1661results about "Glucagons" patented technology

The invention provides a polypeptide or a derivative and a pharmaceutically acceptable salt thereof. The amino acid sequence of the polypeptide is as shown in SEQ ID NO: 1. The polypeptide or the derivative and the pharmaceutically acceptable salt thereof disclosed by the invention can be combined with GLP-1R and GCGR, and is used for effectively activating the GLP-1R and the GCGR. Therefore, the polypeptide or the derivative and the pharmaceutically acceptable salt thereof can be used for detecting GLP-1R and / or GCGR, and can also be used for treating or preventing GLP-1R and / or GCGR related diseases (such as metabolic disorder related diseases, such as obesity, diabetes, fatty liver diseases, non-alcoholic fatty liver diseases and the like).

The invention provides an anti-human serum albumin nano antibody and application thereof, and belongs to the technical field of antibodies. The invention provides an anti-human serum albumin nano antibody HSA-32 with relatively high affinity, and the anti-human serum albumin nano antibody HSA-32 is subjected to humanized transformation, so that three obtained humanized anti-human serum albumin nano antibodies can be used for reducing the immunogenicity of the HSA-32 and improving the immunogenicity of the HSA-32; and high specificity and high affinity of combination with the human serum albumin and stability and solubility of the antibody are maintained. The anti-human serum albumin nano antibody can be used for separation, purification and detection of human serum albumin and separation and purification of polypeptide, protein or antibody drugs fused with the anti-human serum albumin nano antibody. Meanwhile, the anti-human serum albumin nano-antibody can effectively prolong the half-life period of polypeptide, protein and antibody drugs fused with the anti-human serum albumin nano-antibody, and can be used for developing long-acting polypeptide, protein or antibody drugs.

The present invention belongs to the field of biopharmaceutical technology, and specifically relates to a method for preparing GLP-1 or its analogs. The present invention provides a leader peptide, and a fusion protein comprising the leader peptide. The leader peptide or fusion protein is used to prepare a semaglutide precursor polypeptide, and the results show that the yield of the prokaryotic expression fusion protein can reach 26g / L, and the purity of the precursor polypeptide obtained after enzyme cleavage is higher than 95%. The precursor polypeptide is further side-chain modified and connected with a dipeptide to efficiently prepare semaglutide.

The invention discloses a fusion protein and application thereof in preparation of a semeglutide precursor polypeptide, N (N is greater than or equal to 1) semeglutide fragments GLP-1 (11-37) are connected through KR, and then the semeglutide fragments GLP-1 (11-37), enterokinase enzyme cutting sites and fusion protein tags are subjected to fusion tandem expression in sequence, so that the semeglutide precursor polypeptide is obtained. The obtained fusion protein is a fusion protein tag-restriction enzyme cutting site-N-linked semeglutide fragment GLP-1 (11-37), and the fusion protein tag is an albumin affinity peptide (SEQ ID NO.1). The fusion protein is subjected to competent heterologous expression in escherichia coli BL21 (DE3), an engineering strain is constructed, and then fermentation expression is performed by using a shake flask, so that the thallus fermentation density is greatly improved, the expression of a target protein in an inclusion body is promoted, the yield of the target protein is improved, and the method has a good industrial application prospect. The fusion protein can be further successfully recognized by commercial enterokinase, KEX2 enzyme and carboxypeptidase B and cut to release the precursor polypeptide of the semeglutide.

The invention provides a method for synthesizing tirpotide by adopting a solid-liquid combination strategy. The preparation method comprises the following steps: firstly, respectively synthesizing full-protection peptide fragments (1-17) and (18-39), and then carrying out liquid-phase splicing and deprotection to obtain crude peptide; according to the strategy, the synthesis period can be remarkably shortened, large-scale industrial production is facilitated, meanwhile, the synthesis yield is high, and the purity and content of the obtained crude peptide are high.

The invention provides a solid-liquid combined synthesis method of semeglutide, which belongs to the technical field of semeglutide synthesis, and specifically comprises the following steps: segmenting a main chain of the semeglutide into three shorter polypeptide fragments, namely a fragment 4, a fragment 5 and a fragment 2 at sites 7-14, 15-22 and 23-37, and respectively synthesizing at the same time; performing solid-phase assembly on the fragment 4 and the fragment 5 to form 7-22 peptide fragments, namely a fragment 1, and after the fragment 1 is cut off from the resin, forming NHS ester at a C end; after the fragment 2 is assembled, selectively removing a protecting group A11oc on the Lys26, then connecting with a side chain for fatty acid treatment to form a fragment 3, and cutting off the fragment 3 from the resin; and connecting the NHS ester of the fragment 1 with the fragment 3 in a liquid phase reaction system. The crude semeglutide prepared by the synthetic method is relatively high in yield and purity. The refined semeglutide of which the purity is more than 99.3% and the content of single impurity is less than 0.1% can be obtained at high yield through a conventional purification means.

The invention relates to the technical field of chemical synthesis, and particularly provides a preparation method of a semaglutide side chain, which comprises the following steps: step S1, carrying out reaction on octadecanedioic acid mono-tert-butyl ester and trifluoroacetic acid pentafluorophenyl ester to generate a first intermediate; s2, enabling the first intermediate to react with 1-tert-butyl L-glutamic acid to generate a second intermediate; s3, enabling the second intermediate to react with pentafluorophenyl trifluoroacetate to generate a third intermediate; and S4, carrying out an amidation reaction on the third intermediate and 17-amino-10-oxo-3, 6, 12, 15-tetraoxa-9-aza-heptadecanoic acid, so as to generate a semaglutide side chain, and carrying out an amidation reaction on the third intermediate and 17-amino-10-oxo-3, 6, 12, 15-tetraoxa-9-aza-heptadecanoic acid. According to the preparation method disclosed by the embodiment of the invention, the pentafluorophenyl trifluoroacetate is used as the condensation reagent for two times to activate the carboxylic acid, and the amidation reaction is carried out for two times to construct the amido bond after each activation, so that compared with the traditional carbodiimide and urea positive ion type condensation reagent, the chiral racemization in the condensation process can be effectively avoided, and the yield of the product is improved. The condensation chiral racemization impurity can be lower than 0.3%.

The present invention relates to immunoglobulins, such as immunoconjugates, comprising an ActRII receptor antibody and an agonistic GLP-1 molecule, such as agonistic GLP-1 peptide. Such constructs are useful in the treatment of numerous diseases and disorders, including metabolic disorders, such as obesity, diabetes and others.

The invention provides a solid-phase synthesis method of semeglutide, and belongs to the technical field of synthesis of medical intermediates, and the method specifically comprises the following steps: segmenting a semeglutide main chain into three shorter polypeptide fragments, namely a fragment 4, a fragment 5 and a fragment 2, which are 7-14 sites, 15-22 sites and 23-37 sites; and respectively synthesizing and then assembling fragments to obtain a target product. The method specifically comprises the following steps: a) synthesizing a 7-14 peptide fragment and a 15-22 peptide fragment, and assembling the 7-14 peptide fragment and the 15-22 peptide fragment into a 7-22 peptide fragment, namely a fragment 1; b) synthesizing a 23-37 peptide fragment, selectively removing a protecting group A11oc on Lys26, and connecting with a side chain to form a fragment 3; c) connecting the peptide fragment 7-22 with the peptide fragment 23-37, and removing a protecting group to obtain a crude product; and d) separating and purifying to obtain a finished product. According to the invention, the purpose of high-purity, high-yield and low-cost synthesis of the semeglutide bulk drug can be achieved.

The invention discloses a purification method of a GLP-1 analogue crude peptide. The purification method comprises the following steps: treating a sample, preparing a mobile phase, carrying out purification balance, loading the sample, carrying out post-balance and eluting. The purification method aims at single impurities which are difficult to remove near a main peak, solves the problems of high purification difficulty, low recovery rate and high cost in the prior art, also has an obvious removal effect on front and rear impurities which are difficult to remove in crude stemeglutide or crude tilpoitide, reduces the purification difficulty, and has a great application prospect.

The invention discloses a one-pot preparation method of semeglutide, and relates to the field of polypeptide synthesis. The preparation method comprises the following specific steps: 1) taking Arg34GLP-1 (9-37) as a starting material, controlling the pH value of a reaction solution, enabling activated dipeptide Fmoc-His-Aib-OH to react with the starting material to obtain an acylation reaction solution, and directly carrying out next-step reaction without aftertreatment after the reaction is finished; 2) adjusting the pH value of the acylation reaction liquid I, adding a compound 2-tetrapeptide into the acylation reaction liquid I, reacting to obtain acylation reaction liquid II, and directly carrying out next-step reaction without post-treatment after the reaction is finished; (3) adding an alkaline reagent to remove the Fmoc protecting group, settling, centrifuging and drying to obtain a crude product of the semeglutide; 4) separating and purifying the crude product to obtain a refined semeglutide product; the first three steps are carried out in the same reactor. Compared with a conventional liquid-phase synthesis method, the preparation method disclosed by the invention is simpler and more convenient, short in construction period, low in output of waste liquid and waste water, greatly reduced in cost, higher in yield and suitable for large-scale production.

A GLP-1-Fc-FGF21 double-target fusion protein composition and the use thereof. The composition comprises a GLP-1-Fc-FGF21 fusion protein and a buffer solution. In the composition, by means of adding the buffer solution, the generation of visible particles can be effectively controlled, and the stability of the GLP-1-Fc-FGF21 fusion protein is improved, so that the shelf life of a product can be prolonged. The composition can be used for treating and / or preventing metabolic diseases, such as diabetes, hyperlipidemia, obesity and diseases related to fatty liver.

The present application provides protease-resistant Glucagon-Like Peptide 1 (eGLP-1) peptides, and also provides protease-resistant dual agonist peptides against GLP-1R and GIPR, methods of making such peptides, as well as compositions comprising protease-resistant peptides and methods of treatment utilizing such peptides.

Crystalline forms of peptide fragments, methods for their preparation and their use in the preparation of peptides are provided. The crystalline compound of the present invention can be used as an intermediate having improved purity and physical properties for peptide synthesis.

The invention discloses a method for preparing a simeglutide precursor peptide through intracellular enzyme digestion. The method comprises the following steps: (1) connecting a dissolution-promoting tag GST and the simeglutide precursor peptide together through protease specific recognition of amino acid sites to form a fusion protein; (2) integrating the coding nucleotide sequence of the fusion protein, the coding nucleotide sequence of KEN2 protease and the coding nucleotide sequence of carboxypeptidase B into the same expression vector to form a recombinant vector; (3) transforming the recombinant vector into engineering bacteria and carrying out fermentation production; and (4) centrifuging a fermentation product, collecting thalli, and purifying to obtain the precursor peptide powder of the semeglutide. The whole operation process is simpler, more convenient and smoother, tedious recovery steps are omitted, and the preparation and recovery difficulty of the precursor peptide of the semeglutide is remarkably reduced.

The invention provides a preparation method of tiall peptide, and belongs to the technical field of polypeptide synthesis, according to the preparation method, a small amount of lysis solution is adopted for lysis, a small amount of poor solution composed of methyl tert-butyl ether and n-heptane with the volume ratio of 1-2: 1 is used for crude peptide precipitation, precipitated particles are uniform, large and not prone to caking, the crude peptide impurity level is lower, and the purity of the crude peptide is higher. The method has the advantages that the method is simple in process and easy in subsequent purification step, the amount of generated waste liquid is small, the activating agent 7-(1H-pyrrolo [2, 3-b] pyridine-1-yl)-1H-benzo [d] [1, 2, 3] triazole-1-alcohol is used for activating amino acid, the coupling reaction degree is high, and the target peptide yield is high. The preparation method of the tilpotide has the advantages of being simple, economical, capable of achieving mass production, low in impurity content, high in purification yield, high in yield and the like.

Provided herein are compounds having the structure of Formula Ia, wherein the compounds comprise a therapeutic agent having the sequence of SEQ ID NO: 35, and the compounds have an anion:cation molar ratio of from about 1:1 to about 1:7, and methods of using thereof for the treatment of metabolic diseases or disorders, including type 1 diabetes, type 2 diabetes, obesity, overweight, and nonalcoholic steatohepatitis, and for reducing weight in a subject in need thereof.

Linkers for linking a therapeutic agent to other moieties, such as an Fc region of an antibody. A linker having two or more maleimide functional groups, which may be subjected to a nucleophilic conjugate addition reaction. A method for increasing the duration of action of a therapeutic agent by linking a novel linker to the Fc region of an antibody. Antibody drug conjugates having a longer duration of action than the drug alone.

A GLP-1 / GIP dual-targeted polypeptide, containing a first polypeptide having the following amino acid sequence: X1X2X3GT FX4SDY SX5X6X7X8 X9X10X11X12X13 X14FX15X16W LX17X18X19. The GLP-1 / GIP dual-targeted polypeptide can effectively reduce the weight and blood sugar level.

The invention provides a synthesis method of tilpotide. The synthesis method comprises the following steps: carrying out a reaction on Fmoc-Linker-OH, HBTU and a swelling AM Resin to prepare Fmoc-Linker-AM Resin, and carrying out a reaction on the Fmoc-Linker-OH and the HBTU and the swelling AM Resin to prepare the tilpotide. Adding Fmoc-Ser (tBu)-OH and HOBT into a reaction system of the resin and the PIP / DMF, and completely reacting; sequentially completing coupling from the C end to the N end to obtain full-protection peptide resin; adding a cracking reagent into the full-protection peptide resin, reacting, collecting filtrate, cooling, precipitating, and drying to obtain crude peptide; the problem that in the prior art, a linker on MBHA Resin is broken to generate new impurities is solved. The defect of using heavy metal palladium or genotoxic substance hydrazine hydrate is avoided; the problem that [-(Pro-Pro)] deletion impurities are generated by step-by-step coupling of C-terminal Pro36, Pro37 and Pro38 is solved; finished products are saved, and industrial production is facilitated.

The invention discloses a cell and a method for detecting the activity of a GLP-1R / GIPR double-target agonist drug. The invention relates to a cell for co-expressing GLP-1R, GIPR and CRE-miniP-luciferase reporter genes, and provides a method for detecting the biological activity of a GLP-1R / GIPR double-target agonist drug based on the cell, and the method is simple to operate, short in test period, sensitive in reaction, small in variability, strong in specificity and high in accuracy, can be used for release inspection of products, and can be used for detecting the biological activity of the GLP-1R / GIPR double-target agonist drug. The method has important significance on quality control and clinical application of GLP-1R / GIPR double-target agonist drugs.

The invention relates to the field of medicine synthesis, and discloses a GCG-GLP-1 dual agonist compound. The GCG-GLP-1 dual-agonist compound provided by the invention is used for preparing a pharmaceutical composition for treating diseases, and the application of the pharmaceutical composition in preparing a medicine for treating at least one of the following diseases, the diseases include type II diabetes mellitus, impaired glucose tolerance, type I diabetes mellitus, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and / or dyspepsia or gastric ulcer, hepatic fibrosis disease and pulmonary fibrosis disease.

The invention discloses a long-acting polypeptide analogue and application thereof, the polypeptide analogue can act on a GLP-1 receptor, a GIP receptor and a Y2 receptor at the same time, and can play the biological activities of GLP-1, GIP and PYY3-36 at the same time; the PYY3-36 GLP-1 has the treatment effect of GLP-1 on diabetes mellitus, also has the beneficial effect of GIP on glucose and lipid metabolism and appetite inhibition, and also has the beneficial effect of PYY3-36 on appetite inhibition, so that the PYY3-36 GLP-1 has a synergistic effect on glucose, lipid and energy metabolism; the application of the compound in preparation of drugs for treating metabolic syndromes, such as diabetes, obesity and dyslipidemia, has greater potential in preparation of drugs for treating diseases such as diabetes, obesity and dyslipidemia.

The invention provides novel compounds which are peptide hormone analogues, and which are useful in treating disorders such as diabetes and obesity. The compounds of the general sequence recited in the specification possess a tailored profile with regards to potency properties at the GIP receptor. With regard to in vivo properties, administration of example peptides of the invention have been shown, in animal models, to result in increased weight loss. Preferred compounds achieve this without reducing food intake significantly.

The invention relates to the field of medicine synthesis, and discloses a GLP-1 / GIP / GCG tri-agonist compound. The GLP-1 / GIP / GCG three-agonist compound is used for preparing a medicine composition for treating diseases, and the application of the medicine composition in preparing a medicine for treating at least one of the following diseases, namely, the GLP-1 / GIP / GCG three-agonist compound, and the GLP-1 / GIP / GCG three-agonist compound. The diseases include type II diabetes mellitus, impaired glucose tolerance, type I diabetes mellitus, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and / or dyspepsia or gastric ulcer, hepatic fibrosis disease and pulmonary fibrosis disease.

The invention relates to a protein combination for influencing the growth rate of yeast and the expression quantity of foreign proteins. After genes in a peptidase A1 family, a peptidase M1 family, a peptidase M18 family, a peptidase M28 family, a peptidase S54 family, a peptidase S10 family, a peptidase S16 family, a peptidase S14 family and a peptidase S8 family and protein defect related genes are knocked out as transcription activating factors, the growth rate can be increased, and the expression quantity of foreign proteins can be increased.