High-loading Gα ₁₃ -binding EXE peptide nanoparticles prevent thrombosis and protect mice from cardiac ischemia/reperfusion injury

Abstract

Inefficient delivery is a major obstacle to the development of peptide-based drugs targeting the intracellular compartment. We recently showed that selectively inhibiting integrin outside-in signaling using a peptide (mP6) derived from the Gα13-binding ExE motif within the integrin β3 cytoplasmic domain had antithrombotic effects. Here, we engineered lipid-stabilized, high-loading peptide nanoparticles (HLPN), in which a redesigned ExE peptide (M3mP6) constituted up to 70% of the total nanoparticle molarity, allowing efficient in vivo delivery. We observed that M3mP6 HLPN inhibited occlusive thrombosis more potently than a clopidogrel/aspirin combination without adverse effects on hemostasis in rodents. Furthermore, M3mP6 HLPN synergized with P2Y12 receptor inhibitors or the clopidogrel/aspirin combination in preventing thrombosis, without exacerbating hemorrhage. M3mP6 HLPN also inhibited intravascular coagulation more potently than the P2Y12 inhibitor cangrelor. Postischemia injection of M3mP6 HLPN protected the heart from myocardial ischemia–reperfusion injury in a mouse model. This study demonstrates an efficient in vivo peptide delivery strategy for a therapeutic that not only efficaciously prevented thrombosis with minimal bleeding risk but also protected from myocardial ischemia–reperfusion injury in mice.

Targeting the RT loop of Src SH3 in Platelets Prevents Thrombosis without Compromising Hemostasis

“…FeCl 3 ‐induced carotid artery thrombosis model is therefore generally used to test the occlusive thrombosis. [ 7 , 13 , 14 , 48 ]…”

“…[ 29 , 30 , 31 ] As a matter of fact, in in vivo thrombosis models employed in this study, the effect of these antiplatelet drugs at conventional doses was shown to be far below their maximal antithrombotic potential. Aspirin and clopidogrel at low or maintenance doses (4.3 [ 48 ] or 1.25 mg kg −1 , respectively) scarcely affected the occlusive thrombus formation with slightly prolong bleeding time and substantial antithrombotic effect was seen at extremely high doses with severe bleeding (Figure 5a,b ). As for integrilin, a 0.18 mg kg −1 bolus [ 28 ] was designed taking advantage of its rapid onset of action and short plasma half‐life [ 49 ] which is commonly used with benefit especially in patients undergoing PCI.…”

“…FeCl 3 -induced carotid artery thrombosis model is therefore generally used to test the occlusive thrombosis. [7,13,14,48] The mainstay of treatment for thrombosis is antiplatelet therapy. Conventional antiplatelet drugs such as aspirin, clopidogrel and eptifibatide (integrilin) were clinically applied at doses by balancing antithrombotic benefit and bleeding risk which were supposed to be insufficient in terms of platelet inhibition.…”

Gα12 and Gα13: Versatility in Physiology and Pathology

“…Receptor-interacting protein kinase 3 participates in the integrin-Gα 13 signal to promote platelet activation and thrombosis ( Zhang et al, 2017 ). The newly developed high-load ExE peptide nanoparticles, based on the Gα 13 binding ExE motif on the cytoplasmic domain of integrin β 3 , inhibit thrombosis and protect mice from cardiac I/R injury ( Pang et al, 2020 ). This study supports that the integrin-Gα 13 -RhoA-YAP pathway can be targeted for anti-atherosclerosis therapy ( Wang et al, 2016 ).…”

Knowledge-Based Design of 5-Fluororacil Prodrug Liposomal Formulation: Molecular Packing and Interaction Revealed by Interfacial Isotherms and X-ray Scattering Techniques

“…Prodrug and nanoformulations are two effective strategies to prolong drug release, reduce toxicity, and improve bioavailability. Prodrug modification is a chemical approach by either covalently attaching the hydrophilic drug (i.e., peptides) with an alkyl chain to increase drug lipophilicity for efficient passive membrane permeation , or conjugating the drug with a hydrophilic molecule for higher solubility and therefore higher bioavailability (i.e., PEGylation and albumin conjugation with hydrophobic drugs). , Nanoformulation is a physical approach to shift the pharmacokinetic properties of the drug by encapsulating or incorporating the therapeutic compounds with polymers or lipids to form nanostructures. Nanoparticles may promote passive targeting due to the enhanced permeation and retention (EPR) effect and offer additional advantages for active targeting by surface modification.…”