39 results about "Cephalomannine" patented technology

The present invention relates to a yew branch and leaf extract, an extraction method and applications thereof. The yew branch and leaf extract is extracted by adopting branches and leaves of Taxus chinensis var. mairei as raw materials, and comprises cephalomannine and 7-epitaxol, wherein the active ingredients provide an inhabitation effect for squamous cells in skin cancers, and especially provide significant treatment effects for anal squamous cell carcinoma, esophagus squamous carcinoma and tonsil squamous cell carcinoma. According to the present invention, resource advantages of the yew are utilized to deep develop, utilize and research the Taxus chinensis var. mairei; and a yew anti-skin cancer component extraction method with a characteristic of simple operation is provided, and an effect of the component in anti-skin cancer application is determined.

An improved method for the isolation of taxanes of high purity from the crude extract of naturally occurring Taxus canadensis which comprises the steps of: a) filtering the crude extract by liquid column chromatography; b) enriching the taxanes obtained from step a) by precipitation and crystallization; and c) dissolving the taxanes extract in a polar organic solvent mixture comprising an alcohol and a non-alcohol solvent, which solution is then subjected to further separation by liquid column chromatography, and isolating substantially pure paclitaxel from cephalomannine; and all solvent mixtures used may be collected, purified and recycled without separation.

A cephalotmannine derivative, composition containing it, its production and use are disclosed. It can be used to prepare drug-resisting medicine in treatment of tumor.

The invention relates to a method for utilizing enzyme, ultraviolet radiation and oxygen as inductors for the high efficient production of 10-deacetylbaccatin III, baccatin III, cephalomannine and paclitaxel in taxus chinensis branches and leaves by the method of fresh grinding homogenization. The invention takes the fresh branches and leaves of taxus chinensis as raw materials for fresh grinding homogenization, then the enzyme is added, the oxygen is introduced, and a solid is treated with 80 percent ethanol ultrasonic extraction under ultraviolet irradiation. The contents of the 10-deacetylbaccatin III, the baccatin III, the cephalomannine and the paclitaxel in the branches and leaves of the taxus chinensis can be averagely improved by more than 17 percent, 23 percent, 74 percent and 23 percent respectively by the biological induction method. The raw materials used by the invention are the renewable fresh branches and leaves of the taxus chinensis, which can not only ensure no damage to ecological resources, but also can extract the effective components of taxus chinensis and the biological semisynthetic precursor substances with high efficiency. Furthermore, the method has simple and easy operation, which is applicable to industrial application and has great significance for industrial production.

An improved method for the isolation of taxanes of high priority from the crude extract of naturally occurring Taxus canadensis which comprises the steps of: a) filtering the crude extract by liquid column chromatography b) enriching the taxanes obtained from step a) by precipitation and crystalization; c) separating the taxanes obrained from step a) by liquid column chromatography and d) desolving the separate taxanes in a polar organic solvent mixture comprising an alcohol and a non-alcohol solvent, which solution is then subjected to further separation by liquid column chromatography and isolaring substantially pure paclitaxel from cephalomannine; and all solvent mixtures used may be collected, purified and recycled without seperation.

The invention relates to an application of cephalomannine chemical derivation method in taxol preparation, comprising the following steps that: cephalomannine analogues having an alpha-beta unsaturated ketone structure are converted into a single-halogen single-hydroxyl or single-halogen single-alkyl product through addition reaction, and then taxol is obtained through column separation. The process is also suitable for the separation of 10-deacetylated cephalomannine, 10-deacetylated taxol, 10-deacetylated-7-xylose cephalomannine and 10-deacetylated-7-xylose taxol. The application has the advantage of applying an NBS addition method to the separation of taxol and cephalomannine (or analogues of the two) successfully, and using a conventional cheap silica gel column to realize the separation of taxol and cephalomannine so as to achieve the aim of purifying taxol. The method is suitable for the taxol mixtures with different purity and can be used for the separation of other cephalomannine type compounds and analogues. Due to the process of the invention, the taxol with the purity more than 99 percent can be obtained; the overall yield can be up to 80 percent; the overall yield of cephalomannine addition products is over 90 percent.

The present invention separates and purifies taxol by means of column chromatographic process with macroporous resin, such as AB-8, D101, D201 and AK-9 as fixed phase and polar solvent, such as water solution of ethanol or acetone, as flowing phase, with the effluent flow rate is 0.6-1.5 L / hr. The effluent with taxol content lower than cephalotmannine content is concentrated and dried to obtain intermediate taxol product A; and effluent with taxol content higher than cephalotmannine content is concentrated and dried to obtain intermediate taxol product B. The said process can separate and purify coarse taxol product with 9-20% content to obtain intermediate taxol product with 40-60% content in the yield as high as 80%. The process can eliminate water soluble impurity from the coarse product.

The invention discloses a method for separating and purifying paclitaxel industrially and efficiently, and belongs to the field of the separation and purification of compounds. In the method, the paclitaxel is separated by column chromatography, namely the paclitaxel is separated and purified by taking a combination of any three or four of compounds of alkane having 1 to 10 carbon atoms, chlorohydrocarbons, alcohols having 1 to 4 carbon atoms, saturated alkane esters and alkane ketones, and amine compounds as a mobile phase and 200 to 300-mesh ordinary silica gel as a stationary phase by normal phase column chromatography. By selecting a multielement system mixed solvent as the mobile phase, the separation degree of substances which are difficult to separate is increased, and the problem that the paclitaxel cannot be separated from cephalomannine effectively in the conventional separation method is solved. After the paclitaxel is separated and purified by the column chromatography, the purity of paclitaxel crude products serving as raw materials can be improved to be over 98 percent, and according to the condition of the raw materials, the total yield is up to 90 percent only by eluting the paclitaxel crude products once to twice. In the method, the common solvent and silica gel are used, so the cost is low, the method is easy and convenient to operate, the sample volume is large, and industrial production is easy to realize.

The invention discloses a method for efficiently and rapidly extracting paclitaxel, cephalomannine and 10-DABⅢ from yew, which comprises dissolving yew powder in 10-DABⅢ extractant, performing ultrasonic extraction for the first time, and then collecting by centrifugation Centrifugal supernatant Ⅰ, the centrifugal supernatant Ⅰ is eluted after being saturated and adsorbed by the macroporous resin, and the eluate Ⅰ is collected and concentrated to obtain 10-DABⅢ extract. Collect the filter residue and filtrate after ultrasonic extraction; dissolve the filter residue collected in step a in paclitaxel extractant, collect the supernatant II after ultrasonic extraction and centrifugation, and collect the supernatant II after saturated adsorption by macroporous resin The eluate II is concentrated to obtain the extract containing paclitaxel and cephalomannine; it has the characteristics of simple preparation process, production equipment and operation steps, short extraction time, recyclable materials, and long use time.

The invention discloses a method for separating cephalomannine from taxol, which comprises the following steps: a, performing an 1,4-addition reaction, namely dissolving the mixture of taxol and cephalomannine in short-chain fatty alcohol, adding an organic alkali catalyst and stirring at room temperature; b, concentrating the solution, and performing separation by column chromatography to obtain a cephalomannine derivative and taxol; and c, dissolving the cephalomannine derivative, adding acidified active carbon, heating, refluxing, filtering, concentrating the filtrate, and recrystallizing to obtain cephalomannine. When the separation and purification method disclosed by the invention is used, the raw materials are cheap and readily available, the reaction and treatment conditions are mild, the product yield and purity are high, the separation of cephalomannine from taxol is facilitated considerably, and industrialization can be realized conveniently.

The present invention relates to a semi-synthetic process to convert a naturally occurring taxane into a suitable starting material for the synthesis of paclitaxel and related compounds. Specifically, the present invention relates to a process for the conversion of 9-dihydro-13-acetylbaccatin III into a 7-protected baccatin III which can then be used as starting material for the synthesis of taxane derivatives such as paclitaxel, docetaxel, cephalomannine and other taxanes structurally related to baccatin III. The method as described uses a preparative scale technique which is amenable to commercial scale-up.

The present invention is column bromating process of preparing high purity taxol. The coarse taxad extractive containing cephalotmannine is made to react with bromine, the bromide of cephalotmannineand concomitant product are then eliminated through forward filling chromatographic column separation, and through further twice re-crystallization, high purity taxol is prepared in the total yield of 85 %. Taxol with relatively lower purity is transferred to forward silica gel column for further separation. The said process is suitable for large scale production. The reagent and the solvent are recovered for reuse, and this is favorable to environment protection.

The invention discloses a method for co-producing taxane effective ingredients and ethyl alcohol by utilizing Chinese yew. The method for co-producing the taxane effective ingredients and ethyl alcohol by utilizing the Chinese yew comprises the following steps: carrying out enzymolysis on cellulose and hemicellulose under a weak acid condition, fermenting for producing ethyl alcohol, and then extracting the taxane effective ingredients by utilizing the produced ethyl alcohol. The cellulose and hemicellulose in the Chinese yew are hydrolyzed, and fermentation is carried out for producing the ethyl alcohol after the cellulose is hydrolyzed by virtue of an enzyme, so that the taxane effective ingredients can be better dissolved into an ethyl alcohol extracting solution, the taxane effective ingredients such as taxol, cephalomannine and 10-DABIII and the ethyl alcohol can be extracted at the same time, extraction ratio of taxol is more than 99%, the extraction ratio of the cephalomannine is more than 99%, the extraction ratio of the 10-DABIII is more than 99%, content of the taxol in extract is 1-5%, content of the cephalomannine is 1-3%, and content of the 10-DABIII is 1-3%.

The invention provides a method for removing cephalomannine from paclitaxel. The method comprises the following steps: 1) preparation of raw materials: dissolving a cephalomannine-containing paclitaxel crude product into an organic solvent to prepare a solution A, and dissolving potassium permanganate into an organic solvent to prepare a solution B; 2) reaction: reacting the solution A with the solution B, and filtering the solution obtained by reaction; 3) primary crystallization: crystallizing filtrate obtained in step 2); 4) column chromatography: filtering and drying the product obtained in step 3) for column chromatography, collecting paclitaxel-containing effective segments, and performing concentration and drying; 5) secondary crystallization: performing crystallization treatment on a drying product obtained in step 4) to obtain a paclitaxel pure product with paclitaxel content of more than 98.5 weight percent. According to the method, good cephalomannine removal effects are achieved, influence on the paclitaxel is eliminated, the purity and recovery rate of the paclitaxel are both higher, the method can be operated flexibly and conveniently, and is suitable for industrial production, reaction and treatment conditions are mild, and the difficulty of refining steps is lowered.

The present invention relates to taxane bromo-analogues with anticancer activity and preparation method thereof. It uses bromine and taxus crude extract containing cephalotmannine and 7-epi-10-deacetyl cephalotmannine to make addition reaction, then uses normal-phase preparation-type HPLC and reversed-phase semi-preparation HPLC to obtain two new taxane analogues: (2''R,3''S)-dibromo-7-epi-10-deacetyl cephalotmannine and (2''S,3''R)-dibromo-7-epi-10-deacetyl cephalotmannine. As compared with taxol said two analogues have similar and stronger anticancer activity for human mammary cancer cell (MCF-7), human pulmonary carcinoma cell (A549) and human oophoroma cell (A2780), and can be used for preparing anticancer medicine.

The invention provides a method for quickly screening trace taxol active substances in plants. The method comprises the steps that micro-columns filled with unactivated carbon fibers and activated carbon fibers with different surfaces from those of the unactivated carbon fibers are used for carrying out pre-separation on taxus mairei extract, different mobile phases are used for elution, and taxol active substances with different polarities are obtained; by means of an online ionization process, the taxol active substances represented by baccatin III, 10-deacetylation baccatin III, taxol, 7-xylosyl-10 deacetylation taxol, 7-xylosyl-taxol, cephalomannine and 7-epi-10-deacetylation taxol are screened, the whole screening process only needs 5 min, and the screening detection limit reaches the level of 25 ng / g; by means of the method, the material dosage for analysis and background interference are reduced, the detection sensitivity is improved, and the high universality of screening various natural samples with different ingredients is achieved.

The invention discloses a method for preparing a taxane natural product. The taxane natural product is 10-deacetyl cephalomannine. The method includes the steps that 7,10-Di-Troc-docetaxel is dissolved in formic acid, dichloromethane is added for dilution after a reaction is completed, a sodium bicarbonate solution is adopted for neutralization until no bubbles are formed, extraction and liquid separation are carried out, and an organic phase is collected and concentrated to obtain a product I; the product I is dissolved in DMF, 2-methylimidazole is added, TesCl is added dropwise slowly, aftera reaction is completed, water is added for quenching the reaction, ethyl acetate is adopted for extraction, and the organic phase is washed with water and concentrated to obtain a product II; the product II is dissolved in the dichloromethane, tiglic acid and DMAP are added, DIC is added dropwise, after a reaction is completed, a reaction solution is subjected to suction filtration, and a filtrate is diluted with the dichloromethane, washed with water and then concentrated to obtain a product III; the product III is subjected to a concentration process twice, purified by column chromatography, concentrated and dried to obtain the 10-deacetyl cephalomannin. No obvious by-products are generated in each step, the yield is high, the product purity is high, preparation processes of starting materials are mature, the starting materials are easy to obtain, and massive taxane natural products can be prepared.

The present invention relates to taxane bromo-analogues with anticancer activity and preparation method thereof. It uses bromine and taxus crude extract containing cephalotmannine and 7-epi-10-deacetyl cephalotmannine to make addition reaction, then uses normal-phase preparation-type HPLC and reversed-phase semi-preparation HPLC to obtain two new taxane analogues: (2''R,3''S)-dibromo-7-epi-10-deacetyl cephalotmannine and (2''S,3''R)-dibromo-7-epi-10-deacetyl cephalotmannine. As compared with taxol said two analogues have similar and stronger anticancer activity for human mammary cancer cell (MCF-7), human pulmonary carcinoma cell (A549) and human oophoroma cell (A2780), and can be used for preparing anticancer medicine.

The invention relates to a method for separating and purifying paclitaxel and cephalomannine. According to the method disclosed by the invention, a preparative high performance liquid chromatography is adopted, and the chromatographic conditions are that a polystyrene-divinyl benzene copolymer chromatographic column refers to MKF-EP-RP with the diameter of 5-12 microns and the size of (150-600)*(7.0-20.0)mm (I.D.), a mobile phase comprises acetonitrile and water according to a volume ratio of (45-60) to (40-55), the flow velocity is equal to 1.0-3mL / min, the detection wavelength is equal to 229nm and the sample size is equal to 20-100 microL. According to the method disclosed by the invention, the defects such as irreversible adsorption of media such as silica gel and complicated process flow are overcome, and paclitaxel products with the purity of over 98 percent can be obtained by further separation.

The invention discloses a paclitaxel purifying method of a paclitaxel crude product produced by Chinese yew cell culture and belongs to the field of compound purification. With the combination of a crystallization method and a normal phase column-chromatography method, paclitaxel is purified. Firstly, a paclitaxel crude product produced by Chinese yew cell culture is dissolved in an organic solvent with high polarity, and a reverse organic solvent is dropwisely added so as to obtain paclitaxel with purity being more than 85.0%. And then, paclitaxel is further separated and purified by column chromatography. A stationary phase is silica gel of 200 meshes-300 meshes, and a mobile phase is a binary gradient mixed solvent of dichloromethane and ethyl acetate or hexane and acetone. Combining the two methods, the defect that paclitaxel and cephalomannine cannot be separated effectively by a previous separation method is solved, and unknown impurities which are hard to separate can be removed. Thus, a paclitaxel bulk drug which accords with United States Pharmacopeia (USP) standards can be obtained. The method has advantages of good repeatability and reasonable technology, is simple to operate, and is green and safe.

The invention belongs to the medicine technical field and discloses a preparation method of anti-tumour medicine intermediate, namely a method that cephalomannine is utilized to synthesize taxol and taxotere semisynthesis precursor 10-DAB III. Cephalomannine alcoholic solution is contact with hydrazine hydrate and converts the hydrazine hydrate into 10-DAB III. The method can remove C-10 and C-13ester functional group of cephalomannine and has good selectivity. Synthetic product is separated by silicagel column chromatography, petroleum ether-acetone (3:1) is taken as eluent, so as to obtain10-DAB III main section, and then the main section is subject to secondary recrystallization, thus obtaining high purity 10-DAB III. Primary crystallization system is acetone-petroleum ether or normal hexane; and the secondary crystallization system is chloroform-methyl alcohol-normal hexane or petroleum ether or other low polarity solvent. Cephalomannine can be obtained by separation from taxus plant or is byproduct produced in taxol extraction process. The invention has the characteristics of mild reaction condition and higher product purity and yield.

The present invention provides coproducting process of high purity taxol, 10-deacetyl Bakating III and cephalomtaine from alcohol extractive of leaf, bark or branch of taxad. The copreoducing process includes dissolution of the alcohol extractive with butyl acetate, sodium bicarbonate solution reverse extraction to eliminate impurity, decompression evaporation of the organic phase to eliminate solvent, acetonitrile dissolution of the residuum, cooling crystallization to obtain 10-deacetyl Bakating III, purifying crystallization mother liquid through inversed phase chromatography with polymer stuffing as fixed phase to obtain cephalomtanine and taxol, freezing crystallization to separate the effective components from the inversed phase chromatography eluent and forward phase chromatography with silica gel stuffing as fixed phase to obtain taxol product with purity as high as 99.6%.

The invention discloses a paclitaxel purifying method of a paclitaxel crude product produced by Chinese yew cell culture and belongs to the field of compound purification. With the combination of a crystallization method and a normal phase column-chromatography method, paclitaxel is purified. Firstly, a paclitaxel crude product produced by Chinese yew cell culture is dissolved in an organic solvent with high polarity, and a reverse organic solvent is dropwisely added so as to obtain paclitaxel with purity being more than 85.0%. And then, paclitaxel is further separated and purified by column chromatography. A stationary phase is silica gel of 200 meshes-300 meshes, and a mobile phase is a binary gradient mixed solvent of dichloromethane and ethyl acetate or hexane and acetone. Combining the two methods, the defect that paclitaxel and cephalomannine cannot be separated effectively by a previous separation method is solved, and unknown impurities which are hard to separate can be removed. Thus, a paclitaxel bulk drug which accords with United States Pharmacopeia (USP) standards can be obtained. The method has advantages of good repeatability and reasonable technology, is simple to operate, and is green and safe.

The invention discloses a separating and purifying method of taxol synthesized by biological fermentation of nodulisporium sylviforme entophytic fungi as well as precursors of the taxol, relates to a separating and purifying method and aims at solving the problems that taxol in fermentation liquor is low in content and the taxol precursors are low in the recovery rate and low in purity in the traditional method for synthesizing by biological fermentation of entophytic fungi. The separating and purifying method disclosed by the invention comprises the following steps of: activating a bacterial strain and culturing seeds; 2, carrying out fermentation culture; 3, separating and recovering fermentation liquor and mycelium, decoloring the fermentation liquor, and obtaining a sample through extracting and concentrating; 4, carrying out primary chromatography; 5, carrying out secondary chromatography; and 6, separating by using a reversed phase chromatography method to finish separation and purification. According to the separating and purifying method, the content of taxol in the fermentation liquor is 556.80mug / L, the taxol purity is 98 percent, the precursor baccatin purity is 97 percent, and the precursor cephalomannine purity is 98 percent; the separating and purifying method has the characteristics of simple operation process, high product purity and low use amount of a solvent; and meanwhile, the production cost of the taxol is indirectly reduced.

The invention relates to a determining method for a kind of plant yew and the taxol in its cell culture and its homologen. Its characteristic lies in that the chromatographic column is Nova-Pak C18, the mobile phase composed of two chromatographic reagent which is acetonitrile and water, balancing the chromatography system with 30 % acetonitrile and 70 % water.and the cell extractor or the extractor of the cell culture solution of the yew and plant yew enter the system, gradient eluting, the proportion changes from 30:70 to 50:50 and the gradient changing curve is 0.5-0.7. Successively, continuing to elute at the proportion of 50:50 for three minutes; Last, balancing the system for 5 minutes with the proportion of 30:70, the preserval time of taxol is 11.12-11.8 minutes. The invention can determine the yew and its cell culture taxol,10-, simultaneously, easily and quickly. It has the characteristics of accuracy, high sensitivity and economy.

The invention relates to an application of cephalomannine chemical derivation method in taxol preparation, comprising the following steps that: cephalomannine analogues having an alpha-beta unsaturated ketone structure are converted into a single-halogen single-hydroxyl or single-halogen single-alkyl product through addition reaction, and then taxol is obtained through column separation. The process is also suitable for the separation of 10-deacetylated cephalomannine, 10-deacetylated taxol, 10-deacetylated-7-xylose cephalomannine and 10-deacetylated-7-xylose taxol. The application has the advantage of applying an NBS addition method to the separation of taxol and cephalomannine (or analogues of the two) successfully, and using a conventional cheap silica gel column to realize the separation of taxol and cephalomannine so as to achieve the aim of purifying taxol. The method is suitable for the taxol mixtures with different purity and can be used for the separation of other cephalomannine type compounds and analogues. Due to the process of the invention, the taxol with the purity more than 99 percent can be obtained; the overall yield can be up to 80 percent; the overall yield of cephalomannine addition products is over 90 percent.

The invention discloses a method for co-producing taxane effective ingredients and ethyl alcohol by utilizing Chinese yew. The method for co-producing the taxane effective ingredients and ethyl alcohol by utilizing the Chinese yew comprises the following steps: carrying out enzymolysis on cellulose and hemicellulose under a weak acid condition, fermenting for producing ethyl alcohol, and then extracting the taxane effective ingredients by utilizing the produced ethyl alcohol. The cellulose and hemicellulose in the Chinese yew are hydrolyzed, and fermentation is carried out for producing the ethyl alcohol after the cellulose is hydrolyzed by virtue of an enzyme, so that the taxane effective ingredients can be better dissolved into an ethyl alcohol extracting solution, the taxane effective ingredients such as taxol, cephalomannine and 10-DABIII and the ethyl alcohol can be extracted at the same time, extraction ratio of taxol is more than 99%, the extraction ratio of the cephalomannine is more than 99%, the extraction ratio of the 10-DABIII is more than 99%, content of the taxol in extract is 1-5%, content of the cephalomannine is 1-3%, and content of the 10-DABIII is 1-3%.

A method for separating paclitaxel cephalomannin and other related compounds by obtaining a staring material that contains paclitaxel and cephalomannin; dissolving the staring material in any one a number of particularly defined solvents such as butly formate and butly acetate to form a mixture; subjecting the mixture to column chromatography to obtain an eluted fraction of pactitaxel and cephalomannin and a residue ; and separately drying the paclitaxel and cephalomannin fractions to obtain separate crystalline forms of paclitaxel and cephalomannin, respectively.

The invention relates to the field of chemical analysis, in particular to a nuclear-magnetic-resonance-technology-based spatial correlation spectrum (ROESY) test method for cephalomannine. According to the invention, on the basis of the optimized design of a pulse sequence, a spatial correlation spectrum ROESY of cephalomannine is measured by using hydrogen as an observation nucleus. At present, most of spatial correlation spectrums used by instruments are based on the NOESY spectrum and the NOE signals for some medium molecules are zero, so that the spatial correlation conditions can not be characterized by using the NOESY. On the basis of the characteristics of the molecules of cephalomannine, the ROESY spectrum is introduced based on concept changing and the relevant parameters are optimized. Therefore, the defect of weak NOE signal is overcome; and the an effective and simple means for characterizing the spatial structure for medium-sized molecules is provided.