6 results about "Sitagliptina" patented technology

The invention relates to the technical field of bioengineering, in particular to an ester hydrolase mutant and application thereof. According to the present invention, the screened wild-type ester hydrolase derived from Pseudomonas fluorescens is subjected to mutation to obtain the ester hydrolase mutant, the ester hydrolase mutant is applied to the synthesis of the sitagliptin intermediate, and compared with the wild-type ester hydrolase, the ester hydrolase mutant has characteristics of high enzyme activity and high stereoselectivity on the substrate (compound II), the enzyme consumption is significantly reduced, and the yield of the sitagliptin intermediate is significantly improved; the chiral purity of the reaction product (compound I) is greatly improved, and the method has a good industrial application prospect.

The present invention relates to sitagliptin or a pharmaceutically or veterinary acceptable salt thereof for use in the local eye treatment of neovascular retinal diseases. The invention also encompasses pharmaceutical or veterinary compositions for use in the local treatment of these diseases.

The present application relates to the technical field of enzyme engineering, and discloses an amino transferase mutant and application thereof.The present application provides a wild-type amino transferase mutant derived from Aspergillus avenaceus.The amino transferase mutant is obtained by carrying out single-point or multi-point combined mutation on the 20th, 60th, 92nd and 186th positions of the amino acid sequence shown in SEQ ID NO.2, and can directly take the sitagliptin intermediate precursor ketone as a substrate, take isopropylamine as an amino donor, take phosphopyridoxyl as a coenzyme, take dimethyl sulfoxide as a substrate solvent, catalyze the preparation of a sitagliptin intermediate with high optical purity, and the specific enzyme activity reaches 139.3 U / g in a reaction system with a final concentration of 50% DMSO, greatly improves the catalytic efficiency, and the product has high stereoselectivity (the e.e. value of the product reaches 99%), and has a wide industrial application prospect.

This invention relates to a method for preparing sitagliptin phosphate to extend the lifespan of immobilized transaminases, belonging to the field of pharmaceutical synthesis technology. To address the problem of short lifespan of existing immobilized enzymes, this invention provides a method for preparing sitagliptin phosphate to extend the lifespan of immobilized transaminases. The method includes, in the presence of immobilized transaminases, carrying out an enzymatic reaction of sitagliptin precursor ketone and isopropylamine in a non-water-soluble organic solvent. The reaction is terminated after the raw material conversion rate is detected to be ≥75%, the immobilized transaminases are recovered, the filtrate is washed with water to remove acetone, and an aqueous phosphate solution is added to the filtrate for extraction. The aqueous phase is separated to obtain an aqueous sitagliptin phosphate solution, and the solvent is removed. The organic phase is washed with water and directly reused. This invention can significantly shorten the reaction time, reduce the contact time of the immobilized transaminases in a high-concentration acetone system, and avoid excessive soaking time, thereby extending the overall lifespan of the immobilized transaminases.

The invention discloses a preparation method of sitagliptin base, and relates to the technical field of compound preparation, and the preparation method comprises the following steps: S1, adding 75 parts of water and 16 parts of hydrochloric acid into a three-neck bottle, and cooling to 0 DEG C; s2, the pH is adjusted to 7.5, 42.5 parts of a triethanolamine solution, 0.2 part of pyridoxal phosphate and 57 parts of DMSO are added, and the temperature is controlled to be lower than 10 DEG C; s3, adjusting the pH value to 8.5, adding 40 parts of liquid transaminase, and slowly heating to 40-45 DEG C; s4, when the temperature rises to 45 DEG C, 20 parts of diketone is dropwise added, the temperature is controlled to be 45 DEG C, the pH is adjusted to 8.5, and heat preservation is conducted for 24 h; s5, performing suction filtration and extraction, cooling and crystallizing through methyl tert-butyl ether to obtain a white solid, and drying to obtain a finished product sitagliptin base; preparation equipment adopted in the preparation method comprises a closed cylinder, a liquid separation mechanism, an organic phase receiving box, a water phase receiving box and the like, closed control in the liquid separation extraction process is achieved, recycling of the extraction agent is achieved, and the volatilization and dissipation amount of the extraction agent in the preparation process is effectively reduced.

The invention relates to the field of chemical synthesis of medicines, and discloses a synthesis method of sitagliptin, which comprises the following steps: carrying out asymmetric hydroamination reaction on (E)-4-(2, 4, 5-trifluorophenyl)-butyl-2-ethyl olefine acid and an ammonia source at room temperature in the presence of a copper (I) catalyst, a chiral ligand, a hydrogen source, a strong base and a solvent to obtain a chiral hydroamination intermediate; carrying out hydrolysis reaction on ethyl ester in the chiral hydroamination intermediate to obtain a hydrolysis intermediate; the hydrolysis intermediate and 3-(trifluoromethyl)-5, 6, 7, 8-tetrahydro-[1, 2, 4] triazolo [4, 3-a] pyrazine are subjected to an amidation reaction in the presence of a condensation reagent, and an amidation intermediate is obtained; and carrying out debenzylation reaction on the amidation intermediate under the action of a catalyst and hydrogen to obtain sitagliptin. By introducing a brand-new catalytic system, the total yield is increased, and the enantioselectivity is remarkably improved.