60 results about "Phosphoramidite" patented technology

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 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.

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.

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.

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 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 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.

The invention discloses a method for preparing a chiral amine compound through asymmetric hydrogenation of ruthenium catalyzed imine. The chiral ruthenium catalyst is a chiral metal ruthenium complex prepared by reacting metal ruthenium salt with a chiral phosphine-phosphoramidite ester ligand and chiral diamine in a solvent and carrying out column chromatography. Compared with other chiral amine synthesis methods, the method provided by the invention has the characteristics of cheap catalyst, easy preparation of chiral ligand, mild reaction conditions, simple operation, high yield and enantioselectivity and the like, and can be applied to large-scale preparation of chiral amine compounds. The method is also suitable for synthesis of the key intermediate of the herbicide (S)-metolachlor, the yield can reach 92%, the enantioselectivity can reach 90%, and the method has good industrial application prospects.

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.

Embodiments for the synthesis of sensitive oligonucleotides as well as insensitive oligonucleotides are provided. Sulfur-based groups are used for the protection of exo-amino groups of nucleobases, phosphate groups and 2′-OH groups, and as cleavable linker for linking oligonucleotides to a support. Oligonucleotide syntheses are achieved under typical conditions using phosphoramidite chemistry with important modifications. To prevent replacing sulfur-based protecting groups by acyl groups via cap-exchange, special capping agents are used. To retain hydrophobic tag to assist RP HPLC purification, special phosphoramidites are used in the last synthetic cycle. With the sulfur-based groups for protection and linking, oligonucleotide deprotection and cleavage are achieved via oxidation followed by beta-elimination under mild conditions. Therefore, besides for insensitive oligonucleotide synthesis, the embodiments of the invention are capable for the synthesis of oligonucleotide analogs containing sensitive functional groups that cannot survive the harsh conditions used in prior art oligonucleotide synthesis technologies.

The present invention addresses the problem of providing: a phosphoramidite-type capping agent for nucleic acid synthesis, the capping agent having excellent solubility, stability, and capping efficiency; a solution of the capping agent; and a method for producing a nucleic acid using the capping agent. A capping agent for nucleic acid synthesis according to the present invention comprises a compound represented by general formula (1). In general formula (1), Cy forms, together with an adjacent -O-P-O- group, an aliphatic heterocyclic group including a 5- or 6-membered ring and optionally having a substituent, and R1 and R2 each independently represent an aliphatic ring group or an aromatic ring group including a 5- or 6-membered ring and optionally having a substituent.

The application discloses a synthesis method of phosphorodiamidate morpholino oligonucleotide, belongs to the technical field of solid-phase synthesis of phosphorodiamidate morpholino oligonucleotide, and specifically relates to the following steps: adding 9-fluorenylmethyloxycarbonyl-piperazine-succinic acid into Wang resin to mix and react to synthesize 9-fluorenylmethyloxycarbonyl-piperazine-succinic acid-Wang resin, then adding phosphoramidite monomers to perform coupling reaction after deprotection treatment, performing cap treatment after the coupling is completed, then repeatedly performing deprotection treatment, coupling reaction and cap treatment until the coupling of all phosphoramidite monomers is completed, then removing the protecting group of the last phosphoramidite monomer, and then performing cleavage treatment and deprotection treatment of a base protecting group to obtain a phosphorodiamidate morpholino oligonucleotide (PMO) reaction solution. The application provides a synthesis method of phosphorodiamidate morpholino oligonucleotide with high coupling efficiency, low impurity level, high yield and high purity.

The invention discloses synthesis of 3 '-amido-5-nucleotide phosphamide and application of the 3'-amido-5-nucleotide phosphamide in oligonucleotide. The phosphamide modified oligonucleotide comprises at least one of nucleotide structural units shown in a formula I, a formula II, a formula III or a formula IV,..., the formula I, the formula II, the formula III,..., the formula IV. Through specific limitation of a nucleotide phosphamide monomer structure, oligonucleotides are prepared by coupling according to a direction from 5 to 3. Compared with a traditional oligonucleotide molecule, the oligonucleotide molecule provided by the invention contains phosphamide modification and has more excellent effects on pharmaceutical parameters such as enzymatic degradation resistance, stability and the like, in addition, the phosphamide modification can improve the binding performance between the modified oligonucleotide molecule and environmental ions, and the stability of the oligonucleotide molecule is improved. A novel chemical modification strategy is provided for research, development and application of oligonucleotide drugs.

The invention provides a novel chiral phosphine-phosphoramidite ester ligand which is based on an achiral open-chain phenol non-cyclic 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 ligand, a phosphoramidite structural unit in the novel chiral phosphine-phosphoramidite ligand does not contain a chiral element and is of a non-cyclic open-chain structure. The novel ligand is prepared by taking a chiral phosphine / amine intermediate (R)-or (S)-DPPNHMe, a phenol 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.

Provided herein are oligonucleotides comprising a phosphoramidite internucleotide bond having the formula: (I) wherein R1 is isopropyl, isobutyl, sec-butyl, C1-6 haloalkyl, C2-6 hydroxyalkyl, C2-8 alkylene-N (RN) 2, C0-2 alkylene-C3-8 cycloalkyl, 4-10 membered heterocycloalkyl having 1-3 ring heteroatoms selected from O, N and S, or 5-10 membered heteroaryl having 1-3 ring heteroatoms selected from O, N and S, with the proviso that the heterocycloalkyl group or the heteroaryl group is attached to sulfur via a carbocyclic atom, and the cycloalkyl group, the heterocycloalkyl group or the heteroaryl group is substituted with 0, 1, 2 or 3 R2 groups; each R2 is independently halo, CN, N (RN) 2, C1-3 alkyl, C1-3 haloalkyl, C1-3 alkoxy, oxo, CO2RN, or C (O) C1-3 alkyl; and each RN is independently H or a C1-3 alkyl group. Also provided are formulations comprising the oligonucleotides disclosed herein and methods of treating diseases and disorders using the oligonucleotides disclosed herein and formulations thereof.

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.

Disclosed herein are nucleic acids containing a modified internucleotide linkage, which is the 2-propyl (PrON) linkage, to increase durability and exposure of therapeutic oligonucleotides, as well as nucleotides, phosphoramidites, and other intermediate compounds useful for making the same.

The invention relates to the technical field of biological medicine, and particularly discloses synthesis of 5-terminal phosphonic acid nucleotide and application of the 5-terminal phosphonic acid nucleotide in oligonucleotide. The chemical modification strategy comprises the following steps: preparing a 5 '-phosphonite modified nucleotide phosphoramidite monomer and a 5'-phosphonite modified nucleotide phosphoramidite monomer; the modified nucleotide phosphoramidite is used for preparing a target oligonucleotide molecule, carbon-phosphonic acid C-PO32-chemical bond connection is constructed at the 5-terminal of the target oligonucleotide molecule, a 5 '-terminal phosphonic acid structure in a non-natural form is formed and does not belong to a natural substrate of phosphatase, the modified oligonucleotide can resist nuclease degradation, the affinity of the oligonucleotide with specific protein can be improved, and the oligonucleotide can be used for preparing the target oligonucleotide. And the in-vivo biological activity of the siRNA is improved.

The present invention aims to provide a more stable and efficient method for producing oligonucleotide, particularly, oligonucleotide having various functional groups linked to the 3′-terminal and the like. Efficient production of oligonucleotide becomes possible by a production method of an oligonucleotide represented by the formula (Ia-2) (each symbol is as defined in the DESCRIPTION) and having a functional group at 3′-terminal, the method including a step of subjecting an oligonucleic acid with 3′-terminal protected by a silyl-protecting group to 3′-terminal-selective deprotection under desilylation conditions that do not affect protecting groups other than the silyl group, subjecting same to phosphitylation conditions with a phosphoramidite reagent that do not affect protecting groups on the oligonucleic acid to give a 3′-terminal-phosphoramidited oligonucleotide represented by the formula (Ia-1) (each symbol is as defined in the DESCRIPTION), and linking a functional group to 3′-terminal of 3′-terminal phosphoramidited oligonucleotide directly or via a linker, and the like.