UA68355C2 - Method for isolating and purifying taxan analogues, in particular paclitaxel (variants) - Google Patents

Abstract

The method for isolating and purifying the taxan analogues providing for high yield and high purity employs the membrane of the polymeric resin. Such a technique allows one isolating the taxan analogues avoiding the expensive and complicated techniques used at present for this purpose.

Description

Description of the invention

This invention relates to a method of isolation and, in particular, this invention relates to methods of isolation 2 of paclitaxel and related taxanes.

Paclitaxel is a well-known chemotherapy drug for the treatment of various types of metastatic cancer. It has been approved by the FDA (Food and Drug Administration) and the FDA (Health Protection Bureau) for the treatment of ovarian and breast cancer.

This compound is a natural product that was first isolated from the bark of the yew tree, Tahyvs Bhemigiia, and 70 is also found in T. rassaiia, T. maiispiapa, T. uppapepvi, and T. sapadepvi.

The concentration of paclitaxel in various raw materials is usually low, for example, somewhere between 0.0004-0.0195 by weight (m//lm) in yew bark. Such a low concentration makes it very difficult to isolate and purify the compound from raw materials for pharmaceutical use, which has made its production impractical from a commercial level until now. To purify 12 paclitaxel from the crude extract of the raw material, various normal or reverse phase chromatography methods, as well as column, low and high pressure chromatography methods were developed.

The success of low-pressure chromatography depends significantly on the nature of the column. Various problems are caused by the use of silica gel and aluminum oxide, all varieties of which are classic carriers of the stationary phase of separated systems. They form a stable stationary phase with most solvent-containing systems, however, they are strong absorbents and can participate in the separation process as long as they affect the chromatographic behavior and sample separation.

Chromatographic methods were developed for the determination and isolation of paclitaxel from various species of Tachyves based on analytical and preparative chromatography. Methods of allocation, mainly, feasible:.-sya. on a small laboratory scale. their disadvantages are a low level of selectivity and separation and a high cost of production. As a result, there is a serious and unmet need for an economically practical (3) method to separate the valuable antitumor compound paclitaxel from its close analogue cephalomannin, as well as other closely related taxanes.

Methods according to the prior art describe the use of various types of chromatographic procedures for the isolation of paclitaxel and related taxanes, which include normal and reverse phase chromatography on a silica gel or bonded silica gel column. Prior art methods have low yields, high production costs, and use multiple isolation steps that are difficult to adapt to large-scale industrial production. at

A typical prior art method is described in US Pat. No. 5,620,875, issued April 15, 1997, to Neutap et al. The document proposes the isolation of paclitaxel and other taxanes by a multistep process

Hexane extraction and high-performance liquid chromatography (HPLC). The method used is laborious and has only moderate yields of the desired compounds.

U.S. Patent No. 5,670,673, issued September 23, 1997, to Caso, describes the isolation and purification of taxol and its analogs. The method includes the use of reverse-phase liquid chromatography on C18 adsorbent with elution of adsorbed analogues. Despite the advantages of this method, it has limitations in terms of productivity and purity of the compounds obtained. This invention offers a simple separation method based on a polymer resin column that avoids the limitations of existing methods.

Therefore, the object of the present invention is to develop a simple method with a lower cost for the economic isolation and purification of important taxanes, such as 13-acetyl-9-dihydrobaccatin I, 10-deacetylbaccatin I, baccatin I, cephalomannin and paclitaxel.

Me Conventional methods for the isolation of taxanes, including paclitaxel, 13-acetyl-9-dihydrobaccatin Ш, and obaccatin Ш, generally include the steps of extracting the taxanes from the raw material with an alcohol solvent, defatting the extract, and isolating and purifying the individual taxane by chromatography. o One of the tasks of the present invention is the development of a method for isolating and purifying taxane analogues from a source containing taxanes, and the method includes: extraction of the source of taxanes in an organic extractant; sl covering the working absorbent medium with an extractant and loading the medium into the column, while the column contains the absorbent; elution in the first stage with a mixture of organic solvents at a pressure of 68.95-137.90 kPa (10-2 Orvi) to obtain 29 fractions containing taxane compounds;

HF) crystallization of fractions to obtain a solid taxane compound and mother liquor; concentration of mother liquor; and elution in the second stage of the mother liquor with a polar solvent mixture through the polymer resin to obtain at least the second taxane compound. 60 The next task of the present invention is to develop a method for isolating and purifying taxanes from a source containing taxanes, while the method includes: providing a source of taxanes; extracting taxanes from the source into an organic extraction medium to obtain an organic layer containing taxane compounds; because the treatment of the carrier material with an organic layer;

provision of a low-pressure column containing an absorbent; elution in the first stage of an organic solvent through a column for elution of purified taxane fractions; crystallization of taxane fractions to obtain the first taxane analog and mother liquor; elution in a second step of the mother liquor through a polymer resin in a chromatographic column to purify and elute at least the second taxane analogue and the third taxane analogue; and collection of isolated taxane analogs.

The next task of the present invention is to develop a method for isolating and purifying taxanes from a source that contains taxanes, and the method includes: providing a source of taxanes; extracting taxanes from said source into an organic extraction medium to obtain an organic layer containing taxane compounds; treatment of the carrier material with an organic layer; provision of a low-pressure column containing an absorbent; elution in the first stage at a pressure of 68.95-137.90 kPa (10-2 Orzi) of an organic solvent through a column for elution of purified taxane fractions and removal of flavonoid and lignan impurities; crystallization of taxane fractions to obtain the first taxane analog and mother liquor; elution in the second stage at a pressure of 206.zh4kPa (ZOrzi) of the mother solution through polystyrene-divinylbenzene resin In a chromatographic column for purification and elution of at least the second taxane analogue and the third taxane analogue; elution in the third step of the second taxane analogue and the third taxane analogue through a column of normal phase with silica gel to purify the second taxane analogue and the third taxane analogue; and collection of isolated taxane analogs. with

The original source of taxanes was a plant material, Tahiz sapadepviz, which is widely distributed in

Eastern Canada. Takhiz sapadepvziz twigs and needles or their mixtures were used. The material can be fresh or i) dried.

The raw material was ground and extracted with methanol at room temperature for 5 hours and filtered.

The methanol extract was mixed with charcoal and left for one hour at room temperature. Then this yu zo boomish was filtered. The filtrate was concentrated to about 1595 of the original volume by evaporation. A mixture of water:dichloromethane in a volume ratio of 1:11 (M/M) was added to the concentrate to obtain the paclitaxel-enriched part (organic layer). with

The organic layer was concentrated to reduced volume and applied to the support material. In this case, the carrier material was brownmillerite (Seillet 545). co

The coated material was loaded into the upper part of an industrial preparative column, for example, a column measuring 150 mm x 1500 mm, and filled with absorbent AI2O53. The column was eluted with a mixture of hexane and acetone (started with 100.0 and ended with 45:55) under a pressure of 68.95-137.90 kPa ( 10-20 days). Fractions containing taxanes were collected and analyzed by thin layer chromatography (TLC) and then pooled according to the TLC results. "

The collected fractions were then concentrated in vacuo, the obtained material was dissolved in methanol and kept at room temperature overnight. The formed crystal was filtered and washed. methanol, then recrystallized from acetone. 13-acetyl-9-dihydrobaccatin I was obtained in the form of a white i? crystal

The mother liquor was concentrated to dryness and the residue was dissolved in acetone. The acetone solution was mixed with a polymer resin (Oozehtm resin, polystyrene-divinylbenzene). The mixture was evaporated in vacuo to

Gee! remove acetone. The resulting powder was subjected to low-pressure chromatography. An industrial chromatographic column (150 mm x 1500 mm) was filled with Juozhmechtm resin and eluted with acetone:water (45:55, M/M) at a speed

The flow rate is 150O0ml/minute at a working pressure of 206.84 kPa (Zorzi). Fractions with a volume of approximately 2 L each were collected, which were monitored by TLC and HPLC. Fractions containing baccatin II and 10-deacetylbaccatin I were combined and evaporated in vacuo to remove almost all the organic solvent, then diluted with water and extracted with СНоСі. The extract was then concentrated to dryness and the resulting residue was then dissolved in EtOAc sl and purified on a normal phase silica gel column with hexane:EtOAc used as the eluent system (starting at 5:5 and ending at 35:65). In the end, we got baccatin I and 10-deacetylbaccatin I! in the form of a white powder, the purity of which exceeded 98905.

Fractions containing paclitaxel and cephalomannin were combined and evaporated in vacuo to remove substantially all of the acetone, then partitioned between water and CH 2 Cl 2 . The organic layer was separated and evaporated to dryness in vacuo. The residue was dissolved in methanol. About 3095 (m/v) (w/v) of water was added to the solution and the mixture was heated to 60°C for 5 minutes, then kept at room temperature overnight. The crude crystalline solid from the methanol solution was filtered off and dried in vacuum at 6o at 7070-7570 The solid was analyzed by HPLC to consist of about 70905 paclitaxel and 2596 cephalomannin and some other taxanes.

Crude paclitaxel can be processed according to one of three alternative methods. For the first possibility, it was dissolved in acetone and purified by reverse-phase column chromatography. The column was filled with polymer resin (Oiaiop resin, HP2MO) and eluted with acetonitrile:water (45:55). Paclitaxel 65 was obtained in the form of a white crystal, the purity of which exceeded 9995. Cephalomannin was obtained in the form of a white powder, the purity of which exceeded 98905.

Alternatively, for a second possible treatment, paclitaxel and cephalomannin were purified by chemical reaction and then purified by reverse-phase column chromatography on bonded silica gel. A mixture of crude paclitaxel and cephalomannin was dissolved in CH2Cl3 or CHCl3 (1:10, w/w) and reacted with 10 equivalents of bromine in a round-bottom flask in an ice bath (070-527) for approximately 40 minutes. The resulting mixture of paclitaxel and 2",3"-bromocephalomannin was easily separated on a reverse-phase chromatographic column (Civ or phenyl), while the eluting solvent was

SNUSM:NYO (45:55). Paclitaxel was obtained as a white crystal with a purity exceeding 9996. 2",8"-bromocephalomannin was obtained as a mixture of diastereomers which were reacted with activated zinc in 70 acetic acid at room temperature and which were then purified by flash column chromatography to to obtain pure cephalomannin with a purity greater than 9596.

For a third alternative method, paclitaxel and bromocephalomannin were purified on a normal phase silica gel column. Eluting solvent systems can include hexane:E(Ac (starting at 6:4 and ending at 4:6) or SNCl::Me»CO (starting at 8:2 and ending at 65:35), or SNCl2EHUAc at/5 ( starting at 75:25 and ending at 6:4.) As a result of this method, paclitaxel was obtained as a white crystal, the purity of which exceeded 9995, cephalomannin was present in an amount of less than 0.290.

Following the above discussion, the invention will now be described in more detail by way of examples.

Example 1

Approximately 200 kilograms of dry Takhiz sapadepsis twigs and needles were extracted with 1,000 liters of methanol at 80°C in an industrial multifunctional extractor for five hours and then filtered.

The raw material was extracted with 700 liters of methanol at a temperature of 55"C-60"C for another 4 hours and filtered. The filtrate was combined and mixed with 10 kilograms of activated carbon (595 m/m) and kept at room temperature for one hour, then filtered to remove the activated carbon.

The filtrate was then concentrated in vacuo to about 100 liters, then 300 liters of water:dichloromethane sc dv (11) were added. The organic layer was collected and the aqueous solution was extracted twice more with 200 liters of dichloromethane. The dichloromethane solution was combined and evaporated under vacuum until it acquired the consistency of sour cream, then it was diluted with i) 20 liters of acetone.

The acetone solution was applied to 20 kilograms of brownmillerite (SeiShe 545). The coated material was air-dried, and then it was loaded into the column I of three low-pressure industrial chromatographic columns (size: 150 x 15 cm). Each column was filled with 15 kilograms of aluminum oxide absorbent AI»Oz. It should be noted , that the exact size of the columns is not critical. The columns only need to be large enough to accommodate the volume of AI2O3 required for separation. The use of aluminum oxide (AI2O3) is effective for co-absorbing flavonoids and lignans co-extracted from the material sources, they are also difficult to remove, which becomes a problem for the final purity of the paclitaxel product

The columns were eluted with a solvent system of hexane-acetone (starting with 100:0 and ending at 45:55) at a pressure of 68.95-103.42kPa (10-15rvzi) with a flow rate of approximately 150ml/minute.

Fractions containing taxanes were collected and pooled according to TLC results, and then concentrated in vacuo to remove all solvents. The obtained material was dissolved in methanol and kept at room temperature overnight, as a result of which needle-shaped crystals were obtained. The crystals were filtered off and recrystallized from acetone to further obtain white needle-like crystals, which were determined to be c 13-acetyl-9-dihydrobaccatin Her, the purity of which exceeded 9695 (148 grams) (0.074965 on a crude basis).

The mother liquor from the crystallization of 13-acetyl-Udihydrobaccatin I was concentrated to dryness in a vacuum. ;" The residue was dissolved in three liters of acetone. The acetone solution was mixed with 1.5 kilograms of polystyrene-divinylbenzene copolymer resin. The mixture was evaporated to remove the solvent, and the resulting powder was loaded into the top of an industrial low-pressure chromatographic column filled with

Ф copolymer resin of polystyrene-divinylbenzene, and eluted with 4595 solution of acetone in water at a flow rate of 150 ml/minute at an operating pressure of less than 206.84 kPa (ZOorzi). Fractions of 2 liters each were collected and their concentration was controlled by TLC and HPLC. 2) Fractions containing paclitaxel and cephalomannin were combined and evaporated in vacuo to remove most of the acetone, then diluted with deionized water and extracted three times with 2.5 liters of dichloromethane. o The organic layer was concentrated to dryness in vacuo, this residue was dissolved in one liter of methanol. c To the methanol solution was added approximately 3095 (m//y) of water and the mixture was heated to 607°C for several minutes, then kept at room temperature overnight. The crude crystalline solid from the methanol solution was filtered off and dried in a vacuum oven at a temperature of 7070- 7570. The solid consisted of approximately 7090 paclitaxel and 2595 cephalomannin, yielding 31 grams.

Crude paclitaxel (30 g) was dissolved in 200 ml of acetonitrile, diluted with 25 ml of deionized water and crushed in

F) the top of a low-pressure chromatographic column (size: 10x15Osm), filled with polymer resin (biaiysp, macroporous polymethacrylate resin). After crushing the sample, the column was eluted with a step gradient of 35, 40, 45 and 5095 acetonitrile in water. The change of solvent was indicated by the results of TLC and HPLC fractions. in Collected fractions with a volume of approximately 1 l each. they were monitored by TLC and HPLC. The flow rate was 7 bml/minute. Column fractions containing paclitaxel or cephalomannin were combined separately. The two combined solutions were left at a temperature of approximately 5-7C until the crystallization of the individual compound was completed.

The crystals were filtered off separately and both recrystallized from a methanol-water solution at 6570.

Paclitaxel was obtained in the form of white needle-like crystals with a purity exceeding 99905 and in the amount of 18.5 g (0.00990).

Cephalomannin was obtained in the form of white needles with a purity exceeding 98905 and in the amount of 6b.5g

(0.00396).

Fractions containing baccatin I and 10-deacetylbaccatin III from the previous step were combined and evaporated in vacuo to remove virtually all acetone, then partitioned between 3 liters of water and dichloromethane (1:11 M/M). The organic layer was concentrated to dryness and the residue subjected to normal phase chromatography.

The size of the column used was 10.16x122 cm (4"x4); it was packed with silica gel (200-300 mesh). The eluent hexane-ethyl acetate had a step gradient (starting at 50:50 and ending at 35:65). Fractions were collected with a volume of approximately 1000 ml each.Each fraction was controlled by

TSH 70 Fractions containing baccatin I or 10-deacetylbaccatin I were collected separately and combined according to TLC results, then concentrated to dryness.

The part with baccatin IP was dissolved in 200 ml of methanol and kept in the refrigerator overnight.

The formed white crystals were filtered and recrystallized from methanol, resulting in 7.5 g (0.003905) of baccatin PI in the form of white crystals.

Raw 10-deacetylbaccatin PP was dissolved in 15 ml of acetone, and then diluted with 150 ml of hexane. The mixture was kept at room temperature overnight. The formed white crystals were filtered and recrystallized from the same solvent, resulting in 15 g (0.00795) of 10-deacetylbaccatin I in the form of white crystals.

Example 2

Using similar apparatus and methodology as in Example 1, crude paclitaxel was obtained. Crude paclitaxel was dissolved in ZOOml of dichloromethane, which was added to a 1000ml three-neck round-bottom flask. The flask was placed in an ice bath and the solution was stirred with a magnetic stirrer. When the temperature reached about 5°C, a solution of bromine (10 equivalents) in dichloromethane (1:11 M/M) was slowly added with stirring.

The ratio of cephalomannin to bromine was 1:10 mol. high school

Bromination within 5-10 minutes was monitored by TLC. Then the reaction mixture was diluted with 300 ml of dichloromethane and transferred to a separatory funnel after completion of the reaction (completion time was 40-50 minutes). ZB5Oml of 1095 aqueous sodium thiosulfate (Ma»Z»O3z) was added to the reaction mixture in order to absorb the excess volume of bromine. The dichloromethane layer was separated and washed with water and brine, then concentrated to dryness in vacuo. A light brown powder was obtained. yu zo Example Z

The raw material (10 g) from Example 2 was dissolved in 120 ml of acetonitrile, then diluted with 1500 ml of deionized water. at

The solution was squeezed to the top of a reverse-phase chromatographic column (5x100 cm) filled with St.

The column was eluted with acetonitrile:water (45:55) at an operating pressure of 206.84-275.79 kPa (30-4 Org). Speed co

The flow rate was ZOml/minute. Fractions were collected (250 ml each), monitored by TLC and combined. "at

The portion containing paclitaxel was kept overnight; the formed crystals were filtered off and recrystallized from 7095 methanol. Paclitaxel was obtained in the form of white needle-like crystals. Its purity was more than 9995, and the yield was 6.1g.

Fractions containing 2", 3"-bromocephalomannin were combined, partitioned between water and dichloromethane. "

The organic layer was collected and evaporated to dryness in vacuo. The residue was dissolved in 150 ml of methanol, which was diluted with 5 ml of water in order to obtain needle-shaped crystals. The whitish crystal was recrystallized from methanol:water to give 2",3"-bromocephalomannin as white crystals. ;" Two grams of 2",3"-bromocephalomannin were dissolved in 30ml of AsOnH and freshly activated zinc (2g) was added. The mixture was stirred for three hours at room temperature and the reaction was monitored by TLC. After TLC analysis indicated that the reaction was complete, the mixture was partitioned between dichloromethane and 1095 Manceau with

Ge" (Z0Oml, 2:1). The organic layer was washed with water and concentrated to dryness. The crude product was purified by flash chromatography to give cephalomannin as a white powder. bo Example 4 2) The raw material (10 g) from Example 2 was dissolved in 120 ml of dichloromethane. The organic layer was concentrated to 5% bulk and the residue was subjected to normal phase chromatography. The size of the column used was 10.16x122cm (4"x4U. It was filled with silica gel (200-300 mesh). The eluting solvent was dichloromethane:ethyl acetate with a step gradient (starting at 75:25 and ending at 6:4) For the separation of paclitaxel and 2",3"-bromocephalomannin, it is also possible to use the solvent system hexane-ethyl acetate (starting from 6:4 and ending at 4:6b) and dichloromethanesacetone (starting from 8:2 and

Ending at 65:35). Fractions of approximately 500 mL each were collected and each monitored by TLC,

Fractions containing paclitaxel and 2",3"-bromocephalomannin were collected separately and combined according to

F, according to TLC results, was then concentrated to dryness. The part with paclitaxel was dissolved in 100 ml of methanol and diluted with 35 ml of water, then kept in a refrigerator. The formed white crystals were filtered and recrystallized from methanol solution to obtain boron paclitaxel (5.5 g) in the form of white crystals.

Crude 2",3"-bromocephalomannin was dissolved in 50 ml of acetone and then diluted with 50 ml of hexane. The mixture was kept at room temperature overnight. The white crystals formed were filtered and recrystallized from the same solvent to obtain 2",3"-bromocephalomannin as white crystals.

The following examples illustrate the compounds in structural form. b5

: (c) m o

M E on pr-

ОЙ he o. sno no o v - I, o o SN.

Paclitaxel o and o se

Mo. he r ve r: I - O chan: Al he no o not o U -sn, s what o c)

Cephalomannin IS o)

IF) in o o, he (ze)

BUT shen ) | (uh)

TE A o (Te;

ENE z- Kh. o o Na « - s ts o ? H

Bakkatyn ІІІ - ke SN 45 . b 10-deacetylbaccatin IM with K-H o (ee) XX (95) o CH on on y t D sl SNU 0 sane

TE glo no ogor rya o (F) 07 0sn, name) 60 7 13-acetyl-9-dihydrobaccatin III

Despite the above described variants of implementation of the invention, the invention is not limited to them. It will be understood by those skilled in the art that numerous modifications form part of the present invention as long as they do not conflict with the spirit, nature and scope of the described invention and the claims of the present invention.

Claims (20)

The formula of the invention 1. A method of isolating and purifying taxane analogues from a source containing taxanes, which is distinguished by 2 the fact that it includes: extraction of the source of taxanes in an organic extractant; covering the weak absorbent medium with the specified extractant and loading the specified medium into the column containing the absorbent; elution at the first stage with a mixture of organic solvents at a pressure of 68.95-137.90 kPa (10-20 pv) to obtain fractions containing taxane compounds; crystallization of the specified fractions to obtain a solid taxane compound and a mother liquor; concentration of the indicated mother liquor; elution of said 70 mother liquor in the second stage with a mixture of polar solvents Through the polymer resin to obtain at least a second taxane compound. 2. The method according to claim 1, which is characterized by the fact that it additionally includes the stage of extracting lignan and flavonoid impurities in the indicated first stage. 3. The method according to claim 1, which differs in that the indicated source of taxanes includes a plant source. 12 4. The method according to item C, which is characterized by the fact that the indicated plant source is selected from the group consisting of 7/ahuv rgueyo/a, Tahiv rassaia, Tahiv schauspiapa, Tahiv uippapepv and Tahv sapadepv. 5. The method according to claim 3, which differs in that the specified taxanes include at least 13-acetyl-9-dihydrobaccatin III, 10-deacetylbaccatin II, bacatin II, cephalomannin, paclitaxel. 6. The method according to claim 1, which differs in that the specified organic extractant includes dichloromethane. 7. The method according to claim 1, which is characterized by the fact that the specified weak absorption medium includes Seyye 545. 8. The method according to claim 1, which differs in that the specified absorbent includes aluminum oxide. 9. The method according to claim 1, which differs in that the specified mixture of organic solvents in the first stage includes hexane and acetone. 10. A method for isolating and purifying taxane analogues from a source containing taxanes, which differs from 22 in that it includes: providing a source of the specified taxanes; extraction of the indicated taxanes from the indicated Ho) source into an organic extraction medium in order to obtain an organic layer containing taxane compounds; treatment of the carrier material with the specified organic layer; use of a low-pressure column containing an absorbent; elution of the specified column with an organic solvent at the first stage to obtain purified taxane fractions; crystallization of the indicated taxane fractions to obtain the first taxane 30 analog and mother solution; elution of the specified mother solution in the second stage through a polymer resin on a (0) chromatographic column for purification and obtaining at least the second taxane analogue and the third taxane analogue; collecting the separated taxane analogues. o 11. The method according to claim 10, which is characterized by the fact that it additionally includes the stage of extracting lignan and flavonoid impurities in the indicated first stage. 35 12. The method according to claim 10, which is characterized by the fact that it additionally includes the stage of grinding the specified source of ke, taxanes and extraction with methanol. 13. The method according to claim 12, which is characterized by the fact that it additionally includes the stage of mixing the extract with activated carbon, filtering and evaporating to obtain a concentrate. " 14. The method according to claim 13, which is characterized by the fact that the specified carrier material includes Seiye 545. From 50 15. The method according to claim 10, which is characterized by the fact that the indicated organic solvent includes a mixture of acetone and hexane. From" 16. The method according to claim 15, which differs in that the specified organic solvent is passed through the specified column at a pressure of 68.95-137.90 kPa (10-20 rvi). 17. The method according to claim 1, which is characterized by the fact that it additionally includes the stage of collecting and analyzing the specified 45 taxane fractions using thin-layer chromatography. Me 18. A method of isolating and purifying taxane analogues from a source containing taxanes, which differs in that it includes: providing a source of the specified taxanes; extracting the specified taxanes from the specified source into an organic extraction medium in order to obtain an organic layer containing taxane compounds; treatment of the carrier material with the specified organic layer; use of a low-pressure column containing an absorbent layer of 20; elution of the indicated column with an organic solvent at the first stage at a pressure of 68.95-137.90 kPa (10-20 rzi) to obtain purified taxane fractions and extract flavonoid and lignan impurities; sl crystallization of the indicated taxane fractions to obtain the first taxane analog and mother liquor; elution of the specified mother liquor in the second stage through polystyrene-divinylbenzene resin on a chromatographic column at a pressure of 206.84 kPa (30 rvzi) for purification and isolation of at least the second taxane analogue and the third taxane analogue; elution in the third stage of the specified second taxane HF) analogue and the specified third taxane analogue through a normal-phase column with silica gel to purify the specified second taxane analogue and the specified third taxane analogue; assembly of taxane analogs separated by 19. The method according to claim 18, which differs in that the indicated second taxane analog includes baccatin I, and the indicated third taxane analog includes 10-deacetylbaccatin PI. 20. The method according to claim 18, which is characterized by the fact that the specified analogues have a degree of purity of at least 9890. Official Bulletin "Industrial Property". Book 1 "Inventions, useful models, topographies of integrated circuits", 2004, M 8, 15.08.2004. State Department of Intellectual Property of the Ministry of Education and