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The reaction mixture is then diluted with a 0.1 % TFA solution in water/ACN 1 :1 (v/v) and purified using HPLC preparative method 5 to afford 44 mg (77% based on starting compound INT1 ) of intermediate compound (2S,3S,4S,5R,6S)-6-(4-(2-(1-(35- amino-3-glycyl-6,9,12,15,18,21 ,24,27,30,33-decamethyl- 5,8,11 ,14,17,20,23,26,29,32,35-undecaoxo-3,6,9,12,15,18,21 ,24,27,30,33- undecaazapentatriacontyl)-1 H-1 ,2,3-triazol-4-yl)-1-((((1 R,9R)-9-ethyl-5-fluoro-9- hydroxy-4-methyl-10, 13-dioxo-2,3,9,10, 13, 15-hexahydro-1 H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1 ,2-b]quinolin-1-yl)carbamoyl)oxy)ethyl)-2- nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid as a yellow solid