147 results about "Peptide synthesis" patented technology

The invention relates to a compound of formula (I) wherein X is selected from the group consisting of-O-(CH2) n-wherein n is 0 to 4, R1 and R2 are independently selected from the group consisting of C11-C25 alkyl groups, SP is a spacer, in particular selected from the group consisting of saturated C4-C8 alkyl groups, unsaturated C4-C8 alkyl groups and substituted or unsubstituted C6 aromatic compounds, in particular substituted C6 aromatic compounds are alkyl substituted C6 aromatic compounds, Y is a heteroatom, n is 0 to 4, and R1 and R2 are independently selected from the group consisting of C11-C25 alkyl groups, SP is a spacer, in particular selected from the group consisting of saturated C4-C8 alkyl groups, unsaturated C4-C8 alkyl groups, and substituted or unsubstituted C6 aromatic compounds, in particular substituted C6 aromatic compounds are alkyl substituted C6 aromatic compounds. L is a peptide synthesis linker, in particular selected from the group consisting of-O-and-NH-, L is a peptide synthesis linker, and m is 0 or 1, in particular m is 1.

A Solid Phase Peptide Synthesis (SPPS) device and method of using the same for manufacturing peptides is taught herein. The system comprises at least two reactors, each reactor including a quantity of SPPS resin. The reactors are positioned in series. A de-protecting agent is added to the first reactor and then transferred to the second and third reactors, in series, thereby operating to de-protect the protected N-group. Wash solvent is added to the first reactor and then transferred to the second and this operation repeated several times. Likewise, an amino acid activated ester solution is added, in series, to the first, second and third reactors, thereby operating to couple the amino acid to the de-protected N-group. Wash solvent is added to the first reactor and then transferred to the second and this operation repeated several times prior to the next cycle. The use of the reactors in series reduces the overall solvent required. Online LCMS is also used to monitor progress and identity of reactions happening within the solid phase resin particles.

The invention discloses a synthesis method of tripeptide Fmoc-Pro-Pro-Pro-OH, belongs to the technical field of polypeptide synthesis, and particularly relates to Fmoc-Pro-Pro-Pro-OH prepared by a condensation deprotection reaction of a proline reagent and a proline derivative, the proline reagent comprises Fmoc-Pro-OH or Fmoc-Pro-Pro-OH, and the proline derivative is H-Pro-OtBu. An activating reagent is used in the condensation deprotection reaction, and the mass ratio of the usage amount of the Fmoc-Pro-OH to the usage amount of the activating reagent is 1: (0.1-0.9). Alkali is used in the condensation deprotection reaction, the alkali comprises an alcohol amine compound, the alcohol amine compound is prepared through the reaction of ethanolamine and ethyl 3-bromopyruvate, and the synthesis method of the tripeptide Fmoc-Pro-Pro-Pro-OH is high in yield and purity.

The invention relates to the technical field of biological medicine manufacturing, in particular to an environment-friendly and efficient Difelikefalin preparation method. According to the method, a solid-phase peptide synthesis technology is adopted, and the core is that a green mixed solvent system composed of 2-methyltetrahydrofuran and cyclopentyl methyl ether is used for comprehensively replacing traditional toxic solvents such as DMF and DCM. The method comprises the steps of resin swelling, Fmoc deprotection, amino acid coupling, peptide chain cutting, purification and freeze drying, a piperidine / 2-MeTHF solution is adopted in the deprotection, an optimized HATU / DIPEA activation system is adopted in the coupling reaction, a TFA / 2-MeTHF / water mixed solution is adopted in the cutting, and an ethanol-water chromatographic system is adopted in the purification. According to the invention, closed-loop circulation of the solvent is also realized, and 2-MeTHF and CPME are purified through rotary evaporation and reused in the synthesis process. According to the method, high purity and high yield of the product are guaranteed, production toxicity and environmental hazards are remarkably reduced, and the method has excellent industrial application prospects.

The invention relates to the technical field of polypeptide synthesis, and particularly discloses a microwave synthesis method of Angiopep-2. The molecular formula of the Angiopep-2 polypeptide is as shown in formula I in the specification. The method adopts a solid phase polypeptide synthesis strategy and comprises the following steps: (1) coupling Fmoc-Tyr (tBu)-OH to a resin carrier; (2) under the assistance of a microwave synthesizer, sequentially coupling Fmoc protected amino acids from a C end to an N end according to an Angiopep-2 sequence (TFFYGGSRGKRNNFKTEEY, so as to obtain full-protection peptide resin; (3) cracking the resin by using a cracking solution to obtain crude peptide; and (4) purifying through reversed-phase preparative chromatography to obtain the refined peptide. According to the method, the synthesis time is remarkably shortened, the synthesis efficiency is improved, the solvent consumption is reduced, the operation is simple and convenient, the repeatability is high, and the method is suitable for process optimization and large-scale production of Angiopep-2.

The invention discloses a method for synthesizing dipeptide Fmoc-Leu-Aib-OH, and belongs to the field of polypeptide synthesis, the specific preparation method comprises the following steps: under the action of organic alkali and an activating reagent, Fmoc-Leu-OH and H-Aib-OH react to obtain the dipeptide Fmoc-Leu-Aib-OH; the activating reagent comprises at least one of N, N, N ', N'-tetramethyl chloroformamidine hexafluorophosphate, 2-(7-azabenzotriazole)-N, N, N ', N'-tetramethyl urea hexafluorophosphate and benzotriazole-1-yl-oxytripyrrolidinyl phosphorus hexafluorophosphate, and the organic base comprises at least one of N-methylimidazole, N, N-diisopropylethylamine and pyridine. The dipeptide Fmoc-Leu-Aib-OH prepared by the preparation method disclosed by the invention is high in yield, high in purity and high in raw material conversion rate.

The utility model belongs to the field of polypeptide synthesis, and provides a multi-row reaction device for polypeptide synthesis, which comprises a reaction box, a storage component and a swing component, a box cover is hinged on the reaction box, a plurality of rotating shafts are rotatably connected in the reaction box, the storage component is arranged in the middle of the rotating shafts, and a reaction column is arranged in the storage component. A swinging assembly is arranged on the outer side of the reaction box and is used for driving the multiple groups of rotating shafts to rotate to drive the reaction column to swing; the reciprocating driving piece is matched with the T-shaped frame, the T-shaped frame can be driven to drive the row teeth to continuously and transversely reciprocate under the combined action of the motor, the rotating disc and the push column, the row teeth are meshed with the gears to drive the reaction columns to shake left and right, polypeptide synthesis reaction is promoted, reaction efficiency is improved, energy consumption and mechanical abrasion are reduced, and the service life of the reaction columns is prolonged. The push column is adjustably mounted on the turntable through the positioning piece, so that the shaking amplitude of the reaction column can be adjusted as required, and the operability is relatively high.

The invention relates to a chemoenzymatic synthesis method of a high-glucose-type homogeneity mucoprotein core structural domain. The method comprises the following steps: firstly, catalytically synthesizing a key alpha-linked glycosylated amino acid block Fmoc-GalNacalpha-Ser / Thr-OH at a low temperature (-20 DEG C to 0 DEG C); then assembling the building blocks on a repeated polypeptide skeleton rich in proline, serine and threonine at accurate intervals (3-5 amino acid residues) through a solid-phase peptide synthesis technology to form a'sugar chain growth scaffold '; then, a series of high-specificity glycosyl transferases are used for carrying out sequential and directional sugar chain enzymatic extension on the scaffold, and a uniform core structure (such as a core type 3) is constructed; and finally, performing high-efficiency purification by a three-step series method of size exclusion chromatography, ion exchange chromatography and lectin affinity chromatography. The technical bottleneck of inhomogeneity (lt, 70%) of glycoforms in a traditional method is overcome, the mucoprotein core fragment with glycoform homogeneity larger than or equal to 95% and bioactivity highly similar to that of natural mucoprotein can be prepared on a large scale, and the method has wide application prospects in the biomedical fields of mucous membrane protective agents, drug delivery carriers and the like.

This invention relates to the field of peptide synthesis technology, specifically to a type of composition containing long-chain alkane for peptide synthesis and a method for preparing peptides. The composition includes peptide synthesis material A and material B; the general structural formula of peptide synthesis material A is Lca-K-L-AA or L... a -AA; where Lca contains long-chain alkyl and / or long-chain alkenyl groups, L is a polypeptide linker that does not contain long-chain alkyl or long-chain alkenyl groups, and K is an organic fragment or group used to link Lca and L; L a The material B is a polypeptide linker containing long-chain alkyl and / or long-chain alkenyl groups, where AA is an amino acid residue or a peptide containing fewer than 10 amino acid residues; the structure of material B contains long-chain alkyl and / or long-chain alkenyl groups. The compositions of this invention are applied to the synthesis of polypeptides, enabling the use of soluble carbon-based supports for solid-phase polypeptide synthesis.

The invention relates to the technical field of polypeptide synthesis, discloses a semaglutide large-fragment coupling process based on an SPPS-LPPS mixing method, and aims to solve the problems that an SPPS fragment is easy to fold and the LPPS coupling efficiency is low when semaglutide is synthesized by an existing SPPS-LPPS mixing method. SPPS is adopted for solid-phase synthesis of a semaglutide 1-21 site fragment I and a semaglutide 22-31 site fragment II, and the fragment quality is guaranteed through optimization of a solid-phase carrier, staged conformation washing and differential activation; according to the method, peptide chain folding and activating agent hydrolysis can be inhibited, the coupling efficiency and the fragment utilization rate can be improved, the purity of a coupling intermediate can be guaranteed, meanwhile, the process stability and economical efficiency can be enhanced, and the industrial production requirements of semaglutide can be met.

The present invention relates to processes for obtaining glucagon-like-peptide-2 (GLP-2) analogues, such as glepaglutide. In particular, the processes described herein use a multi-step purification method of GLP-2 analogues synthesized by solid phase peptide synthesis (SPPS).

The present invention is directed to a method of peptide synthesis, the method comprising reacting a compound of formula (I), or a salt thereof, with a carbodiimide reagent to form an O-acylisourea intermediate; reacting the O-acylisourea intermediate with an additive to form an activated ester; and reacting the activated ester with an amino-containing moiety, which is an amino acid, peptide, or salt thereof, containing an amino group, wherein the amino group forms an amide bond with the carbonyl marked with an * in formula (I), wherein the compound of formula (I) has the structure: TIFF2026501761000047.tif61170 formula (I) where R 1 and R 2 is as defined in the present disclosure. The present invention is also directed to compounds of formula (III) or salts thereof as defined in the present disclosure.

The application discloses a kind of double S-alkyl isothiourea derivatives, its preparation method and application, belong to biochemical technical field.The double S-alkyl isothiourea derivative is the compound shown in general formula 1 or its pharmaceutically acceptable salt, in formula, R is saturated alkane or unsaturated alkane;X is any one of i, ii and iii;I is branched C1-C 20 Saturated alkane or straight-chain C1-C 20 Saturated alkane;II is unsaturated alkane;III is heteroatom-substituted alkane.The application further discloses a preparation method and application of the above-mentioned double S-alkyl isothiourea derivative, a stapled peptide and a preparation method thereof.The double S-alkyl isothiourea derivative of the application can be used for stapled peptide synthesis, and the obtained stapled peptide containing guanidyl structure has the advantages of good water solubility and strong membrane permeability in addition to various advantages of traditional all-carbon hydrogen stapled peptide in polypeptide structure modification.

The invention relates to the technical field of polypeptide preparation, in particular to a solid-phase polypeptide synthesis system and method. The system comprises a flow path switcher, an injection pump, a reagent converging part, a pre-activation reaction module and a coupling reaction module, wherein the flow path switcher comprises a multi-channel switching valve. When the system disclosed by the invention is used for synthesizing the polypeptide, the reaction efficiency is remarkably improved, the use equivalent of amino acid is reduced, the yield and purity of the polypeptide are improved, and long-difficult peptides and modified peptides can be synthesized on a relatively large scale.

This invention provides a class of D-configuration oncolytic peptide-camptothecin conjugates, their preparation methods, and applications, belonging to the fields of peptide preparation and biomedical technology. The steps are as follows: D-configuration peptides are synthesized using a solid-phase peptide synthesis method based on 9-fluorenemethoxycarbonyl. Then, the synthesized D-configuration peptides are covalently linked to a small molecule antitumor drug via a linker group. The D-configuration oncolytic peptide-camptothecin conjugates are obtained through peptide cleavage, separation and purification, and freeze-drying. The D-configuration oncolytic peptide-camptothecin conjugates significantly improve the solubility of camptothecin, exhibiting advantages such as high enzymatic stability, long half-life, and stronger inhibitory effect on tumor cell proliferation. Furthermore, this invention enables spatiotemporal monitoring of the release of camptothecin conjugates, thus possessing good practical application value.

Object of the present invention are compounds of formula (1) as protecting groups to perform liquid phase peptide synthesis and a versatile, sustainable and cost-effective industrial process for liquid phase peptide synthesis.

The present invention provides a polypeptide ligase mutant and a method for preparing a polypeptide. The polypeptide ligase mutant comprises: (a) a protein mutated from the amino acid sequence shown in SEQ ID NO: 1, the mutation comprising a mutation occurring at the S307 site; or (b) a protein having 70% or more identity with the amino acid sequence defined in (a) and having polypeptide ligase activity. The present invention can solve the problem in the prior art of poor activity of polypeptide ligases, and is applicable to the field of polypeptide synthesis.

The present invention relates to a Bsmoc protected lysine salt (1 X ) of formula 1 X , for example the hydrochloride salt (n = 1, HY = hydrochloric acid). The present invention also relates to a method for preparing a Bsmoc protected lysine derivative, which method employs a Bsmoc protected lysine salt of formula 1 X and comprises the step of reacting the epsilon-amino group of lysine with an activated carboxylic acid derivative. Some of the obtained Bsmoc protected lysine derivatives with side chain modifications are used as starting materials in solid phase peptide synthesis.

The present invention relates to methods for performing solid phase peptide synthesis, automated parallel solid phase peptide synthesis, and devices designed to perform such methods. According to the invention, ultrasound at a frequency greater than 25 kHz is applied at least intermittently during the process to the reaction medium in which the solid phase peptide synthesis is carried out.

This invention discloses an interchain disulfide bond Cys A24 -Cys B23 H2Relaxin derivatives with thioether bonds as alternatives and their synthetic methods, wherein the structural formula of the H2Relaxin derivatives is shown below: This invention first utilizes diaminodioic acid molecules to synthesize interchain disulfide bonds (Cys) in a single solid-phase process using an N-fluorenemethyloxycarbonyl (Fmoc) solid-phase polypeptide synthesis method. A24 -Cys B23 The folded refolding precursor of H2Relaxin derivatives with thioether bonds as substitutes was then efficiently obtained via one-step oxidative folding and refolding to yield interchain disulfide bonds (Cys). A24 -Cys B23 The H2Relaxin derivative is replaced by a thioether bond.

The invention provides a polypeptide synthesis platform. The platform comprises a pulse microfluidic system, an ionic liquid assisted supercritical CO2 system, a polypeptide synthesis system and a control system, the pulse microfluidic system provides a reactor, the ionic liquid assisted supercritical CO2 system provides a reaction solvent, the polypeptide synthesis system provides a reactant, and the control system provides reaction conditions. The platform integrates a pulse flow micro-fluidic technology, a pressure-resistant micro-fluidic chip, an ionic liquid-assisted supercritical CO2 system and the like, realizes automatic production, remarkably improves the efficiency, yield and purity of polypeptide synthesis, reduces solvent consumption and waste generation, realizes an efficient and green polypeptide synthesis process, is suitable for various polypeptides, and has wide application prospects. The method is especially suitable for synthesis of long-chain and special peptides which cannot be effectively solved in the prior art.

This utility model discloses an inert gas protective stirring system for solid-phase peptide synthesis to prevent aggregation, comprising: a main body and a sealing mechanism. The sealing mechanism includes a stirring tank, a stirring cover installed at the upper end of the stirring tank, a lower abutment block installed at the upper end of the stirring tank, and a sealing strip connected to the upper end of the lower abutment block; and a disassembly mechanism located inside the sealing mechanism. This utility model provides an inert gas protective stirring system for solid-phase peptide synthesis to prevent aggregation. By installing a sealing mechanism inside the main body of the device, it achieves the effect of sealing and isolation while facilitating disassembly and cleaning. By installing a disassembly mechanism inside the sealing mechanism, the threaded block at one end of the stirring rod can be quickly unscrewed and disassembled.

The invention provides a synthesis method of canagliptin, and belongs to the technical field of polypeptide synthesizing.The method comprises the steps that firstly, a peptide fragment I, a peptide fragment II, a peptide fragment III and a peptide fragment IV are subjected to solid-phase synthesis through an Fmoc method, then all the fragments are condensed in sequence, then splitting decomposition is conducted to remove side chain protecting groups, and crude linear canagliptin peptide is obtained; the preparation method comprises the following steps: taking 7-(trifluoromethyl)-1H-[1, 2, 3] triazolo [4, 5-e] triazolo [1, 5-a] pyrazine-1-ol as a condensation system in a coupling reaction system for solid-phase synthesis of a peptide fragment by an Fmoc method, and then carrying out cyclization, separation and purification, salt conversion, concentration and freeze-drying to obtain a canagliptin finished product. The invention also uses DIC / 7-(trifluoromethyl)-1H-[1, 2, 3] triazolo [4, 5-e] triazolo [1, 5-a] pyrazine-1-ol as a condensation system. The synthesis method of the canagliflozin has the advantages that the reaction rate is high, the cost is low, large-scale production is easy, the obtained crude product is beneficial to purification, the purity and yield are high, the amount of waste liquid is small, and energy conservation and environmental protection are achieved.

The invention relates to the technical field of polypeptide synthesis and purification, in particular to a purification method of tirpotide, which comprises the following steps: dissolving a tirpotide crude product in an ammonia water solution, and filtering to obtain a solution containing tirpotide; the method comprises the following steps: carrying out first purification on a solution containing tilpotitide by taking a reverse silica gel filler as a stationary phase, a polar organic solvent as a mobile phase A, an ammonium trifluoroacetate aqueous solution as a mobile phase B and purified water as a mobile phase C, so as to obtain a component of which the single impurity content is less than 0.5%; carrying out secondary purification on the obtained component by taking a reverse silica gel filler as a stationary phase, a polar organic solvent as a mobile phase A, an ammonium formate aqueous solution as a mobile phase B and purified water as a mobile phase C, so as to obtain a component of which the single impurity is less than 0.1%; and concentrating, filtering and drying the obtained components. The method has the advantages of short purification period, high impurity removal efficiency, high recovery rate and low environmental protection cost.

The invention relates to an amino acid monomer based on 3, 5-dinitropyridine-2-sulfenyl protection and a synthetic method of polypeptide of the amino acid monomer. The monomer has the following structural formula, according to the preparation method, a corresponding polypeptide is synthesized from a 3, 5-dinitropyridine-2-sulfenyl protected amino acid monomer through a solid-phase synthesis method under a mild weakly acidic deprotection condition (2M 2-mercaptopyridine and an N, N-dimethylformamide solution of 1M pyridine hydrochloride), so that the generation of by-products caused by adopting an alkali (piperidine) deprotection condition in a current polypeptide solid-phase synthesis technology is avoided. The solid-phase synthesis method of the polypeptide based on the 3, 5-dinitropyridine-2-sulfenyl, disclosed by the invention, has a wide application prospect in the field of polypeptide synthesis.

A method and apparatus for online UV monitoring of an automated synthesis reaction. The apparatus includes a module having a fluid cell, the operating parameters of which remain substantially constant not only during the monitoring process but also during the time between subsequent processes. The module includes a separable housing structure that accommodates a UV light source and photodetector integrated with a fluid cell that is substantially unaffected by temperature. A portion of the cell is formed within a cell chassis and defined by a slot complemented by an inlet and an outlet, the inlet and outlet being sized so that no bubbles and / or stagnant fluid are present in the cell during the time the liquid reactants to be measured are delivered to the cell through the inlet. The method is configured to determine the progress and completion of the reaction, as well as modifications to the reaction time and the number of reaction iterations, in real time.

The invention provides a preparation method and application of a multi-target enzymatic self-assembly fluorescence activated nanoprobe, and belongs to the field of pharmacy. According to the method, an Fmoc solid-phase peptide synthesis strategy is adopted, an N end is connected with a fluorescence quencher Dabcyl, a C end is marked with fluorescein, a near C end is a self-assembly sequence Y (pY) YG capable of generating self-assembly behavior through dephosphorylation of alkaline phosphatase, and a middle section is a cleavage sequence KGGFLGK capable of being cleaved into a fluorescent'switch 'by cathepsin B; the near N end is a functional polypeptide F-pY-LyP-1 of a targeting sequence CGNKRTRGC or LyP-1 which can be highly combined with a p32 receptor on the surface of a foam cell. The fluorescence activated nanoprobe accurately reaches a plaque part under the synergistic effect of multiple target points and is self-assembled into spherical nanoparticles, so that the aggregation and retention effects of the probe in the plaque are improved, the non-specific signal interference is reduced, and the early atherosclerotic plaque is more accurately identified.

The invention provides an ultrasonic-assisted polypeptide C-terminal amidation method and application thereof. The method comprises the following steps: mixing a substrate polypeptide, a reducing agent and a buffer solution to obtain a reaction system; performing ultrasonic treatment on the reaction system for reaction to obtain a polypeptide C-terminal amidated product; the reducing agent comprises one or more of tris (2-carbonyl ethyl) phosphorus hydrochloride, triphenylphosphine tri-m-sodium sulfonate or triphenylphosphine; the structures of the substrate polypeptide and the amino acid residue at the C end of the substrate polypeptide are as shown in a formula I, and the structures of the product and the amino acid residue at the C end of the product are as shown in a formula II, r2 is a sequence composed of other amino acids except the amino acid at the C terminal in the substrate polypeptide; r is a sequence composed of other amino acids except the amino acid at the C terminal in the product; x1 is SH or SeH; and X2 is OH or NH2. The problem that in the prior art, the large-scale synthesis cost of polypeptide C-terminal amidation is too high can be solved, and the method is suitable for the field of polypeptide synthesis.

The invention discloses a high-throughput screening method for a polypeptide modified drug preparation system based on glass bead immobilization, which comprises the following steps: S1, gradually coupling amino acid according to a polypeptide solid-phase synthesis method by taking glass beads with active functional groups on the surfaces as solid-phase carriers to obtain glass bead immobilized polypeptide; s2, adding a modifier into the glass bead immobilized polypeptide obtained in the step S1, and reacting; at least one of the modifiers contains a fluorescence labeling group; and S3, detecting a fluorescence signal of the product obtained in the step S2. According to the screening method, excellent physical and chemical and optical characteristics of the glass beads are utilized, the glass beads are used as carriers for polypeptide synthesis modification, and meanwhile, fluorescence signals are introduced into detection of solid-phase polypeptide modification reaction, so that high-throughput screening of a polypeptide modified drug preparation system is realized; the screening efficiency of the polypeptide modified medicine and the preparation conditions and method thereof is effectively improved.

The invention belongs to the field of polypeptide synthesis reaction, and particularly relates to a polypeptide synthesis reaction device for biological medicine manufacturing and an operation method thereof.The polypeptide synthesis reaction device comprises a base, a side plate is fixedly connected to the top of the base, a supporting plate is slidably connected to the inner wall of the side plate, and a hydraulic cylinder is fixedly connected to the position, located below the supporting plate, in the side plate; according to the polypeptide synthesis reaction device for biological medicine manufacturing and the operation method of the polypeptide synthesis reaction device, the baffle is inserted into the sliding groove, the bottom of the reaction tank is blocked, the baffle in the sliding groove is pulled out, so that the bottom of the reaction tank is not blocked, polypeptide synthesis reactants in the reaction tank directly fall down, the polypeptide synthesis reactants are convenient to take, and the operation efficiency is improved. The reaction tank is convenient to clean, the reaction tank after discharging is controlled to move downwards, the reaction tank is vibrated downwards multiple times through cooperation of a plurality of protruding plates and clamping blocks, polypeptide synthesis reactants adhering to the interior of the reaction tank can be conveniently shaken off, and waste caused by the fact that the polypeptide synthesis reactants adhere to the interior of the reaction tank is avoided.