Conjugation of pleuromutilin is an attractive strategy for development of novel antibiotics and the fight against multi-resistant bacteria as the class is associated with low rates of resistance and cross-resistance development. Herein, the preparation of 35 novel (+)-pleuromutilin conjugates is reported. Their design was based on a synthetically more efficient benzyl-adaption of a potent lead, but still relied on the Cu(I)-catalysed alkyne-azide [3+2] cycloaddition for conjugation onto pleuromutilin. Their antibacterial activity was evaluated against the multi-resistant Staphylococcus 2 aureus strain USA300 for which they displayed moderate to excellent activity. Compound 35, bearing a para-benzyladenine substituent proved particularly potent against USA300 as well as additional strains of MRSA and displayed as importantly no cytotoxicity in four mammalian cell lines. Structure-activity relationship analysis revealed that the purine 6-amino is essential for high potency, likely due to strong hydrogen bonding with the RNA backbone of C2469 as suggested by a molecular model based on the MM-GBSA approach. 9 Alkylation of the nucleobases (thymine, adenine, 6-chloropurine and 2-amino-chloropurine) was achieved in the presence of an inorganic base (K2CO3 or NaH) in DMF. The yields for 68-78 were mainly deteriorated by lack of regioselectivity, i.e. formation of the N3 isomer (thymines) or N7 isomer (purines), which in turn also complicated their subsequent separation during Flash Chromatography. The strategy was overall still more efficient and straightforward than our previous approach.The chloropurins 74-78 were converted via nucleophilic aromatic substitution with various amines in anhydrous EtOH to grant 79-86, generally in excellent yields. Compound 87 was prepared via guanidination in accordance with a procedure optimized by Česnek and coworkers. 35 The primary benzylamines 89 and 96-97 were synthesized via two strategies (Scheme 3); the para analogue 89 via standard Gabriel synthesis, which proved highly efficient, simultaneously also granting the phthalimide 88. The second strategy involved reduction of ethynylbenzonitriles with LiAlH4, 36 which also proved efficient, granting the nitriles 94 and 95 as well. Scheme 3.Reagents and conditions: (a) K-phthalimide, DMF, rt.; (b) H2N-NH2  H2O 50-60% w/w, EtOH, 78 o C; (a) Trimethylsilylacetylene, Pd(PPh3)4, CuI, Anh. Piperidine 80 o C; (d) K2CO3, MeOH, rt.; (e) i) 1 M LiAlH4 in THF, THF, -10 o C → rt. ii) H2O, NaOH (sat.).