87 results about "Phosphoramidite" patented technology

Methods featuring phosphoryl guanidinium based backbone and morpholino with phosphoramidite chemistry result in neutral antisense oligonucleotides. The PGMO is composed of a morpholino backbone linked to a phosphoryl guanidinium internucleotide (PG) linkage. These oligonucleotides can be used to treat cancer, autoimmune diseases, and other rare diseases.

The present disclosure, among other things, provides technologies for oligonucleotide synthesis. In some embodiments, the present disclosure provides phosphoramidites and methods for synthesis thereof. In some embodiments, provided methods provides higher yields and / or purities. In some embodiments, provided methods remove byproducts without contact with an aqueous solution.

The present invention relates to 5′-morpholino amidite monomers as 5′-CE / 5′-tBu phosphoramidite morpholino monomers (2) and process chemistry for the efficient synthesis of N-Trityl or monomethoxytrityl (MMTr)-protected 5′-morpholino amidite monomers and their use in the synthesis of phosphorodiamidate morpholino oligonucleotides (PMO), thiophosphoramidate morpholino oligonucleotides (TMO) and their chimeras which can be used for antisense technology or in general oligonucleotides related research.

The invention provides a preparation method of a phosphoramidite compound. The preparation method comprises the following steps: (1) mixing a nitrogen-containing compound and an acid-binding agent with a first solvent to obtain a first mixed solution; mixing phosphorus trichloride with a second solvent to obtain a second mixed solution; mixing aromatic diphenol with a third solvent to obtain a third mixed solution; (2) pumping the first mixed solution and the second mixed solution obtained in the step (1) into a first micro-mixer, and then enabling the first mixed solution and the second mixed solution to enter a first micro-channel reactor for reaction to obtain a reaction solution containing monochlorophosphoramidite; (3) enabling the reaction solution containing the monochlorophosphoramidite obtained in the step (2) and the third mixed solution obtained in the step (1) to enter a second micro-mixer, and then enabling the mixture to enter a second micro-channel reactor for reaction to obtain a phosphoramidite compound; the acid-binding agent is a polyether tertiary amine compound. The preparation method is short in reaction time and high in product yield.

The invention relates to the technical field of biological medicines, and particularly discloses synthesis and application of oligonucleotide with a spacer group embedded between 5-terminal oxygen and thiophosphoric acid. The chemical modification strategy comprises the following steps: preparing a modified nucleotide phosphoramidite monomer in which a spacer group is embedded between O5'and thiophosphoric acid P (V); performing solid-phase synthesis on the modified nucleotide phosphoramidite monomer to construct a target oligonucleotide molecule; a spacer group is embedded between oxygen at the 5 '-terminal of the oligonucleotide and thiophosphoric acid P (V) to form a 5'-terminal thiophosphoric acid structure; as the terminal modified thiophosphoric acid belongs to a non-phosphatase substrate, the terminal modified thiophosphoric acid can resist exonuclease degradation and improve the biological activity of siRNA after being modified.

The invention discloses a method for detecting 5 '(E)-VPA phosphoramidite monomer related substances, and belongs to the technical field of liquid chromatography detection. Related substances comprise 5 '-(E)-VP-2'-OMe-A (Bz), 5 '(Z)-VPA, 5'-(E)-VP-2 '-OMe-A-CE-P and 5' (E)-VPA-OX, and chromatographic detection conditions comprise that the column temperature is 23-27 DEG C, a stationary phase of a chromatographic column is octadecyl bonded silica gel, a mobile phase A is an ammonium acetate solution, a mobile phase B is acetonitrile, gradient elution is performed, and the like. According to the method, four related substances in the 5 '(E)-VPA phosphoramidite monomer are effectively separated through specific liquid chromatography conditions, the system adaptability, specificity, linearity and range, detection limit, quantitation limit and accuracy verification of the method all meet acceptable standards of pharmacopoeia, and detection of multiple related substances can be completed within a short time; and a reliable means is provided for detecting and controlling the quality of the 5 '(E)-VPA phosphoramidite monomer.

The invention discloses a method for preparing a chiral benzhydrol compound through asymmetric hydrogenation of o-amido benzophenone under catalysis of rhodium. The chiral rhodium catalyst is generated in situ from a metal rhodium salt and a chiral phosphine-phosphoramidite ligand in various solvents. Compared with other methods for synthesizing chiral benzhydrol, the method has the characteristics of cheap catalyst, easiness in preparation of chiral ligand, mild reaction conditions, simplicity and convenience in operation, high yield, high enantioselectivity and the like, and can be applied to large-scale preparation of chiral benzhydrol compounds. The method is also suitable for efficient preparation of the rice plant growth regulation inhibitor (S)-Inabenide, the yield can reach 92%, the enantioselectivity can reach 96%, and the method has good industrial application prospects.

An object is to provide an amidite monomer of diaminopurine and / or an amidite monomer of a thiouracil derivative suitable for use in synthesis of an acyclic single-stranded polynucleotide comprising diaminopurine and / or a thiouracil derivative by a phosphoramidite method. This object is achieved by an amidite monomer of diaminopurine and / or an amidite monomer of a thiouracil derivative protected with a specific protecting group removable by a base.

The invention belongs to the technical field of medicinal chemistry, and relates to a novel small nucleic acid drug intermediate and a preparation method thereof, the intermediate is a 2 '-FANA-dU phosphoramidite monomer, the preparation method comprises the following steps: 1, reacting a compound a with a 33% hydrogen bromide-acetic acid solution in dichloromethane, and performing post-treatment to obtain a compound b; 2, reacting uracil and the like to generate a white solid, and reacting the white solid with the compound b to obtain a compound c; 3, reacting the compound c in methanol and ammonia water, crystallizing and drying to obtain a compound d; 4, the compound d reacts with 4, 4 '-dimethoxytriphenylchloromethane, and a compound e is obtained through treatment; 5, the compound e reacts with anhydrous diisopropyl imidazole and a 2-cyanoethyl-N, N, N ', N'-tetraisopropyl phosphoramidite reagent, and the 2 '-FANA-dU phosphoramidite monomer is obtained.The 2'-FANA-dU phosphoramidite monomer prepared through the method can modify siRNA, the biological activity of the siRNA can be reserved or even enhanced, and the biological stability of the siRNA can be remarkably improved.

The invention provides a device for DNA solid-phase synthesis and a preparation method of the device. The device comprises a solid-phase synthesis carrier and a micropore array which is arranged on the solid-phase synthesis carrier and is provided with a wrinkled inner wall. The pore diameter of the micropores is 70-90 [mu] m, the pore depth is 5-6 [mu] m, the inner wall roughness is 0.5-5 [mu] m, the distribution density of the micropores on the surface of the solid-phase synthesis carrier is greater than 1111 / mm < 2 >, and the reaction contact surface provided by the solid-phase synthesis carrier is further expanded; meanwhile, hydrophilic treatment and synthesis starting point grafting are carried out on the inner walls of the micropores, hydrophobic membrane coating is carried out outside the micropores, the synthesis density is increased, and the error rate of the synthesis process is reduced. According to the solid-phase synthesis device, the reaction contact area is further enlarged while the synthesis accuracy is ensured, the coupling density is increased, and a high-yield synthesis device is provided for solid-phase synthesis of DNA by a phosphoramidite method.

This method for producing an oligonucleotide comprises a step for reacting a phosphoramidite compound represented by formula (II), or an optical isomer thereof, and a compound represented by formula (III). (In formula (II), ring A, B, X, Y, Z, R1, R2, R5, R6, R7, R8, R9, R10, p, and m are as defined in the description. In formula (III), B, X, Y, Z, R9, R12, and q are as defined in the description.)

The invention discloses synthesis of 5 '-amino-3-nucleotide phosphamide and application of the 5'-amino-3-nucleotide phosphamide in oligonucleotide. The phosphamide modified oligonucleotide comprises a nucleotide structural unit shown as a formula I or a formula II,..., the formula I; ... formula II; through specific limitation of a nucleotide phosphamide monomer structure, the prepared oligonucleotide contains a phosphamide structural unit; compared with a traditional oligonucleotide molecule, the oligonucleotide molecule with the phosphamide modified nucleotide structure has more excellent effects on pharmaceutical parameters such as enzymatic degradation resistance, stability and the like, and in addition, phosphamide modification can improve the binding performance between the modified oligonucleotide molecule and environmental ions; a novel chemical modification strategy is provided for research, development and application of oligonucleotide drugs.

The invention relates to the technical field of biological medicine, and particularly discloses synthesis and application of oligonucleotide with threose nucleic acid 3-oxygen at the tail end indirectly connected with thiophosphoric acid. The chemical modification strategy comprises the following steps: O < 3->-oxygen of threose nucleic acid is connected with thiophosphoric acid through a spacer group, and O < 2->-hydroxyl is combined with a nucleotide monomer of phosphoramidite; carrying out solid-phase synthesis on the threose nucleic acid O2 '-phosphoramidite monomer to construct an oligonucleotide molecule; the oxygen of the threose nucleic acid at the terminal of the oligonucleotide is linked to the thiophosphoric acid through a spacer group, and since the type of phosphonic acid does not belong to a substrate of phosphatase, the modified oligonucleotide can resist exonuclease degradation and improve its in vivo biological activity.

Disclosed are methods for making targeting ligand-conjugated nucleotide phosphoramidites known as adem-A-GalNAc phosphoramidites, which can be used in the synthesis of oligonucleotides, such as therapeutic oligonucleotides.

Methods are disclosed for making a 4′-phosphate analog phosphoramidite known as MeMOP, which can be used in the synthesis of oligonucleotides such as therapeutic oligonucleotides.

The application provides an imidazole-phosphoramidite bidentate ligand, a synthesis method thereof, a metal catalyst prepared from the imidazole-phosphoramidite bidentate ligand and application of the metal catalyst, and relates to the technical field of catalyst preparation.The imidazole-phosphoramidite bidentate ligand developed by the application has stable chemical properties and is not easy to be oxidized, and solves the problems of poor stability of an imidazole-based bidentate ligand, harsh use and storage conditions in the prior art, and the synthesis method is simple, and the total yield can reach 59%; the imidazole-phosphoramidite bidentate ligand has strong coordination ability, the metal nickel catalyst prepared from the imidazole-phosphoramidite bidentate ligand has high catalytic activity, and in a carbon dioxide carboxylation reaction, the metal nickel catalyst exhibits excellent reaction activity, and is a ligand with very good industrial application prospect.

To provide a method for synthesizing an oligonucleotide by using a segment-type amidite.SOLUTION: A method for producing an oligonucleotide comprises performing at least one coupling step for coupling a nucleoside phosphoramidite to a hydroxyl group or a thiol group at the 3' or 5' of a nucleotide or a nucleoside in the presence of an activator, wherein, in the at least one coupling step, the nucleoside phosphoramidite is (a) a nucleoside phosphoramidite having two or more nucleoside portions or (b) a nucleoside phosphoramidite having at least one nucleoside portion and a linker portion, and the activator has a structure represented by formula (1), wherein in formula (1), X represents an organic base, or by formula (2), wherein in formula (2), R1 and R2 are each independently selected from the group consisting of H, straight chain or branched chain C1-7 alkyl groups and optionally substituted aromatic groups.SELECTED DRAWING: None

The invention provides a novel chiral phosphine-phosphoramidite ester ligand which is based on an achiral methylene bisphenol phosphoramidite ester structural unit and only has skeleton single-center chirality, and a preparation method and application thereof. Different from a traditional chiral phosphine-phosphoramidite ester ligand, the novel chiral phosphine-phosphoramidite ester ligand has the advantage that a phosphoramidite ester structural unit in the novel chiral phosphine-phosphoramidite ester ligand does not have a chiral element. The novel ligand is prepared by taking a chiral phosphine / amine intermediate (R)-or (S)-DPPNHMe, a methylene bisphenol compound and PC13 as raw materials under a mild condition. The novel chiral phosphine-phosphoramidite ligand provided by the invention has the characteristics of cheap and easily available raw materials, simple synthesis, stable properties, adjustable space structure and electronic properties, excellent activity and enantioselectivity in catalysis of asymmetric hydrogenation reaction, and the like.

The application provides a nucleoside dimer, a preparation method thereof and application in DNA synthesis. The nucleoside dimer of the application adopts a mixed skeleton of methyl phosphate and beta-cyanoethyl phosphoramidite, a new preparation method of the nucleoside dimer is obtained by optimizing a synthesis method and a purification process, and a foundation for high-quality DNA synthesis is laid. On this basis, the nucleoside dimer with the new skeleton is applied to DNA synthesis, an oligonucleotide fragment synthesis method suitable for the dimer is established, and an oligonucleotide fragment with a length of 100 nt is synthesized. Fidelity experiment shows that the error rate of the DNA fragment synthesized by using the dimer is only 3.75 errors / Kb, and high-fidelity DNA synthesis is realized. The preparation method of the nucleoside dimer with the new skeleton can realize large-scale and high-purity preparation, the nucleoside dimer can play an important role in improving the length and fidelity of oligonucleotide synthesis, and has practical value.

The invention discloses a synthesis process of chiral UNA phosphoramidite, phosphoryl chloride and phosphoramidite and application of the chiral UNA phosphoramidite, phosphoryl chloride and phosphoramidite in oligonucleotides, and particularly designs preparation and synthesis of (2S)-UNA monomer phosphoramidite, phosphoryl chloride and phosphoramidite. And the method can also be used for solid-phase synthesized oligonucleotides in the conventional 3-to-5 direction, and the influence of introduction of monomers with different chirality to design sites on the structure, the biological activity and the like is researched.

The present invention provides a method for efficiently producing a nucleic acid oligomer, particularly a method for efficiently oxidizing a nucleic acid precursor that has a phosphite triester bond. Specifically, the present invention provides a method which is for producing, via the phosphoramidite method, a nucleic acid compound that has, at the 5' terminal, a nucleotide represented by formula (I) and in which a nucleic acid that has at least one phosphorothioate bond is produced, said method comprising a step for reacting: a precursor that has, at the 5' terminal, a phosphite triester bond represented by formula (II); an aromatic hydrocarbon or an aromatic heterocyclic compound that has a conjugate acid pKa of less than 5; and an oxidizing solution that contains iodine, pyridine, and water. [The substituents in formulas (I) and (II) are as defined in the description.]

The present invention belongs to the technical field of biochemistry, and specifically relates to a phosphoramidite lipid molecule and a use thereof in an RNAi molecule. In the present invention, a series of lipid molecules containing phosphoramidite are prepared and reacted with oligonucleotides to prepare an RNAi molecule. The RNAi molecule can effectively target extrahepatic tissues, while not interfering with the effect of inhibiting a target gene. The lipid molecule prepared in the present invention has good prospects in extrahepatic delivery vectors, and the RNAi molecule prepared in the present invention has important value for the research and development and clinical application of RNAi drugs.

The invention relates to the technical field of biological medicines, and particularly discloses application of nucleotide with oxygen and phosphoramidite embedded spacer groups, threose nucleic acid TNA and ethylene glycol ribose GNA in oligonucleotides. The chemical modification strategy comprises the following steps: embedding nucleotide phosphoramidite of a spacer group between O3'and P (III), TNA phosphoramidite and GNA phosphoramidite to construct a target oligonucleotide molecule through solid-phase synthesis; the oligonucleotide comprises a nucleic acid unit with a spacer group embedded between O3'and phosphate P (V) and TNA or GNA, and the former can be directly coupled with the TNA or GNA and can also be linked with the TNA or GNA at an interval of one or more nucleotide units; the prepared and synthesized oligonucleotide contains a nucleotide unit of an O3 '-spacer group-P (V), can release O2'-hydroxyl, and is closer to the structure of natural nucleic acid. By finely adjusting the space structure of the modified oligonucleotide, the stability of the modified oligonucleotide drug molecule can be effectively improved, and the pharmacokinetic (PK) and pharmacodynamic (PD) performance can be optimized at the same time.

The embodiment of the present application provides a nucleic acid synthesis method. The method comprises connecting a linker to a solid phase carrier; synthesizing a first nucleic acid sequence by using the solid phase carrier connected with the linker, connecting the linker to the 5' end of the first nucleic acid sequence, and synthesizing a second nucleic acid sequence based on the linker to obtain a synthesis product; and performing cleavage treatment on the synthesis product to obtain a first nucleic acid product and a second nucleic acid product; wherein the linker comprises a phosphoramidite group; and the linker is connected to the solid phase carrier by a phosphoramidite triester method. Compared with the prior art, the present application has at least one of the following beneficial effects: reducing the production cycle and time cost of the nucleic acid synthesis carrier, realizing the synthesis of multiple sequences on a single synthesis carrier, improving the synthesis flux and sequence diversity, and improving the stability of the synthesis carrier.

Disclosed is a catalyst composition comprising: (a) a transition metal; and (b) a diphosphoramidite ligand having the structure of formulas (I), (II), or (III): The catalyst composition is particularly useful for the hydroformylation of olefins to form aldehydes.

The application discloses a phosphoramidite ligand with a carbazole structure and a preparation method and application thereof. The structural formula of the phosphoramidite ligand is shown in the following formula: wherein R1 or R3 is independently selected from hydrogen or a phenyl group, and R2 is selected from hydrogen or a methyl group. The carbazole structure is introduced into the SPHENOL type phosphoramidite ligand, compared with a traditional imino stilbene structure, the carbazole structure makes the N-terminal modification of the phosphoramidite ligand simpler and easier, different substituents are added on different positions of the carbazole structure, and a series of different SPHENOL type phosphoramidite ligands can be derived, which is a new attempt to enrich the SPHENOL type phosphoramidite ligand. The phosphoramidite ligand has a regulating effect on the formation of chirality in a catalytic reaction, and sometimes shows different performance from traditional phosphoramidite in the chiral regulation, has unique advantages, and therefore, on the basis of the traditional phosphoramidite ligand, the application range of the ligand is expanded.

The invention discloses a gRNA (guide Ribonucleic Acid) synthesis method which comprises the following steps: step 1, solid-phase synthesis circulation: loading a solid-phase carrier into a synthesis column, then carrying out four-step circulation of deprotection, coupling, oxidation and cap reaction, and sequentially connecting nucleotide monomers to form a gRNA chain; step 2, post-treatment: after solid-phase synthesis is completed, cutting the gRNA strand from a solid-phase carrier and removing a protecting group, and then purifying and freeze-drying to obtain a gRNA product; wherein the nucleotide monomer is a phosphoramidite monomer with a protecting group at the 2'position of a sugar ring, and in the coupling reaction, the phosphoramidite monomer and an activating agent are mixed and activated in an organic solvent and then react with free hydroxyl on a solid-phase carrier. According to the method provided by the invention, the steric hindrance of molecules in the coupling process is effectively reduced, the coupling efficiency is greatly improved by selecting a proper activating agent and optimizing the adding proportion of the activating agent, the purity and the yield of the gRNA crude product are further improved, the HPLC (High Performance Liquid Chromatography) purity of the crude product is greater than 30%, and the large-scale production and development of the gRNA are facilitated.

Click-labeled uridine bases, nucleosides, and phosphoramidites are provided, including improved methods of synthesis, oligonucleotides comprising the click-labeled nucleosides, methods of synthesizing spin-labeled oligonucleotides using click-labeled nucleotides, and spin-labeled oligonucleotides comprising click-labeled nucleosides.

The present invention discloses a rapid liquid chromatography analysis method for phosphoramidite monomers, comprising the following steps: preparing a sample solution: weighing a sample and placing it in a volumetric flask, dissolving and then constant volume to obtain a sample solution; preparing mobile phase A and mobile phase B, filtering through a microporous membrane, and ultrasonically degassing the sample solution, and then detecting the phosphoramidite purity using reversed-phase high-performance liquid chromatography; and calculating the purity results using an area normalization method. The present invention adopts reversed-phase high-performance liquid chromatography for analysis, which has the advantages of short analysis time, good component peak separation effect, high resolution, high analytical accuracy, good reproducibility, and simple operation, thereby achieving effective control of the quality of the phosphoramidite.

The invention relates to the technical field of biological medicine, and particularly discloses synthesis and application of oligonucleotide with a spacer group embedded in C4-oxygen and thiophosphoric acid at the tail end. The chemical modification strategy comprises the following steps: connecting C4-oxygen of nucleotide with thiophosphoric acid through a spacer group Y, and combining O3-hydroxyl with a nucleotide monomer of phosphoramidite; carrying out solid-phase synthesis on the nucleotide phosphoramidite to construct an oligonucleotide molecule; c4-oxygen at the tail end of the oligonucleotide is connected with thiophosphoric acid through a spacer group Y, the formed O4-Y-PSO22 < 2-> structure is phosphonic acid in a non-natural form and does not belong to a substrate of phosphatase, and the modified oligonucleotide can resist exonuclease degradation and improve the biological activity of the oligonucleotide.