431 results about "Stereoselectivity" patented technology

The invention discloses a dihydrodaidzein reductase mutant and application of the dihydrodaidzein reductase mutant in synthesis of (3S, 4R)-tetrahydrodaidzein, and belongs to the technical field of biology. According to the mutant, wild-type dihydrodaidzein reductase DHDR is taken as a parent, single-point mutation or combined mutation is carried out, and a DHDR mutant is obtained; the parent site of the single point mutation is one of the 120th site, the 122nd site, the 170th site, the 178th site, the 214th site, the 219th site, the 220th site or the 220th site; the combined mutation adopts two modes: one mode is that a single-point mutation DHDR mutant is taken as a parent, mutation is carried out, and a combined mutation mutant I is obtained; and 2, taking the DHDR combined mutation mutant I as a parent, and performing mutation to obtain a combined mutation mutant II. The DHDR mutant disclosed by the invention is used for synthesizing (3S, 4R)-tetrahydrodaidzein by reducing dihydrodaidzein, and asymmetric reduction with high activity and high stereoselectivity can be realized.

The invention discloses an aldehyde reductase mutant and application thereof in synthesis of a dexmethylphenidate hydrochloride intermediate, and belongs to the field of molecular biology and enzyme engineering. The aldehyde reductase mutant, polynucleotide for coding the mutant, and the recombinant expression vector can express the aldehyde reductase mutant and are used for constructing a recombinant cell or a recombinant strain for expressing the aldehyde reductase mutant. The provided aldehyde reductase mutant can catalyze 2-phenyl-2-((R)-piperidine-2)-acetaldehyde into (R)-2-phenyl-2-((R)-piperidine-2)-1-ethanol, especially improves the stereoselectivity of (R)-2-phenyl-2-((R)-piperidine-2)-1-ethanol, solves the problems of strict conditions, complex reaction and high cost in the existing synthesis method, and has a wide application prospect in the field of synthesis of (R)-2-phenyl-2-((R)-piperidine-2)-1-ethanol. Wide application prospects are realized.

The invention discloses a carbonyl reductase mutant as well as a preparation method and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. According to the invention, wild-type carbonyl reductase of Stachybotrys chlorohallonata is taken as an evolution template, and a plurality of mutants with catalytic activity, thermal stability and substrate spectrum obviously superior to those of the wild-type carbonyl reductase are obtained through an enzyme engineering technology. The carbonyl reductase mutant can catalyze various prochiral carbonyl compounds to obtain chiral alcohol with high stereoselectivity (ee is greater than 99%). The carbonyl reductase mutant provides a novel, efficient and environment-friendly technical route for industrial production of chiral alcohol products, and has very high industrial application potential.

The invention discloses an enzyme catalytic synthesis method of a luliconazole chiral intermediate, which comprises the following steps: adding a recombinant escherichia coli wet cell for expressing alcohol dehydrogenase, a recombinant escherichia coli wet cell for expressing formate dehydrogenase, a buffer solution, a substrate, a cosolvent, ammonium formate and a coenzyme into a reaction container, reacting at 25-35 DEG C for 2-24 hours, extracting, separating, and carrying out rotary evaporation, thereby obtaining the luliconazole chiral intermediate. The luliconazole chiral intermediate is obtained; the substrate is 2, 2 ', 4'-trichloroacetophenone; the alcohol dehydrogenase is a mutant of AKR3. The method is mild in reaction condition, environmentally friendly, high in regioselectivity and stereoselectivity, high in conversion rate, high in chiral purity, small in enzyme dosage, low in preparation cost and suitable for industrial production.

A pharmaceutical composition comprises a GPCR agonist fusion protein in which a GPCR agonist peptide is covalently coupled to albumin via a linker in a manner that is resistant to a retro-Michael addition. Advantageously, compositions presented herein avoid decoupling of the agonist form the albumin while retaining the agonist in a steric relationship to the albumin that allows for effective binding and activation of the GPCR while also enabling gp60-mediated transcytosis and FcRn-mediated albumin recycling. These properties enable ultra-low dosages for the GPCR agonist fusion protein to give a therapeutic effect while substantially reducing or even entirely avoiding adverse effects otherwise commonly associated with unbound agonists. Such retro-Michael resistant composition is generally achieved by conformational modification of the albumin, resulting in stereoselective coupling of the linker to the albumin.

A pharmaceutical composition comprises a GPCR agonist fusion protein wherein the GPCR agonist peptide is covalently conjugated to an albumin via a linker in a manner that is resistant to a reverse Michael addition. Advantageously, the compositions presented herein avoid uncoupling of the agonist to the albumin while maintaining the agonist and the albumin in a spatial relationship that allows efficient binding and activation of the GPCR while also enabling gp60-mediated endocytosis transport and FcRn-mediated albumin recirculation. These characteristics enable ultra-low doses of GPCR agonist fusion proteins to produce therapeutic effects while significantly reducing or even completely avoiding side effects that would otherwise be typically associated with unbound agonists. Such reverse Michael resistant compositions are typically achieved by conformationally modifying an albumin, resulting in a stereoselective coupling of a linker to the albumin.

The application discloses a method for preparing a CGRP receptor antagonistic intermediate, and belongs to the field of synthesis of medical intermediates. Compound 1 is used as a raw material, reacts with hydroxylamine hydrochloride in an organic solvent to generate an intermediate 2, and then reacts in the presence of a reducing agent to obtain a remikiren intermediate 3. The application is characterized in that a large steric hindrance silicon group is used as the R substituent in the compound 1, so that high stereoselectivity is ensured in the reduction reaction. The key intermediate obtained by using the method has high chiral purity, and the operation is simple, so that the industrial production of the product in the later stage can be realized.

The invention provides immobilized ketoreductase, an immobilization method and application of the immobilized ketoreductase in preparation of a key intermediate of bentonatate, the amino acid sequence of the ketoreductase is as shown in SEQ ID NO.2, preferably a ketoreductase mutant is as shown in SEQ ID NO.16, and the immobilized ketoreductase comprises resin and corresponding ketoreductase. The ketoreductase mutant and the immobilized ketoreductase prepared from the ketoreductase mutant are very high in enzyme activity during ketoreduction reaction, and when the ketoreductase mutant is used for preparing a key intermediate of bentonamate, a solvent is pure water, so that the ketoreductase mutant and the immobilized ketoreductase prepared from the ketoreductase mutant are environment-friendly. The immobilized transaminase is high in conversion rate, good in stereoselectivity, good in stability, reusable and simpler to operate, so that the production cost is reduced, and industrial production is facilitated.

To provide a novel stereoselective method for producing a cyclic dinucleotide derivative that can be used for an antibody-immunostimulator conjugate, and a production intermediate thereof, and to provide methods for producing a cyclic dinucleotide-linker and antibody-immunostimulator conjugate using the production method.SOLUTION: The present invention relates to a method that makes it possible to produce, in large amounts and with high yield, a cyclic dinucleotide derivative having a desired steric configuration, by using an optically active phosphitylation agent twice, during coupling and cyclization.SELECTED DRAWING: None

The invention relates to a carbonyl reductase mutant based on cosubstrate catalytic ability optimization and application thereof, the mutant is constructed on the basis of a carbonyl reductase BaSDRX amino acid sequence as shown in SEQ ID No.2, the nucleotide sequence of the carbonyl reductase is as shown in SEQ ID No.1 in a sequence table, and the nucleotide sequence of the carbonyl reductase is as shown in SEQ ID No.2 in the sequence table. The mutant is single-point mutation or multi-point combined mutation on the basis of an amino acid sequence of carbonyl reductase BaSDRX as shown in SEQ ID No.2: threonine Thr133 at the 133 site, arginine Arg137 at the 137 site and glutamic acid Glu142 at the 142 site; when the mutant enzyme or a recombinant cell containing the mutant enzyme is applied to synthesis of (S)-1-(2, 6-difluorophenyl) ethanol through asymmetric reduction of 2, 6-difluoroacetophenone, high catalytic activity and high stereoselectivity are achieved, and high-optical-purity (S)-1-(2, 6-difluorophenyl) ethanol (eet; therefore, the method has a good industrial application and development prospect.

The invention relates to a synthesis method of milobalin or acid salt thereof, and belongs to the technical field of medicine synthesis. In order to solve the problem that chiral resolution needs to be adopted in the prior art, the invention provides a synthesis method of miloabalin or an acid salt thereof, which comprises the following steps: under the action of an organic strong base, carrying out Michael addition reaction on a compound shown as a formula III and acetonitrile in a non-polar organic solvent at a low temperature of-50 DEG C or below to obtain an intermediate product; under the alkaline condition, the intermediate product is subjected to an oxidation reaction and a hydrolysis reaction under the action of an oxidizing agent, after the reaction is finished, an intermediate product is obtained through acidification, and the intermediate product is subjected to a high-temperature decarboxylation reaction in an aprotic solvent to obtain a compound of a formula VI; and carrying out Hofmann degradation reaction on the compound of formula VI to obtain the compound of formula I miloabalin or miloabalin acid salt. The method has the advantages of high stereoselectivity, no need of chiral resolution operation, avoidance of the problem of large material loss caused by resolution, high chiral purity and high yield.

The invention relates to the technical field of biology, in particular to a carbonyl reductase mutant, a gene, an engineering bacterium and application. The amino acid sequence of the carbonyl reductase mutant is shown as SEQ ID NO: 1, the catalytic efficiency of asymmetric reductive hydrogenation of prochiral ketone compounds is improved, the stereoselectivity of chiral hydroxyl compounds formed through catalysis is improved, and the synthesis efficiency of (S)-1-(5-fluoro-2-iodophenyl) ethanol is improved.

The invention provides a method for synthesizing N-methylated polypeptide, which comprises the following steps: dissolving a carboxylic acid compound, N-methyl amino-acid ester hydrochloride or N-methyl amino-acid ester, an alkaline substance and pivaloic anhydride in an organic solvent, reacting at 15-80 DEG C for 3-10 hours, and post-treating the obtained reaction liquid to obtain an N-methyl dipeptide compound, the method comprises the following steps: removing a protecting group in an N-methyl dipeptide compound to obtain a free-state N-methyl dipeptide compound, and condensing the free-state N-methyl dipeptide compound and amino acid or N-methyl amino-acid ester protected by amino to obtain the N-methylated polypeptide, the mixed anhydride intermediate is formed in situ, and the reaction is completed in one step; according to the method, trimethylacetic anhydride is used as a condensing agent, the structure is simple, the reaction is safe, cheap and non-toxic, a large amount of nitrogen-containing byproducts are not generated in the reaction, and purification is convenient; the method is high in stereoselectivity, and racemization is avoided; according to the method, environment-friendly solvents such as ethyl acetate can be used, the reaction condition is mild, and the reaction time is short.

The application discloses a carbonyl reductase mutant and application thereof in preparation of (R)-8-chloro-6-hydroxyoctanoate compounds. The application relates to a mutant of carbonyl reductase with improved catalytic performance and stereoselectivity after rational molecular modification, a recombinant expression plasmid containing the mutant gene of the enzyme, a recombinant expression transformant, a preparation method of the recombinant enzyme, and application of the recombinant enzyme or the recombinant cell as a catalyst in asymmetric reduction of 8-chloro-6-carbonyloctanoate compounds (I) to prepare (R)-8-chloro-6-hydroxyoctanoate compounds (II), in particular, in an intermediate (R)-8-chloro-6-hydroxyoctanoate methyl ester of thioctic acid. The disclosed carbonyl reductase SsCR mutant has the advantages of high reaction efficiency and high catalytic activity, and exhibits the advantages of high substrate concentration, low catalyst consumption and short reaction time in the process of catalytic preparation of the compound (II).

The application discloses a kind of generation (4S) 4-hydroxyisophorone monooxygenase mutant and preparation method thereof, the monooxygenase is cytochrome P450 BM3 Monooxygenase from bacillus megaterium. The cytochrome P450 BM3 Monooxygenase mutant has high stereoselectivity and high reaction activity to catalyze isophorone hydroxylation, mutant R47L / Y51F / F81W / A191T / N239H / I259V / A276T / A328I / A330S / L353I / I401L / L437LL catalyzes isophorone to obtain (4S) 4-hydroxyisophorone diastereomeric excess rate can reach 95%, yield can reach 96%; the cytochrome P450 BM3 Monooxygenase mutant catalyzes isophorone synthesis (4S) 4-hydroxyisophorone process is simple, environmental protection, and industrialization prospect is good.

The present application discloses a chiral dienyl-substituted carboxylate compound and an asymmetric preparation method thereof, wherein the chiral dienyl-substituted carboxylate compound is an optically active compound with the structure shown in the following formula I, including stereoisomers with the same chemical formula: in the formula, * represents a chiral carbon atom; the substituent R is selected from hydrogen, C 1‑10 alkyl or aryl; Ar is an aromatic ring or a substituted aromatic ring; R 1 , R 2 , R 3 , R 4 , R 5 are independently selected from hydrogen, halogen, C 1‑10 alkyl or alkoxy; R 6 is independently selected from C 1‑10 alkyl or aryl; R 7 is independently selected from C 1‑10 alkyl or aryl. The present application applies a chiral isothiourea catalytic electrochemical synthesis system to precisely and quickly synthesize the chiral dienyl-substituted carboxylate compound in a high yield and high stereoselectivity and in a gram-scale. The preparation method of the present application is simple, and has biomedical practicability and industrial application prospect.

The invention provides an R-type transaminase mutant and application thereof, and compared with an amino acid sequence of wild-type transaminase as shown in SEQ ID NO.1, the mutant has mutation of a 62nd site, a 152nd site and a 155th site. The mutant has high catalytic activity and excellent stereoselectivity on a heterocyclic ketone substrate, and can be used for efficiently preparing R-configuration heterocyclic chiral amine.

The application relates to a chiral NSiN ligand based on a phenyloxazoline skeleton, a preparation method and application thereof, wherein the chiral NSiN ligand is prepared from a chiral phenyloxazoline bromide with R or S configuration through lithiation and silylation. The chiral NSiN ligand can be used in transition metal catalyzed asymmetric synthesis, and is used in catalyzing an allylic sulfonylation reaction with a cobalt catalyst, and shows high reaction activity and high stereoselectivity.

The invention belongs to the technical field of organic synthesis. The invention particularly relates to a synthesis method of chiral beta-diaryl ethanol. The physiological activity, the pharmacological action and the material performance of the chiral molecule are directly determined by the spatial configuration of the chiral molecule. Therefore, efficient construction of chiral compounds is always the core research direction of organic synthesis and medicinal chemistry. Wherein the chiral beta-diaryl ethanol is a kind of key intermediates with irreplaceable application value. According to the invention, the Suzuki-Miyaura type asymmetric ring-opening coupling reaction of nickel catalyzed aryl epoxy and aryl boric acid with high stereoselectivity is realized, and the asymmetric ring-opening coupling reaction is used for synthesizing chiral beta-diaryl alcohol compounds. The method is wide in substrate compatibility and raw material source, mild in reaction condition, free of exogenous alkali and convenient to operate. An aryl epoxy compound and arylboronic acid are used as raw materials, diethylene glycol dimethyl ether nickel bromide is used as a catalyst, chiral biimidazoline is used as a ligand, under the promotion of sodium iodide, reaction is performed in an ethanol solvent according to the following reaction formula at the temperature of 50 DEG C for 10 hours, and the chiral beta-diaryl ethanol compound with the high ee value is obtained.

The application discloses a kind of using cobalt catalyst by unreported double migration coupling strategy, realizes migration asymmetric nitrogen hetero Nozaki-Hiyama-Kishi (aza-NHK) reaction, and the selective preparation α-chiral (E)-allyl amine starting from simple and easy to obtain ortho halogen phenyl ethylene or corresponding E / Z-alkyl bromide or its mixture belongs to the field of organic chemistry and medicinal chemistry.The application under the action of metal cobalt source, ligand, organic additive, inorganic additive etc., makes cyclic sulfonamide and ortho alkenyl substituted aryl halide or alkenyl halide react in organic solvent, to obtain a series of polysubstituted olefins with excellent regioselectivity, stereoselectivity and good yield, which can be used as active drug molecules and important synthetic intermediates.The application uses cheap transition metal cobalt as catalyst, and uses ortho alkenyl substituted aryl iodine and cyclic sulfonamide as raw material, and the reaction condition is warm, simple operation, and good functional group compatibility.

The invention belongs to the technical field of biosynthesis, and particularly relates to a method for preparing chiral alcohol through asymmetric reduction of large steric hindrance rigid ketone based on aldehyde ketoreductase and a mutant thereof. Comprising the following steps: adding a recombinant Escherichia coli wet cell for expressing aldehyde ketoreductase, a recombinant Escherichia coli wet cell for expressing formate dehydrogenase, a buffer solution, a substrate, a cosolvent, ammonium formate and coenzyme into a reaction container, reacting at a certain temperature, and after the reaction is completed, extracting, separating and carrying out rotary evaporation to obtain a product alcohol. The biological catalysis method is mild in reaction condition, environmentally friendly and high in regioselectivity and stereoselectivity, chiral resolution and heavy metal residues in products are avoided, and the defects of a chemical method are exactly overcome.

Disclosed is a method for preparing (S)-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid and derivatives thereof, comprising: taking a racemate of a compound represented by Formula (I) or a racemate of a salt of the compound represented by Formula (I) as a substrate, and making a R-isomer of the compound represented by Formula (I) in the substrate react under the catalysis of oxidative dehydrogenase to generate imino acid represented by formula (II); and converting the imino acid represented by Formula (II) into an S-isomer of the compound represented by Formula (I) in the presence of pipecolic acid reductase and a coenzyme capable of supplying hydrogen anions. The process has mild reaction conditions, strong stereoselectivity, high reaction efficiency, and high conversion rate.

The present invention provides a method for stereoselectively reacting a xyloside stereoisomer mixture. The method includes a step for bringing a reaction mixture that contains a cyclic esterified intermediate compound, in which a cyclic ester of boronic acid or boric acid is formed at the 2-position and the 4-position of the xylose moiety of each of α-xyloside and β-xyloside, into contact with silica gel. The step includes returning the cyclic esterified intermediate compound corresponding to α-xyloside to the α-xyloside form before cyclic esterification, and maintaining the cyclic esterified form of the cyclic esterified intermediate compound corresponding to β-xyloside.

The invention discloses a diketopiperazine heterodimer as well as a biosynthesis method and application thereof, and belongs to the technical field of bioengineering. The biosynthesis method of the diketopiperazine heterodimer comprises the following steps: S1, culturing a recombinant cell containing P450 dimerization enzyme or a mutant thereof, and performing induced expression to obtain a whole-cell culture; and S2, adding a cyclic dipeptide substrate containing tryptophan into the whole-cell culture, carrying out a catalytic reaction, and separating and purifying the obtained product to obtain the diketopiperazine heterodimer. According to the invention, the P450 dimerization enzyme or the mutant thereof is used for catalyzing the biosynthesis of the tryptophan-containing diketopiperazine into the diketopiperazine heterodimer with different regioselectivity / stereoselectivity, the biosynthesis method is green and environment-friendly, the efficiency is high, and the variety of the diketopiperazine heterodimer is greatly expanded.

The present invention relates to a use of an enzyme belonging to the aldolase group and comprising a polypeptide having at least 30% identity with an amino acid sequence selected from SEQ ID NO: 1 to SEQ ID NO: 4, for stereoselectively preparing D-tagatose-1,6-bisphosphate (TBP) or D-tagatose-1-phosphate. The present invention also relates to a method for preparing D-tagatose-1,6-bisphosphate (TBP) or D-tagatose-1-phosphate, comprising a step of reacting, in a suitable reaction medium, dihydroxyacetone phosphate (DHAP) or at least an isomer thereof or analog thereof, and optionally D-glyceraldehyde, with an enzyme belonging to the aldolase group, said enzyme comprising a polypeptide having at least 30% identity with an amino acid sequence selected from SEQ ID NO: 1 to SEQ ID NO: 4. Finally, the present invention relates to a process for preparing D-tagatose.

The invention discloses a chiral 1, 4-diazabicyclo [3.1. 1] heptane derivative as well as a synthesis method and application of the chiral 1, 4-diazabicyclo [3.1. 1] heptane derivative. According to the invention, a 1-azabicyclo [1.1. 0] butane derivative is taken as a raw material, under the catalytic action of a chiral iridium complex, the 1-azabicyclo [1.1. 1] butane derivative and an N-allyl carbonate derivative are subjected to [3 + 2] cycloaddition reaction, a series of chiral 1, 4-diazabicyclo [3.1. 1] heptane skeletons with diversified structures are constructed with high stereoselectivity, and the obtained product has the potential of further derivatization. The method has the advantages of easily available raw materials, mild reaction conditions, simplicity and convenience in operation, good functional group compatibility and the like.

The invention relates to the technical field of biological engineering and biological catalysis, in particular to two alcohol dehydrogenase mutants and application thereof. The invention discloses a pair of alcohol dehydrogenase mutants CpAR2-M3 (F85A / F192L / E209A) and CcPAR-M1 (V158L) with opposite stereoselectivity, wherein the corresponding amino acid sequences of the alcohol dehydrogenase mutants CpAR2-M3 (F85A / F192L / E209A) and CcPAR-M1 (V158L) are respectively shown as SEQ ID No.5 and SEQ ID No.6. The invention further discloses a preparation method of the alcohol dehydrogenase mutants. The method can be applied to asymmetric reduction of hemiacetal compounds, synthesis of corresponding key intermediate axial chiral biaryl benzyl alcohol compounds with complementary stereoselectivity is achieved, the highest yield can reach 95%, the highest stereoselectivity can reach 99% e.e., and the method is suitable for industrial production. A green, efficient and high-stereoselectivity synthesis strategy is provided for subsequent enantiocomplementary synthesis of the compounds.

This invention discloses a migration C(sp²)-H / C(sp³)-H cross-coupling reaction achieved using metal / photoredox catalysis, belonging to the fields of organic chemistry and medicinal chemistry. Under the action of a nickel source, photocatalyst, ligands, base, and additives, this invention enables the reaction of alkanes with ortho-alkenyl-substituted aryl halides or ortho-pyridine-substituted aryl halides in an organic solvent under light irradiation, yielding a series of polysubstituted olefins or polysubstituted aromatic compounds with excellent regioselectivity, stereoselectivity, and good yields, which can serve as active pharmaceutical molecules. This invention uses inexpensive transition metal nickel as a catalyst, readily available alkanes and haloaromatics as raw materials, and features mild and simple conditions, good functional group compatibility, and easy operation.

The invention discloses an application of halogenase in catalytic synthesis of 2 '-halogenated nucleoside. The halogenase is any one of NSH1, NSH2, NSH3, NSH4 and NSH5, and a substrate used for catalysis is any one of 2 '-deoxyguanosine, 2'-deoxyinosine and 2 '-deoxyadenosine. The biosynthesis method provided by the invention has the advantages of high reaction stereoselectivity, high product yield, simple halogen source, easily controlled operation process, low production cost, green and environment-friendly reaction, high catalytic activity and the like, and a new method is provided for synthesis of the 2 '-halide.

The application discloses an esterase with stereoselectivity and application thereof, and the esterase is: a) an esterase comprising a sequence as shown in SEQ ID NO: 2 or a sequence as shown in SEQ ID NO: 2; or b) a mutant based on SEQ ID NO: 2 comprising one or more than two mutations, and the esterase is used as a catalyst in the application of asymmetric resolution preparation of chiral oxacycloalkane carboxylate, has good substrate tolerance, high optical purity (ee s The reaction condition is mild, the environment is friendly, the operation is simple, the industrial amplification is easy, and the esterase has a good industrial application development prospect.