Most peptides target extracellular molecules

The extracellular molecular target is mainly G protein coupled receptor (GPCR). GPCR family is the largest receptor family, and about 800-1000 members have been identified. GPCR plays an extremely important role in the development of modern medicine. About 50% of modern medicine is targeted at GPCR. All of these GPCRs have seven transmembrane domains. GPCR signal generally interacts with these GPCRs through extracellular ligands, causing conformational changes of GPCR, and regulating various signal pathways downstream of GPCR by activating triplet G protein. Some receptors of GPCR family are abnormally expressed in specific tissues and cells, regulating normal or abnormal physiological functions of human body, which are potential targets of drug development. Some GPCR ligands are small peptides, and the modification of these peptides has become one of the most important directions in peptide drug development.

Less than 10% target intracellular molecules

1、 Preparation of polypeptide drugs

Extraction method

A considerable part of polypeptide drugs are extracted from plants, such as insulin from the pancreas of pigs. The purity of polypeptide obtained by extraction method is low, and the content of polypeptide in organism is very small, so it is easy to introduce animal pathogenic bacteria or virus 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. Whether natural polypeptides or synthetic polypeptides produced by organisms, there are different amino acids connected by amide bonds in a certain order. The amide bond is formed by the removal of one amino acid's amino group from the carboxyl group of another amino acid. Therefore, the emphasis of peptide synthesis lies in the activation or protection of amino and carboxyl groups at the right place and time. The synthesis of polypeptide includes three steps: Step 1, protect the active part which does not participate in the reaction; step 2, activate the carboxyl group as the active intermediate; step 3, deprotect the protected group. The peptide was purified by HPLC.

(2) Liquid phase synthesis of polypeptide drugs

The liquid-phase synthesis of polypeptide is mainly carried out in solution, and there are two strategies: Step-by-Step synthesis and fragment combination. These two strategies are often used in combination. First, some short peptide fragments were synthesized by step synthesis. The peptide fragment obtained in the previous step is then connected to the target peptide by the method of fragment combination. The method of liquid phase synthesis is convenient and high purity, which is suitable for the situation of short target polypeptide and large amount of synthesis.

(3) Solid phase synthesis of polypeptide drugs

The N-terminal of amino acids is fixed on the insoluble resin, and then the amino acids are condensed successively on the resin. Solid phase method has become a common technology in peptide and protein synthesis. Especially in the synthesis of long-chain polypeptide or protein, the solid-phase synthesis method shows the advantages that the classical liquid-phase synthesis method can not compare.

Gene recombination technology

Many living things in nature can produce active peptides, such as insulin in mammals. However, the extraction of active polypeptides from driven plants requires a lot of raw materials, which is expensive and not green enough. Using gene technology to produce natural active polypeptides solves this problem. Recombinant technology is to construct the gene sequence of polypeptide on the vector to form recombinant DNA expression vector, and to express, extract and purify polypeptide molecules in prokaryotic or eukaryotic cells. This method is suitable for the preparation of target peptides with more than 50 amino acids and is easy to obtain. With the improvement of the technology of peptide production by genetic engineering, the development and clinical application of peptide drugs by genetic engineering have been accelerated.

Preparation of polypeptide drugs by enzymatic degradation

Organisms contain a large number of proteins, some active peptides may be a certain sequence in the protein, if the relatively easy protein can be degraded into the required peptide molecules (may be many), it can also play a cost saving effect. In recent years, some scholars have used enzymolysis to synthesize peptides. Enzyme degradation often needs to find the enzyme that catalyzes the decomposition reaction at a specific structure. It can efficiently play a role in all the same structures of proteins. However, it is difficult to separate and purify a series of peptides, so it is not suitable to synthesize a single peptide. In addition, due to the strong specificity of enzymes, sometimes it is necessary to cut off multiple structural units, then it is necessary to find a variety of enzymes, which increases the application difficulty and scope of the method.