Design of an allosterically modulated doxycycline and doxorubicin drug-binding protein

Abstract

The allosteric interplay between distant functional sites present in a single protein provides for one of the most important regulatory mechanisms in biological systems. While the design of ligand-binding sites into proteins remains challenging, this holds even truer for the coupling of a newly engineered binding site to an allosteric mechanism that regulates the ligand affinity. Here it is shown how computational design algorithms enabled the introduction of doxycycline- and doxorubicin-binding sites into the serine proteinase inhibitor (serpin) family member α1-antichymotrypsin. Further engineering allowed exploitation of the proteinase-triggered serpin-typical S-to-R transition to modulate the ligand affinities. These design variants follow strategies observed in naturally occurring plasma globulins that allow for the targeted delivery of hormones in the blood. By analogy, we propose that the variants described in the present study could be further developed to allow for the delivery of the antibiotic doxycycline and the anticancer compound doxorubicin to tissues/locations that express specific proteinases, such as bacterial infection sites or tumor cells secreting matrix metalloproteinases.

Pharmacokinetic/Pharmacodynamic Integration of Doxycycline Against Mycoplasma hyopneumoniae in an In Vitro Model

“…First, delayed growth of M. hyopneumoniae was observed, and M. hyopneumoniae did not recover instantaneously in the logarithmic growth phase upon transfer to the new culture environment. Second, the antibacterial mechanism of doxycycline is to inhibit protein synthesis, and it acted as a long-acting bacteriostatic agent (Schmidt et al, 2018; Chukwudi and Good, 2019). Third, due to the slow growth of M. hyopneumoniae , bacteriostatic agents (e.g., doxycycline) need longer to elicit their effects.…”

Doxorubicin loaded ferritin nanoparticles for ferroptosis enhanced targeted killing of cancer cells

“…[15][16][17] To reduce the side effects of DOX in vivo, various formulations of DOX have been investigated in clinical or preclinical elds including encapsulated in nanomaterial, such as liposome, protein and PLGA nanoparticles. [18][19][20] Nanomaterials have shown to great potential in encapsulating and transporting drugs, penetrating cell membranes and releasing drugs in tumor cells through the enhanced permeability and retention (EPR) effect, 21 which augment the accumulation of drugs in tumor cells but reduce that in normal cells. 22 Ferritin (Fn), a universal intracellular protein that stores iron and releases it in a controlled fashion, was designed to encapsulate DOX for targeting delivery drugs to transferrin receptor 1 (TFR1) overexpressed tumor cells, 23,24 human leukemia (HL-60), human colorectal carcinoma (Lovo), human breast adenocarcinoma (MDA-MB-468), human liver carcinoma (Hep G2), human cervical adenocarcinoma (HeLa), human breast (MCF-7), human small cell lung carcinoma (NCI-H69).…”

Biological Profile of Two Gentiana lutea L. Metabolites Using Computational Approaches and In Vitro Tests

“…Moreover, the binding pocket contains many hydrophobic residues, which are ideal for accommodating a molecule such as doxycycline. Binding mode of doxycycline with alpha-1-antichymotrypsin (5OM2) [41]. Pink arrows are hydrogen bonds, the green line represents a π-π interaction, and the red-to-blue line represents an ionic bridge.…”

“…Figure 2.Binding mode of doxycycline with alpha-1-antichymotrypsin (5OM2)[41]. Pink arrows are hydrogen bonds, the green line represents a π-π interaction, and the red-to-blue line represents an ionic bridge.…”