Acylation of neomycin B (I) with N-alpha-carbobenzoxy-N-omega-nitro-L-arginine (II) by means of EDC affords the protected neomycin B hexa-amide (III). Removal of the N-carbobenzoxy and N-nitro protecting groups of (III) is then achieved by catalytic hydrogenation over Pd/C, to furnish the title compound.