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Multi channel peptide synthesizer assists in Alzheimer's drug development
Date: 2025-01-08Read: 0
Human β - amyloid protein 1-42 (Figure 1) is the main component of brain deposition plaques in Alzheimer's disease (AD) patients. The daily research on AD pathology, diagnostic tools, and potential drugs requires an uninterrupted supply of beta amyloid protein. However, according to reports, due to the high hydrophobicity of the C-terminus and resin aggregation caused by sequence pre deposition, β - amyloid protein (1-42) is difficult to prepare through traditional solid-phase peptide synthesis (SPPS). The latest progress in the automatic synthesis of β - amyloid protein includes infrared heating and new coupling reagent combinations, which can better synthesize difficult hydrophobic peptides with shorter cycle times. This article is in Symphony®The X 24 channel peptide synthesizer was used to explore the synthesis efficiency of β - amyloid protein under different synthesis methods in parallel.
Using Rink Amide ChemMatrix ™ (0.47 mmol/g) and R TentaGel ™ Resin (0.19 mmol/g), PurePep ™ Reagent: HDMI/OxymaPure ™ Or HDMC/OxymaPure and DIPEA with pre loaded Fmoc amino acids (1:1:1:2), in Symphony®On the X synthesizer, synthesize peptides under conditions of 10 fold excess and a scale of 50 µ mol. Two synthesis methods were tested:
Method A:Unprotection for 2 x 3 min, coupling for 2 x 5 min, 25 ° C.
Method B:Unprotection for 2 x 30 s, 90 ° C, coupling for 2 x 1 min, 90 ° C, infrared heating.
The Symphony X peptide synthesizer has flexible functions and can simultaneously test 5 independent conditions for β - amyloid protein synthesis. HDMA (Table 1) and HDMC (Table 2) were each combined with OxymAPure reagent, and Rink ChemMatrix or R TentaGel resin were used to synthesize β - amyloid protein using the slower method A. The purity of the crude product obtained was similar. The highest purity comes from the combination of HDMA/OxymaPure reagent and Rink Amide ChemMatrix resin (Figure 2).
After increasing the temperature (90 ° C) and accelerating the reaction (Method B), the crude purity of HDMA/OxymaPure/DIPEA reagent combined with Rink Amide ChemMatrix resin was 33.9%. The synthesis purity of the combination of accelerated method+high temperature+HDMA is similar to that of the slow method (Table 1).
Table 1Synthesize β - amyloid protein using HDMA/OxymaPure/DIPEA reagents according to method A, and obtain the purity and yield of the crude product.
Table 2Synthesize β - amyloid protein using HDMC/OxymaPure/DIPEA reagent according to method A, and obtain the purity and yield of the crude product.
12- ß - amyloid protein -4.940
min
1
Using Symphony X peptide synthesizer, 24 synthesis conditions can be evaluated simultaneously within 24 hours (room temperature) or 17 hours (90 ℃), and automatic peptide chain cleavage can be completed on the instrument
2
Six different synthesis conditions (temperature, resin, and coupling reagent combination) were evaluated, and the optimal synthesis conditions for the purity and yield of hydrophobic peptides were preliminarily determined
3
Using the novel peptide reagent HDMA, β - amyloid protein (1-42) - NH was successfully synthesized on the Symphony X peptide synthesizer by rapid method and infrared heating (90 ° C)2
4
On this basis, further optimization of the synthesis of hydrophobic long-chain peptides (such as β - amyloid protein) can be achieved, including shortening the coupling time and using new coupling reagent combinations
