148 results about "Methotrexate" patented technology

Belonging to the technical field of bioengineering, the invention relates to corynebacterium glutamicum and a production method of alpha-ketoglutarate through fermentation of corynebacterium glutamicum. The bacterial strain of the invention is corynebacterium glutamicum GKG-047, with a preservation number of CGMCC No. 5481. With a strain of glutamic acid producing strain GDK-9 as a parent, the bacterial strain of the invention is a mutant strain that is screened out through mutagenesis and sensitive to methotrexate. Under the circumstance of a limited nitrogen source supply, the bacterial strain can accumulate alpha-ketoglutarate. During fermentation, the bacterial strain is characterized by aerobic fermentation, fast bacterium growth, and rapid acid production rate. After fermentation of the bacterial strain for 32h in a fermentation tank of 10L, the maximum output of alpha-ketoglutarate can be 47.2g/L. The method provided in the invention has the advantages of simple and easily controllable fermentation process, and low production cost of alpha-ketoglutarate, thus being in favor of the popularization and application of industrial production.

The invention discloses a method for simultaneously detecting multiple anti-tumor drugs in a blood sample. A pretreated sample to be detected is detected by adopting ultra-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The pretreatment process comprises the following steps: adding serum into a mixed solution of methanol and acetonitrile, oscillating and centrifuging,taking out the centrifuged supernatant, drying, dissolving the dried powder into a methanol aqueous solution, and filtering to obtain a sample to be detected. The method can be used for simultaneously detecting 13 kinds of anti-tumor drugs such as methotrexate, 5-fluorouracil, apatinib, busulfan, carboplatin, cyclophosphamide, docetaxel, gemcitabine, imatinib, illinotecan, lenalidomide, oxaliplatin, paclitaxel and the like in blood.

The invention discloses pH sensitive-type Fe3O4 coated LDH methotrexate-loaded nano drug particles. According to the invention, LDH is directly added in a preparation system of Fe3O4 nanoparticles, while the Fe3O4 nanoparticles are formed, at the same time, the surface of the Fe3O4 nanoparticles is coated with LDH through a self-assembling mode, a Fe3O4 coated LDH carrier is formed, and MTX can effectively loaded on the Fe3O4 coated LDH carrier through a subject and object exchange technology. The employed LDH can increase the biocompatibility of the Fe3O4 nanoparticles, and a special structure of LDH can greatly increase the load capacity of an anticancer medicine. The prepared Fe3O4 coated LDH-MTX has excellent medicine slow release performance, has obvious inhibition effect on tumor cells, has good antineoplastic effect, and has large application potential in the field of targeting tumour treatment.

The invention relates to a methotrexate detection reagent as well as a preparation method and a detection method thereof, and specifically relates to a methotrexate homogenous enzyme immunoassay reagent as well as a preparation method and a detection method thereof. The methotrexate homogenous enzyme immunoassay reagent comprises an anti-methotrexate specific antibody, and an indication reagent for detecting an anti-methotrexate specific antibody-methotrexate compound, wherein the anti-methotrexate specific antibody is obtained from immune animals with methotrexate immunogen. The methotrexate homogenous enzyme immunoassay reagent disclosed by the invention has the following beneficial effects: the methotrexate immunogen is high in specificity and immunogenicity, and the prepared anti-methotrexate specific antibody is high in specificity and valence, and free from any cross reaction with 62 common medicines; the homogenous enzyme immunoassay reagent containing the anti-methotrexate specific antibody is capable of conveniently, rapidly and accurately determining the content of methotrexate in a sample and measuring a plurality of samples on a fully-automatic biochemical analyser to realize high-flux rapid measurement for methotrexate, is high in accuracy and high in specificity, and is capable of greatly improving the accuracy and the detection efficiency.

The invention discloses a method for preparing a nanometer capsule and a method for preparing a nanometer capsule composite microsphere. The method for preparing the nanometer capsule comprises the following: step 1, preparing a methotrexate alkaline solution and a PLGA-CH2Cl2 solution; step 2, injecting the methotrexate alkaline solution into the PLGA-CH2Cl2 solution to obtain a W/O colostrum after ultrasonic emulsification; step 3, adding the colostrum into sodium cholate to obtain a W/O/W multiple emulsion after the ultrasonic emulsification; and step 4, placing the obtained multiple emulsion in the sodium cholate to be evaporated through decompression to remove CH2Cl2 so as to obtain a nanometer capsule particle suspension. The method for preparing the nanometer capsule composite microsphere comprises the following: step a, preparing a nanometer capsule; and step b, preparing the nanometer capsule composite microsphere. The nanometer capsule particle prepared by the invention has the characteristics of even size, higher drug loading and encapsulation efficiency, peak value drug release, and smooth drug release.

This invention relates to a novel process of manufacture of nanoparticles of substantially water insoluble materials from emulsions. The emulsions have the ability to form a single liquid phase upon dilution of the external phase, instantly producing dispersible solid nanoparticles. The formed nanoparticles have average diameter of about 10 to 200 nm and are suitable for drug delivery and targeting of water insoluble therapeutic or diagnostic agents. Examples of such agents are methotrexate, progesterone, testosterone, prednisolone, and ibuprofen. Such agents can be used in a wide range of therapeutic and diagnostic treatments including treatment for cancer, hormonal therapy, and pain management.

The invention provides a targeted fluorescent liposome based on low-frequency ultrasonic response, a preparation method of the liposome and an application, and particularly provides an active targeting medicine carrying fluorescent acoustic liposome which comprises a bimolecular lamellar lipid membrane, medicines and fluorescent agents, wherein the bimolecular lamellar lipid membrane comprises dipalmitoyl phosphatidyl choline (DPPC), cholesterol (CHO), polyethylene glycol modified distearoyl phosphoethanolamine (DSPE-PEG) and (DSPE-PEG-iRGD), and the acoustic liposome wraps the medicines and the fluorescent agents. The medicines are selected from methotrexate (MTX), the fluorescent agents are selected from indocyanine green (ICG), and polyethylene glycol (PEG) in the polyethylene glycol modified distearoyl phosphoethanolamine (DSPE-PEG) is selected from PEG1000-PEG3000.

The invention relates to a preparation method of a methotrexate and lecithin compound, which comprises the following steps: dissolving methotrexate and lecithin in an organic solvent A, carrying out mixed reaction at 20-40 DEG C for 12-24h, removing the organic solvent A in the mixed liquor after mixed reaction to obtain a dried product, and then purifying the dried product in which the organic solvent A is removed by adopting an organic solvent B, so as to obtain the methotrexate and lecithin compound. The invention also relates to a preparation method of nano-particles of the methotrexate and lecithin compound as well as a preparation method of a targeted sustained release preparation of the methotrexate and lecithin compound. The nano-particles of the methotrexate and lecithin compound, prepared by the method provided by the invention, have particle sizes of 100-500nm, good stability and better tumor targeting and sustained release characteristics. The targeted sustained release preparation of the methotrexate and lecithin compound, prepared by the method provided by the invention, has the advantages of targeting property, long circulation, low toxicity and good curative effect.

Gel nanoparticles for encapsulating and delivering a pharmaceutical compound to a patient. The nanoparticles are formed from N-trimethyl chitosan and polysialic acid, preferably in the presence of sodium tripolyphosphate. A ratio of polysialic acid to N-trimethyl chitosan of about 0.5 to 1 produces nanoparticles having diameter of about 100 nm (plus or minus 25 nm) and a zero potential above 30 milivolts that can stability contain a pharmaceutical compound, such as methotrexate, for delivery to a patient.

The invention provides a preparation method for conductive superparamagnetic nanometer gamma-ferric-oxide/polyaniline-methotrexate and relates to the technical field of chemical synthesis, in particular to a method for synthesizing the novel gamma-ferric-oxide/polyaniline-methotrexate. The preparation method prepares the nanometer gamma-ferric-oxide/polyaniline-methotrexate with conductive superparamagnetism by a doping method. In the preparation method, a doping mechanism of polyaniline is adopted, and methotrexate is doped into the conductive superparamagnetic nanometer gamma-ferric-oxide/polyaniline. The method for fixing medicaments has the unique advantage of avoiding influence on the treatment group of the fixed methotrexate. The gamma-ferric-oxide/polyaniline-methotrexate can suspend in water for three months or even longer. The method is simple and economic; the conductive superparamagnetic nanometer gamma-ferric-oxide/polyaniline-methotrexate has good electrochemical activity and good superparamagnetism under a nearly neutral environment, can be regulated and controlled to a specific position under the control of an external magnetic field and is expected to be an ideal targeted medicament, and to treat tumors in human bodies.

The invention discloses nano-scale mesoporous silicon dioxide-methotrexate-mitoxantrone nanoparticles. Its preparation method is disclosed, that is, amino functionalized mesoporous silica nanoparticles (MSNN) are prepared by modifying amino groups on the surface of mesoporous silica nanoparticles, and then the amino groups of MSNN are covalently combined with MTX to form methotrexate ‑Amino-modified mesoporous silica nanoparticles (MSNN‑MTX), and finally load mitoxantrone into the nanopores of MSNN‑MTX to obtain nanoscale mesoporous silica‑methotrexate/mitoxantrone Quinone nanoparticles (MSNN‑MTX/MIT). Its antitumor effect is disclosed. Compared with the combined application of conventional MTX and MIT, MSNN‑MTX/MIT significantly prolongs the survival time of S180 mice, improves the curative effect and reduces systemic toxicity. Therefore, the present invention discloses the application of MSNN-MTX/MIT in the preparation of antitumor drugs combined with MTX and MIT. The MSNN-MTX/MIT of the present invention has a good clinical application prospect.

The invention belongs to the field of material synthesis and biological medicine, and relates to a dual-drug controlled release system with pH and glutathione dual response and a preparation method thereof.The dual-drug controlled release system is constructed by adopting mesoporous silica, and sodium hyaluronate serves as a gating material to block pore channels of the mesoporous silica; Mesoporous channels of the mesoporous silica are loaded with cytarabine. The mesoporous silica, the chitosan and the sodium carboxymethyl cellulose form a gel system, and methotrexate is loaded in the gel system. The preparation method has the beneficial effects that the prepared pH and glutathione dual-response dual-drug controlled release system can load two anti-cancer drugs at the same time, and realizes the successive release of the two drugs in a subacid tumor environment and a high-concentration glutathione environment. The double-drug controlled release system is simple to prepare and high in biocompatibility, and can be widely applied to the field of biological medicines.

The invention discloses a preparation method of a multi-component nanometer medicine with a collaborative treatment effect and high drug loading. The preparation method comprises the following steps of: 1) dissolving three hydrophobic cancer-fighting medicines, namely, methotrexate, 10-hydroxycamptothecine and taxol which are in a mass ratio of 1: 1: 1, into dimethylsulfoxide; ultrasonically treating for 5 minutes to produce mixed solution with concentration of 3mg/mL; 2) dropwise adding 400 micro-millimeters of the mixed solution into ultrapure water; and magnetically stirring to obtain suspension liquid of the multi-component nanometer medicine; and 3) adding 300 micro-millimetres of polymaleic anhydride 18 carbene-polyethylene glycol with concentration of 1mg/mL to the suspension liquid of the multi-component nanometer medicine; ultrasonically dispersing for 5 minutes; and standing for 3 minutes. The preparation method is simple, efficient, high in repeatability, excellent in controllability and high in universality, and provides experimental bases for preparing the nanometer medicine applied to multi-medicine collaborative treatment in the cancer field.

The invention pertains to methods for treating ectopic pregnancy. More particularly, the present invention relates to methods for treating unruptured ectopic pregnancy using a non-surgical method comprising the administration of an EGFR inhibitor alone or in combination with an anti-metabolite e.g. methotrexate (MTX). The methodology is potentially applicable to treatment of unruptured ectopic pregnancies of all sizes.

The present invention relates to a recombinant expression vector for an animal cell containing a dihydrofolate reductase (DHFR) coding nucleotide sequence operatively linked to a DHFR promoter, to an animal cell line transformed by the vector, and to a method for preparing a target protein using the same. As compared with existing animal cell expression vectors, the vector of the present invention enables an effective screening of a cell line clone in which foreign genes are amplified together with DHFR genes even at a much lower methotrexate concentration. The present invention exhibits excellent effects in cell line preparation as high-productivity cell lines can be ensured in a short time through the use of a lower concentration of methotrexate in the process of protein production cell line establishment.

The invention discloses a pH responding dual-drug-loading system, a preparation method and application thereof. Combining calcium carbonate nano microspheres, sodium alginate and sodium carboxymethylcellulose together to prepare a pH responding dual-drug-loading system. The preparation method comprises the following steps: preparing anti-cancer drug/calcium carbonate nano microspheres, and preparing an anti-cancer drug/calcium carbonate/analgesic drug/sodium carboxymethylcellulose/calcium ion/sodium alginate dual-drug-loading system. The pH responding dual-drug-loading system prepared by the invention can simultaneously load an anti-cancer drug and an analgesic drug, and realizes independent release of the two drugs in different pH environments in vivo. In the dual-drug-loading system, a short-term release of the analgesic drug aspirin can be achieved, and a long-term and continuous release of the anti-cancer drug methotrexate can be achieved.

A crosslinking reaction and a reaction for reducing molecular weight that take place in a hyaluronic acid-methotrexate conjugate upon irradiation with light are suppressed to improve the photostability of the conjugate. Specifically, a substance having a quenching effect and/or a radical scaveng effect is added to a pharmaceutical composition containing the hyaluronic acid-methotrexate conjugate.