阅读说明：本技术 特拉万星的纯化方法 (Purification method of telavancin ) 是由 贺元康 王艺 于 2019-11-27 设计创作，主要内容包括：本发明涉及一种特拉万星的纯化方法。包括如下步骤：将特拉万星粗品加入溶剂中,充分溶解后过滤并保留滤液,得到上样液；将上样液上样至已装填有填料的分离柱,用以分离上样液中的目标组分和杂质组分,上样之后的填料中保留有目标组分；采用洗脱液对填料中的目标组分进行洗脱,得到目标流出液,洗脱液为有机溶剂与水的混合液或者有机溶剂；以及收集目标流出液,即得纯化后的特拉万星。本发明的上述特拉万星的纯化方法无需经过萃取、柱层析、结晶等步骤,工艺简单,生产成本较低,整体有利于应用。经试验证明,采用本发明的特拉万星的纯化方法得到的特拉万星纯品的纯度大于99％,单杂小于0.5％,周期短,具有较好的可行性与实用性,适合工业化生产。(The invention relates to a purification method of telavancin. The method comprises the following steps: adding the crude telavancin into a solvent, fully dissolving, filtering and retaining filtrate to obtain a sample solution; loading the sample loading liquid to a separation column filled with a filler to separate a target component and an impurity component in the sample loading liquid, wherein the target component is remained in the filler after the sample loading; eluting the target component in the filler by using an eluent to obtain a target effluent liquid, wherein the eluent is a mixed liquid of an organic solvent and water or the organic solvent; and collecting the target effluent to obtain the purified telavancin. The purification method of telavancin does not need steps of extraction, column chromatography, crystallization and the like, has simple process and lower production cost, and is integrally beneficial to application. Tests prove that the purity of the purified telavancin product obtained by the purifying method of telavancin is more than 99%, the single impurity content is less than 0.5%, the period is short, and the method has good feasibility and practicability and is suitable for industrial production.)

1. A method for purifying telavancin is characterized by comprising the following steps:

adding the crude telavancin into a solvent, fully dissolving, filtering and retaining filtrate to obtain a sample solution;

loading the sample loading liquid to a separation column filled with a filler to separate a target component and an impurity component in the sample loading liquid, wherein the target component is remained in the filler after the sample loading;

eluting the target component in the filler by adopting an eluent to obtain a target effluent, wherein the eluent is a mixed solution of an organic solvent and water or an organic solvent; and

and collecting the target effluent to obtain the purified telavancin.

2. The method for purifying telavancin according to claim 1, wherein the crude telavancin is added to a solvent, wherein the solvent is one or more of an aqueous methanol solution containing 0.05-0.5% by volume of organic acid and an aqueous acetonitrile solution containing 0.05-0.5% by volume of organic acid; wherein the organic acid is one or more of trifluoroacetic acid, formic acid and acetic acid; the volume fraction of methanol or acetonitrile in the solvent is 20-50 percent respectively.

3. A purification process of telavancin according to claim 1, wherein the filler is silica gel-based, and decaoctaalkylsilane and divinylbenzene are bonded to the silica gel, wherein the mass ratio of octadecylsilane to divinylbenzene is (1-6): 1.

4. A process for the purification of telavancin according to claim 3, wherein the filler has a particle size of 10 to 50 μm and a pore size of 10 to 50 μm

5. The method for purifying telavancin according to claim 1, wherein a mass ratio of the solute in the sample liquid to the filler is 1:100 to 11:100 in an operation of loading the sample liquid into a separation column filled with the filler.

6. The method of purifying telavancin according to claim 1, further comprising, prior to the step of loading the loading solution onto a packed separation column: the separation column filled with the packing is equilibrated with an eluent.

7. The method for purifying telavancin according to claim 1 or 6, wherein the eluent is one or more of an aqueous methanol solution containing 0.05-0.5% by volume of organic acid and an aqueous acetonitrile solution containing 0.05-0.5% by volume of organic acid; wherein the organic acid is one or more of trifluoroacetic acid, formic acid and acetic acid; the volume fraction of methanol or acetonitrile in the solvent is 20-50 percent respectively.

8. The method of claim 1, wherein the elution of the target component with an eluent is carried out at a flow rate of 100cm/h to 500 cm/h.

9. The method of purifying telavancin according to claim 1, further comprising, after the step of collecting the target effluent in stages: and detecting the target effluent, and freeze-drying the qualified sample after detection.

10. The method for purifying telavancin according to claim 1, wherein the purity of the crude telavancin is 70% to 90%.

Technical Field

The invention relates to the field of medicines, in particular to a method for purifying telavancin.

Background

Telavancin (Telavancin) is a novel lipoglycopeptide antibiotic developed by schwann (Theravance) and Astellas (Astellas) pharmaceutical, japan. The chemical name is N-3' -2- (decylamino) ethyl-29- [ (N-phosphonomethyl) aminomethyl ] vancomycin hydrochloride. Marketed in the united states in 2009, it was indicated for adult complex skin infections caused by gram-positive bacteria. The european union has increased usage in 2011 for the treatment of adult hospital-acquired pneumonia and ventilator-associated pneumonia that have been diagnosed or suspected to be caused by methicillin-resistant staphylococcus aureus (MRSA). Telavancin is the second generation of semi-synthetic lipoglycopeptide antibiotics, and the parent drug core of telavancin is vancomycin. Compared with vancomycin, telavancin is formed by connecting a hydrophobic group, namely decylaminoethyl, to vancomycin sugar amine, so that the affinity of the vancomycin sugar amine to a bacterial cell membrane is favorably increased, the membrane anchoring is favorably realized, the half-life period is prolonged, and the antibacterial activity to enterococcus is improved; and the acyl position of the 7 th amino acid is connected with the amino phosphate, so that the water solubility is increased, the metabolism and the clearance of tissues in vivo are promoted, the renal toxicity is reduced, and the pharmacokinetic property of the medicament in vivo is improved. As a novel lipoglycopeptide antibiotic, the unique action mechanism and good clinical treatment effect of telavancin can be expected to have wide market prospect.

Telavancin is not pure enough and further purification is required. However, the conventional purification method of telavancin is complicated, requires extraction, column chromatography, crystallization and other steps, is time-consuming, leads to high cost, is difficult to industrially amplify and produce, and is not beneficial to application.

Disclosure of Invention

Therefore, it is necessary to provide a purification method of telavancin with simple process and high purity for the problem of complicated purification steps of telavancin.

A method for purifying telavancin, comprising the following steps:

adding the crude telavancin into a solvent, fully dissolving, filtering and retaining filtrate to obtain a sample solution;

loading the sample loading liquid to a separation column filled with a filler to separate a target component and an impurity component in the sample loading liquid, wherein the target component is remained in the filler after the sample loading;

eluting the target component in the filler by adopting an eluent to obtain a target effluent, wherein the eluent is a mixed solution of an organic solvent and water or an organic solvent; and collecting the target effluent to obtain the purified telavancin.

Compared with the traditional purification method of telavancin, the purification method of telavancin only needs to load the sample loading solution to a separation column, then elute and reserve the target effluent, and collect to obtain the purified telavancin. Without the steps of extraction, column chromatography, crystallization and the like. Therefore, the purification method of telavancin has the advantages of simple process, low production cost and overall benefit for application. In addition, tests prove that the purity of the purified telavancin product obtained by the purification method of telavancin is more than 99%, the single impurity content is less than 0.5%, the period is short, and the method has good feasibility and practicability and is very suitable for industrial production.

In one embodiment, the crude telavancin is added into a solvent, wherein the solvent is one or more of a methanol aqueous solution containing 0.05-0.5% by volume of organic acid and an acetonitrile aqueous solution containing 0.05-0.5% by volume of organic acid; wherein the organic acid is one or more of trifluoroacetic acid, formic acid and acetic acid; the volume fraction of methanol or acetonitrile in the solvent is 20-50 percent respectively.

In one embodiment, the filler is silica gel as a matrix, and decaoctasilane and divinylbenzene are bonded on the silica gel, wherein the mass ratio of octadecylsilane to divinylbenzene is (1-6): 1.

In one embodiment, the filler has a particle size of 10 to 50 μm and a pore size of

In one embodiment, in the operation of loading the loading solution to the separation column filled with the filler, the mass ratio of the solute in the loading solution to the filler is 1: 100-11: 100.

In one embodiment, the step of loading the loading solution to the separation column filled with the filler further comprises the following steps: the separation column filled with the packing is equilibrated with an eluent.

In one embodiment, the eluent is one or more of methanol aqueous solution containing 0.05-0.5% of organic acid volume fraction and acetonitrile aqueous solution containing 0.05-0.5% of organic acid volume fraction; wherein the organic acid is one or more of trifluoroacetic acid, formic acid and acetic acid; the volume fraction of methanol or acetonitrile in the solvent is 20-50 percent respectively.

In one embodiment, the target component is eluted by an eluent at a flow rate of 100cm/h to 500 cm/h.

In one embodiment, the step of collecting the target effluent in stages further comprises the steps of: and detecting the target effluent, and freeze-drying the qualified sample after detection.

In one embodiment, the crude telavancin is 70% to 90% pure.

Drawings

FIG. 1 is a flow diagram of a process for purifying telavancin in accordance with one embodiment of the present invention;

FIG. 2 is a high performance liquid chromatogram of example 1 of the present invention after purification of crude Telavancin.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Referring to fig. 1, the method for purifying telavancin according to an embodiment of the present invention includes the following steps:

s10, adding the crude telavancin into the solvent, fully dissolving, filtering and retaining the filtrate to obtain a sample solution.

Preferably, the purity of the crude telavancin is 70% to 90%. That is to say, the purification method of telavancin can be applied to crude telavancin products with the purity of 70% -90%, and tests prove that the purification effect is good.

Preferably, in the operation of adding the crude telavancin into the solvent, the solvent is one or more of methanol aqueous solution containing 0.05-0.5% of organic acid by volume fraction and acetonitrile aqueous solution containing 0.05-0.5% of organic acid by volume fraction; wherein the organic acid is one or more of trifluoroacetic acid, formic acid and acetic acid; the volume fraction of methanol or acetonitrile in the solvent is 20-50 percent respectively. The solvents are easy to obtain, have low cost, can not react with telavancin or generate adverse effects, and are easy to remove subsequently.

And S20, loading the sample loading liquid obtained in the step S10 to a separation column filled with the filler, so as to separate the target component and the impurity component in the sample loading liquid, wherein the target component is remained in the filler after loading.

Wherein the separation column is a preparative liquid chromatography column. The filler is special for telavancin.

Preferably, the filler is silica gel as a matrix, and decaoctasilane and divinylbenzene are bonded on the silica gel, wherein the mass ratio of the octadecylsilane to the divinylbenzene is (1-6): 1.

More preferably, the filler has a particle size of 10 to 50 μm and a pore size of

Preferably, in the operation of loading the loading liquid to the separation column filled with the filler, the mass ratio of the solute in the loading liquid to the filler is 1: 100-11: 100.

Preferably, the step of loading the loading liquid to the separation column filled with the filler further comprises the following steps: and (4) balancing the separation column filled with the filler by using eluent, and starting to sample after the base line is leveled.

And S30, eluting the target component in the filler obtained in the step S20 by using an eluent to obtain a target effluent, wherein the eluent is a mixed solution of an organic solvent and water or the organic solvent.

Preferably, the eluent is one or more of methanol aqueous solution containing 0.05-0.5% of organic acid volume fraction and acetonitrile aqueous solution containing 0.05-0.5% of organic acid volume fraction; wherein the organic acid is one or more of trifluoroacetic acid, formic acid and acetic acid; the volume fraction of methanol or acetonitrile in the solvent is 20-50 percent respectively. These types of eluents are readily available, low cost, do not react with telavancin or otherwise adversely affect, and are subsequently easily removable.

Preferably, in the operation of eluting the target component by adopting the eluent, the flow rate of the eluent is 100 cm/h-500 cm/h; more preferably, the flow rate of the eluent is 200 cm/h-400 cm/h; most preferably, the elution flow rate is 300 cm/h.

S40, collecting the target effluent obtained in the step S30 to obtain the purified telavancin.

Preferably, the step of collecting the target effluent in stages further comprises the following steps: detecting the target effluent liquid, and freeze-drying the qualified sample after detection.

Compared with the traditional purification method of telavancin, the purification method of telavancin only needs to load the sample loading solution to a separation column, then elute and reserve the target effluent, and collect to obtain the purified telavancin. Without the steps of extraction, column chromatography, crystallization and the like. Therefore, the purification method of telavancin has the advantages of simple process, low production cost and overall benefit for application. In addition, tests prove that the purity of the purified telavancin product obtained by the purification method of telavancin is more than 99%, the single impurity content is less than 0.5%, the period is short, and the method has good feasibility and practicability and is very suitable for industrial production.

The purification process of telavancin according to the present invention will be further described with reference to specific examples and comparative examples.