Landry DW

References (9)

Title : Efficient Cocaine Degradation by Cocaine Esterase-Loaded Red Blood Cells - Rossi_2020_Front.Physiol_11_573492
Author(s) : Rossi L , Pierige F , Agostini M , Bigini N , Termopoli V , Cai Y , Zheng F , Zhan CG , Landry DW , Magnani M
Ref : Front Physiol , 11 :573492 , 2020
Abstract : Recombinant bacterial cocaine esterase (CocE) represents a potential protein therapeutic for cocaine use disorder treatment. Unfortunately, the native enzyme was highly unstable and the corresponding mutagenized derivatives, RBP-8000 and E196-301, although improving in vitro thermo-stability and in vivo half-life, were a partial solution to the problem. For cocaine use disorder treatment, an efficient cocaine-metabolizing enzyme with a longer residence time in circulation would be needed. We investigated in vitro the possibility of developing red blood cells (RBCs) loaded with RBP-8000 and E196-301 as a biocompatible system to metabolize cocaine for a longer period of time. RBP 8000 stability within human RBCs is limited (approximately 50% residual activity after 1 h at 37degC) and not different as for the free enzyme, while both free and encapsulated E196-301 showed a greater thermo-stability. By reducing cellular glutathione content during the loading procedure, in order to preserve the disulfide bonds opportunely created to stabilize the enzyme dimer structure, it was possible to produce an encapsulated protein maintaining 100% stability at least after 4 h at 37degC. Moreover, E196-301-loaded RBCs were efficiently able to degrade cocaine in a time- and concentration-dependent manner. The same stability results were obtained when murine RBCs were used paving the way to preclinical investigations. Thus, our in vitro data show that E196-301-loaded RBCs could act as efficient bioreactors in degrading cocaine to non-toxic metabolites to be possibly considered in substance-use disorder treatments. This approach should now be investigated in a preclinical model of cocaine use disorder to evaluate if further protein modifications are needed to further improve long term enzyme stability.
ESTHER : Rossi_2020_Front.Physiol_11_573492
PubMedSearch : Rossi_2020_Front.Physiol_11_573492
PubMedID: 33013487

Title : New therapeutic approaches and novel alternatives for organophosphate toxicity - Katz_2018_Toxicol.Lett_291_1
Author(s) : Katz FS , Pecic S , Schneider L , Zhu Z , Hastings A , Luzac M , MacDonald J , Landry DW , Stojanovic MN
Ref : Toxicol Lett , 291 :1 , 2018
Abstract : Organophosphate compounds (OPCs) are commonly used as pesticides and were developed as nerve agents for chemical warfare. Exposure to OPCs results in toxicity due to their covalent binding and inhibition of acetylcholinesterase (AChE). Treatment for toxicity due to OPC exposure has been largely focused on the reactivation of AChE by oxime-based compounds via direct nucleophilic attack on the phosphorous center. However, due to the disadvantages to existing oxime-based reactivators for treatment of OPC poisoning, we considered non-oxime mechanisms of reactivation. A high throughput screen of compound libraries was performed to discover previously unidentified reactivation compounds, followed by studies on their analogs. In the process, we discovered multiple non-oxime classes of compounds, the most robust of which we have already reported [1]. Herein, we report other classes of compounds we identified in our screen that are efficient at reactivation. During biochemical characterization, we also found some compounds with other activities that may inspire novel therapeutic approaches to OPC toxicity. Specifically, we found compounds that [1] increase the rate of substrate hydrolysis by AChE and, [2] protect the enzyme from inhibition by OPC. Further, we discovered that a subset of reactivator compounds recover activity from both AChE and the related enzyme butyrylcholinesterase (BuChE). We now report these compounds, their activities and discuss how each relates to therapeutic approaches that would provide alternatives to traditional oxime-based reactivation.
ESTHER : Katz_2018_Toxicol.Lett_291_1
PubMedSearch : Katz_2018_Toxicol.Lett_291_1
PubMedID: 29614332

Title : Cover Picture: Discovery of New Classes of Compounds that Reactivate Acetylcholinesterase Inhibited by Organophosphates (ChemBioChem 15\/2015) - Katz_2015_Chembiochem_16_2113
Author(s) : Katz FS , Pecic S , Tran TH , Trakht I , Schneider L , Zhu Z , Ton-That L , Luzac M , Zlatanic V , Damera S , MacDonald J , Landry DW , Tong L , Stojanovic MN
Ref : Chembiochem , 16 :2113 , 2015
Abstract : The cover picture shows that a mouse exposed to otherwise lethal doses of an organophosphorous agent survives if it is treated with compounds that we recently identified as reactivators of organopsphate-inhibited acetylcholinesterase (AChE). We have demonstrated that survival correlates with reactivation of AChE activity across a panel of tissues, including brain tissues across the blood-brain barrier. We have also determined the co-crystal of one of these reactivators bound to organophosphate-inhibited AChE and found that it binds in a unique manner to the enzyme, forming a dimer within the active site. These compounds were also efficient at reactivating the human form of AChE, thus indicating that their protective effects could be translated for human use. Molecular graphics for the cover were performed with the UCSF Chimera package. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIGMS P41-GM103311).
ESTHER : Katz_2015_Chembiochem_16_2113
PubMedSearch : Katz_2015_Chembiochem_16_2113
PubMedID: 26444304

Title : Discovery of New Classes of Compounds that Reactivate Acetylcholinesterase Inhibited by Organophosphates - Katz_2015_Chembiochem_16_2205
Author(s) : Katz FS , Pecic S , Tran TH , Trakht I , Schneider L , Zhu Z , Ton-That L , Luzac M , Zlatanic V , Damera S , MacDonald J , Landry DW , Tong L , Stojanovic MN
Ref : Chembiochem , 16 :2205 , 2015
Abstract : Acetylcholinesterase (AChE) that has been covalently inhibited by organophosphate compounds (OPCs), such as nerve agents and pesticides, has traditionally been reactivated by using nucleophilic oximes. There is, however, a clearly recognized need for new classes of compounds with the ability to reactivate inhibited AChE with improved in vivo efficacy. Here we describe our discovery of new functional groups-Mannich phenols and general bases-that are capable of reactivating OPC-inhibited AChE more efficiently than standard oximes and we describe the cooperative mechanism by which these functionalities are delivered to the active site. These discoveries, supported by preliminary in vivo results and crystallographic data, significantly broaden the available approaches for reactivation of AChE.
ESTHER : Katz_2015_Chembiochem_16_2205
PubMedSearch : Katz_2015_Chembiochem_16_2205
PubMedID: 26350723
Gene_locus related to this paper: mouse-ACHE

Title : Design, synthesis and evaluation of non-urea inhibitors of soluble epoxide hydrolase - Pecic_2012_Bioorg.Med.Chem.Lett_22_601
Author(s) : Pecic S , Deng SX , Morisseau C , Hammock BD , Landry DW
Ref : Bioorganic & Medicinal Chemistry Lett , 22 :601 , 2012
Abstract : Inhibition of soluble epoxide hydrolase (sEH) has been proposed as a new pharmaceutical approach for treating hypertension and vascular inflammation. The most potent sEH inhibitors reported in literature to date are urea derivatives. However, these compounds have limited pharmacokinetic profiles. We investigated non-urea amide derivatives as sEH inhibitors and identified a potent human sEH inhibitor 14-34 having potency comparable to urea-based inhibitors.
ESTHER : Pecic_2012_Bioorg.Med.Chem.Lett_22_601
PubMedSearch : Pecic_2012_Bioorg.Med.Chem.Lett_22_601
PubMedID: 22079754

Title : Thermostable variants of cocaine esterase for long-time protection against cocaine toxicity - Gao_2009_Mol.Pharmacol_75_318
Author(s) : Gao D , Narasimhan DL , MacDonald J , Brim R , Ko MC , Landry DW , Woods JH , Sunahara RK , Zhan CG
Ref : Molecular Pharmacology , 75 :318 , 2009
Abstract : Enhancing cocaine metabolism by administration of cocaine esterase (CocE) has been recognized as a promising treatment strategy for cocaine overdose and addiction, because CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine yet identified. A major obstacle to the clinical application of CocE is the thermoinstability of native CocE with a half-life of only a few minutes at physiological temperature (37 degrees C). Here we report thermostable variants of CocE developed through rational design using a novel computational approach followed by in vitro and in vivo studies. This integrated computational-experimental effort has yielded a CocE variant with a approximately 30-fold increase in plasma half-life both in vitro and in vivo. The novel design strategy can be used to develop thermostable mutants of any protein.
ESTHER : Gao_2009_Mol.Pharmacol_75_318
PubMedSearch : Gao_2009_Mol.Pharmacol_75_318
PubMedID: 18987161

Title : First-principle studies of intermolecular and intramolecular catalysis of protonated cocaine - Zhan_2005_J.Comput.Chem_26_980
Author(s) : Zhan CG , Deng SX , Skiba JG , Hayes BA , Tschampel SM , Shields GC , Landry DW
Ref : J Comput Chem , 26 :980 , 2005
Abstract : We have performed a series of first-principles electronic structure calculations to examine the reaction pathways and the corresponding free energy barriers for the ester hydrolysis of protonated cocaine in its chair and boat conformations. The calculated free energy barriers for the benzoyl ester hydrolysis of protonated chair cocaine are close to the corresponding barriers calculated for the benzoyl ester hydrolysis of neutral cocaine. However, the free energy barrier calculated for the methyl ester hydrolysis of protonated cocaine in its chair conformation is significantly lower than for the methyl ester hydrolysis of neutral cocaine and for the dominant pathway of the benzoyl ester hydrolysis of protonated cocaine. The significant decrease of the free energy barrier, approximately 4 kcal/mol, is attributed to the intramolecular acid catalysis of the methyl ester hydrolysis of protonated cocaine, because the transition state structure is stabilized by the strong hydrogen bond between the carbonyl oxygen of the methyl ester moiety and the protonated tropane N. The relative magnitudes of the free energy barriers calculated for different pathways of the ester hydrolysis of protonated chair cocaine are consistent with the experimental kinetic data for cocaine hydrolysis under physiologic conditions. Similar intramolecular acid catalysis also occurs for the benzoyl ester hydrolysis of (protonated) boat cocaine in the physiologic condition, although the contribution of the intramolecular hydrogen bonding to transition state stabilization is negligible. Nonetheless, the predictability of the intramolecular hydrogen bonding could be useful in generating antibody-based catalysts that recruit cocaine to the boat conformation and an analog that elicited antibodies to approximate the protonated tropane N and the benzoyl O more closely than the natural boat conformer might increase the contribution from hydrogen bonding. Such a stable analog of the transition state for intramolecular catalysis of cocaine benzoyl-ester hydrolysis was synthesized and used to successfully elicit a number of anticocaine catalytic antibodies.
ESTHER : Zhan_2005_J.Comput.Chem_26_980
PubMedSearch : Zhan_2005_J.Comput.Chem_26_980
PubMedID: 15880781

Title : Crystallographic and biochemical analysis of cocaine-degrading antibody 15A10 - Larsen_2004_Biochemistry_43_8067
Author(s) : Larsen NA , de Prada P , Deng SX , Mittal A , Braskett M , Zhu X , Wilson IA , Landry DW
Ref : Biochemistry , 43 :8067 , 2004
Abstract : Catalytic antibody 15A10 hydrolyzes the benzoyl ester of cocaine to form the nonpsychoactive metabolites benzoic acid and ecgonine methylester. Here, we report biochemical and structural studies that characterize the catalytic mechanism. The crystal structure of the cocaine-hydrolyzing monoclonal antibody (mAb) 15A10 has been determined at 2.35 A resolution. The binding pocket is fairly shallow and mainly hydrophobic but with a cluster of three hydrogen-bond donating residues (TrpL96, AsnH33, and TyrH35). Computational docking of the transition state analogue (TSA) indicates that these residues are appropriately positioned to coordinate the phosphonate moiety of the TSA and, hence, form an oxyanion hole. Tyrosine modification of the antibody with tetranitromethane reduced hydrolytic activity to background level. The contribution from these and other residues to catalysis and TSA binding was explored by site-directed mutagenesis of 15A10 expressed in a single chain fragment variable (scFv) format. The TyrH35Phe mutant had 4-fold reduced activity, and TrpL96Ala, TrpL96His, and AsnH33Ala mutants were all inactive. Comparison with an esterolytic antibody D2.3 revealed a similar arrangement of tryptophan, asparagine, and tyrosine residues in the oxyanion hole that stabilizes the transition state for ester hydrolysis. Furthermore, the crystal structure of the bacterial cocaine esterase (cocE) also showed that the cocE employs a tyrosine hydroxyl in the oxyanion hole. Thus, the biochemical and structural data are consistent with the catalytic antibody providing oxyanion stabilization as its major contribution to catalysis.
ESTHER : Larsen_2004_Biochemistry_43_8067
PubMedSearch : Larsen_2004_Biochemistry_43_8067
PubMedID: 15209502

Title : Fundamental reaction mechanism for cocaine hydrolysis in human butyrylcholinesterase - Zhan_2003_J.Am.Chem.Soc_125_2462
Author(s) : Zhan CG , Zheng F , Landry DW
Ref : Journal of the American Chemical Society , 125 :2462 , 2003
Abstract : Butyrylcholinesterase (BChE)-cocaine binding and the fundamental pathway for BChE-catalyzed hydrolysis of cocaine have been studied by molecular modeling, molecular dynamics (MD) simulations, and ab initio calculations. Modeling and simulations indicate that the structures of the prereactive BChE/substrate complexes for (-)-cocaine and (+)-cocaine are all similar to that of the corresponding prereactive BChE/butyrylcholine (BCh) complex. The overall binding of BChE with (-)-cocaine and (+)-cocaine is also similar to that proposed with butyrylthiocholine and succinyldithiocholine, i.e., (-)- or (+)-cocaine first slides down the substrate-binding gorge to bind to Trp-82 and stands vertically in the gorge between Asp-70 and Trp-82 (nonprereactive complex) and then rotates to a position in the catalytic site within a favorable distance for nucleophilic attack and hydrolysis by Ser-198 (prereactive complex). In the prereactive complex, cocaine lies horizontally at the bottom of the gorge. The fundamental catalytic hydrolysis pathway, consisting of acylation and deacylation stages similar to those for ester hydrolysis by other serine hydrolases, was proposed on the basis of the simulated prereactive complex and confirmed theoretically by ab initio reaction coordinate calculations. Both the acylation and deacylation follow a double-proton-transfer mechanism. The calculated energetic results show that within the chemical reaction process the highest energy barrier and Gibbs free energy barrier are all associated with the first step of deacylation. The calculated ratio of the rate constant (k(cat)) for the catalytic hydrolysis to that (k(0)) for the spontaneous hydrolysis is approximately 9.0 x 10(7). The estimated k(cat)/k(0) value of approximately 9.0 x 10(7) is in excellent agreement with the experimentally derived k(cat)/k(0) value of approximately 7.2 x 10(7) for (+)-cocaine, whereas it is approximately 2000 times larger than the experimentally derived k(cat)/k(0) value of approximately 4.4 x 10(4) for (-)-cocaine. All of the results suggest that the rate-determining step of the BChE-catalyzed hydrolysis of (+)-cocaine is the first step of deacylation, whereas for (-)-cocaine the change from the nonprereactive complex to the prereactive complex is rate-determining and has a Gibbs free energy barrier higher than that for the first step of deacylation by approximately 4 kcal/mol. A further analysis of the structural changes from the nonprereactive complex to the prereactive complex reveals specific amino acid residues hindering the structural changes, providing initial clues for the rational design of BChE mutants with improved catalytic activity for (-)-cocaine.
ESTHER : Zhan_2003_J.Am.Chem.Soc_125_2462
PubMedSearch : Zhan_2003_J.Am.Chem.Soc_125_2462
PubMedID: 12603134