Solid Phase Fragment Synthesis of Thymosin α1

Abstract

The invention discloses a synthesis technology of thymosin alpha1, which comprises the following steps: synthesizing multiple segments according to the difficult-synthesis sequence of thymosin alpha1, wherein a solid-phase segment condensation method is adopted for the area of difficult sequences so as to improve the synthesis quality and shorten the synthesis time, and a conventional step-by-step condensation method is adopted for the area of easy-synthesis sequences; obtaining all-protected thymosin alpha1 peptide resin on Wang Resin; cracking the peptide resin to obtain crude peptide by taking TFA / m-cresol or TFA / thioanisole / EDT / anisole as a cracking reagent; dissolving the crude peptide and filtering; and performing preparative reversed phase HPLC purification, desalting, rotary evaporation and freeze drying to obtain a finished product of thymosin alpha1 of which the purity can exceed 99%. According to the technology disclosed by the invention, the purity of the synthesized crude peptide can reach about 80%, the purification is easy, and the total yield can reach 30%. The total yield and product quality of thymosin alpha1 produced by the method disclosed by the invention are both much remarkably improved than those of other methods reported in documents; and moreover, the production cycle can be shortened by 40-50%, the production cost is reduced, and the method is suitable for large-scale preparation of high-purity thymosin alpha1.

Description

technical field [0001] The invention relates to the field of chemical synthesis of polypeptide drugs, in particular to a method for synthesizing solid-phase fragments of thymosin α1. Background technique [0002] thymosin a 1 It is an important polypeptide immunoregulatory factor secreted by the thymus, which can significantly enhance immune function, and is recognized as the most effective drug for specifically regulating immune imbalance and enhancing immunity. thymosin a 1 Originally extracted from calf thymus. thymosin a 1 It consists of 28 amino acid residues, with a molecular weight of 3108, an isoelectric point of 4.2, and an acetylated structure at the N-terminal. thymosin a 1 Promote the maturation of T lymphocytes by stimulating peripheral blood lymphocyte mitogens, increase the levels of interferon α, interferon γ, interleukin 2, interleukin 3 and other lymphokines secreted by T cells after antigen or mitogen activation, and increase T cells Surface lymphoki...

AI Technical Summary

Problems solved by technology

thymosin alpha 1 The 1, 8, and 9 positions of the sequence contain 3 Ser, and the 6, 12, and 13 positions contain 3 Thr. Compared with the classic Fmoc / tBu stepwise condensation process, this method can increase thymosin α to a certain extent. 1 Synthetic quality, but thymosin alpha 1 The most difficult sequence interval to synthesize (Lys 17 to Glu 24 The sequence between them can form a ~ helical structure) does not contain Thr or Ser, and the preparation process of Thr or Ser pseudoproline dipeptide is complicated and expensive

Therefore, it is difficult to use this process to prepare cost-competitive high-quality adenosin α on a large scale 1 API

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