108 results about "Deoxyuridine" patented technology

The invention relates to the field of molecular biology and discloses mutant A type DNA (deoxyribonucleic acid) polymerase, and an encoding gene and an application of the mutant A type DNA polymerase. The mutant A type DNA polymerase is generated by amino acid site mutation of conservational motif of A type DNA polymerase in a dNTP (deoxyribonucleoside triphosphate) bond zone. Compared with the A type DNA polymerase not modified or mutated, the mutant A type DNA polymerase has increased dUTP (deoxyuridine triphosphokinase) doping speed; a dUTP doping effect of the mutant A type DNA polymerase is obviously better than control A type DNA polymerase; therefore, the mutant A type DNA polymerase is more applicable to some nucleic acid amplification systems using dUTP to substitute dTTP (deoxy- thymidine triphosphate) and allows the system to avoid nucleic acid amplification product contamination and not to lose amplification efficiency of a target product at the same time; and the mutant A type DNA polymerase meets application requirements of multiple PCR (polymerase chain reaction) fields of food, animal quarantine, human disease screening and the like, a forensic medicine field and a scientific research.

This invention relates to pharmaceutical formulations and formulation strategies of protides (phosphoramidate derivatives of nucleosides) and, in particular, protides useful in the treatment of cancer, such as NUC-3373 (5-fluoro-2′-deoxyuridine-5′-O-[1-naphthyl (benzoxy-L-alaninyl)] phosphate) and NUC-7738 (3′-deoxyadenosine-5′-O-[phenyl(benzyloxy-L-alaninyl)] phosphate). In particular, the invention relates to formulations that comprise a polar aprotic solvent, for example, dimethyl acetamide (DMA).

Purine N⁹-β-D-nucleosides containing 3-amino-3-deoxy-β-D-ribofaranose, 3-amino-2,3-dideoxy-β-D-ribofuranose, and 2-amino-2-deoxy-β-D-ribofuranose as sugar moieties are synthesized by biocatalytic transglycosylation of purine bases and the respective 3′-amino-3′-deoxyuridine, 3′-amino-3′-deoxythymidine and 2′-amino-2′-deoxyuridine as donors of the carbohydrate moiety, and the cells of Escherichia coli as a biocatalyst or glutaraldehyde (GA) treated cells of Escherichia coli as a biocatalyst or a mixture of thymidine (uridine) phosphorylase and purine nucleoside phosphorylase.

A 2′-deoxyuridine derivative of formula I is bioavailable and thermally stable and forms a gel in water at a low concentration; and, can be employed as a drug delivery vehicle:

A method of detecting N-glycosylase activity in a sample involves incubating an oligodeoxyribonucleotide substrate containing a deoxyadenosine or deoxyuridine residue with the sample to be tested such that the N-glycosylase, if present, hydrolyzes the deoxyadenosine or deoxyuridine residue to result in an N-glycosylase product having an abasic site. A primer is annealed to the N-glycosylase product, and the primer is extended with a DNA polymerase, such as Taq DNA polymerase, that pauses at abasic sites. The resulting extension products are melted from the N-glycosylase product, allowed to form hairpins due to self-complementarity, and further extended in the presence of labeled precursors to result in labeled products. Extension products synthesized from undigested substrate as template do not result in labeled products. Thus, detection of labeled products results in detection of N-glycosylase activity. Oligodeoxyribonucleotide substrates, primer, and positive controls and a kit for N-glycosylase assay are also disclosed.

The invention relates to a chemical method for detecting how deoxythymidine is oxidized and mutated into five-position aldehyde-group deoxidizing uridine, which includes: using 4-methoxy group o-phenylenediamine or o-phenylenediamine as a detection reagent, using dithiothreitol as an antioxidant reagent, and dissolving the detection reagent and the antioxidant reagent into 10mM of acetic acid buffer liquid with the potential of hydrogen (pH) of 4.5 to form a detection sample; and leading the detection sample to react for 3 hours under aerobic room temperature condition and using a fluorophotometric detector to detect a blank sample and the detection sample, wherein through the detection, users can find out that when the 4-methoxy group o-phenylenediamine is used as the detection reagent, the peak value shifts to 475nm, and when the o-phenylenediamine is used as the detection reagent, the peak value shifts to 430nm. The fluorescence detection method generates compounds capable of generating fluorescence by using the 4-methoxy group o-phenylenediamine or the o-phenylenediamine and 5-aldehyde group deoxyuridine which are easy to achieve and are arranged in acetic acid buffer liquid so that existence of oxidation and mutation of the deoxythymidine can be detected in fluorescence mode.

The invention discloses a preparation method of trifluridine. The method comprises the specific steps of: adding 2'-deoxyuridine and trifluoromethyl sulfinate sodium to a reaction solvent, stirring and cooling to - 5 to - 3 DEG C, introducing of nitrogen for protection, stirring to dissolve, dropwise adding tert-butyl hydroperoxide, controlling the temperature at less than 5 DEG C, heating to 60-65 DEG C for reaction, and conducting posttreatment after the reaction to obtain a finished product. The method provided by the invention has mild reaction conditions, simple operation, little side reaction, short reaction time, great reduction of feeding amount of tert-butyl hydroperoxide, great saving of the production cost, and high yield and high purity of the product, and is especially applicable to industrial production, and has significance to the quality control of medicine and clinical curative effect.

The invention relates to a novel virus double-fluorescence labeling method based on nucleic acid and protein biosynthesis, and belongs to the field of chemistry and biomedicine. The method comprises specific steps as follows: a methionine analogue methionine azide and a deoxythymidine analogue ethylene deoxyuridine are synthesized; viruses are added to host cells for infection, methionine azide and ethylene deoxyuridine are added respectively, vinyl derivation of viral nucleic acid and azido derivation of protein are realized with biosynthesis processes of nucleic acid and protein; double-fluorescence labeling of viral nucleic acid and protein is naturally realized through two biological orthogonal reactions including a Diels-Alder reaction of tetrazine-oefin and a copper-free catalytic click chemical reaction of azido-cyclooctyne. The labeling method is simple, convenient and reliable, can be applied to all DNA (deoxyribonucleic acid) viruses including single-stranded DNA viruses, double-stranded DNA viruses, enveloped viruses and non-enveloped viruses, and is a universal virus fluorescence labeling method on an absolute basis.

The invention relates to the field of pharmaceutical synthesis and particularly relates to a preparation method of 3'-deoxyuridine. The method comprises the steps: by adopting a compound 3 as a raw material, firstly protecting amino through acetic anhydride to obtain a compound 4, obtaining a compound 5 under the action of acetyl bromide, reducing through a hypophosphite system to obtain a compound 6; removing deacetylated amino under the action of high-pressure water vapor and an organic solvent to obtain a compound 8 or removing N-acetyl to obtain a compound 7; and finally removing all acetyl to obtain a mixture of 3'-deoxyuridine and 3'-deoxycytidine; separating and purifying to obtain 3'-deoxyuridine crystal and 3'-deoxycytidine crystal separately, or directly removing all acetyl through the compound 6 to obtain the 3'-deoxycytidine. Available natural products are taken as initial raw materials, so that the method is simple in operation and convenient to purify, and industrial large-scale production is extremely easy to implement.

The invention provides a method for detecting recombinant human keratinocyte growth factors (rhKGF-1). The method includes steps of 1), cultivating keratinocytes; 2) inoculating the keratinocytes and controlling the density of the keratinocytes; 3), treating rhKGF-1 samples, carrying gradient dilution on the concentration of the rhKGF-1 samples and adding 5-acetenyl-2'-deoxyuridine (EdU) into the rhKGF-1 samples with various rhKGF-1 concentration dilution degrees to prepare loading samples with the series of rhKGF-1 concentration dilution degrees; 4), washing the keratinocytes in keratinocyte plates obtained at the step 2) and removing cultivation media; 5), loading the loading samples obtained at the step 3) into keratinocyte cultivation plates obtained at the step 4) and cultivating and incubating the keratinocytes; 6), solidifying and permeabilizing the keratinocytes, and then carrying out 'Click-iT' reaction on the EdU; 7), arranging the keratinocytes treated at the step 6) in a multifunctional microplate reader, reading the fluorescence intensity (RFU) and computing the biological activity of the rhKGF-1. The method has the advantages of low detection background, high sensitivity, specificity and accuracy, good repeatability and the like. Besides, the method is particularly applicable to detecting the biological activity of the rhKGF-1 and establishing quality standards.

The invention discloses an aromatic acid ester derivative of 5-fluoro-2'-deoxyuridine and a synthesis method thereof. The synthesis method comprises the following steps: adding the 5-fluoro-2'-deoxyuridine with the concentration of 1.0-40mg/ml and an acyl donor to a non-aqueous medium, wherein, mol ratio of the 5-fluoro-2'-deoxyuridine to the acyl donor is 1:1.1-30; adding enzyme at a ratio of 0.1-10:1 based on the weight of the 5-fluoro-2'-deoxyuridine; performing an acylation reaction at the temperature of 20-60 DEG C and the normal pressure, and at an oscillation speed of 100-300rpm; and separating to obtain the aromatic acid ester derivative of 5-fluoro-2'-deoxyuridine. The method helps synthesize various aromatic acid ester derivatives of 5-fluoro-2'-deoxyuridine with novel structures and potential pharmaceutical value. The method has the advantages of mild condition, environmental protection, high regioselectivity of the reaction, simple and controllable reaction process, easy product separation and the like.

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 invention relates to the synthesis process of 5-trifluoromethyl-2'-dexoy uridine. The compound is prepared with 2'-dexoy uridine as initial material and through iodizing, hydroxyl protection, trifluoromethylation and deprotection. The said process of the present invention has easy-to-obtain material, simple synthesis and no pollution.

A process for an injection from frozen powder of doxifluridine applying to mainline and its preparation method, said injection comprise 5 parts of doxifluridine, 0.1-100 parts of excipient or 0.1-100 parts of cyclodextrin.; said product can be dissolved quickly when meeting water and convenient for clinical use.

The present invention discloses a new crystal form of trifluridine, and a preparation method thereof, and particularly relates to a new 5-trifluoromethyl-2'-deoxyuridine crystal form represented by a formula (I), and a preparation method thereof, wherein the characteristic peaks of the crystal form in the X-ray powder diffraction spectrum are positioned at (a) 7.3 DEG, (b) 10.0 DEG, (c) 14.5 DEG and (d) 23.4 DEG C by adopting 2[theta] (2[theta]+/-0.2 DEG) to represent. According to the present invention, the crystal form has characteristics of stable characteristics and good repeatability, and is suitable for drug development. The formula I is defined in the specification.

The invention discloses a synthesis method of 2'-deoxy-beta-uridine, and belongs to the technical field of medicinal chemistry. The synthesis method comprises the following steps: preparing 1-chloro-2-deoxy-3, 5-O-bis(4-chlorobenzoyl)-alpha-D- red-furanose from 2'-deoxyribose by a one-pot methylation, acylation and chlorine substitution method, then coupling with 2, 4-bis(trimethylsilyloxy)pyrimidine, and finally deacylating and recrystallizing to obtain the 2'-deoxy-beta-uridine. By the synthesis method, the defects that when uridylic acid is used as a raw material, stereoisomers are difficult to control and the quality is relatively poor are overcome, the reaction condition is simple, the raw materials are easy to obtain, the product is high in yield and purity and the energy consumptionis reduced, so that the industrial application prospect is greatly improved.