a) Coupling of Fmoc-Gly-OH with Resin in a solvent and a base by solid phase peptide synthesis to provide Fmoc-Gly-Resin, b) treating Fmoc-Gly-Resin with a deprotecting agent to provide H-Gly-Resin, c) coupling of Fmoc-Arg(pbf)-OH to H-Gly-Resin in a solvent optionally in presence of a coupling agent and/or a base to provide Fmoc-Arg(Pbf)-Gly-Resin, d) treating Fmoc-Arg(Pbf)-Gly-Resin with a deprotecting agent to provide H-Arg(Pbf)- Gly-Resin, e) repeating the coupling and deprotection steps with remaining amino acids Fmoc-Gly- OH, Fmoc-Arg(pbf)-OH, Fmoc-Val-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc- Ala-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Ala-OH, Fmoc-Ala-OH, Fmoc-Gln(Trt)-OH, Fmoc-Gly-OH, Fmoc-Glu(OtBu)- OH, Fmoc-Leu-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Val-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc- Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Gly-OH, Fmoc-Glu(OtBu)-OH by linear approach as per the amino acid sequence in Semaglutide backbone in a solvent optionally in presence of a coupling agent and/or a base
Step 3: C18 column was washed with Acetonitrile: Water and then equilibrated with aqueous ammonium acetate
Recombinant vs Chemical : Chemical synthesis is far more common: most (31 / 38) of the recently approved small peptide drugs are chemically synthesized
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Purity by HPLC: 90.15%
