Fmoc gly oh

The GCPF peptide was chemically synthesized using Fmoc solid-phase peptide synthesis (SPPS). Briefly, Fmoc-Arg(OtBu)-Wang resin used as a solid-phase material was sequentially coupled to the protected amino acids: Fmoc-Gly-OH, Fmoc-Asn(Trt)-OH, Fmoc-Phe-OH, Fmoc-Arg(tBu)-OH, Fmoc-Trp-OH, Fmoc-Pro-OH and Fmoc-Arg(tBu)-OH (Sigma, USA). The coupling reactions were performed with HOBt/DIC (1.1/1.5, v/v). After all reactions were complete, the synthesized peptide was cleaved from the resin with the lysate solution (TFA/phenol/water/thioanisole/EDT, 92:2.5:2.5:1.5:1.5, v/v/v/v). Then, the purity of synthesized polypeptide was determined using high-performance liquid chromatography (HPLC) on an Amethyst C18-H column (150 mm×4.6 mm i.d., 5 μm, Sepax, USA) with a gradient elution using buffer A (0.1% trifluoroacetic acid (TFA) in 100% water (v/v)) and buffer B (a mixture of 0.1% TFA in 80% acetonitrile and 20% water (v/v)) as the mobile phase at a flow-rate of 1.0 mL/min and UV detection at 220 nm. The gradient started from 28% and ended at 48% of buffer B. Then, the purity of the synthesized polypeptide was determined confirmed to be more than 97.5%.

Fmoc-Gly-OH (≥98.0%), Fmoc-Gln(Trt)-OH
(≥98.0%), Fmoc-Trp-OH (≥97.0%), Fmoc-Trp(Boc)-OH (≥97.0%),
Fmoc-Pro-OH (≥99.0%), Fmoc-Tyr(tBu)-OH (≥98.0%),
DIC (99%), Oxyma (97%), NMP (≥99.0%), toluene (≥99.5%),
piperidine (99%), piperazine (99%), and DBU (98%) were purchased from
Sigma-Aldrich; 2-Cl-trityl chloride resin (100–200 mesh, 1%
DVB, 1.0–1.6 mmol/g), Fmoc-His(Trt)-OH (≥98.0%), and
Fmoc-Asn(Trt)-OH were procured from Iris Biotech GmbH; Fmoc-Cys[(CH₂)₃COOtBu]-OH (≥98.5%) was obtained from
Flamma, Dalian HonKai Chemical Development and Fmoc–N-4-F-Bn-Gly-OH (≥98.5%) was purchased from PolyPeptide
Group; Fmoc-Arg(Pbf)-OH (≥98.5%) was purchase from Flamma;
Fmoc-Sar-OH (≥98.0%) was procured from Fluorochem; DMF (≥99.5%)
was purchased from Merck; and Isolute C18 was procured from Biotage.

Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ala-OH,
Fmoc-Pro-OH, Sieber amide resin, DMF, piperidine, oxyma, N,N′-diisopropylcarbodiimide (DIC), trifluoroacetic
acid (TFA), and dichloromethane (DCM) were purchased from Sigma Aldrich.
The chemicals used did not go through any purification.