Inhibition of Arterial Thrombus Formation by Blocking Exposed Collagen Surface Using LWWNSYY-Poly(l-Glutamic Acid) Nanoconjugate

Abstract

Exposed collagen surface on diseased blood vessel wall is a trigger of platelet adhesion and subsequent thrombus formation, which is associated with many serious diseases such as myocardial infarction and stroke. Various antithrombotic agents have been developed, but are usually targeted on blood components such as platelet, which suffered from the risk of bleeding due to interference with hemostasis. In contrast, blocking the exposed collagen surface would prevent thrombus formation without the risk of bleeding. In the present study, an antithrombotic nanoconjugate (LWWNSYY-poly glutamic acid, L7-PGA) targeting collagen surface was designed by immobilizing heptapeptide LWWNSYY, a biomimetic inhibitor designed in our previous work, on poly(L-glutamic acid). Successful binding of L7-PGA on the collagen surface was confirmed by a negative ΔG of −5.99 ± 0.26 kcal/mol. L7-PGA was found to effectively inhibit platelet adhesion on the collagen surface, with a reduced IC 50 of only 1/5 of that of free LWWNSYY. The inhibition of thrombus formation by L7-PGA was also validated in vivo by a reduction of 31.2% in the weight of thrombus. These results highlight L7-PGA as an effective inhibitor of arterial thrombus formation via blocking exposed collagen surface, which would be helpful for the development of novel antithrombotic nanomedicine.

Development of antithrombotic peptides based on the molecular interactions between von Willebrand factor and GPIbα

“…1). 46 Briefly, microtiter HB 96-well plates (Costar, Corning Inc., USA) were coated with 100 mL of I7, I16 and I17 (100 mg mL À1 ) dissolved in 0.05 M CBS (15 mM Na 2 CO 3 and 35 mM NaHCO 3 ) for 12 h at 4 1C, and blanks were coated with 100 mL of CBS. Reagents without adhesion were discarded, and the wells were washed with 300 mL of Wash Buffer (0.02 M PBS containing 0.05% Tween-20, pH 7.4) three times.…”

“…The collagen peptide was dissolved in PBS at a concentration of 0.1 mM, 46 where 1 L of PBS contains 8 g of NaCl, 0.2 g of KCl, 1.44 g of Na 2 HPO 4 and 0.24 g of KH 2 PO 4 (pH7.2). I7 and I17 were dissolved in 900 mL of PRP at a concentration of 0.3 mM.…”

“…As reported in our previous work, the water solubility of an inhibitor is very important for its actual use. 46,55 The fluorescence emission spectra of 200-500 nm were scanned at an excitation wavelength of 280 nm for different concentrations of inhibitors to evaluate the water solubility of the inhibitor, as shown in Fig. S4 (ESI †).…”

“…This strategy was established and successfully used to examine the protein interface in various systems including a collagen-GPIba complex. [41][42][43][44][45][46] Inhibitors targeting the vWF A1 domain were then designed based on the molecular interactions, followed by the screening and experimental validations.…”

Biting Innovations of Mosquito-Based Biomaterials and Medical Devices

“…Existing methods to address the detrimental events associated with blood-biomaterial interaction include the use of polymeric and non-polymeric coatings that either elute or incorporate grafted anticoagulant agents [58,59]. One of the most common therapeutic agents that is administered intravenously to prevent clot formation is heparin [60].…”