13 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 application provides a transaminase, the amino acid sequence of which is shown as SEQ ID NO. 3, which can catalyze the conversion of (2Z)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7-(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-one into sitagliptin in a methanol solution reaction system.

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.

The present invention relates to a pharmaceutical composition for treatment of Parkinson's disease, containing sitagliptin as an active ingredient. It has been identified that, among patients with Parkinson's disease with comorbid diabetes, a sitagliptin-administered group exhibited less dopaminergic neuron loss at the time of diagnosis in comparison to a non-administered group, and in comparison even to a group without diabetes, the loss of dopaminergic neurons was milder. It has been also identified that oral administration of the sitagliptin in an animal model of Parkinson's disease improves intestinal inflammation and microbial groups, alleviates deposition of α-synuclein in the intestine, and alleviates deposition of α-synuclein in brain tissue, and thus the sitagliptin is provided as a novel treatment for Parkinson's disease on the basis of the gut-brain axis.

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.

Disclosed are compositions, including pharmaceutical compositions, wherein the compositions comprise sitagliptin or a pharmaceutically acceptable salt and / or hydrate thereof in combination with an antioxidant.

The present invention relates to a kind of ω-transaminase mutant, coding gene, carrier containing coding gene, genetic engineering bacteria, and its application in microbial catalysis preparation sitagliptin intermediate.The ω-transaminase mutant is obtained by single mutation or multi-point combined mutation of amino acid sequence 275th, 115th and 97th shown in SEQ ID NO.2.The present invention provides a transaminase mutant (biocatalyst) derived from a kind of Aspergillus fungus Lante (Aspergillus lentulus), with sitagliptin intermediate precursor ketone (such as: 1-piperidine-4-(2,4,5-trifluorophenyl)-1,3-dibutyl ketone) as substrate, while isopropylamine is amino donor, pyridoxal phosphate is coenzyme, dimethyl sulfoxide is cosolvent, biocatalytic reaction, separation and purification are carried out to prepare high optical purity sitagliptin intermediate, the total yield of the method reaches 66.7% (including conversion yield and separation and purification yield), and product ee value reaches 99% (stereoselectivity is high), with good application prospect.

Provided is an omega-transaminase mutant acquired by a single-point mutation or multi-point mutation at positions 275, 115, and 97 of the amino acid sequence set forth in SEQ ID NO. 2. The transaminase mutant is derived from Aspergillus lentulus. It catalyzes bioreactions with a ketone precursor of a sitagliptin intermediate as the substrate, isopropylamine as the amino donor, pyridoxal phosphate as the coenzyme, and a protonic polar solvent as the cosolvent, thus separating and purifying sitagliptin or the sitagliptin intermediate with high optical purity.

The invention relates to the technical field of synthesis of sitagliptin intermediates, and discloses a preparation method of a sitagliptin intermediate. According to the preparation method, ethyl trifluoroacetate is subjected to a new intermediate IV (5-(iodometyl)-2-(triflurometyl)-1, 3, 4-oxadiazole, 5-iodomethyl-2-trifluoromethyl-1, 3, 4-oxadiazole) which does not obtain a CAS number, so that the yield of the intermediate N-[(2Z)-piperazine-2-subunit]-2, 2, 2-trifluoroacethydrazide can be effectively improved, and the product yield is further improved. The preparation method of the novel intermediate IV is mild in reaction condition, convenient to operate and easy to industrially amplify, and has obvious economic benefits and social benefits.

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.

The present invention provides a transaminase and a use for preparing sitagliptin or an intermediate thereof. The amino acid sequence of the transaminase comprises one, two or three amino acid residue differences at residue positions compared to the amino acid sequence shown in SEQ ID NO:1. The present invention also provides a nucleic acid encoding the transaminase, a recombinant expression vector including the nucleic acid, and a transformant comprising the nucleic acid or the recombinant expression vector, and their applications. The present invention also provides an enzyme preparation comprising the transaminase or a combination of the transaminase. The present invention also provides a method for preparing (R)-3-amino-1-morpholine-4-(2,4,5-trifluorophenyl)-1-butanone, the method using the transaminase, with high conversion rate, good stability, high stereoselectivity, thereby reducing production costs, and being conducive to the progressive effect of industrialized production.