Conformational dynamics plays an important role in enzyme catalysis, allosteric regulation of protein functions and assembly of macromolecular complexes. Despite these well-established roles, such information has yet to be exploited for drug design. Here we show by nuclear magnetic resonance spectroscopy that inhibitors of LpxC—an essential enzyme of the lipid A biosynthetic pathway in Gram-negative bacteria and a validated novel antibiotic target—access alternative, minor population states in solution in addition to the ligand conformation observed in crystal structures. These conformations collectively delineate an inhibitor envelope that is invisible to crystallography, but is dynamically accessible by small molecules in solution. Drug design exploiting such a hidden inhibitor envelope has led to the development of potent antibiotics with inhibition constants in the single-digit picomolar range. The principle of the cryptic inhibitor envelope approach may be broadly applicable to other lead optimization campaigns to yield improved therapeutics.

SUMMARY LpxC is an essential enzyme in the lipid A biosynthetic pathway in Gram-negative bacteria. Several promising antimicrobial lead compounds targeting LpxC have been reported, though they typically display a large variation in potency against different Gram-negative pathogens. We report that inhibitors with a diacetylene scaffold effectively overcome the resistance caused by sequence variation in the LpxC substrate-binding passage. Compound binding is captured in complex with representative LpxC orthologs, and structural analysis reveals large conformational differences that mostly reflect inherent molecular features of distinct LpxC orthologs, whereas ligand-induced structural adaptations occur at a smaller scale. These observations highlight the need for a molecular understanding of inherent structural features and conformational plasticity of LpxC enzymes for optimizing LpxC inhibitors as broad-spectrum antibiotics against Gram-negative infections.

Compounds inhibiting LpxC in the lipid A biosynthetic pathway are promising leads for novel antibiotics against multidrug-resistant Gram-negative pathogens. We report the syntheses and structural and biochemical characterizations of LpxC inhibitors based on a diphenyl-diacetylene (1,4-diphenyl-1,3-butadiyne) threonylhydroxamate scaffold. These studies provide a molecular interpretation for the differential antibiotic activities of compounds with a substituted distal phenyl ring as well as the absolute stereochemical requirement at the C2, but not C3, position of the threonyl group.

Hyperpolarized magnetic resonance imaging is a powerful technique enabling real time monitoring of metabolites at concentration levels not accessible by standard MRI techniques. A considerable challenge this technique faces is the T1 decay of the hyperpolarization upon injection into the system under study. Here we show that AnA’nXX’ spin systems such as 13C2-1,2-diphenylacetylene (13C2-DPA) sustain long-lived polarization for both 13C and 1H spins with decay constants of almost 4.5 min at high magnetic fields of up to 16.44 T without spin-locking; the T1 of proton polarization is only 3.8 s. Therefore, storage of the proton polarization in a 13C2-singlet state causes a 69 fold extension of the spin lifetime. Notably, this extension is demonstrated with proton-only pulse sequences, which can be readily implemented on standard clinical scanners.

Photocatalysis is intensely employed to remove refractory organic pollutants in water but suffers from low efficiency due to the rapid recombination of photogenerated electrons and holes. Here, carbon-vacancy-modified g-C3N4 (VC-C3N4) is prepared via a handy two-step calcination method and first applied in the photocatalytic removal of bisphenol A (BPA). Compared to pristine g-C3N4, the photocatalytic degradation activity of VC-C3N4 for BPA is largely enhanced, in which the kinetic constant (k) of BPA degradation is 1.65 times that of pristine g-C3N4. The enhanced photocatalytic performance of VC-C3N4 is ascribed to the critical role of carbon vacancies. On the one hand, carbon vacancies serve as the reservoir of photogenerated electrons to inhibit the recombination of photogenerated holes and electrons. On the other hand, carbon vacancies as conversion centers transfer trapped photogenerated electrons to absorbed O2 for the generation of an abundant superoxide radical (•O2 –), which takes a dominant effect in the photocatalytic degradation process.