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Using preloaded Fmoc-Val-Wang resin (491 mg, 0.30 mmol) as solid phase, each coupling used 4 fold amino acid excess, and HBTU, HOBT as activators and DIEA as base in a 1:1:1:3 ratio. Fmoc deprotection was performed using 25% perperidine in DMF solution. After the NILS sequence (Pro-Lys-Lys-Lys-Arg-Lys- Val) was completed, a Lys(Mtt) residue was coupled to the end. One sixth of the resin was placed in a 25 mL fritted glass tube, swollen with DMF. A 3-fold excess of N-Boc-Baa (157 mg, 0.15 mM) dissolved in DMF/ DCM (9 mL, 2:1) was activated with PyBOP/HOBt/DIEA (1:1:1:3) for 2 minutes, then mixed with the resin in the fritted glass tube, and shaken on an automated shaker for 1 day at room temperature. The resin was washed thoroughly with DMF and DCM to remove any unreacted N-Boc-Baa. The resin was washed with DCM for complete removal of DMF. To achieve the maximum cleavage of Mtt protecting group, the resin was shrunk with MeOH twice. When the resin was treated with 1% TFA and 5% TIPS in
-
-1 flow rate. The elution time was 37 min. After purification 59.1 mg (50.6%) was recovered.
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