Genome targeting methods enable cost-effective capture of specific subsets of the genome for sequencing. We present here an automated, highly scalable method for carrying out the Solution Hybrid Selection capture approach that provides a dramatic increase in scale and throughput of sequence-ready libraries produced. Significant process improvements and a series of in-process quality control checkpoints are also added. These process improvements can also be used in a manual version of the protocol.

A series of novel macrocyclic taxoids 6 and 6′ bearing 16-, 17-, and 18-membered rings connecting the substituents at the C-2 and C-3′ positions were designed and synthesized. The syntheses of these macrocycles 6 and 6′ were accomplished using the highly efficient ruthenium-catalyzed ring-closing metathesis (RCM) of taxoid-ω,ω′-dienes 7 in the key step. Taxoid-ω,ω′-dienes 7 were obtained through the ring-opening coupling of 4-alkenyl-β-lactams 9 with 2-alkenoylbaccatins 8 in good to high yields. Although various novel pentacyclic macrocycles 6 and 6′ were successfully synthesized, there were cases in which the desired RCM did not proceed. The scope and limitation of RCM in its application to highly functionalized complex substrates are discussed. All macrocyclic taxoids 6 and 6′ were found to be cytotoxic, with some of them exhibiting submicromolar IC 50 values against a human breast cancer cell line.

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