and a cross-linking agent in the process of simultaneously participating in the preparation of a molecularly imprinted polymer, and can obtain a large number of pores by means of the corrosion effect of hydrofluoric acid on silicon oxide, thereby improving the utilization rate of paclitaxel binding sites.

In some embodiments of the present invention, paclitaxel is used as a template molecule, and the prepared molecularly imprinted material has a spatial site with the same structure as paclitaxel, and has a higher adsorption capacity for paclitaxel, wherein paclitaxel belongs to one of taxanes, and other taxane molecules have a similar structure to paclitaxel and also have a higher adsorption capacity. Similarly, the template molecule can also be 10-deacetylbaccatin III, baccatin III, cephalomannine and 7-epi-taxol.

The hollow porous medium for efficiently and specifically separating the taxane provided by the invention adopts the molecularly imprinted polymer as a solid phase extraction separation enrichment material, and can effectively improve the separation and purification efficiency of the paclitaxel and various taxanes.

The following are specific examples:

example 1

A preparation method of a porous medium for efficiently and specifically separating taxane comprises the following steps:

(1) silica gel activation and modification treatment

Weighing 10g of amorphous silica gel, adding the amorphous silica gel into 200mL of 20 wt% dilute nitric acid solution, stirring, filtering, washing with pure water to be neutral, and drying to obtain activated amorphous silica gel particles P1A; 10g P1A was added to 200mL of toluene solution, and 0.33mL of vinyltrimethoxysilane was added and the reaction was carried out for 24 hours to obtain modified silica gel P1B.

(2) Surface molecular imprinting technique treatment

0.05mmol of paclitaxel and 0.5mmol of functional monomer methacrylic acid are weighed, fully dissolved in 20mL of dichloromethane and ultrasonically mixed for 10 min. 1g of the above P1B, 1mmol of crosslinking agent ethylene glycol dimethacrylate, 0.05mmol of octaphenyl-POSS and 20mg of initiator AIBN were added, and after thorough mixing, ultrasound was performed for 10min and nitrogen was degassed for 30 min. Putting the round-bottom flask into a magnetic stirrer, reacting for 6h under the condition of 40 ℃ and stirring speed of 100rpm, then eluting with a methanol and acetic acid mixed solution (V: V ═ 9:1) by adopting a mode of combining ultrasound and Soxhlet extraction, recovering template molecules, and washing residual acetic acid with methanol to obtain the surface molecular imprinting polymeric filler P1C with the molecular imprinting binding sites.

20mL of 5% hydrofluoric acid solution was added to the above P1C, magnetically stirred for 6 hours, filtered, washed with pure water to neutrality, and dried to obtain a porous core-shell structured filler P1D.

The SEM and TEM of the P1D material are shown in fig. 2 (a) and fig. 2 (b), respectively, and FTIR results of the material are shown in fig. 6, wherein the core collapse inside can be seen by scanning electron microscopy, and disappearance of the inner silica gel structure can be confirmed by transmission electron microscopy. Meanwhile, the internal existence of SI-O bonds or the obvious reduction of SI-O bonds can be further judged by FTIR, so that the obtained filler can be seen to have an obvious shell-core structure.

(3) Use of fillers

Filling a filler P1D into a normal-phase medium-low pressure column chromatography system, eluting by using dichloromethane and methanol according to different gradients of 40:1, 30:1, 20:1 and 10:1 in volume ratio, wherein the volume of an elution solvent is 3 times that of the column, collecting in batches, detecting by using a high performance liquid chromatography, and combining the same components to obtain various high-purity taxane products.

Meanwhile, a normal-phase medium-low pressure column chromatography system is filled, an initial sample (wherein the purity of the paclitaxel is 15.1%, the purity of the cephalomannine is 12.7%, and the purity of the 7-table 10-deacetyl paclitaxel is 8.9%) can be subjected to gradient elution, a plurality of high-purity taxane products can be synchronously obtained, the purity of the paclitaxel is successfully improved to 65.4%, the purity of the cephalomannine is improved to 46.1%, the purity of the 7-table 10-deacetyl paclitaxel is improved to 36.2%, and the concentration and the improvement multiple of the sample before and after treatment are shown in table 1.

TABLE 1 purification of crude taxane by column chromatography system packed in P1D

Example 2

(1) silica gel activation and modification treatment

Weighing 10g of amorphous silica gel, adding the amorphous silica gel into 200mL of 35 wt% dilute nitric acid solution, stirring, filtering, washing with pure water to be neutral, and drying to obtain activated amorphous silica gel particles P2A; 10g of 2A was added to 200mL of a toluene solution, and 0.67mL of methacryloxypropyltrimethoxysilane was added thereto and reacted for 24 hours to obtain a modified silica gel P2B.

(2) Surface molecular imprinting technique treatment

Weighing 1g P2B, adding 0.05mmol of paclitaxel and 0.4mmol of functional monomer 2-vinylpyridine, fully dissolving in 20mL of toluene, and ultrasonically mixing for 10 min. Adding 2mmol of cross-linking agent maleated rosin acrylate, 0.2mmol of octavinyl-POSS, adding 40mg of initiator AIBN, fully mixing, performing ultrasonic treatment for 10min, and degassing for 30min by nitrogen. Putting the round-bottom flask into a magnetic stirrer, reacting for 6h under the condition of 40 ℃ and stirring speed of 100rpm, then eluting with a methanol and acetic acid mixed solution (V: V ═ 9:1) by adopting a mode of combining ultrasound and Soxhlet extraction, recovering template molecules, and washing residual acetic acid with methanol to obtain the surface molecular imprinting polymeric filler P2C with the molecular imprinting binding sites.

20mL of 10% hydrofluoric acid solution was added to the P2C solution, magnetically stirred for 6 hours, filtered, washed with pure water to neutrality, and dried to obtain a porous core-shell structured filler P2D.

The SEM and TEM of the P2D material are shown in fig. 3 (a) and fig. 3 (b), respectively, and FTIR results of the material are shown in fig. 6, and it can be seen that the obtained filler has a distinct core-shell structure.

(3) Use of fillers

Filling a filler P2D into a normal-phase medium-low pressure column chromatography system, eluting by using dichloromethane and methanol according to different gradients of 40:1, 30:1, 20:1 and 10:1 in volume ratio, collecting elution solvents in batches, detecting by using a high performance liquid chromatography, and combining the same components to obtain various high-purity taxane products.

Meanwhile, a normal-phase medium-low pressure column chromatography system is filled, an initial sample (wherein the purity of the paclitaxel is 15.1%, the purity of the cephalomannine is 12.7%, and the purity of the 7-table 10-deacetyl paclitaxel is 8.9%) can be subjected to gradient elution, a plurality of high-purity taxane products can be synchronously obtained, the purity of the paclitaxel is successfully improved to 67.4%, the purity of the cephalomannine is improved to 49.2%, the purity of the 7-table 10-deacetyl paclitaxel is improved to 36.7%, and the concentration and the improvement multiple of the sample before and after treatment are shown in table 2.

TABLE 2 purification of crude taxane by column chromatography system packed in P2D

Example 3

(1) silica gel activation and modification treatment

Weighing 10g of amorphous silica gel, adding the amorphous silica gel into 200mL of 50 wt% dilute nitric acid solution, stirring, filtering, washing with pure water to be neutral, and drying to obtain activated amorphous silica gel particles P3A; 10g P3A was added to 200mL of toluene solution, and 1mL of vinyltris (2-methoxyethoxy) silane was added to the solution to react for 24 hours to obtain modified silica gel P3B.

(2) Surface molecular imprinting technique treatment

Weighing 1g P3B, adding 0.2mmol of paclitaxel and 0.8mmol of functional monomer 4-vinylpyridine, fully dissolving in 20mL of toluene, and ultrasonically mixing for 10 min. Adding 4mmol of cross-linking agent trimethoxy propane trimethacrylate, 0.5mmol of octaamino-POSS, adding 50mg of initiator AIBN, fully mixing, performing ultrasonic treatment for 10min, and degassing for 30min by nitrogen. Putting the round-bottom flask into a magnetic stirrer, reacting for 6h under the condition of 40 ℃ and stirring speed of 100rpm, then eluting with a methanol and acetic acid mixed solution (V: V ═ 9:1) by adopting a mode of combining ultrasound and Soxhlet extraction, recovering template molecules, and washing residual acetic acid with methanol to obtain the surface molecular imprinting polymeric filler P3C with the molecular imprinting binding sites.

20mL of 20% hydrofluoric acid solution was added to the above P3C, magnetically stirred for 6 hours, filtered, washed with pure water to neutrality, and dried to obtain a porous core-shell structured filler P3D.

The SEM and TEM of the P3D material are shown in FIG. 4 (a) and FIG. 4 (b), respectively, and FTIR results of the material are shown in FIG. 6, and it can be seen that the obtained filler has a distinct core-shell structure

(3) Use of fillers

Filling a filler P3D into a normal-phase medium-low pressure column chromatography system, eluting by using dichloromethane and methanol according to different gradients of 40:1, 30:1, 20:1 and 10:1 in volume ratio, collecting elution solvents in batches, detecting by using a high performance liquid chromatography, and combining the same components to obtain various high-purity taxane products.

Meanwhile, a normal-phase medium-low pressure column chromatography system is filled, an initial sample (wherein the purity of the paclitaxel is 15.1%, the purity of the cephalomannine is 12.7%, and the purity of the 7-table 10-deacetyl paclitaxel is 8.9%) can be subjected to gradient elution, a plurality of high-purity taxane products can be synchronously obtained, the purity of the paclitaxel is successfully improved to 62.1%, the purity of the cephalomannine is improved to 41.2%, the purity of the 7-table 10-deacetyl paclitaxel is improved to 31.6%, and the like, and the concentration and the improvement times of the sample before and after treatment are shown in table 3.

TABLE 3 purification of crude taxane by column chromatography system packed in P3D

Comparative example 1

The other conditions were the same as in example 3, except that no caged silsesquioxane was added in the step (2), and the porous shell-core structured filler P4D was prepared according to the same method and procedure.

The SEM and TEM of the P4D material are shown in fig. 5 (a) and fig. 5 (b), respectively, and FTIR results of the material are shown in fig. 6, and it can be seen that the obtained filler has a distinct core-shell structure.

(3) Use of fillers

Filling a filler P4D into a normal-phase medium-low pressure column chromatography system, eluting by using dichloromethane and methanol according to different gradients of 40:1, 30:1, 20:1 and 10:1 in volume ratio, collecting elution solvents in batches, detecting by using a high performance liquid chromatography, and combining the same components to obtain various high-purity taxane products.

Meanwhile, a normal-phase medium-low pressure column chromatography system is filled, an initial sample (wherein the purity of the paclitaxel is 15.1%, the purity of the cephalomannine is 12.7%, and the purity of the 7-table 10-deacetyl paclitaxel is 8.9%) can be subjected to gradient elution, a plurality of high-purity taxane products can be synchronously obtained, the purity of the paclitaxel is successfully improved to 46.1%, the purity of the cephalomannine is improved to 32.6%, the purity of the 7-table 10-deacetyl paclitaxel is improved to 21.5%, and the like, and the concentration and the improvement times of the sample before and after treatment are shown in table 4.

TABLE 4 purification of crude taxane by column chromatography system packed in P4D

As can be seen from the results in tables 1, 2, 3 and 4, in examples 1 to 3, when the caged silsesquioxane reagent is introduced into the bulk polymerization system, compared to comparative example 1 without introducing the caged silsesquioxane reagent, the purity of different kinds of taxane molecules is greatly improved when the prepared molecularly imprinted polymer filler is used for separating and enriching taxane products, and the purity of paclitaxel can be improved to 67.4% by using the molecularly imprinted polymer filler prepared in the embodiment of the present invention, which is usually achieved by adding an additional column chromatography process or crystallization process if the paclitaxel concentration needs to be improved from 56% to 65% in the prior art.

It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims

1. A preparation method of a hollow porous medium for separating and enriching taxane is characterized by comprising the following steps:

2. The preparation method according to claim 1, wherein the silanization reagent in the step (1) is one or more of vinyldimethylethoxysilane, vinyltrimethoxysilane, vinyltriethoxysilane, methacryloxypropyltrimethoxysilane and vinyltris (2-methoxyethoxy) silane, and the mass ratio of the silanization reagent to the acid-activated silica gel particles is 1:30 to 1: 10.

3. The preparation method according to claim 1, wherein the porogen in step (2) is one or more of dichloromethane, chloroform and toluene.

4. The preparation method according to claim 1, wherein the template molecule in the step (2) is 0.16 to 0.64 parts by weight, the functional monomer is 0.12 to 0.48 parts by weight, the crosslinking agent is 0.8 to 2 parts by weight, the cage-like silsesquioxane agent is 0.16 to 0.8 parts by weight, and the initiator is 0.04 to 0.2 parts by weight.

5. The method of claim 4, wherein the template molecule of step (2) is one or more of 10-deacetylbaccatin III, baccatin III, paclitaxel, cephalomannine, and 7-epi-paclitaxel.

6. The method of claim 1, wherein the functional monomer of step (2) is one or more of methacrylic acid, 2-vinylpyridine, 4-vinylpyridine and acrylamide.

7. The method according to claim 1, wherein the crosslinking agent in the step (2) is one or more of ethylene glycol dimethacrylate, maleated rosin acrylate, trimethoxypropane trimethacrylate and divinylbenzene; the mass ratio of the modified silica gel particles to the cross-linking agent is 1: 1-1: 4, and preferably 1: 1.5-1: 2.5.

8. The method of claim 1, wherein the caged silsesquioxane reagent of step (2) is one or more of octavinyl-POSS, phenyl-POSS, and amino-POSS.

9. A hollow porous medium produced by the production method according to any one of claims 1 to 8.

10. Use of the hollow porous medium of claim 9 for column chromatography to enrich for a taxane.