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Polypeptide drug substance
Targets of peptide drugs
Most peptides target extracellular molecules
The main extracellular molecular targets are G protein coupled receptor (GPCR). The GPCR family is the largest receptor family with 800-1000 members. GPCR plays an important role in the development of modern drugs. About 50% of modern drugs are targeted at GPCR. The common feature of these GPCRs is that they all have seven transmembrane domains. GPCR signal usually interacts with these GPCRs through extracellular ligands, causing conformational changes of GPCR, and regulating various signal pathways downstream of GPCR by activating G protein of triplet. Some receptors of GPCR family are expressed abnormally in specific tissues and cells, which regulate normal or abnormal physiological functions of human body, and are potential targets of drug development. Some GPCR ligands are small molecular peptides, and the modification of these peptides has become one of the most important directions of peptide drug development.
Less than 10% of them targeted intracellular molecules
Preparation of polypeptide drugs
Extraction method
A considerable number of peptide drugs are extracted from animals and plants, such as insulin from pig pancreas. The purity of peptides obtained by extraction method is low, and the content of polypeptides in organism is very small, so it is easy to introduce animal pathogenic bacteria or viruses in the extraction process, which limits its application. Therefore, biological peptide extraction technology has been gradually replaced by chemical synthesis or gene recombination technology.
Chemical synthesis
(1) Protective agents for peptide synthesis
Polypeptides are composed of amino acids. Both natural peptides and synthetic peptides produced by organisms have different amino acids connected by amide bond in a certain order. The amide bond is formed by the removal of one amino acid amino group and another amino acid carboxyl group. Therefore, the key point of the chemical synthesis of polypeptides is to activate or protect the amino and carboxyl groups at the appropriate position and time. The synthesis of polypeptide includes three steps: Step 1, protecting the active part not involved in the reaction; step 2, activating the carboxyl group as the active intermediate; step 3, deprotection of the protected group. The peptide was purified by HPLC.
(2) Liquid phase synthesis of polypeptide drugs
The liquid-phase synthesis of polypeptides is mainly carried out in solution, and there are two strategies: Step-by-Step synthesis and fragment combination. The two strategies are often used in combination. Some short polypeptide fragments were synthesized by step-by-step synthesis. Then, the peptide fragment obtained in the previous step is connected to the target peptide by the method of fragment combination. Liquid phase synthesis method is convenient, rapid and high purity. It is suitable for the situation that the target peptide is short and needs large amount of synthesis.
(3) Solid phase synthesis of peptide drugs
Solid phase synthesis is to fix the N-terminal of amino acids on insoluble resin, and then condense amino acids on the resin in turn. Solid phase method has become a common technique in peptide and protein synthesis. Especially in the synthesis of long chain polypeptides or proteins, the solid-phase synthesis method has more advantages than the classical liquid-phase method.
Gene recombination technology
Many organisms in nature can produce active peptides, such as mammalian insulin. However, the extraction of bioactive peptides from plants and animals requires a lot of raw materials, which is expensive and not green enough. Using gene technology to produce natural active polypeptide solves this problem. Recombinant technology is to construct the gene sequence of polypeptide on the vector to form recombinant DNA expression vector, and then to express, extract and purify polypeptide in prokaryotic or eukaryotic cells. This method is suitable for the preparation of peptides with more than 50 amino acids and is easy to obtain. With the improvement of the technology of producing peptides by genetic engineering, the development and clinical application of genetic engineering peptide drugs are accelerated.
Preparation of peptide drugs by enzymatic degradation
Organisms contain a large number of proteins, and some active peptides may be some sequences in proteins. If the more easily available proteins can be degraded into the required polypeptide molecules (possibly many kinds), the cost can also be saved. In recent years, some scholars use enzymatic hydrolysis to synthesize peptides. Enzymatic degradation often needs to find enzymes that catalyze the decomposition reaction at specific structures, which can efficiently function at all the same structures in proteins. However, a series of peptides are often obtained by this method, which is difficult to separate and purify, so it is not suitable for the synthesis of single peptide. In addition, due to the strong specificity of enzymes, it is sometimes necessary to cut off multiple structural units, so it is necessary to search for a variety of enzymes, which increases the difficulty and scope of application of this method.
Detection methods of polypeptide drugs
Reversed phase high performance liquid chromatography
capillary electrophoresis
High performance liquid chromatography mass spectrometry
High performance molecular exclusion chromatography
How many companies are there in peptide api manufacturer in china? The peptide api market is very promising, and the world is encouraging the development of peptide business. There is a peptide api list on the website Biofda.com, which contains various specifications of peptide APIs for customers to choose from. Shengnuo Technology is a peptide api manufacturer located in Chengdu, a city in southwest China. Not only peptide APIs, but also carnosine custom suppliers and cosmetic peptide suppliers
There are many peptide apis manufacture in China, but they are all small-scale companies. The China peptide company such as Sinotech is a leading company in China and has a very high position.
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