49 results about "Epothilones" patented technology

The present invention describes polymer nanoparticles with a cationic surface potential, in which both hydrophobic and hydrophilic pharmaceutically active substances can be encapsulated. The hydrophilic and thus water-soluble substances are encapsulated in the particle core by co-precipitation through ionic complexing with a charged polymer. Both therapeutic agents and diagnostic agents can be used as pharmaceutically active substances for encapsulation. The cationic particle surface permits stable, electrostatic surface modification with partially oppositely charged compounds, which can contain target-specific ligands for improving passive and active targeting.

The invention relates to a new process for concentrating epothilones in culture media, a new process for the production of epothilones, a new process for separating epothilones A and B and a new strain obtained by mutagenesis for the production of epothilones, as well as aspects related thereto. New crystal forms of epothilone B are also described.

The invention relates to a preparation method of epothilones B. In particularly, the invention provides a technique method to prepare epothilones B through adopting 100 to 500 liters fermentor to ferment and to separate epothilones B in large scale. The invention also relates to the preparation and application of epothilones B in preparation of gleevec resistance chronic myeloid leukemia treatment drug.

The present invention provides compounds of formula (I): as described generally and in classes and subclasses herein. The present invention additionally provides pharmaceutical compositions comprising compounds of formula (I) and provides methods of treating cancer comprising administering a compound of formula (I).

The invention relates to a new process for concentrating epothilones in culture media, a new process for the production of epothilones, a new process for separating epothilones A and B and a new strain obtained by mutagenesis for the production of epothilones, as well as aspects related thereto. New crystal forms of epothilone B are also described.

Formulations comprising one or more epothilones together with a pharmaceutically acceptable carrier.

The invention discloses a production process for high-density fermentation of sprangium cellulosum and separation coupling of an epothilone product. In the invention, the high-density fermentation of the sprangium cellulosum is realized by feeding carbon and nitrogen source substances and precursors; and at the same time, a polymeric adsorbent which can be repeatedly used can be used for separating the epothilone product and the high-density fermentation and product separation coupling are realized, thus feedback inhibition of the epothilone on the sprangium cellulosum is removed and output of the epothilone is increased. The invention has the technical characteristics of cost conservation, simple process, convenience in operation and high production efficiency.

Recombinant host cells of the suborder Cystobacterineae, genus Stizmatella, containing recombinant expression vectors that encode heterologous epothilone polyketide synthase (PKS) genes and can produce epothilones at high levels are provided.

Recombinant nucleic acids that encode all or a portion of the epothilone polyketide synthase (PKS) are used to express recombinant PKS genes in host cells for the production of epothilones, epothilone derivatives, and polyketides that are useful as cancer chemotherapeutics, fungicides, and immunosuppressants.

A process for extracting Epophilone from fermented liquid of myxobacterium includes mixed resin adsorption, solid and liquid extractions, molecular sieve chromatography, crystallizing and effiicent liquid-phase separation. Its advantages are high purity and extraction rate (more than 80%). The said Epophilone is an antineoplastic compound.

The invention relates to a desorption process for epothilones, especially epothilone A and / or epothilone B, from resins and new production, work-up or purification processes or manufacturing methods for epothilones comprising said desorption process, as well as the use of certain solvents for the desorption of epothilones from resins.

An anticancer composition containing neovascular inhibitor and epothilone and able to be made into slow-release implant or injection is composed of the slow-release microballs and solvent. Said slow-release microball consists of active anticancer component (neovascular inhibitor and / or epothilone) and slow-release additives (p (LAEG-EOP), p (DAPG-EOP), etc). Said solvent is an ordinary solvent or the special solvent containing the suspending aid.

Nucleic acid molecules are isolated from Sorangium cellulosum that encode polypeptides necessary for the biosynthesis of epothilone. Disclosed are methods for the production of epothilone in recombinant hosts transformed with the genes of the invention. In this manner, epothilone can be produced in quantities large enough to enable their purification and use in pharmaceutical formulations such as those for the treatment of cancer.

A method for synthesizing an organometallic compound, which is usefully utilized as a catalyst for manufacturing polyolefins by ring-opening metathesis polymerization of an olefin having strain in a molecule such as dicyclopentadiene or synthesizing epothilones by ring-dosing metathesis reaction, by an efficient and low cost synthesis method using a starting material which is easily available due to relatively simple chemical structure, and further without any possibility of coexistence of a vinylhetero compound or a vinyl compound exchanged which tends to accompany as an impurity in the system in conventional methods, characterized by reacting a starting material comprising a zero-valent transition metal complex (A) or a polyvalent transition metal complex (A′), a compound (B) or (B′) shown by the following general formula (1) or (4), respectively, and a neutral ligand (C) or (C′), in one step under non-reducing condition or reducing condition. R1Y1CR2X12  (1) R4Y2CR5X22  (4)

The present invention concerns methods for the production of pharmaceutical formulations of Epothilones suitable for being administered parenterally, such as intravenously.

The invention discloses an Epothilone freeze-dried compound which mainly contains the Epothilone, a solubilizer and a freeze-dried excipient; wherein, a preferable solubilizer is Carmowax 12-hydroxyl stearate. The product not only is simple in technique and is stable in quality, but also is low in side effect.

The invention discloses a method for absorbing and fixing sorangium cellulosum for fermentation based on porous ceramics and belongs to the field of microbial fermentation pharmacy. The method comprises the steps that the prepared porous ceramics are processed by fermentation liquid to be dried for stand-by application; after sorangium cellulosum is cultivated through an M26 solid and a liquid medium respectively, bacterium suspension is formed; and after dilution, the bacterium suspension is inoculated in the fermentation liquid, the processed porous ceramics are added into the fermentation liquid, and shake cultivation is performed. The method for absorbing and fixing an epothilone production bacterial strain based on kieselguhr-based porous ceramics is provided by the invention, when epothilone B is in the fermentation production, sorangium cellulosum, which is the production bacterial strain of the epothilone B, is fixed, so that a solid attachment surface can be provided for a slime bacteria cell growing in a liquid environment, the method has the advantages that the fermentation production yield can be increased, the production cost of epothilone which is an anti-tumor medicine is lowered, and the operation steps are simple and easy to implement, therefore, the method can be amplified to the industrial production and establishes an experimental basis for the industrial production of epothilone.

A slowly-release anticancer medicine in the form of injection or implant is disclosed. Said slowly-release injection is composed of a special solvent containing suspending aid and the slowly-release microballs consisting anticance medicine, iterstitial hydrolyte and slowly-releasing auxiliary. Said anticancer medicine is chosen from Epothilone A, Epothilone B, etc. Said interstitial hydrolyte is chosen from collagenase, relaxin, etc. Said slowly-releasing auxiliary is chosen from p(BHET-EOP / TC), p(LAEG-EOP), polyethanediol, etc.

The invention relates to a membrane extraction separation technique of epothilone, which comprises the following steps: filtering an epothilone a and b fermentation liquid through an organic membrane to obtain an organic membrane filtrate, and filtering the organic membrane filtrate through a ceramic membrane to obtain a ceramic membrane filtrate; eluting with a gel chromatographic column, sectionally collecting the eluates subjected to HPLC (high performance liquid chromatography) detection, concentrating the eluates through a nanofiltration membrane, eluting the concentrated solution by normal-phase silica gel chromatography, sectionally collecting the eluates subjected to HPLC detection, and crystallizing the collected eluates at low temperature to obtain the epothilone a and b crystal. The molecular weight cutoff of the organic membrane is 5000-6000. The molecular weight cutoff of the ceramic membrane is 1000-1500. The molecular weight cutoff of the nanofiltration membrane is 500-800. The crystallization temperature is -30 to -10 DEG C. The normal-phase chromatography is utilized to separate the epothilone a and b in the concentrated solution, thereby reducing the consumption of the organic solvent, simplifying the extraction process and lowering the cost.

The invention belongs to the technical field of biology, and particularly discloses a pretreatment method of epothilone B fermentation liquor. An inorganic salt is added into the fermentation liquor,so that resin and thalli are thoroughly separated, only the resin phase remains after the thalli are removed, the subsequent extract is reduced to 1 / 3 of the original mixture, an organic solvent for further extraction is saved, and impurities in the thalli are prevented from being introduced into the subsequent purifying process; and impurities with the retention time similar to that of epothiloneB are removed from the source. In addition, the separated resin phase does not need to be centrifuged or filtered by a plate frame and is directly loaded into a column without blocking a sieve plate,so that the column passing time is saved. According to the pretreatment method of the epothilone B fermentation liquor, the thalli and the resin are well separated, subsequent extraction steps are simplified, the usage amount of subsequent extraction organic solvents is reduced, the extraction efficiency and yield are improved, the universality is high, and the industrialization cost is low.

The invention provides a method for preparing epothilone B based on coupling of separation and fermentation of a molecularly imprinted membrane. The method comprises the following steps: carrying out early fermentation to obtain a seed culture solution, carrying out fermentation culture on the seed culture solution in a fermentation tank, filtering a fermentation solution by utilizing the molecularly imprinted membrane in the fermentation process, eluting a substance obtained through filtration, collecting the eluent and finally concentrating the eluent, thus obtaining epothilone B. The method has the beneficial effects that epothilone B is produced through fermentation by utilizing the fermentation tank and the specific adsorption separation and the fermentation process of the molecularly imprinted membrane are coupled, namely, the metabolite epothilone B is timely separated while producing epothilones B through fermentation, so that the concentration of epothilone B in the fermentation tank is maintained at a lower level, thus reducing feedback inhibition and product toxicity and increasing the fermentation yield; meanwhile, epothilone B is specifically separated by utilizing the molecularly imprinted membrane but other components are not separated to reduce the costs of downstream separation and purification; the method is beneficial for aftertreatment of the fermentation solution and refining of the products and has extensive application prospect in the industrial circles.

The invention relates to the treatment of a proliferative disease, especially according to certain treatment regimens, with an epothilone, especially with epothilone A and more preferably epothilone B; as well as to the treatment of certain cancers with such an epothilone.

The invention relates to a preparation method of epothilones B. In particularly, the invention provides a technique method to prepare epothilones B through adopting 100 to 500 liters fermentor to ferment and to separate epothilones B in large scale. The invention also relates to the preparation and application of epothilones B in preparation of gleevec resistance chronic myeloid leukemia treatment drug.

A method of treating a hyperproliferative disorder, including a cancer, in a subject in need of such treatment, comprising administering to said subject a pharmaceutical combination containing a treatment effective amount of: (a) a vitamin A derivative (i.e., a retinoid), or a pharmaceutically acceptable salt thereof, and an inhibitor of microtubule structure or function; or (b) a combination containing fenretinide (i.e., N-(4-hydrophenyl) retinamide, 4-HPR) and ABT-751 (i.e., N-[2-[(4-hydroxyphenyl)amino]-3-pyridinyl]-4-methoxybenzenesulfonamide). Vitamin A derivatives that may be useful for this invention according to (a) include, but are not limited to, all-trans-retinoic acid, 13-cis-retinoic acid, and fenretinide. Microtubule inhibitors that may be useful for this invention according to (a) include, but are not limited to, inhibitors of the Vinca binding domain (e.g., vincristine, vinblastine, vinorelbine, and cryptophycin 52), inhibitors of the Taxane domain (e.g., paclitaxel, docetaxel, and epothilones), and inhibitors of the colchicine site (e.g., colchicine, ABT-751, CI-980, and combretastatin). A preferred retinoid according to (a) is fenretinide. A preferred microtubule inhibitor according to (b) is ABT-751.

The present invention provides compounds of formula (I):as described generally and in classes and subclasses herein. The present invention additionally provides pharmaceutical compositions comprising compounds of formula (I) and provides methods of treating cancer comprising administering a compound of formula (I).

The invention discloses sorangium cellulosum highly producing epothilone D as well as a construction method and application of the sorangium cellulosum. The invention discloses construction of a recombinant sorangium cellulosum So ce M4 strain with an epoK gene knocked out by a TALEN technology for the first time. Tests prove that by knocking out the epoK gene in the sorangium cellulosum So ce M4strain by the TALEN technology, the yield of the epothilone D can be significantly increased, and meanwhile, the yield of epothilone B can be reduced, so that directed biosynthesis of epothilone in the sorangium cellulosum can be realized. The invention discloses the increase of the yield of the epothilone produced by the sorangium cellulosum by the TALEN for the first time, the TALEN technology is applied to increase of the yield of the epothilone D produced by the sorangium cellulosum, biosynthesis of the epothilone is promoted while the development of a sorangium cellulosum genetic engineering modification technology is promoted, and the application of the epothilone D in antitumor aspects is enlarged, which has quite important significance in development of biomedicine in China.

The invention discloses an additive for increasing the fermentative yield of epothilone, which is indoleacetic acid, indole methyl formate or indole, and the added final concentration of the additive in epothilone liquid fermentation medium is 20 plus / minus 3Mu mol / L. An experiment proves that the additive disclosed by the invention can promote thalli to absorb and utilize nutrient substances and enhance the secondary metabolism of sprangium cellulosums, so that the total amount of epothilone is increased by 2 to 19 percent on average in comparison with the total amount of epothilone not added with the additive. The additive provided by the invention reduces fermentation cost, increases the yield of epothilone, and provides another approach for epothilone production.

The invention relates to epothilone analog represented by formula (I) wherein (i) R2 is absent or oxygen; ''a'' can be either a single or double bond; ''b'' can be either absent or a single bond; and ''c'' can be either absent or a single bond, with the proviso that if R2 is oxygen then ''b'' and ''c'' are both a single bond and ''a'' is a single bond; if R2 is absent then ''b'' and ''c'' are absent and ''a'' is a double bond; and if ''a'' is a double bond, then R2, ''b'' and ''c'' are absent; R3 is a radical selected from the group consisting of hydrogen; lower alkyl; -CH=CH2; -CCH; -CH2F; -CH2Cl; -CH2-OH; -CH2-O-(C1-C6-alkyl); and -CH2-S-(C1-C6-alkyl); R4 and R5 are independently selected from hydrogen, methyl or a protecting group; and R1 is as defined in the specification, or a salt of a compound of formula (I) where a salt-forming group is present. A further aspect of the invention is related to the synthesis of epothilone E. These compounds have inter alia microtubuli depolymerization inhibiting activity and are e.g. useful against proliferative diseases.

This invention provides the use of an Epothilone, which shows an average distribution coefficient between plasma and brain of 0.3 to 1.5 in the mouse intravenous bolus injection assay, for the preparation of a medicament for the treatment of a brain disease associated with proliferative processes.

The invention relates to the field of microbial fermentation, in particular to a preparation method of a defoamer for fermentative production of epothilone; the invention firstly synthesizes di(ethylene glycol) ethyl ether methacrylate modified silicone oil, and then mixes it with a phosphoric acid The phosphorylated polyether-modified aminosilicone oil was mixed with solid powder and reacted to prepare a defoamer suitable for the fermentation of Cystobacter celluliformis. The phosphorylated polyether-modified aminosilicone oil had a branched structure and excellent Shear resistance, suitable for the environment in the fermentation broth; in addition, the addition of polyether structure is beneficial to increase the hydrophilicity of the defoamer, increase the dispersibility of the defoamer in the fermentation broth, and is conducive to enhancing the defoaming Foam-inhibiting ability; the defoamer of the present invention has a promoting effect on the growth of C. cellulosum and the production of epothilone, and is a defoamer product suitable for the fermentation liquid of C. cellulosum.