113 results about "Modified nucleosides" patented technology

Described herein are modified RNA molecules where a 3′-stabilizing region is covalently attached to the RNA, and where the 3′-stabilizing region comprises one or more modified nucleosides. Methods of synthesizing said RNAs are also provided herein.

The invention provides a nucleoside phosphoramidite compound as shown in a formula I. The invention further provides a preparation method of the nucleoside phosphoramidite compound. The nucleoside phosphoramidite compound disclosed by the invention can be introduced into the head and tail ends and / or the middle of oligonucleotide, the stability of the oligonucleotide is enhanced while the biological function of the oligonucleotide is enhanced or basically maintained, and the continuous acting time is prolonged.

The present disclosure relates to a nucleoside analogue and a use thereof. The nucleoside analogue is a compound shown in formula (Ia) or a prodrug thereof. The nucleoside analogue can enhance targeted delivery of oligonucleotide drugs to nervous system cells, thereby increasing the inhibition rate of the oligonucleotide drugs delivered in a targeted manner on expression of specific genes in the nervous system cells, or enabling the oligonucleotide drugs delivered in a targeted manner to achieve the purpose of preventing and / or treating pathological conditions or diseases caused by abnormal expression of the specific genes in the nervous system cells. The present disclosure also relates to a nucleotide analogue-containing double-stranded oligonucleotide and a use thereof. The double-stranded oligonucleotide comprises a sense strand and an antisense strand, each strand has 17-25 modified and / or unmodified nucleotides, and the antisense strand and the sense strand are complementary to form a duplex region; the sense strand and / or the antisense strand contains at least one nucleotide analogue represented by the structure of formula (100), or a tautomer, or stereoisomer, or pharmaceutically acceptable salt thereof; the double-stranded oligonucleotide can effectively treat and / or prevent pathological conditions or diseases caused by abnormal expression of specific genes in nervous system cells.

The invention discloses base-modified mRNA (messenger Ribonucleic Acid) and application thereof in a cell-free protein synthesis system, and belongs to the technical field of nucleic acid modification. Natural nucleoside triphosphate is completely replaced by different types of basic group modified nucleoside triphosphate, mRNA with different chemical modifications is obtained through an in-vitro transcription method and purification, and the mRNA is put into an in-vitro cell-free protein synthesis system for reaction. A reaction element recombination system-PURE system based on prokaryote Escherichia coli purified protein and an eukaryote CFPS system taking rabbit reticulated red lysate as a material are mainly selected to detect the corresponding translation efficiency of mRNA modified by different bases in two cell-free protein synthesis systems respectively. Different regulatory activities may be shown in different cell-free protein synthesis systems when different modified nucleotides are doped into mRNA, base modified triphosphoric acid capable of improving the translation efficiency of an in-vitro protein synthesis system is screened out, and the stability of mRNA in the cell-free protein synthesis system and the corresponding protein expression level are improved.

Provided are a double‑stranded RNA composition for modulating DGAT2 expression, a conjugate thereof, a pharmaceutical composition comprising same, and a use thereof. The double-stranded RNA comprises a sense strand and an antisense strand, and each nucleotide in the double-stranded RNA is independently a modified or unmodified nucleotide; the sense strand comprises one nucleotide sequence selected from the nucleotide sequences shown in SEQ ID NOs: 1-86 or a nucleotide sequence having no more than 3 nucleotide differences from the above sequences, and the antisense strand comprises one nucleotide sequence selected from the nucleotide sequences shown in SEQ ID NOs: 87-172 or a nucleotide sequence having no more than 5 nucleotide differences from the above sequences. The double-stranded RNA, the conjugate thereof and the pharmaceutical composition comprising same can be delivered to the liver by means of small nucleic acid delivery technology, and are expected to provide a safer and more effective therapeutic plan for patients with MASH-associated metabolic diseases.

This disclosure relates to the modified 3' region elements that confer improved therapeutic properties including, but not limited to, increased stability, increased expression, and / or a reduced innate immune response when introduced into a population of cells, even in the absence of a 5'-cap. The disclosure also provides polynucleotides (e.g., RNA molecules) containing modified 3' regions. In some embodiments of the disclosure, the modified 3' regions contain modified adenosines (e.g., 2'-0-methoxy- adenosine, 2'-fluoro-adenosine, or locked nucleic acid (LNA)-adenosine) and / or a modified thymidine (e.g., inverted deoxythymidine). In some embodiments of the disclosure, the modified 3' region may contain one or more modified internucleoside linkages (e.g., a phosphorothioate linkage) or modified nucleobase (e.g., N1-methyl-pseudouridine). The polynucleotides of the disclosure may be used to express a desired polypeptide in a subject, such as a subject having a disease or condition associated with a deficiency in the corresponding endogenous polypeptide.

Provided herein are RNA molecules (e.g. self-amplifying messenger ribonucleic acids (samRNA)) comprising modified nucleotides, such as N1-methyl-pseudouridine.

A single-stranded oligonucleotide having a length of 16-30 nucleotides. The single-stranded oligonucleotide and complement component C3 (CC3) mRNA have sufficient complementarity to mediate RNAi effect; each nucleotide in the single-stranded oligonucleotide is a modified or unmodified nucleotide, at least one nucleotide in the single-stranded oligonucleotide is a nucleotide X, and the at least one nucleotide is a fluoro-modified nucleotide; moreover, according to a direction from the 5' end to the 3' end, at least one nucleotide X is located after an eighth nucleotide and is spaced apart from the eighth nucleotide in the single-stranded oligonucleotide by 4-7 nucleotides; and each nucleotide X is a deoxynucleotide or an unmodified nucleotide. The present invention further relates to a double-stranded oligonucleotide comprising the single-stranded oligonucleotide as an antisense strand, an oligonucleotide conjugate, and a pharmaceutical composition.

A single-stranded oligonucleotide having a length of 16-30 nucleotides. The composition of the single-stranded oligonucleotide enables the single-stranded oligonucleotide to inhibit the expression of target mRNA by means of an RNAi mechanism. Each nucleotide in the single-stranded oligonucleotide is independently a modified or unmodified nucleotide, wherein at least one nucleotide in the single-stranded oligonucleotide is a nucleotide X; at least one nucleotide is a fluoro-modified nucleotide; and, in the 5' to 3' direction, the 13th nucleotide of the single-stranded oligonucleotide is a substituted alkoxy-modified nucleotide, the 14th nucleotide of the single-stranded oligonucleotide is a nucleotide X, and each of the 15th nucleotide and all subsequent nucleotides of the single-stranded oligonucleotide is independently a modified nucleotide. A double-stranded oligonucleotide comprising the single-stranded oligonucleotide as an antisense strand, an oligonucleotide conjugate and a pharmaceutical composition.

Disclosed herein are improved methods and compositions of de novo synthesis of nucleic acids by cyclic extension using nucleotides that have an N-linked or O-linked scar or protecting group, and subsequent removal of the scar or protecting group. During polymerase-nucleotide conjugate-based polynucleotide synthesis or after the synthesis is completed, nucleobases having a scar from linker cleavage can be converted back into native form by removal of the scars from the nucleobases. Secondary structure formation in the nascent chain, which may inhibit extension reactions, is suppressed by the use of monomers with protected nucleobases that prevent Watson-Crick base pairing and / or other structures. After the synthesis is completed, the nucleobases can be converted back into native form by palladium-based removal of the protecting groups.

The present invention relates to the detection of modifications in polynucleotides by binding an affinity binding element to a nucleotide sequence. The affinity binding element may include a compound that binds to a modified nucleotide base and a nucleotide sequence complementary to a target nucleotide sequence, an oligonucleotide primer complementary to the target nucleotide sequence, or a nucleotide sequence complementary to an oligonucleotide linker. The present invention provides methods for detecting one or more modifications in a nucleotide sequence. The invention also provides affinity binding elements, and optionally, oligonucleotide primers and / or oligonucleotide probes, and methods of using the affinity binding elements in assays to detect modified bases. The methods of the invention enable the detection of modified nucleotides associated with cancer, psychiatric disorders or metabolic diseases.

Provided herein is an antisense oligonucleotide for targeting long non-coding RNA maternally expressed gene 3 (IncRNA Meg3), wherein the oligonucleotide comprises or consists of one of the following sequences in the 5' to 3' direction: (I) C G C dC dG dC dC dA dT dA dT dC dT C C C (SEQ I D NO: 1 ); (ii) C G G dA dC dA dA dA dA dC dT dG dG T T G (SEQ ID NO: 2); (ill) A A G dA dG dT dC dT dC dC dT dC dC dT T A A (SEQ ID NO: 3); (iv) G A T dT dA dG dC dC dC dT dG dT dG dT T C A (SEQ ID NO: 4); (v) C G G dC dA dC dA dA dG dA dG dC dC dA A A G (SEQ ID NO: 5); (vi) T O T dC dC dT dC dC dT dT dA dA dG dC C C C (SEQ ID NO: 6); or (vii) G T C dT dC dC dT dC dC dT dT dA dA dG C C C (SEQ ID NO: 7); wherein dA, dT, dG and dC are deoxyribonucleotides, or wherein dC is 5-methyl-2'-deoxycytidine; and wherein A, T, G and C are modified or unmodified nucleotide building blocks.

Disclosed herein are modified RNAs comprising one or more one or more modified nucleotides at position +1 to position +6 with reference to a 5' terminus of the RNA, and methods of making the same. Also provided are compositions comprising one or more of the modified RNAs provided herein, and methods of using said compositions for therapeutic applications.

Disclosed are a modified cytidine compound, i.e. a new derivative cytidine generated by adding a guanidyl at position 4 of a cytidine, and a nucleic acid containing the modified compound, for example, RNA. The nucleic acid containing the modified cytidine, especially mRNA, can significantly increase the expression quantity of the mRNA in vivo.

A nucleoside phosphoramidite compound, the compound being a 2'-phosphoramidite group, 4'-modified TNA structured compound. The nucleoside phosphoramidite compound can be used to incorporate into the first and / or last and / or intermediate of an oligonucleotide such that the modified oligonucleotide hybridizes to a portion of a target mRNA, thereby causing a loss or down-regulation of normal function of the target mRNA.

This invention provides the application of a class of nucleoside compounds that effectively inhibit CCHFV replication, particularly the application of 2'-α-fluoro-2'-β-methyl modified nucleoside compounds in the preparation of drugs for treating CCHF disease. Studies have found that 2'-α-fluoro-2'-β-methyl-CTP can specifically and efficiently inhibit CCHFV L protein polymerase through an immediate chain termination mechanism, but it cannot be incorporated into the L protein polymerases of Lassa virus (LASV) and Rift Valley fever virus (RVFV), which belong to the same order Arenavirales but different families as CCHFV. Further introduction of this modification into ATP, UTP, and GTP also exhibits good immediate chain termination activity. This invention provides a new direction for the preparation of anti-CCHFV nucleoside drugs and demonstrates its feasibility.

The present invention relates to 5'-modified nucleoside analogs and oligonucleotides produced therefrom, and more specifically, to modified nucleosides and their analogs that can be used to be incorporated into the oligonucleotide terminus, which can be bound to double-stranded oligonucleotides (short interfering RNA) or single-stranded oligonucleotides (e.g., antisense oligonucleotides). The oligonucleotides provided herein are expected to result in the loss of normal function of the target RNA by hybridizing to a portion of the target RNA.

The invention belongs to the field of biological medicine, and relates to siRNA for inhibiting MAPT gene expression and a conjugate and application thereof. The siRNA comprises a positive-sense strand and an antisense strand, the positive-sense strand comprises a nucleotide sequence I, and the antisense strand comprises a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reversely complementary to form a double-stranded region; the nucleotide sequence I is basically consistent with a first section of nucleotide sequence, and the first section of nucleotide sequence is a section of nucleotide sequence with the length of at least 19 nucleotides in mRNA expressed by the MAPT gene. The siRNA and the conjugate thereof provided by the invention can specifically target the liver, complementarily pair with a liver MAPT mRNA sequence and induce MAPT mRNA degradation, so that synthesis of MAPT in the liver is inhibited, Tau protein is reduced, and neurodegenerative diseases can be effectively prevented and / or treated.

This invention provides an siRNA for inhibiting MSTN gene expression, its conjugates, and their applications. The siRNA comprises a sense strand and an antisense strand, wherein the sense strand comprises nucleotide sequence I, and the antisense strand comprises nucleotide sequence II; each nucleotide in nucleotide sequence I and nucleotide sequence II is a modified or unmodified nucleotide. The siRNA, its conjugates, and the pharmaceutical composition provided by this invention exhibit strong inhibitory activity against the MSTN gene, significantly reducing the expression level of MSTN mRNA, and have low drug toxicity.

The technology described herein relates to 5 '-modified nucleosides, nucleotides, oligonucleotides and double stranded RNAs, such as siRNAs, as well as kits comprising them and their use for inhibition of target genes.

Disclosed in the present invention are a modified nucleoside and nucleoside monomer, an oligonucleotide containing the modified nucleoside monomer and the use thereof. According to the present invention, lipophilic / hydrophilic groups, fragments with a pharmacological activity and receptor-targeting properties, etc., are introduced to modify a nucleoside, thereby obtaining a modified nucleoside and a phosphoramidite monomer thereof. The modified nucleoside has significantly improved physicochemical properties and bioavailability. According to the present invention, the modified nucleoside is further incorporated into an oligonucleotide, such that the cellular free uptake efficiency of an oligonucleotide drug can be significantly enhanced, and a target mRNA can be silenced. The present invention holds promise for achieving efficient delivery of a modified nucleic acid drug to various organs and tissues for the treatment of relevant indications.

The invention provides a novel nucleic acid polymerase which can be used for a single molecule sequencing technology. The polymerase belongs to a Phi29 polymerase family, but the sequence homology of the polymerase is lower than that of other reported polymerase for a sequencing technology by 60%. Through characterization identification, the polymerase shows good comprehensive performance in the aspects of thermal stability, extension capacity at normal temperature, extension proceeding, utilization capacity / chain substitution function catalytic kinetics for modified nucleotide substrates, electrolyte resistance and the like, and is suitable for a single-molecule sequencing technology based on a scheme of'sequencing while synthesizing '. The invention also provides a beneficial mutant strain of the wild type polymerase, which shows better dynamic characteristics and can improve the sequencing accuracy and flux.

The present invention provides compounds, methods, and pharmaceutical compositions for reducing the amount or activity of PLP1 RNA in cells or subjects, and, in some cases, the amount of proteolipid protein 1 in cells or subjects. [Solution] The present invention provides an oligomeric compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides, wherein the nucleic acid base sequence of the modified oligonucleotide is at least 85% complementary to equal-length portions of the PLP1 nucleic acid, and the modified oligonucleotide comprises at least one modification selected from modified sugar moieties and modified nucleoside-to-nucleoside bonds. Such compounds, methods, and pharmaceutical compositions are useful for improving at least one symptom or characteristic of leukodystrophy.

The invention belongs to the field of biological medicine, and relates to siRNA, a conjugate thereof and application of the siRNA and the conjugate in related diseases such as blood fat. The siRNA comprises a positive-sense strand and an antisense strand, the positive-sense strand comprises a nucleotide sequence I, and the antisense strand comprises a nucleotide sequence II; each nucleotide in the nucleotide sequence I and the nucleotide sequence II is modified or unmodified nucleotide; the nucleotide sequence I and the nucleotide sequence II are at least partially reversely complementary to form a double-stranded region; the nucleotide sequence I is basically consistent with the first nucleotide sequence, and the first nucleotide sequence is a nucleotide sequence with the length of at least 19 nucleotides in mRNA expressed by the PCSK9 gene. According to the present invention, the siRNA and the conjugate thereof can specifically target the liver, can be complementarily paired with the liver PCSK9 mRNA sequence, and can induce the PCSK9 mRNA degradation, such that the synthesis of the PCSK9 in the liver can be inhibited, the PCSK9 serum level can be durably and significantly reduced, and the related diseases caused by excessive PCSK9 can be effectively prevented and / or treated.

The present disclosure generally relates to methods for the enzymatic synthesis of nucleotide triphosphates. In particular, the methods include a one-pot preparation of a modified nucleotide triphosphate starting from 3´-vinyl blocked nucleoside monophosphate.

The present disclosure provides a single-stranded oligonucleotide, wherein the single-stranded oligonucleotide has a length of 16-30 nucleotides and can inhibit the expression of APOE4 mRNA by the mechanism of RNA interference (RNAi); wherein each nucleotide in the single- stranded oligonucleotide independently of one another is a modified or unmodified nucleotide; and wherein at least one nucleotide is a nucleotide X, and at least one nucleotide is a fluoro modified nucleotide; and in a 5' to 3' direction, in the single-stranded oligonucleotide, the 13th nucleotide is a substituted alkoxy modified nucleotide; the 14th nucleotide is a nucleotide X; and each of the 15th nucleotide and all the subsequent nucleotides independently of one another is a modified nucleotide; and each nucleotide X is independently a deoxyribonucleotide or an unmodified nucleotide. The present disclosure also provides a double-stranded oligonucleotide, an oligonucleotide conjugate and a pharmaceutical composition comprising the single-stranded oligonucleotide as an antisense strand.

Provided herein are systems and methods for analyzing nucleic acids. The present disclosure relates in aspects to methods of detecting an RNA molecule suspected of having a modified nucleotide. The method can further comprise detecting a location of the modified nucleotide on the RNA molecule.