236 results about "Uracil formation" patented technology

Provided are compounds represented by: X is O, S or NR6, R1 is H or (CH2)mR5, R2, R2', R3 and R3' are independently NO2, N3 or (CH2)mR5, OH R4 is H, OR6, SR6, NR6R6a, CN, C(O)OR6, C(O)NR6R6a, R6, OR7 or (CH2)nR7, R5 is H, halo, OR6, SR6, NR6R6a, CN, C(O)OR6, C(O)NR6R6a, R6, OR7 or (CH2)mR7, R6 and R6a are individually H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl or substituted aryl, R7 is: R8 is H, F, SR9 or OR9, R9 is H, alkyl, alkenyl, alkynyl, aryl or hydroxyprotecting group, Y is H, CH3 or (CH2)mR5, Z is O or S W is CH2, CF2, CHF or O, m is 0-4, B is adenine, guanine, cytosine, uracil, thymine, modified purines and pyrimidines substituted pyridines, five membered heterocycles substituted by at least one of amines, substituted amines, amides, substituted amides, esters, halogens, alkyls, ethers; and pharmaceutically acceptable salts thereof and prodrugs thereof. These ring systems may be substituted.

The present invention provides novel nucleoside and nucleotide derivatives that are useful agonists or antagonists of P1 or P2 receptors. For example, the present invention provides a compound of formula A-M, wherein A is modified adenine or uracil and M is a constrained cycloalkyl group. The adenine or uracil is bonded to the constrained cycloakyl group. The compounds of the present invention are useful in the treatment or prevention of various diseases including airway diseases (through A2B, A3, P2Y2 receptors), cancer (through A3, P2 receptors), cardiac arrhythmias (through A1 receptors), cardiac ischemia (through A1, A3 receptors), epilepsy (through A1, P2X receptors), and Huntington's Disease (through A2A receptors).

The present invention provides uracil-DNA glycosylase (UDG) gene originating from Psychrobacter sp. HJ147, and amino acid sequences deduced from the gene; expression and purification of Psp HJ147 UDG gene in Escherichia coli; and characterization of UDG obtained therefrom, and the use thereof in a polymerase chain reaction (PCR). The UDG according to the present invention has a specific activity of excising uracil bases in a uracil-containing DNA substrates at a low temperature, and is easily heat-inactivated. It thus can effectively eliminate cross contamination and carry-over contamination of PCR templates often occurring after a PCR process using dUTP. Therefore, it is useful for increasing preciseness (elimination of false positives), purity and amplification efficiency of PCR.

A novel nucleotide derivative, in case of existing as a member of a single-stranded sequence, undergoing a change in the fluorescent signal intendity depending on the corresponding base type in the partner strand with which the single-stranded sequence is hybridized, and which is a thymin / uracil derivative (1) emitting light most intensely when a confronting base in the partner strand with which the single-stranded nucleotide sequence is hybridized is adenine; a cytosine derivative (2) emitting light most intensely when the confronting base is guanine; an adenine derivative (3) emitting light most intensely when the confronting base is cytosine; a guanine derivative (4) emitting light most intensely when the confronting base is cytosine or thymine / uracil; and an adrnine derivative (5) emitting light most intensely when the confronting base is thymine / uracil / .

This invention provides a nucleotide analogue comprising (i) a base selected from the group consisting of adenine, guanine, cytosine, thymine and uracil, (ii) a deoxyribose, (iii) an allyl moiety bound to the 3′-oxygen of the deoxyribose and (iv) a fluorophore bound to the base via an allyl linker, and methods of nucleic acid sequencing employing the nucleotide analogue.

Methods and preparations for treating disorders of the eye and / or causing posterior vitreous disconnection or disinsertion. Preparations containing a) urea, b) urea derivatives (e.g., hydroxyurea, thiourea), c) a non-steroidal anti-inflamatory agents, d) antmetabolites, e) urea, urea derivatives, non-enzymatic proteins, nucleosides, nucleotides and their derivatives (e.g., adenine, adenosine, cytosine, cytadine, guanine, guanitadine, guanidinium, thymidine, thimitadine, uradine, uracil, cystine), uric acid, calcium acetal salicylate, ammonium sulfate or other compound capable of causing non-enzymatic dissolution of the hyaloid membrane or e) any of the possible combinations thereof, are administered to the eye in therapeutically effective amounts.

The invention is related to the detection of a methylated cytosine in a nucleic acid wherein guanidinium hydrogen sulfite is used for the preparation of a solution containing guanidinium ions and sulfite ions and subsequent modification of the nucleic acid. Thereby, a non-methylated cytosine is converted to uracil. The invention further discloses kits for performing the methods of the invention.

A methylation state information about CpG island in the promoter region of the gene associated with primary liver cancer, its use and the method and reagent kit for detecting primary liver cancer aredisclosed. Said reagent kit contains the methylation-specific restriction endonuclease and the promoter CpG island-specific primer pair of the gene associated with liver cancer, or the reagent for transforming the methylation cytosine to uracil and the primer for the promoter CpG island of the gene associated with liver cancer. Said genes are also disclosed.

The present invention relates to a method for treating a cancer comprising orally administering a composition containing α,α,α-trifluorothymidine (FTD) and 5-chloro-6-(1-(2-iminopyrrolidinyl)methyl)uracil hydrochloride in a molar ratio of 1:0.5 at a dose of 20 to 80 mg / m2 / day in terms of FTD in 2 to 4 divided portions per to patients in need of the treatment.

The invention discloses a method and application for improving application efficiency of a gene targeting technique in aspergillus terreus, and belongs to the technical field of gene engineering. The method comprises the following steps: firstly, by taking Aspergillus terreus as an initial bacterium, knocking off a ku80 gene or an lig4 gene so as to increase the exogenous DNA homologous recombination probability of a strain; secondly, establishing a pyrG gene deletion uracil auxotroph stain, establishing a inheritance conversion system based on a pyrG gene as a screening tag; and finally, cutting off the screening tag by using a Cre / LoxP specific binding site recombinant system, thereby obtaining a uracil auxotroph stain which can be applied to genetic modification again. By adopting the method disclosed by the invention, an efficient aspergillus terreus gene targeting platform can be established, the method has the advantages that high homologous recombination efficiency can be achieved, the bidirectional screening of the conversion system can be achieved, a screening tag cutting method is simple and feasible, the screening tag can be recycled, and the like, and basic support can be provided for efficient genetic modification of aspergillus terreus by using the gene targeting technique.

In the present invention, a sequence segment conforming to the following rules (a) to (d) is searched from the base sequences of a target gene of RNA interference and, based on the search results, siRNA capable of causing RNAi is designed, synthesized, etc.: (a) The 3′ end base is adenine, thymine, or uracil, (b) The 5′ end base is guanine or cytosine, (c) A 7-base sequence from the 3′ end is rich in one or more types of bases selected from the group consisting of adenine, thymine, and uracil, and (d) The number of bases is within a range that allows RNA interference to occur without causing cytotoxicity.

The present invention concerns a method for the detection of cytosine methylation in DNA samples, wherein the following steps are conducted: (a) a genomic DNA sample, which comprises the DNA to be investigated and background DNA, is chemically treated in such a way that all of the unmethylated cytosine bases are converted to uracil, whereas the 5-methylcytosine bases remain unchanged; (b) the chemically treated DNA sample is amplified with the use of at least 1 primer oligonucleotide as well as a polymerase, whereby the DNA to be investigated is preferred as the template over the background DNA, and (c) the amplified products are analyzed and the methylation status in the DNA to be investigated is concluded from the presence of an amplified product and / or from the analysis of additional positions.

The invention discloses a preparation method of a ribofuranose phosphate derivative. The preparation method comprises preparation steps as follows: L-alanine isopropyl ester hydrochloride, phenol dichlorophosphate and substituted phenol are taken as starting materials and have a docking reaction under the action of alkali; (2R)-2-deoxy-2-difluoro-2-methyl-D-erythropentonic acid GAMMA-lactone and 3,5-dibenzoate reduce carbonyl in a dichloromethane or ether solvent into an alcoholic hydroxyl group under the action of a strong reducing agent; an intermediate with a formula 2-1 has a reaction with paratoluensulfonyl chloride under the action of alkali to obtain p-toluenesulfonates; an intermediate with a formula 2-2 and a benzoyl cytosine derivative have a docking reaction under the action of a condensing agent; an intermediate with a formula 2-3 converts cytosine into uracil under the action of organic acid; benzoyl protection for an intermediate with a formula 2-4 is released under the action of an alkaline agent; an intermediate with a formula 2-5 and an intermediate with a formula 1 are docked under the action of a Grignard reagent to obtain Sofosbuvir.

Particular aspects of the present invention provide a method for quantification of two different variations of a DNA sequence. Particularly, the invention relates to a quantification of methylated DNA, and for this purpose, the test DNA is converted so that cytosine is converted to uracil, while 5-methylcytosine remains unchanged. The converted DNA is amplified by means of a real-time PCR, wherein two labeled real-time probe types are utilized: one specific for methylated DNA; and one for unmethylated DNA. Preferably, the degree of methylation of the test DNA is calculated from the ratio of the signal intensities of the probes or from the Ct values. The inventive methods have substantial utility for diagnosis and prognosis of cancer and other disorders associated with altered or characteristic DNA methylation status, as well as having substantial utility for analysis of SNPs, allelic expression, and prediction of drug response, drug interactions, among other uses.

The invention specifically discloses a new crystalline form of 5-chlorine-6-(1-(2-iminopyrrolidyl)methyl)uracil hydrochloride, and discloses a preparation method thereof. The crystalline form has the advantages of good stability, high crystallinity, easiness in preparation and the like, and can be used as a thymidine phosphorylase inhibitor for preparing colorectal cancer treating drugs.

The invention relates to a method for analyzing cytosine methylations in DNA sequences, according to which non-methylated cytosines are first converted into uracil while 5-methylcytosine remains unmodified. The DNA is then amplified by means of a polymerase and at least one primer whose 5 end is connected to a probe via a linker. The probe is intramolecularly hybridized onto the amplified products in accordance with the methylation state of the DNA, hybridization being detectable via different detection systems. The inventive method is particularly suitable for diagnosing and predicting cancer diseases and other diseases associated with a modification of the methylation state as well as for predicting undesired effects of medicaments.

The invention discloses a method for detecting thymine DNA glycosylase activity based on a cyclophorase-remediation mediated double-signal amplification strategy. A fluorescent method for detecting the thymine DNA glycosylase activity in real time can be achieved through circular rolling circle index amplification triggered by thymine DNA glycosylase and assisted by enzyme remediation and circular cutting catalyzed and mediated by endonuclease IV. As efficiency of circular rolling circle amplification assisted by uracil excision and efficiency of circular cutting based on endonuclease IV are both high and index amplification mediated by uracil can greatly inhibit non-specific amplification, the method has very high detection flexibility; detection shows that lower detection limit of the method to thymine DNA glycosylase is 5.6*10<-7>U / mu L; meanwhile, the method disclosed by the invention utilizes self repair characteristics of the thymine DNA glycosylase and uracil DNA glycosylase to enable the whole repairing reaction to be performed strictly according to a natural repairing mechanism, so that the method disclosed by the scheme of the invention has very high specificity.

Presence or absence of methylation of C located on 5' side of G in a sample DNA comprising a target sequence containing a dinucleotide consisting of C that may be methylated and a nucleotide on the 3' side of the C (CpN dinucleotide) is determined based on a result of hybridization performed for a plurality of capture oligonucleotides immobilized on a base material and including at least an oligonucleotide having a nucleotide sequence complimentary to or identical to a nucleotide sequence corresponding to the target sequence in which C other than C in the CpN dinucleotide is replaced with T and an oligonucleotide having a nucleotide sequence complimentary to or identical to a nucleotide sequence corresponding to the target sequence in which all of C's are replaced with T's and the sample DNA in which cytosines not methylated are converted into uracils by deamination or an amplification product thereof.

Methods for fragmenting and labeling DNA in a single reaction volume and incubation step using a uracil DNA glycosylase, an apurinic / apyrimidinic endonuclease, and a terminal transferase are disclosed. In a preferred embodiment the UDG, AP and TdT activities are first mixed together to form an enzyme mixture and then the enzyme mixture is mixed with the uracil containing DNA. The fragmentation and labeling reactions thus take place simultaneously as part of the same reaction. The methods may be used in a variety of applications where fragmenting and end-labeling single or double stranded DNA is desired.

The present invention exploits the discovery that amounts of uracil and thymine metabolites, especially β-aminoisobutyric acid, in various bodily fluids, especially urine, are correlated with the occurrence of epilepsy when compared to matched control subjects. Analytical and diagnostic protocols, including a novel high performance liquid chromatography system, for use in the invention are disclosed.

Systems, kits and methods are disclosed for assessing epigenetic regulation of genome function via deoxyribonucleic acid (DNA) methylation status. The systems, kits and methods rely on a covert then capture concept in which unmethylated cytosine residues in a nucleic acid sequence are first converted to uracil residues and then captured for subsequent analysis. The systems, kits and method use a solution-phase capture probe pool having a mixture of at least three (3) types of capture probes. One type is “wobble” probes, in which some cytosine residues in a nucleic acid sequence are randomly assumed to be unmethylated and thus converted to uracil residues, while other cytosine residues are assumed to be methylated and thus conserved as cytosine residues. Moreover, each type of probe can include a mixture of probes that bind / hybridize to one or the other strand of a nucleic acid sequence of interest, thereby improving sequencing depth and reliability.

The invention discloses an establishing method for a lung cancer characteristic metabolite fingerprint spectrum in urine. The gas chromatography and mass spectrum combined technology is adopted to detect metabolic profiles of urine samples of lung cancer patients and healthy donors, and is combined with the biological method of a support vector machine to build a smoker lung cancer metabolite fingerprint spectrum and a non-smoker lung cancer metabolite fingerprint spectrum, wherein the smoker lung cancer metabolite fingerprint spectrum comprises oxalic acid, phosphoric acid, uracil, threonine, 5-pidolic acid, citric acid and galactose or comprises phosphoric acid, uric acid, citric acid and oxalic acid; and the non-smoker lung cancer metabolite fingerprint spectrum comprises alanine, oxalic acid, phosphoric acid, uracil, serine, threonine, 5-pidolic acid, ribose, aconitate, citric acid, galactose, tyrosine, palmitic acid and stearic acid. The method can detect only needing the urine samples, satisfies the requirements of effectiveness, simplicity and feasibility in the operation aspect, and is suitable for large-scale application.

The present invention relates to the discovery that certain beta -L-dioxolane nucleoside analogs which contain a uracil base, and preferably, a 5-halosubstituted uracil base, exhibit unexpectedly high activity against Epstein-Barr virus (EBV), Varciella-Zoster virus (VZV) and Herpes Virus 8 (HV-8). In particular, the compounds according to the present invention show potent inhibition of the replication of the virus (viral growth) in combination with very low toxicity to the host cells (i.e., animal or human tissue). Compounds are useful for treating EBV, VZV and HV-8 infections in humans.

The invention discloses a composition for modifying a nucleotide sequence, a method and application, and relates to the technical field of gene editing. The composition comprises a first vector and asecond vector, wherein the first vector is provided with the following expression elements, such as a cytosine deaminase expression element, an adenine deaminase expression element and a mutant Cas enzyme expression element; the second vector is provided with the following expression elements, such as a gRNA (guide ribonucleic acid) expression element and a uracil glycosidase inhibitor expressionelement. The composition is used for modifying the C bases at the 1st to 16th sites at the upstream of PAM (protospacer adjacent motif) of the target nucleotide sequence, so that the transformation ofC / G to T / A is realized. Compared with the existing gene editing technique, the composition and the method have the advantages that the working window is broader; the DSB (double-strand break) and indels (insertion / deletion mutations) are not introduced, the off-target effect is very low, the safety is higher, and the like.

The invention discloses a method of detecting the UDG activity by enzyme-mediated two-step serial signal amplification based on excision repair. The method comprises the following steps: (1) adding a UDG active substrate in an excision reaction buffer solution to carry out excision repair reaction, excising a uracil base of the UDG active substrate, and leaving an abasic site; and (2) adding an excision repair reaction product in an amplification reaction buffer solution to carry out enzyme-assisted two-step serial signal amplification reaction, namely inducing strand displacement reaction and index amplification reaction, generating strengthened fluorescence signals in a circulation manner, and determining the UDG activity through weakness of the fluorescence signals. According to the method provided by the invention, ultrahigh sensitive detection to the activity of uracil DNA glycosylase (UDG) is realized by utilizing high amplification efficiency of constant-temperature index amplification reaction and specific circulation digestion of ribonuclease H.

Disclosed are compounds having the structure: wherein, R1 is H, NH2, NH—(C1-C4)alkyl, or N—[(C1-C4)alkyl]2; R2 and R3 are each independently H, (C1-C4)alkyl, or wherein R4 is H, (C1-C4)alkyl, halogen, hydroxy, (C1-C10)alkoxy, cyano, nitro, —NR5R6, or —OCONR7R8; wherein R5 and R6 are each independently H, or a substituted or unsubstituted (C1-C4)alkyl; and wherein R7 and R8 are each independently H, or substituted or unsubstituted (C1-C4)alkyl, or (C1-C10)aryl; and wherein only one of R2 and R3 is H, enantiomers, tautomers, and pharmaceutically acceptable salts of the compounds, pharmaceutical compositions containing such compounds or salts, and processes for their preparation. The subject invention also provides methods of alleviating symptoms of neurologic and inflammatory disorders, methods of preventing oxidation of lipids, proteins, or deoxyribonucleic acids on a cellular level, and methods of protecting human red blood cells from lysis by O2 radicals.

The present invention relates to a method for treating a cancer comprising orally administering a composition containing α,α,α-trifluorothymidine (FTD) and 5-chloro-6-(1-(2-iminopyrrolidinyl)methyl)uracil hydrochloride in a molar ratio of 1:0.5 at a dose of 20 to 80 mg / m2 / day in terms of FTD in 2 to 4 divided portions per to patients in need of the treatment.

An improved process and intermediate compounds (N) for the preparation of 2-(N,N-disubstituted)amino-4-(perfluoroalkyl)-1,3-oxazin-6-one compounds having structural formula (I) and an improved process for the preparation of 6-(perfluoroalkyl)uracil compounds having structural formula (V).