14 results about "Dihydrofolic acid" patented technology

A conjugate provided. The conjugate is a compound represented by Formula (I), or the conjugate is a stereoisomer, tautomer, homologue, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of the compound represented by Formula (I):where A is a linker, and B is a dihydrofolate reductase inhibitor; R1, R2, R3, R4, and R5 are independently selected from H or C1-6 alkyl; and n1, n2, and n3 are each an integer selected from 0 to 6.

The present disclosure relates to providing an engineered CHO cell line wherein the two essential metabolic genes are knocked out. Particularly, the present invention relates to a double knockout CHO cell line (DHFR− / − and GS− / −) with disrupted Dihydrofolate Reductase (DHFR) and Glutamine Synthetase (GS) genomic loci. The double knockout CHO cell line (DHFR− / − and GS− / −) being suitable for expression of monoclonal antibodies, dimeric therapeutic proteins, Fab, single chain fragments, or the like. The present disclosure also provides method of generation of a double knock out CHO cell line (DHFR− / − and GS− / −) using gene selection and / or manipulating techniques such as CRISPR / Cas9 system, Zinc Finger Nuclease, TALEN, or the like. The present disclosure further provides method of selection of clones and production of therapeutic proteins of interest with increased titre.

The present application relates to an assay method using affinity crosslinking of DNA-linked ligands to proteins to enable small molecule screening and determination of apparent affinity constants of ligands to the proteins. This method has been applied to determine 96 compounds' dissociation constants to a protein target simultaneously, and directly determine a compound's IC50 against five protein targets concurrently in crude cell lysates. Additionally, this approach was used to screen a Library of Pharmacologically Active Compounds (LOPAC) library against dihydrofolate reductase (eDHFR), enabling the discovery of a novel eDHFR inhibitor (IC50=7.9 μM). An assay kit and the method of uses are within the scope of this disclosure.

This invention belongs to the fields of pharmaceutical raw material preparation and compound preparation, specifically relating to a method for preparing calcium folinate. The method includes: irradiating folic acid in a solvent with low light intensity in the presence of a photocatalyst and hydrogen donor I to achieve a first-stage hydrogen transfer reduction; after the concentration of dihydrofolate in the system reaches its peak, adding hydrogen donor II for high-intensity irradiation to achieve a second-stage hydrogen transfer reduction; then stopping the irradiation and sequentially performing a formylation reaction and a calcium salt precipitation reaction to obtain calcium folinate. The method provided by this invention not only enables a one-pot reaction but also improves the yield of calcium folinate, showing broad application prospects.

The invention relates to a method for synthesizing levofolinic acid based on enzymatic cascade reaction, and belongs to the technical field of bioengineering and pharmacy. The reaction system provided by the invention comprises dihydrofolate reductase, tetrahydrofolate formyltransferase and an NAD (P) dependent coenzyme circulation system. The method comprises the following steps: taking folic acid as a substrate, enabling the folic acid to be subjected to asymmetric hydrogenation reaction under the catalysis of dihydrofolate reductase (DHFR) to obtain an intermediate compound L-tetrahydrofolic acid, and then catalyzing the intermediate compound L-tetrahydrofolic acid by tetrahydrofolate formyltransferase (FTCD) (taking N-formyl-L-glutamic acid as a formyl donor) to obtain optically pure L-5-formyl tetrahydrofolic acid (namely levofolinate). The method for preparing the levofolinic acid does not need to separate an intermediate product, does not need to split, is high in yield, and has great application value and market prospect.

The invention relates to a coding gene of tetrahydrofolate methyltransferase, the tetrahydrofolate methyltransferase, an engineering bacterium and application of the tetrahydrofolate methyltransferase in preparation of L-5-methyltetrahydrofolate, and belongs to the technical field of bioengineering. The coding gene of the tetrahydrofolate methyltransferase provided by the invention is derived from alpha proteus, an engineering bacterium based on the coding gene, the tetrahydrofolate methyltransferase produced by the engineering bacterium, and a method for producing L-5-methyltetrahydrofolate by combining the tetrahydrofolate methyltransferase with dihydrofolate reductase LbuDHFR and alcohol dehydrogenase LbADH. And the concentration of the substrate is obviously improved, the substrate conversion rate is high, and the catalyst has higher catalytic activity and shows good application value.

The invention relates to the technical field of medicine, in particular to a pyrrole quinazoline diamine derivative and a preparation method and application thereof, the preparation method of the pyrrole quinazoline diamine derivative is mild in reaction condition, the reaction process is easy to operate, and used reagents are low in price and easy to obtain. The compound has excellent broad-spectrum antibacterial activity, bactericidal ability and drug resistance, and has good biological safety to cells. Cell membrane permeability experiments and transmission electron microscopes show that the compound can destroy bacterial cell membranes, so that intracellular substances leak to cause bacterial death; a dihydrofolate reductase inhibition experiment shows that the compound has high affinity with e-DHFR and can effectively inhibit enzyme activity. The pyrroloquinazoline diamine derivative compound provided by the invention effectively inhibits the growth of bacteria and resists the generation of drug resistance through the double-target action of bacterial DHFR activity inhibition and bacterial cell membrane destruction.

The application belongs to the technical field of anti-infective drugs, and discloses 7H-pyrrolo[3,2-f]quinazoline derivatives and anti-infective applications thereof. Specifically, it relates to 1,3-diamino-7H-pyrrolo[3,2-f]quinazoline heterocyclic compounds, a preparation method of the compounds and the use of the compounds in the preparation of anti-infective drugs. The compounds contain an aromatic fused ring structure, and are connected to the 1,3-diamino-7H-pyrrolo[3,2-f]quinazoline skeleton through different carbon chains. Compared with existing dihydrofolate reductase inhibitors, the compounds have better antibacterial activity and can be used as anti-infective drugs.

In one aspect, compounds and associated compositions are described herein for the treatment of various bacterial infections, including antibiotic resistant infections. In some embodiments, an antibacterial composition described herein comprises a dihydrofolate reductase inhibitor of Formula (I) and / or a salt thereof, and potentiator of the dihydrofolate reductase inhibitor of Formula (I). Moreover, in some embodiments, the antibacterial composition further comprises a thymine component, thereby enabling the antibacterial composition to selectively target bacterial species incapable of utilizing exogenous thymine.

The present application belongs to the technical field of veterinary nutritional supplements or feed additives, and particularly relates to a rumen-protected folic acid preparation, a preparation method thereof and application thereof in improving lactation performance of ruminants. The rumen-protected folic acid preparation prepared by the present application is double-coated, and the degradation rate thereof in the rumen can be as low as 12.5% in 12 hours, and the release rate thereof in the small intestine can be as high as 71.7%, effectively solving the technical problem that more than 95% of ordinary folic acid is degraded in the rumen, and significantly improving the absorption and utilization rate of 5-methyltetrahydrofolic acid (5-MTHF) in the small intestine. In addition, ordinary folic acid needs to be reduced by dihydrofolic acid reductase (DHFR) and methylene tetrahydrofolic acid reductase (MTHFR) in two steps to generate active 5-MTHF, and the metabolic efficiency is limited; the present application directly supplements 5-MTHF, bypasses the metabolic bottleneck, and has a more direct and efficient effect, and thus can significantly improve the lactation performance and milk quality of ruminants.

This invention belongs to the field of enzyme engineering technology, specifically relating to proteins with tetrahydrofolate-dependent demethylase activity and their application in the synthesis of L-5-methyltetrahydrofolate. This invention utilizes two proteins with tetrahydrofolate-dependent demethylase activity, namely *Sphingosine monocytogenes* (…). Sphingomonas paucimobilis Vanillic acid O-demethylase and Amorpha fruticosa intermediate rhizobia (from) Mesorhizobium amorphae The aminomethyltransferase derived from 6S-THF uses inexpensive vanillic acid or dicamba as methyl donors to directly methylate 6S-THF to synthesize L-5-MTHF. Combined with highly active dihydrofolate reductase and NADP cofactor regeneration system, this method effectively achieves efficient and green synthesis of L-5-methyltetrahydrofolate, and has good industrialization prospects.

The application relates to the field of drug synthesis and discloses a one-pot enzyme method for preparing L-5-MTHF. The method provided by the application adds dihydrofolate reductase and tetrahydrofolate methyltransferase into folic acid, so that the folic acid is subjected to asymmetric hydrogenation reaction under the catalysis of dihydrofolate reductase (DHFR), an intermediate compound L-tetrahydrofolic acid is obtained, and then the L-tetrahydrofolic acid is subjected to catalysis by tetrahydrofolate methyltransferase (DmdA) to obtain L-5-MTHF with optical purity. The one-pot enzyme method can avoid the separation step of an intermediate product, meanwhile, the produced intermediate is converted into the product L-5-MTHF at the moment, the product is used immediately, and the problem of product inhibition is avoided. The method for preparing L-5-MTHF has high yield, great application value and market prospect.

The present disclosure relates to mutant polypeptides derived from Escherichia coli dihydrofolate reductase (DHFR) which can be fused to a polypeptide of interest for efficient conditional modulation of its activity. Also disclosed are polynucleotides encoding such mutant polypeptides, vectors comprising such polynucleotides, and the use of such polypeptides, polynucleotides and vectors for treating a disorder.