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. 2025 Mar:114:107257.
doi: 10.1016/j.ultsonch.2025.107257. Epub 2025 Feb 7.

Sustainable Ultrasound-Assisted Solid-Phase peptide synthesis (SUS-SPPS): Less Waste, more efficiency

Salvatore Mottola  1 , Alessandra Del Bene  1 , Vincenzo Mazzarella  1 , Roberto Cutolo  1 , Ida Boccino  2 , Francesco Merlino  3 , Sandro Cosconati  1 , Salvatore Di Maro  4 , Anna Messere  5
Affiliations

Sustainable Ultrasound-Assisted Solid-Phase peptide synthesis (SUS-SPPS): Less Waste, more efficiency

Salvatore Mottola et al. Ultrason Sonochem. 2025 Mar.

Abstract

The integration of low-frequency ultrasound with Solid-Phase Peptide Synthesis (SPPS) was explored to establish a Sustainable Ultrasound-assisted Solid-Phase Peptide Synthesis (SUS-SPPS) method. This innovative approach significantly reduces solvent consumption, washing steps, time, and reagent usage compared to conventional manual SPPS protocols. The SUS-SPPS method exploits ultrasound at every stage of synthesis and work-up, reducing the process to just two steps. The first step sequentially combines Fmoc-amino acid coupling, capping of unreacted amino groups, and Fmoc deprotection into a single operation, while the second one consists of a single washing procedure. Moreover, we demonstrated that the method is compatible with various resin types, including Rink-amide, Wang, and Cl-Trt resins, and facilitates the efficient synthesis of peptides of varying lengths (up to 20-mers) and compositions, including those traditionally considered "difficult sequences", with excellent yields and purity. Notably, SUS-SPPS reduces solvent usage per coupling cycle by 83-88%, marking a significant breakthrough in sustainable peptide synthesis.

Keywords: Green approach; Minimized solvent consumption; Solid-Phase Synthesis; Sonochemistry; Sustainable Peptide Synthesis; Ultrasound.

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Conflict of interest statement

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

None
Graphical abstract
Fig. 1
Fig. 1
Schematic overview of the conventional SPPS process.
Fig. 2
Fig. 2
Schematic overview of the SUS-SPPS process.
Fig. 3
Fig. 3
Optimization study for the SUS-SPPS of the model 5-mer peptide (Fmoc-KFRFD-NH2). Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 4
Fig. 4
Unwanted and Desired Effects of Adding Piperidine at the End of the Coupling Step.
Fig. 5
Fig. 5
SUS-SPPS of peptide Kisspeptin10 (YNWNSFGLRF). Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 6
Fig. 6
SUS-SPPS of peptide PAMP1-20 (ARLDVASEFRKKWNKWALSR). Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 7
Fig. 7
SUS-SPPS of peptide Aib-Enk, (YAib-Aib-FL). Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 8
Fig. 8
SUS-SPPS of peptide ACP65−74 (VQAAIDYING). Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 9
Fig. 9
SUS-SPPS of peptide Angiotensin-I (DRVYIHPFHL-COOH) on Wang resin. Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 10
Fig. 10
SUS-SPPS of peptide Leu-Arg-Lys-Tyr-Arg-Phe on TRT-Cl resin. Peptide synthesis was attained in triplicate and the resulting crude purities are expressed as a percentage (mean values ± standard error of measurement (SEM), N = 3).
Fig. 11
Fig. 11
Solvent waste during the synthesis of Fmoc-KFRFD-NH2 on 0.5 mmol scale. A: standard protocol, B: SUS-SPPS protocol b (Table 2).
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