Study On The Synthetic Process Of GHK Copper Peptide

Objective: GHK copper peptide is a compound formed by the complexation of copper ion and GHK tripeptide with Glycyl-L-Histidyl-L-Lysine-Copper as its specific composition.It is widely used in many fields,such as blood circulation improvement,anti-oxidation,anti-inflammatory,neurotrophic factors’ synthesis promotion and cancer treatment.Most of the reported GHK copper peptide synthesis methods require the application of resins,and it is difficult to obtain GHK copper peptides with high yield and stable parameters such as acetic acid content,which are not suitable for large-scale industrial production.Therefore,this paper focuses on the synthetic process of GHK copper peptide,and is committed to developing a low-cost,simple and easy-to-use GHK copper peptide synthetic method.This method meets the requirements of acetic acid content to be stable and controllable,and meanwhile suitable for large-scale production.Method: Glycine,L-histidine and L-lysine were the starting materials,tert-butoxycarbonyl(Boc-)was used to selectively protect glycine and lysine,and trityl(Trt-)was used to selectively protect histidine.In this way,Boc-glycine,Trt-histidine and Boc-lysine are generated respectively.N-hydroxysuccinimide and N,N-dicyclohexylcarbodiimide system are used to connect the peptide,Boc-glycine and Trt-histidine are firstly connected to obtain a protected dipeptide before connecting with Boc-lysine to obtain protected tripeptide.Select trifluoroacetic acid to carry out deprotection,remove Boc group and Trt group,obtain GHK tripeptide trifluoroacetate,then according to the solubility property of the substance,remove trifluoroacetate through acid-base neutralization to obtain GHK tripeptide,which is therefore complexed with copper acetate and copper hydroxide to eventually obtain GHK copper peptide.Results: In this paper,the liquid phase synthetic method was adopted to avoid the use of resin.Through amino acid protection,peptide attachment,deprotection and complexation in a total of 10 steps,the important parameters such as acetic acid content were effectively controlled,and the total yield was 41%.The preparation of GHK copper peptide was completed.The liquid phase purity of the final product was99.63%,the GHK content was 72.1%,the acetic acid content was 13.3%,and the copper content was 11.14%.Conclusion: The synthetic process designed in this paper is easy to operate,and the reaction conditions are mild.As the column chromatography separation and purification is avoided,the liquid phase purity of GHK copper peptide is managed to be ≥99%,the acetic acid content ≤15%,and the copper content is in the range of8%-16%.The industrial production target of GHK has good process stability,which lays a foundation for the industrialization of GHK copper peptide liquid-phase synthesis.