Synthesis and biochemical evaluation of cephalosporin analogues equipped with chemical tethers.

Lisa M Miller,Ivan Gyulev,Thomas F Krauss,Anne-Kathrin Duhme-Klair,Reyme Herman,Gavin H Thomas

doi:10.1039/d0ra04893c

Lisa M Miller, Ivan Gyulev + Show 4 more

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https://doi.org/10.1039/d0ra04893c

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Abstract

Molecular probes typically require structural modifications to allow for the immobilisation or bioconjugation with a desired substrate but the effects of these changes are often not evaluated. Here, we set out to determine the effects of attaching functional handles to a first-generation cephalosporin. A series of cephalexin derivatives was prepared, equipped with chemical tethers suitable for the site-selective conjugation of antibiotics to functionalised surfaces. The tethers were positioned remotely from the β-lactam ring to ensure minimal effect to the antibiotic's pharmacophore. Herein, the activity of the modified antibiotics was evaluated for binding to the therapeutic target, the penicillin binding proteins, and shown to maintain binding interactions. In addition, the deactivation of the modified drugs by four β-lactamases (TEM-1, CTX-M-15, AmpC, NDM-1) was investigated and the effect of the tethers on the catalytic efficiencies determined. CTX-M-15 was found to favour hydrolysis of the parent antibiotic without a tether, whereas AmpC and NDM-1 were found to favour the modified analogues. Furthermore, the antimicrobial activity of the derivatives was evaluated to investigate the effect of the structural modifications on the antimicrobial activity of the parent drug, cephalexin.

Highlights

The controlled functionalisation of surfaces is imperative for the preparation of functional materials
To a solution of (6R,7R)-7-[(2R)-2-(5-{[(tert-butoxy)carbonyl] amino}pentanamido)-2-phenylacetamido]-3-methyl-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid [23] (100 mg, 0.183 mmol, 1 equiv.) in DCM (4 mL, 0.04 M) was added and 0.1 mL triethyl silane followed by 0.5 mL TFA
The residue was azeotroped with DCM (3 Â 10 mL) to afford a cream solid. This solid was triturated with diethyl ether to afford (6R,7R)-7-[(2R)2-(5-aminopentanamido)-2-phenylacetamido]-3-methyl-8-oxo5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid [10] (77 mg, 94%) as a cream solid. 1H NMR (400 MHz, DMSO-d6) d 9.31 (d, J 1⁄4 8.4 Hz, 1H), 8.57 (d, J 1⁄4 8.2 Hz, 1H), 7.67, 7.47–7.41 (m, 2H), 7.35–7.26 (m, 3H), 5.69 (d, J 1⁄4 8.2 Hz, 1H), 5.62, 4.96 (d, J 1⁄4 4.7 Hz, 1H), 3.47 (d, J 1⁄4 18.8 Hz, 1H), 3.28 (d, J 1⁄4 18.2 Hz, 1H), 2.77, 2.31–2.22 (m, 2H), 1.98 (s, 3H), 1.59–1.48 (m, 4H). 13C NMR (101 MHz, DMSO-d6) d 171.6, 170.9, 164.1, 163.5, 138.3, 129.6, 128.2, 127.6, 127.1, 122.7, 58.4, 57.2, 55.5, 38.7, 34.1, 28.9, 26.7, 22.1, 19.4

Summary

Introduction

The controlled functionalisation of surfaces is imperative for the preparation of functional materials. The organic layer was collected and concentrated to a cream solid, which was the puri ed by trituration from diethyl ether to afford (6R,7R)-3-methyl-8-oxo-7-[(2R)-2-(pent-4-ynamido)2-phenylacetamido]-5-thia-1-azabicyclo[4.2.0] oct-2-ene-2-carboxylic acid [4] (277 mg, 70% yield) as a white solid. The reaction mixture was concentrated under reduced pressure and triturated using 5% DCM/ diethyl ether to afford (6R,7R)-7-[(2R)-2-[5-(1,2-dithiolan-3-yl) pentanamido]-2-phenylacetamido]3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid [5] (190 mg, 55%) as a white solid.

Results

Conclusion