31 results about "Uridine phosphorylase" patented technology

Methods are provided for using methotrexate (MTX) in which reduced host toxicity is observed. Aspects of the methods include administering to a subject an effective amount of MTX in conjunction with a MTX toxicity-reducing adjuvant, such as a 2,2′-anhydropyrimidine, a derivative thereof or a uridine phosphorylase inhibitor. Also provided are compositions that find use in practicing embodiments of the invention. The methods and compositions find use in a variety of applications, including the treatment of a variety of different disease conditions.

Pyrimidine nucleotide precursors including acyl derivatives of cytidine, uridine, and orotate, and uridine phosphorylase inhibitors, and their use in enhancing resistance to sepsis or systemic inflammation are disclosed.

Pyrimidine nucleotide precursors including acyl derivatives of cytidine, uridine, and orotate, and uridine phosphorylase inhibitors, and their use in enhancing resistance to sepsis or systemic inflammation are disclosed.

The use of at least one compound of formula I or II or pharmaceutically acceptable salts thereof, for preparing a pharmaceutical composition that acts by inhibiting the phosphorylase uridine enzyme, and the use of the compounds for preparing a pharmaceutical composition that acts by inhibiting the human phosphorylase uridine enzyme, which can be optionally used in combination with at least one antineoplastic, wherein the inhibition increases the effectiveness of antineoplastic and decreases the side effects caused by the administration of antineoplastics.

Methods of treating a subject for a liver condition, e.g., NAFLD, NASH, and / or DILI, are provided. Aspects of the methods include administering to the subject an effective amount of a UPase inhibitor, optionally in combination with a uridine active agent (e.g., uridine (UR), a UR pro-drug or a UR mimetic), such as supplemental uridine, to treat the subject for the liver condition. Also provided are compositions for use in practicing the subject methods.

The invention provides a genetic engineering bacterium capable of producing uridine at high yield as well as a building method and application thereof. The genetic engineering bacterium is characterized in that pyrimidine nucleoside operons pyrBCAKDFE with the nucleotide sequence shown as SEQ ID NO:1 is integrated on a genome of colon bacillus; the starting is realized by a strong promoter P[trc];a uridine synthesis path is reconstituted; the self PRPP synthetase coding gene prsA on the genome is subjected to dual copying, and the starting is realized by the strong promoter P[trc]; meanwhile,the activity of udk, udp, and rihA, rihB and rihC is lacked; the thrA activity is lacked; the argF activity is lacked. The genetic engineering bacterium is applied to fermentation production of uridine; 40 to 67g / L of uridine can be produced after the fermentation is performed for 40 to 70h in a 5L fermentation tank; the maximum production intensity can reach 1.5g / (L*h); the glucoside conversionrate is 15 to 25 percent; the genetic engineering bacterium belongs to the highest level for producing the uridine by the fermentation method reported in the prior art.

The invention provides a method for preparing 2'-deoxyuridine by a chemical-biological enzyme method in combination. A 'crystallization induction asymmetric transformation' technology is adopted to synthetize and obtain single alpha-configuration nucleoside analogues medicine intermediate 2-deoxy-alpha-D-ribose-1-phosphate; the intermediate is utilized as a substrate; uracil is added, and 2'-deoxyuridine is synthetized and obtained by biotransformation under the action of uridine phosphorylase. By applying the chemical-biological combination technology disclosed by the invention, the advantages of a chemical method and a biological method are combined; the defects of the chemical method and the biological method are avoided; the yield is high; the method is suitable for industrial production; enough supply can be obtained by the 'crystallization induction asymmetric transformation' technology; 2-deoxy-alpha-D-ribose-1-phosphate intermediate with single configuration does not need to be separated and purified; the biotransformation process is finished by catalysis with efficient and specific uridine phosphorylase in one step; the specificity is strong; the conversion rate is high; the condition is mild and the environment is friendly.

A fermentation medium comprises: NaCl (3-12 g / L), glucose (10-25 g / L), extract yeast (5-15 g / L), peptone (2-15 g / L), K2HPO4-H2O (1 / 3) (2-5 g / L), uridine (7-25 mmol / L), inosine (15-30 mmol / L) and double distilled water. A fermentation method based on the fermentation medium comprises: streaking, inoculating and culturing lactobacillus brevis for 12-48 h, preserving at 4 DEG C for a standby application; taking and dissolving a certain amount of extract yeast in double distilled water, adjusting pH at 6.0-8.0, disinfecting at 121 DEG C for 15-20 min, and obtaining an activating solution containing 1-10% by mass of extract yeast; picking lactobacillus brevis, inoculating to the activating solution, activating at 30-40 DEG C for 4-12 h, and obtaining activated lactobacillus brevis thalluses; inoculating the lactobacillus brevis thalluses to the fermentation medium with an inoculation volume of 1-10% by volume; fermenting for 4-12 h with a medium temperature of 30-40 DEG C and a shaking-table rotating speed of 90-180 r / min, and obtaining a fermentation liquor; centrifuging the fermentation liquor for 10-30 min with a speed of 3000-5000 r / min, in a centrifuge tube obtaining a deposition which is the fermented whole-cell lactobacillus brevis wet thallus, and taking the fermented whole-cell lactobacillus brevis wet thallus as a uridine phosphorylase source to carry out nucleosides fermentation.

A method for producing cladribine (2-chloro-2′ deoxyadenosine) comprising the steps of: a) reaction of 2-deoxyuridine with 2-chloroadenine, in the presence of uridine phosphorylase (UPase) and purine nucleoside phosphorylase (PNPase) in an aqueous reaction medium possibly containing up to 40% v / v of an aprotic dipolar solvent, to obtain cladribine dissolved in said reaction medium; b) isolation of the cladribine by precipitation by means of concentration and alkalinisation of the reaction medium up to pH 11.5-12.5.

The present disclosure relates to a genetically engineered strain with high production of uridine and its construction method and application. The strain was constructed as follows: heterologously expressing pyrimidine nucleoside operon sequence pyrBCAKDFE (SEQ ID NO:1) on the genome of E coli prompted by strong promoter Ptrc to reconstruct the pathway of uridine synthesis; overexpressing the autologous prsA gene coding PRPP synthase by integration of another copy of prsA gene promoted by strong promoter Ptrc on the genome; deficiency of uridine kinase, uridine phosphorylase, ribonucleoside hydrolase, homoserine dehydrogenase I and ornithine carbamoyltransferase. When the bacteria was used for producing uridine, 40-67 g / L uridine could be obtained in a 5 L fermentor after fermentation for 40-70 h using the technical scheme provided by the disclosure with the maximum productivity of 0.15-0.25 g uridine / g glucose and 1.5 g / L / h respectively which is the highest level of fermentative producing uridine reported at present.