Targeted thrombolysis by using c-RGD-modified N,N,N-Trimethyl Chitosan nanoparticles loaded with lumbrokinase

Abstract

Lumbrokinase (LK) has strong fibrinolytic and thrombolytic activities, but it has a short half-life, can be easily inactivated, and may cause hemorrhage as a side effect. This study develops a potential thrombolytic therapy by fabricating N,N,N-Trimethyl Chitosan (TMC) nanoparticles modified with the cyclic Arg-Gly-Asp-Phe-Lys peptide (c-RGD) and loaded with LK (i.e. c-RGD-LK-NPs). The binding of c-RGD to platelet membrane GPIIb/IIIa receptors is expected to enable targeted delivery of the c-RGD-conjugated TMC to the thrombus. The synthesized c-RGD-LK-NPs had a mean particle size of 232.0 nm, zeta potential of 19.8 mV, entrapment efficiency of 52.7% ± 2.5%, and loading efficiency of 17.4% ± 0.65%. Transmission electron microscopy showed that they were generally spherical. The c-RGD-LK-NPs gave a cumulative in vitro LK release of 80.6% over 8 h, and the activity of LK was close to 80%, indicating that the nanoparticles protected the activity of LK. In vitro blood clot lysis assays were carried out and in vivo thrombolysis effect was tested in Sprague-Dawley rats carotid artery thrombus model. In all cases, the c-RGD-LK-NPs showed superior performance compared with the free LK and the unmodified TMC nanoparticles loaded with LK. The c-RGD-LK-NPs reagent is expected to be potentially useful in treating thromboembolic diseases.

Nanobiotechnology approaches for cardiovascular diseases: site-specific targeting of drugs and nanoparticles for atherothrombosis

“…Using a cyclic RGD peptide (c-RGD) with better in vivo stability, Liao et al created targeted chitosan NPs that were coated with lumbrokinase to dissolve fibrin-rich thrombi in Sprague–Dawley rats [ 152 ]. This anti-thrombotic drug stimulate adenylate cyclase, increase cAMP level, thereby limiting the increase in intracellular Ca 2+ , and suppressing the expression of GPIIb/IIIa and P-selectin [ 153 ].…”

Potensi Lumbrokinase Dengan Carrier N,n,n-Trimetil Kitosan Klorida (Tmc) Menggunakan Ligand C-RGD Sebagai Agen Fibrinolitik Dan Anti-Platelet Terbarukan

“…[17] Akan tetapi, kitosan hanya dapat larut dalam larutan asam sehingga bermasalah dengan pengangkutan protein obat. [19] N,N,N-Trimetil Kitosan Klorida (TMC) adalah turunan kuartener kitosan yang memiliki kelarutan yang baik dalam rentang pH yang luas.…”

“…c-RGD dapat digunakan sebagai ligand dan hasil penelitian menunjukkan bahwa siklik-RGD berikatan pada platelet aktif lebih banyak dibanding dengan linear-RGD. [19] Tulisan ini akan menjabarkan mengenai kitosan yang sudah dimodifikasi menjadi TMC dan ditambahkan ligand c-RGD sebagai komponen yang meningkatkan potensi dari lumbrokinase bekerja pada reseptor GPIIb/IIIa. Oleh karena itu, agen fibrinolitik ini dapat bekerja lebih spesifik.…”

Nano-Medicine for Thrombosis: A Precise Diagnosis and Treatment Strategy

“…The contrast agent was applied to detect platelet-rich thrombi, and its selectivity to the thrombus area with rapid speed was confirmed. (2) The cyclic Arg-Gly-Asp (cRGD) peptide was reported to target the activated platelets due to the surface-overexpressed GPIIb/IIIa receptor [45]. The cRGD peptide was decorated on the surface with Fe 3 O 4 -poly(lactic-co-glycolic acid) nanoparticles (Fe 3 O 4 @PLGA) [46].…”