Structure–Activity Relationships for the N-Me- Versus N-H-Amide Modification to Macrocyclic ent-Verticilide Antiarrhythmics

Smith, Abigail N.; Thorpe, Madelaine P.; Blackwell, Daniel J.; Batiste, Suzanne M.; Hopkins, Corey R.; Schley, Nathan D.; Knollmann, Björn C.; Johnston, Jeffrey N.

doi:10.1021/acsmedchemlett.2c00377

Cited by 6 publications

(5 citation statements)

References 26 publications

(35 reference statements)

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“…Figure summarizes these findings; no inhibition of RyR2 (lack of activity) is reflected as a value of 1 (dotted line). The relative activity of Series 1 compounds was reported previously, finding that introduction of one or more N –H amide(s) to the structure leads to loss of measurable activity . In that study, we noted that the 18-membered oligomer was more forgiving.…”

Section: Resultsmentioning

confidence: 99%

“…The relative activity of Series 1 compounds was reported previously, finding that introduction of one or more N−H amide(s) to the structure leads to loss of measurable activity. 35 In that study, we noted that the 18-membered oligomer was more forgiving. Returning to the 24-membered ring oligomers studied here, Series 2 showed a significant trend, with introduction of a single N−H amide as a replacement for an ester (2.1) producing an analog with modest potency.…”

Section: Final Steps Of Analog Synthesis (Ho)mentioning

confidence: 99%

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Backbone-Determined Antiarrhythmic Structure–Activity Relationships for a Mirror Image, Oligomeric Depsipeptide Natural Product

Thorpe,

Blackwell,

Knollmann

et al. 2024

J. Med. Chem.

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Cyclic oligomeric depsipeptides (COD) are a structural class within naturally occurring compounds with a wide range of biological activity. Verticilide is a COD (24-membered ring) that was identified by its inhibition of insect ryanodine receptor (RyR). We have since found that the enantiomer of verticilide (entverticilide, 1) is a potent inhibitor of mammalian RyR2, a cardiac calcium channel, and therefore a potential antiarrhythmic agent. Oddly, nat-verticilide does not inhibit RyR2. To further develop entverticilide as an antiarrhythmic, we explored potential SAR through systematic modification of the ester's functionality to both N−H and N−Me amides. The syntheses of these ent-verticilide-inspired analogs are detailed using a monomer-based platform enabled by enantioselective catalysis. Two analogs among 23 exhibited measurable reduction of calcium sparks in a functional assay of RyR2 activity. These findings illustrate the value of natural product-inspired therapeutic development, but the less-studied approach where the non-natural enantiomeric series harbors important SAR.

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Section: Resultsmentioning

confidence: 99%

Section: Final Steps Of Analog Synthesis (Ho)mentioning

confidence: 99%

Backbone-Determined Antiarrhythmic Structure–Activity Relationships for a Mirror Image, Oligomeric Depsipeptide Natural Product

Thorpe,

Blackwell,

Knollmann

et al. 2024

J. Med. Chem.

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“…The synthesis of AF568-labeled ent -verticilide (Figure , analog 3 ) was planned from an alkyne-modified ent -verticilide (Figure , analog 2 ), providing a modest structural change that was not expected to affect activity against RyR2 . Two syntheses of ent -verticilide have been reported by us, ,− but the oligomeric nature of 1 simplifies the didepsipeptide monomer synthesis and assembly.…”

Section: Resultsmentioning

confidence: 99%

RyR2 Binding of an Antiarrhythmic Cyclic Depsipeptide Mapped Using Confocal Fluorescence Lifetime Detection of FRET

Šeflová,

Schwarz,

Smith

et al. 2023

ACS Chem. Biol.

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Hyperactivity of cardiac sarcoplasmic reticulum (SR) ryanodine receptor (RyR2) Ca 2+ -release channels contributes to heart failure and arrhythmias. Reducing the RyR2 activity, particularly during cardiac relaxation (diastole), is a desirable therapeutic goal. We previously reported that the unnatural enantiomer (ent) of an insect-RyR activator, verticilide, inhibits porcine and mouse RyR2 at diastolic (nanomolar) Ca 2+ and has in vivo efficacy against atrial and ventricular arrhythmia. To determine the ent-verticilide structural mode of action on RyR2 and guide its further development via medicinal chemistry structure−activity relationship studies, here, we used fluorescence lifetime (FLT)measurements of Forster resonance energy transfer (FRET) in HEK293 cells expressing human RyR2. For these studies, we used an RyR-specific FRET molecular-toolkit and computational methods for trilateration (i.e., using distances to locate a point of interest). Multiexponential analysis of FLT-FRET measurements between four donor-labeled FKBP12.6 variants and acceptorlabeled ent-verticilide yielded distance relationships placing the acceptor probe at two candidate loci within the RyR2 cryo-EM map. One locus is within the Ry12 domain (at the corner periphery of the RyR2 tetrameric complex). The other locus is sandwiched at the interface between helical domain 1 and the SPRY3 domain. These findings document RyR2-target engagement by ent-verticilide, reveal new insight into the mechanism of action of this new class of RyR2-targeting drug candidate, and can serve as input in future computational determinations of the ent-verticilide binding site on RyR2 that will inform structure−activity studies for lead optimization.

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“…2 analog 3) was planned from an alkynemodified ent-verticilide (Fig. 2, analog 2), providing a modest structural change that was not expected to affect activity against RyR2 (22). Two syntheses of ent-verticilide have been reported by us (7,(23)(24)(25)(26) but the oligomeric nature of 1 simplifies the didepsipeptide monomer synthesis and assembly.…”

Section: Resultsmentioning

confidence: 99%

RyR2 Binding of an Antiarrhythmic Cyclic Depsipeptide Mapped Using Confocal Fluorescence Lifetime Detection of FRET

Šeflová

Schwarz

Smith

et al. 2023

Preprint

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Hyperactivity of cardiac sarcoplasmic reticulum (SR) ryanodine receptor (RyR2) Ca2+-release channels contributes to heart failure and arrhythmias. Reducing RyR2 activity, particularly during cardiac relaxation (diastole), is a desirable therapeutic goal. We previously reported that the unnatural enantiomer (ent) of an insect-RyR activator, verticilide, inhibits porcine and mouse RyR2 at diastolic (nanomolar) Ca2+and has in vivo efficacy against atrial and ventricular arrhythmia. To determine theent-verticilide structural mode of action on RyR2 and guide its further development via medicinal chemistry structure-activity relationship studies, here we used fluorescence lifetime (FLT)-measurements of Förster resonance energy transfer (FRET) in HEK293 cells expressing human RyR2. For these studies, we used an RyR-specific FRET molecular-toolkit and computational methods for trilateration (i.e., using distances to locate a point of interest). Multi-exponential analysis of FLT-FRET measurements between four donor-labeled FKBP12.6 variants and acceptor-labeledent-verticilide, yielded distance relationships placing the acceptor probe at two candidate loci within the RyR2 cryo-EM map. One locus is within the Ry12 domain (at the corner periphery of the RyR2 tetrameric complex). The other locus is sandwiched at the interface between helical domain 1 and the SPRY3 domain. These findings document RyR2-target engagement byent-verticilide, reveal new insight into the mechanism of action of this new class of RyR2-targeting drug candidate, and can serve as input in future computational determinations of theent-verticilide binding site on RyR2 that will inform structure-activity studies for lead optimization.

show abstract

Structure–Activity Relationships for the N-Me- Versus N-H-Amide Modification to Macrocyclic ent-Verticilide Antiarrhythmics

Cited by 6 publications

References 26 publications

Backbone-Determined Antiarrhythmic Structure–Activity Relationships for a Mirror Image, Oligomeric Depsipeptide Natural Product

Backbone-Determined Antiarrhythmic Structure–Activity Relationships for a Mirror Image, Oligomeric Depsipeptide Natural Product

RyR2 Binding of an Antiarrhythmic Cyclic Depsipeptide Mapped Using Confocal Fluorescence Lifetime Detection of FRET

RyR2 Binding of an Antiarrhythmic Cyclic Depsipeptide Mapped Using Confocal Fluorescence Lifetime Detection of FRET

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