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[12] The 520IU dose provides meaningful support without excessive intake that could lead to soft tissue calcification

Step 3: Preparation of H-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)- Val-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Gly-Gln(Trt)-Ala-Ala-Lys(Boc)- Glu(OtBu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(Pbf)-Gly-Arg(Pbf)-Gly-Resin (3-31) Fmoc-Arg(Pbf)-OH (18 gm) was dissolved in DMF (125 ml) at 25-30C

The present inventors when carried out the reaction of (3-31) amino acid fragment of Semaglutide having the amino acid sequence Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser- Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly with PGi-His(PG)-Aib-OH and further converted the obtained compound to Semaglutide surprisingly found that the said process efficiently controlled the formation of D-His, Des- His, Des-Aib, Di-Aib, Des-Aib-His impurities in the Semaglutide API which is highly advantageous on industrial scale especially for complex peptide molecules like Semaglutide

The run time in the RP-HPLC purification processes of the present invention is about 50 min, or about 60 min, or about 70 min, or about 80 min, or about 90 min, or about 100 min, or about 110 min, or about 120 min, or about 140 min, or about 160 min, or about 180 min, or about 200 min, or about 220 min, or about 240 min, or about 260 min, or about 280 min, or about 300 min, or about 320 min, or about 340 min, or about 360 min or about 380 min or about 400 min

Kanamycin sulphate solution was added in Erlenmeyer flasks containing pre seed medium