Development and Testing of Thrombolytics in Stroke

다국어 초록 (Multilingual Abstract)

Despite recent advances in recanalization therapy, mechanical thrombectomy will never be a treatment for every ischemic stroke because access to mechanical thrombectomy is still limited in many countries. Moreover, many ischemic strokes are caused by occlusion of cerebral arteries that cannot be reached by intra-arterial catheters. Reperfusion using thrombolytic agents will therefore remain an important therapy for hyperacute ischemic stroke. However, thrombolytic drugs have shown limited efficacy and notable hemorrhagic complication rates, leaving room for improvement. A comprehensive understanding of basic and clinical research pipelines as well as the current status of thrombolytic therapy will help facilitate the development of new thrombolytics. Compared with alteplase, an ideal thrombolytic agent is expected to provide faster reperfusion in more patients; prevent re-occlusions; have higher fibrin specificity for selective activation of clot-bound plasminogen to decrease bleeding complications; be retained in the blood for a longer time to minimize dosage and allow administration as a single bolus; be more resistant to inhibitors; and be less antigenic for repetitive usage. Here, we review the currently available thrombolytics, strategies for the development of new clot-dissolving substances, and the assessment of thrombolytic efficacies in vitro and in vivo.

참고문헌 (Reference)

1 Armstead WM, "tPA-S481A prevents neurotoxicity of endogenous tPA in traumatic brain injury" 29 : 1794-1802, 2012

2 Lee SH, "tPA regulates neurite outgrowth by phosphorylation of LRP5/6in neural progenitor cells" 49 : 199-215, 2014

3 Jong S. Kim, "tPA Helpers in the Treatment of Acute Ischemic Stroke: Are They Ready for Clinical Use?" 대한뇌졸중학회 21 (21): 160-174, 2019

4 Niego B, "t-PA-specific modulation of a human blood-brain barrier model involves plasmin-mediated activation of the Rho kinase pathway in astrocytes" 119 : 4752-4761, 2012

5 Lee IJ, "Zebrafish model of photochemical thrombosis for translational research and thrombolytic screening in vivo" 10 : 494-502, 2017

6 Gething MJ, "Variants of human tissue-type plasminogen activator that lack specific structural domains of the heavy chain" 7 : 2731-2740, 1988

7 Kadir RRA, "Urokinase plasminogen activator : a potential thrombolytic agent for ischaemic stroke" 40 : 347-355, 2020

8 Fossum S, "Urokinase and non-urokinase fibrinolytic activity in protease-inhibitor-deprived plasma, assayed by a fibrin micro-plate method" 32 : 119-121, 1996

9 Laing ST, "Ultrasound-enhanced thrombolytic effect of tissue plasminogen activator-loaded echogenic liposomes in an in vivo rabbit aorta thrombus model-brief report" 31 : 1357-1359, 2011

10 Alexandrov AV, "Ultrasound-enhanced systemic thrombolysis for acute ischemic stroke" 351 : 2170-2178, 2004

11 Narita M, "Two receptor systems are involved in the plasma clearance of tissue-type plasminogen activator(t-PA)in vivo" 96 : 1164-1168, 1995

12 Yaghi S, "Treatment and outcome of hemorrhagic transformation after intravenous alteplase in acute ischemic stroke : a scientific statement for healthcare professionals from the American Heart Association/American Stroke Association" 48 : e343-e361, 2017

13 Tseng MT, "Transendothelial migration of ferric ion in FeCl3 injured murine common carotid artery" 118 : 275-280, 2006

14 Collen D, "Tissue-type plasminogen activator : a historical perspective and personal account" 2 : 541-546, 2004

15 Su EJ, "Tissue plasminogen activator-mediated PDGF signaling and neurovascular coupling in stroke" 7 (7): 155-158, 2009

16 Correa F, "Tissue plasminogen activator prevents white matter damage following stroke" 208 : 1229-1242, 2011

17 Pawlak R, "Tissue plasminogen activator in the amygdala is critical for stress-induced anxiety-like behavior" 6 : 168-174, 2003

18 National Institute of Neurological Disorders, "Tissue plasminogen activator for acute ischemic stroke" 333 : 1581-1587, 1995

19 Louessard M, "Tissue plasminogen activator expression is restricted to subsets of excitatory pyramidal glutamatergic neurons" 53 : 5000-5012, 2016

20 Baranes D, "Tissue plasminogen activator contributes to the late phase of LTP and to synaptic growth in the hippocampal mossy fiber pathway" 21 : 813-825, 1998

21 Karatas H, "Thrombotic distal middle cerebral artery occlusion produced by topical FeCl(3)application : a novel model suitable for intravital microscopy and thrombolysis studies" 31 : 1452-1460, 2011

22 Suzuki S, "Thrombolytic properties of a novel modified human tissue-type plasminogen activator(E6010) : a bolus injection of E6010 has equivalent potency of lysing young and aged canine coronary thrombi" 17 : 738-746, 1991

23 Collen D, "Thrombolytic and pharmacokinetic properties of human tissue-type plasminogen activator variants, obtained by deletion and/or duplication of structural/functional domains, in a hamster pulmonary embolism model" 65 : 174-180, 1991

24 Agnelli G, "Thrombolytic and haemorrhagic effects of bolus doses of tissue-type plasminogen activator and a hybrid plasminogen activator with prolonged plasma half-life(K2tu-PA : CGP 42935)" 70 : 294-300, 1993

25 Spiecker M, "Thrombolysis with saruplase versus streptokinase in acute myocardial infarction : five-year results of the PRIMI trial" 138 : 518-524, 1999

26 Hao CH, "Thrombolysis with rhPro-UK 3 to 6 hours after embolic stroke in rat" 41 : 1034-1042, 2019

27 Hacke W, "Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke" 359 : 1317-1329, 2008

28 Gurewich V, "Thrombolysis vs. bleeding from hemostatic sites by a prourokinase mutant compared with tissue plasminogen activator" 4 : 1559-1565, 2006

29 Donahue SM, "Thromboelastography: a tool for measuring hypercoagulability, hypocoagulability, and fibrinolysis" 15 : 9-16, 2005

30 Jovin TG, "Thrombectomy within 8 hours after symptom onset in ischemic stroke" 372 : 2296-2306, 2015

31 Albers GW, "Thrombectomy for stroke at 6 to 16hours with selection by perfusion imaging" 378 : 708-718, 2018

32 Sila CA, "Therapy for acute ischemic stroke : the door opens : interpreting the NINDS rt-PA stroke study" 63 : 77-79, 1996

33 Chakrabarti R, "The ‘fibrinolytic potential’ as a simple measure of spontaneous fibrinolysis" 15 : 228-230, 1962

34 Kim JY, "The use of PEGylated liposomes to prolong circulation lifetimes of tissue plasminogen activator" 30 : 5751-5756, 2009

35 Bell WR, "The urokinase-streptokinase pulmonary embolism trial(phase II)results" 50 : 1070-1071, 1974

36 Sidenius N, "The urokinase plasminogen activator system in cancer : recent advances and implication for prognosis and therapy" 22 : 205-222, 2003

37 Collen D, "The tissue-type plasminogen activator story" 29 : 1151-1155, 2009

38 Ny T, "The structure of the human tissue-type plasminogen activator gene : correlation of intron and exon structures to functional and structural domains" 81 : 5355-5359, 1984

39 Shih AY, "The smallest stroke : occlusion of one penetrating vessel leads to infarction and a cognitive deficit" 16 : 55-63, 2013

40 Thiebaut AM, "The role of plasminogen activators in stroke treatment : fibrinolysis and beyond" 17 : 1121-1132, 2018

41 Marsh NA, "The rapid fibrin plate : a method for plasminogen activator assay" 38 : 545-551, 1977

42 Nicole O, "The proteolytic activity of tissue-plasminogen activator enhances NMDA receptor-mediated signaling" 7 : 59-64, 2001

43 Cartwright T, "The plasminogen activator of vampire bat saliva" 43 : 317-326, 1974

44 Krätzschmar J, "The plasminogen activator family from the salivary gland of the vampire bat Desmodus rotundus : cloning and expression" 105 : 229-237, 1991

45 Riedel CH, "The importance of size : successful recanalization by intravenous thrombolysis in acute anterior stroke depends on thrombus length" 42 : 1775-1777, 2011

46 Van de Werf FJ, "The ideal fibrinolytic : can drug design improve clinical results" 20 : 1452-1458, 1999

47 Tillett WS, "The fibrinolytic activity of hemolytic streptococci" 58 : 485-502, 1933

48 Astrup T, "The fibrin plate method for estimating fibrinolytic activity" 40 : 346-351, 1952

49 Blix S, "The effectiveness of activators in clot lysis, with special reference to fibrinolytic therapy: a new method for determination of preformed clot lysis" 386 : 1-24, 1962

50 Hacke W, "The desmoteplase in acute ischemic stroke trial(DIAS) : a phase II MRI-based 9-hour window acute stroke thrombolysis trial with intravenous desmoteplase" 36 : 66-73, 2005

51 Millar WT, "The comparison of solid phase and fibrin plate methods for the measurement of plasminogen activators" 30 : 431-439, 1983

52 IST-3 collaborative group, "The benefits and harms of intravenous thrombolysis with recombinant tissue plasminogen activator within 6 h of acute ischaemic stroke(the third international stroke trial [IST-3]) : a randomised controlled trial" 379 : 2352-2363, 2012

53 Huang X, "Tenecteplase versus alteplase in stroke thrombolysis : an individual patient data meta-analysis of randomized controlled trials" 11 : 534-543, 2016

54 Logallo N, "Tenecteplase versus alteplase for management of acute ischaemic stroke (NOR-TEST): a phase 3, randomised, open-label, blinded endpoint trial" 16 : 781-788, 2017

55 Campbell BCV, "Tenecteplase versus alteplase before thrombectomy for ischemic stroke" 378 : 1573-1582, 2018

56 Campbell BC, "Tenecteplase versus alteplase before endovascular thrombectomy (EXTEND-IA TNK): a multicenter, randomized, controlled study" 13 : 328-334, 2018

57 Hill MD, "Tenecteplase knocking on the door" 49 : 2276-2277, 2018

58 Bivard A, "Tenecteplase in ischemic stroke offers improved recanalization : analysis of 2 trials" 89 : 62-67, 2017

59 Coutts SB, "Tenecteplase for the treatment of acute ischemic stroke : a review of completed and ongoing randomized controlled trials" 13 : 885-892, 2018

60 Saxena V, "Temperature-sensitive liposome-mediated delivery of thrombolytic agents" 31 : 67-73, 2015

61 Jin R, "Taurine reduces tPA(tissue-type plasminogen activator)-induced hemorrhage and microvascular thrombosis after embolic stroke in rat" 49 : 1708-1718, 2018

62 Korin N, "Targeted drug delivery to flow-obstructed blood vessels using mechanically activated nanotherapeutics" 72 : 119-122, 2015

63 van Bruggen N, "T2-and diffusion-weighted magnetic resonance imaging of a focal ischemic lesion in rat brain" 23 : 576-582, 1992

64 Roychoudhury PK, "Synthesis, regulation and production of urokinase using mammalian cell culture: a comprehensive review" 24 : 514-528, 2006

65 Jolkkonen J, "Subacute hemorrhage and resolution of edema in Rose Bengal stroke model in rats coincides with improved sensorimotor functions" 428 : 99-102, 2007

66 Xu Y, "Structure-based antigenic epitope and PEGylation improve the efficacy of staphylokinase" 16 : 197-, 2017

67 Bringmann P, "Structural features mediating fibrin selectivity of vampire bat plasminogen activators" 270 : 25596-25603, 1995

68 Mican J, "Structural biology and protein engineering of thrombolytics" 17 : 917-938, 2019

69 Zhang Y, "Streptococcus uberis plasminogen activator(SUPA)activates human plasminogen through novel species-specific and fibrin-targeted mechanisms" 287 : 19171-19176, 2012

70 Lijnen HR, "Strategies for the improvement of thrombolytic agents" 66 : 88-110, 1991

71 Radcliffe R, "Stimulation of tissue plasminogen activator by denatured proteins and fibrin clots : a possible additional role for plasminogen activator" 211 : 750-761, 1981

72 Khursade PS, "Stenotrophomonas maltophilia Gd2 : a potential and novel isolate for fibrinolytic enzyme production" 26 : 1567-1575, 2019

73 Lack CH, "Staphylokinase; an activator of plasma protease" 161 : 559-, 1948

74 Collen D, "Staphylokinase: a potent, uniquely fibrin-selective thrombolytic agent" 4 : 279-284, 1998

75 Chiou JF, "Staphylokinase-annexin XI chimera exhibited efficient in vitro thrombolytic activities" 71 : 1122-1129, 2007

76 Zamanlu M, "Spectrophotometric analysis of thrombolytic activity : SATA assay" 8 : 31-38, 2018

77 Bader KB, "Sonothrombolysis" 880 : 339-362, 2016

78 Zhang L, "Sonic hedgehog signaling pathway mediates cerebrolysin-improved neurological function after stroke" 44 : 1965-1972, 2013

79 Andrzejewska A, "Single-cell, high-throughput analysis of cell docking to vessel wall" 39 : 2308-2320, 2019

80 Ishikawa A, "Single and repeated intravenous toxicity studies of pamiteplase(genetical recombination)in rats and monkeys" 22 : 117-133, 1997

81 Liu S, "Simple quantitative measurement based on DWI to objectively judge DWI-FLAIR mismatch in a canine stroke model" 21 : 348-354, 2015

82 Mehta BP, "Should clot composition affect choice of endovascular therapy" 79 : S63-67, 2012

83 Jiwon Kim, "Short‐Term Cessation of Dabigatran Causes a Paradoxical Prothrombotic State" Wiley 89 (89): 444-458, 2021

84 Griffin MT, "Shear-induced platelet aggregation : 3D-grayscale microfluidics for repeatable and localized occlusive thrombosis" 13 : 054106-, 2019

85 Kuge Y, "Serial changes in cerebral blood flow and flow-metabolism uncoupling in primates with acute thromboembolic stroke" 21 : 202-210, 2001

86 Jahan R, "Selective middle cerebral artery occlusion in the rabbit : technique and characterization with pathologic findings and multimodal MRI" 313 : 6-12, 2019

87 Al-Ahmady ZS, "Selective liposomal transport through blood brain barrier disruption in ischemic stroke reveals two distinct therapeutic opportunities" 13 : 12470-12486, 2019

88 Raigani M, "Scale up and pharmacokinetic study of a novel mutated chimeric tissue plasminogen activator(mt-PA)in rats" 7 : 43028-, 2017

89 Smith WS, "Safety and efficacy of mechanical embolectomy in acute ischemic stroke : results of the MERCI trial" 36 : 1432-1438, 2005

90 Albers GW, "Safety and efficacy of desmoteplase given 3-9 h after ischaemic stroke in patients with occlusion or high-grade stenosis in major cerebral arteries (DIAS-3): a double-blind, randomised, placebo-controlled phase 3 trial" 14 : 575-584, 2015

91 Jander S, "Role of NMDA receptor signaling in the regulation of inflammatory gene expression after focal brain ischemia" 109 : 181-187, 2000

92 Papadopoulos N, "Review of protocols used in ultrasound thrombolysis" 26 : 2447-2469, 2017

93 Mohammadi E, "Reteplase: structure, function, and production" 8 : 19-, 2019

94 Ciciliano JC, "Resolving the multifaceted mechanisms of the ferric chloride thrombosis model using an interdisciplinary microfluidic approach" 126 : 817-824, 2015

95 Urano T, "Regulation of plasminogen activation on cell surfaces and fibrin" 16 : 1487-1497, 2018

96 Jensen JK, "Receptor-associated protein(RAP)has two high-affinity binding sites for the low-density lipoprotein receptor-related protein(LRP) : consequences for the chaperone functions of RAP" 421 : 273-282, 2009

97 Kurz KD, "Rat model of arterial thrombosis induced by ferric chloride" 60 : 269-280, 1990

98 Goyal M, "Randomized assessment of rapid endovascular treatment of ischemic stroke" 372 : 1019-1030, 2015

99 Hume M, "Radioactive(I125)human plasma clots for assay of thrombolytic activity" 63 : 699-702, 1964

100 Kim DE, "Quantitative imaging of cerebral thromboemboli in vivo : the effects of tissue-type plasminogen activator" 48 : 1376-1385, 2017

101 Dolmer K, "Quantitative dissection of the binding contributions of ligand lysines of the receptor-associated protein(RAP)to the low density lipoprotein receptor-related protein(LRP1)" 288 : 24081-24090, 2013

102 Rijken DC, "Purification and partial characterization of plasminogen activator from human uterine tissue" 580 : 140-153, 1979

103 Hu Y, "Purification and characterization of a novel, highly potent fibrinolytic enzyme from Bacillus subtilis DC27 screened from Douchi, a traditional Chinese fermented soybean food" 9 : 9235-, 2019

104 Hu X, "Progressive and reproducible focal cortical ischemia with or without late spontaneous reperfusion generated by a ring-shaped, laser-driven photothrombotic lesion in rats" 7 : 76-85, 2001

105 Angles-Cano E, "Production of monoclonal antibodies to the high fibrin-affinity, tissue-type plasminogen activator of human plasma: demonstration of its endothelial origin by immunolocalization" 66 : 913-920, 1985

106 Bolliger D, "Principles and practice of thromboelastography in clinical coagulation management and transfusion practice" 26 : 1-13, 2012

107 Sánchez MC, "Pregnancy zone protein-tissue-type plasminogen activator complexes bind to low-density lipoprotein receptor-related protein(LRP)" 389 : 218-222, 2001

108 Qian C, "Precise characterization of the penumbra revealed by MRI : a modified photothrombotic stroke model study" 11 : e0153756-, 2016

109 Huang L, "Polyacrylic acid-coated nanoparticles loaded with recombinant tissue plasminogen activator for the treatment of mice with ischemic stroke" 516 : 565-570, 2019

110 Sun H, "Plasminogen is a critical host pathogenicity factor for group A streptococcal infection" 305 : 1283-1286, 2004

111 Horn IR, "Plasminogen activator inhibitor 1 contains a cryptic high affinity receptor binding site that is exposed upon complex formation with tissue-type plasminogen activator" 80 : 822-828, 1998

112 Barlow GH, "Plasma urokinase levels measured by chromogenic assay after infusions of tissue culture or urinary source material" 18 : 431-437, 1980

113 Dietrich WD, "Photochemically induced cortical infarction in the rat. 1. Time course of hemodynamic consequences" 6 : 184-194, 1986

114 van Duinen V, "Perfused 3D angiogenic sprouting in a high-throughput in vitro platform" 22 : 157-165, 2019

115 Rehan M, "Penta-l-lysine potentiates fibrin-independent activity of human tissue plasminogen activator" 119 : 13271-13277, 2015

116 Sawhney P, "PEGylation of truncated streptokinase leads to formulation of a useful drug with ameliorated attributes" 11 : e0155831-, 2016

117 Goulay R, "Optimized tPA : a non-neurotoxic fibrinolytic agent for the drainage of intracerebral hemorrhages" 38 : 1180-1189, 2018

118 Esmaeeli-Nadimi A, "Opening the window : ischemic postconditioning reduces the hyperemic response of delayed tissue plasminogen activator and extends its therapeutic time window in an embolic stroke model" 764 : 55-62, 2015

119 Bonnard T, "Novel thrombolytic drug based on thrombin cleavable microplasminogen coupled to a single-chain antibody specific for activated GPIIb/IIIa" 6 : e004535-, 2017

120 He J, "Novel recombinant thrombolytic and antithrombotic staphylokinase variants with an RGD motif at their N-termini" 50 : 17-23, 2008

121 Schroeter M, "Non-invasive induction of focal cerebral ischemia in mice by photothrombosis of cortical microvessels : characterization of inflammatory responses" 117 : 43-49, 2002

122 Lapchak PA, "Neuroprotective effects of the spin trap agent disodium-[(tert-butylimino)methyl]benzene-1,3-disulfonate N-oxide (generic NXY-059) in a rabbit small clot embolic stroke model: combination studies with the thrombolytic tissue plasminogen activator" 33 : 1411-1415, 2002

123 Lee KY, "Neuroprotective effect of asiatic acid in rat model of focal embolic stroke" 37 : 1397-1401, 2014

124 Seeds NW, "Neuronal migration is retarded in mice lacking the tissue plasminogen activator gene" 96 : 14118-14123, 1999

125 Chen H, "Nattokinase : a promising alternative in prevention and treatment of cardiovascular diseases" 13 : 1-, 2018

126 Partoazar A, "Nanoliposome containing cyclosporine A reduced neuroinflammation responses and improved neurological activities in cerebral ischemia/reperfusion in rat" 31 : 185-193, 2017

127 Kanazawa M, "Multiple therapeutic effects of progranulin on experimental acute ischaemic stroke" 138 : 1932-1948, 2015

128 Atochin DN, "Mouse model of microembolic stroke and reperfusion" 35 : 2177-2182, 2004

129 Orset C, "Mouse model of in situ thromboembolic stroke and reperfusion" 38 : 2771-2778, 2007

130 Kusch A, "Monocyte-expressed urokinase regulates human vascular smooth muscle cell migration in a coculture model" 383 : 217-221, 2002

131 Costa PF, "Mimicking arterial thrombosis in a 3D-printed microfluidic in vitro vascular model based on computed tomography angiography data" 17 : 2785-2792, 2017

132 Lapchak PA, "Microplasmin : a novel thrombolytic that improves behavioral outcome after embolic strokes in rabbits" 33 : 2279-2284, 2002

133 Loyau S, "Microfluidic modeling of thrombolysis : effect of antiplatelet and anticoagulant agents on tPA(tissue-type plasminogen activator)-induced fibrinolysis" 38 : 2626-2637, 2018

134 Krishnamurthy A, "Methods available to assess therapeutic potential of fibrinolytic enzymes of microbial origin : a review" 9 : 10-, 2018

135 Bhatia R, "Low rates of acute recanalization with intravenous recombinant tissue plasminogen activator in ischemic stroke : real-world experience and a call for action" 41 : 2254-2258, 2010

136 Bruch GE, "Liposomes for drug delivery in stroke" 152 : 246-256, 2019

137 Koudelka S, "Liposomal nanocarriers for plasminogen activators" 227 : 45-57, 2016

138 Miao P, "Laser speckle contrast imaging of cerebral blood flow in freely moving animals" 16 : 090502-, 2011

139 Gauberti M, "Lack of secondary microthrombosis after thrombin-induced stroke in mice and non-human primates" 12 : 409-414, 2014

140 Parimalam SS, "Lab-on-a-chip for the development of pro-/anti-angiogenic nanomedicines to treat brain diseases" 20 : 6126-, 2019

141 Hoylaerts M, "Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin" 257 : 2912-2919, 1982

142 Hekman CM, "Kinetic analysis of the interactions between plasminogen activator inhibitor 1 and both urokinase and tissue plasminogen activator" 262 : 199-210, 1988

143 Verheijen JH, "Involvement of finger domain and kringle 2 domain of tissue-type plasminogen activator in fibrin binding and stimulation of activity by fibrin" 5 : 3525-3530, 1986

144 Chen K, "Intrinsic biotaxi solution based on blood cell membrane cloaking enables fullerenol thrombolysis in vivo" 12 : 14958-14970, 2020

145 Hacke W, "Intravenous desmoteplase in patients with acute ischaemic stroke selected by MRI perfusion–diffusion weighted imaging or perfusion CT (DIAS-2): a prospective, randomised, double-blind, placebo-controlled study" 8 : 141-150, 2009

146 Tennant SN, "Intracoronary thrombolysis in patients with acute myocardial infarction : comparison of the efficacy of urokinase with streptokinase" 69 : 756-760, 1984

147 Sakharov DV, "Interactions between staphylokinase, plasmin(ogen), and fibrin. Staphylokinase discriminates between free plasminogen and plasminogen bound to partially degraded fibrin" 271 : 27912-27918, 1996

148 Higgins DL, "Interaction of one-chain and twochain tissue plasminogen activator with intact and plasmin-degraded fibrin" 26 : 7786-7791, 1987

149 Kuiper J, "Interaction of mutants of tissue-type plasminogen activator with liver cells : effect of domain deletions" 313 : 775-780, 1996

150 Lehmann J, "Inflammatory cell recruitment after experimental thromboembolic stroke in rats" 279 : 139-154, 2014

151 Watson BD, "Induction of reproducible brain infarction by photochemically initiated thrombosis" 17 : 497-504, 1985

152 Lu H, "Induction and imaging of photothrombotic stroke in conscious and freely moving rats" 19 : 96013-, 2014

153 Salman-Kesner N, "In-vitro assessment of the thrombolytic efficacy of therapeutic ultrasound" 178 : 63-68, 2019

154 Kleinschnitz C, "In vivo monitoring of macrophage infiltration in experimental ischemic brain lesions by magnetic resonance imaging" 23 : 1356-1361, 2003

155 Cole ES, "In vivo clearance of tissue plasminogen activator : the complex role of sites of glycosylation and level of sialylation" 7 : 15-22, 1993

156 Silva PEDCE, "In vitro thrombolytic activity of a purified fibrinolytic enzyme from Chlorella vulgaris" 1092 : 524-529, 2018

157 Agren A, "In vitro combinations of red blood cell, plasma and platelet components evaluated by thromboelastography" 12 : 491-496, 2014

158 Ghaheh HS, "Improving the solubility, activity, and stability of reteplase using in silico design of new variants" 14 : 359-368, 2019

159 Stewart RJ, "Identification of the mechanism responsible for the increased fibrin specificity of TNK-tissue plasminogen activator relative to tissue plasminogen activator" 275 : 10112-10120, 2000

160 Tsurupa G, "Identification and characterization of novel tPA-and plasminogen-binding sites within fibrin(ogen)alpha C-domains" 40 : 801-808, 2001

161 He J, "Identification and characterization of Harobin, a novel fibrino(geno)lytic serine protease from a sea snake (Lapemis hardwickii)" 581 : 2965-2973, 2007

162 Kim DE, "Hyperacute direct thrombus imaging using computed tomography and gold nanoparticles" 73 : 617-625, 2013

163 Medcalf RL, "Human synovial fibroblasts produce urokinase-type plasminogen activator" 29 : 1397-1401, 1986

164 Pierpaoli C, "Histopathologic correlates of abnormal water diffusion in cerebral ischemia : diffusion-weighted MR imaging and light and electron microscopic study" 189 : 439-448, 1993

165 Lassen M, "Heat denaturation of plasminogen in the fibrin plate method" 27 : 371-376, 1953

166 GBD 2016 Stroke Collaborators, "Global, regional, and national burden of stroke, 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016" 18 : 439-458, 2019

167 Beech JS, "Further characterisation of a thromboembolic model of stroke in the rat" 895 : 18-24, 2001

168 Zhang L, "Focal embolic cerebral ischemia in the rat" 10 : 539-547, 2015

169 Nieuwenhuizen W, "Fluorogenic substrates for sensitive and differential estimation of urokinase and tissue plasminogen activator" 7 : 146-149, 1978

170 Rathore YS, "First structural model of full-length human tissue-plasminogen activator : a SAXS data-based modeling study" 116 : 496-502, 2012

171 Rijken DC, "Fibrinolytic properties of one-chain and two-chain human extrinsic(tissue-type)plasminogen activator" 257 : 2920-2925, 1982

172 Narasimhan MK, "Fibrinolytic enzyme production by newly isolated Bacillus cereus SRM001 with enhanced in-vitro blood clot lysis potential" 61 : 157-164, 2015

173 Marini C, "Fibrinolytic effects of urokinase and heparin in acute pulmonary embolism : a randomized clinical trial" 54 : 162-173, 1988

174 Macfarlane RG, "Fibrinolytic activity of normal urine" 159 : 779-, 1947

175 Medcalf RL, "Fibrinolysis : from blood to the brain" 15 : 2089-2098, 2017

176 Tsurupa G, "Fibrinogen alpha C domains contain cryptic plasminogen and tPA binding sites" 936 : 328-330, 2001

177 Franke AE, "Expression of human plasminogen activator inhibitor type-1 (PAI-1) in Escherichia coli as a soluble protein comprised of active and latent forms. Isolation and crystallization of latent PAI-1" 1037 : 16-23, 1990

178 Saadatirad A, "Expression of a novel chimeric-truncated tPA in Pichia pastoris with improved biochemical properties" 56 : 1143-1150, 2014

179 Davami F, "Expression of a novel chimeric truncated t-PA in CHO cells based on in silico experiments" 2010 : 108159-, 2010

180 Vanacek P, "Exploration of enzyme diversity by integrating bioinformatics with expression analysis and biochemical characterization" 8 : 2402-2412, 2018

181 Kito G, "Experimental thromboembolic stroke in cynomolgus monkey" 105 : 45-53, 2001

182 Fan NK, "Experimental and imaging techniques for examining fibrin clot structures in normal and diseased states" 98 : e52019-, 2015

183 Lee VM, "Evolution of photochemically induced focal cerebral ischemia in the rat : magnetic resonance imaging and histology" 27 : 2110-2118, 1996

184 Burgos AM, "Evidence that tenecteplase is noninferior to alteplase for acute ischemic stroke : meta-analysis of 5 randomized trials" 50 : 2156-2162, 2019

185 Chmielewska J, "Evidence for a rapid inhibitor to tissue plasminogen activator in plasma" 31 : 427-436, 1983

186 Zhu YQ, "Evaluation of neurovascular function in mouse cortex using multispectral optical imaging after ischemic stroke" 99 : 2943-2946, 2019

187 den Heijer P, "Evaluation of a weight-adjusted single-bolus plasminogen activator in patients with myocardial infarction: a double-blind, randomized angiographic trial of lanoteplase versus alteplase" 98 : 2117-2125, 1998

188 Wind T, "Epitope mapping for four monoclonal antibodies against human plasminogen activator inhibitor type-1 : implications for antibody-mediated PAI-1-neutralization and vitronectin-binding" 268 : 1095-1106, 2001

189 Taheri MN, "Engineering, expression and purification of a chimeric fibrin-specific streptokinase" 128 : 14-21, 2016

190 Absar S, "Engineering of plasminogen activators for targeting to thrombus and heightening thrombolytic efficacy" 13 : 1545-1556, 2015

191 Wu XC, "Engineering of plasmin-resistant forms of streptokinase and their production in Bacillus subtilis : streptokinase with longer functional half-life" 64 : 824-829, 1998

192 Babbal A, "Engineering of deglycosylated and plasmin resistant variants of recombinant streptokinase in Pichia pastoris" 102 : 10561-10577, 2018

193 Li Z, "Engineering of Harobin for enhanced fibrinolytic activity obtained by random and site-directed mutagenesis" 129 : 162-172, 2017

194 Collen D, "Engineered staphylokinase variants with reduced immunogenicity" 12 (12): 59-65, 1998

195 Xu J, "Engineered nanoplatelets for targeted delivery of plasminogen activators to reverse thrombus in multiple mouse thrombosis models" 32 : e1905145-, 2020

196 Amar DN, "Endothelial cell activation in an embolic ischemia-reperfusion injury microfluidic model" 10 : 857-, 2019

197 Jin R, "Embolic middle cerebral artery occlusion(MCAO)for ischemic stroke with homologous blood clots in rats" 91 : 51956-, 2014

198 Ren M, "Embolic middle cerebral artery occlusion model using thrombin and fibrinogen composed clots in rat" 211 : 296-304, 2012

199 Van Hulle F, "Efficacy of urokinase lock to treat thrombotic dysfunction of tunneled hemodialysis catheters : a retrospective cohort study" 20 : 60-69, 2019

200 Takano K, "Efficacy of intra-arterial and intravenous prourokinase in an embolic stroke model evaluated by diffusion-perfusion magnetic resonance imaging" 50 : 870-875, 1998

201 Orset C, "Efficacy of alteplase in a mouse model of acute ischemic stroke : a retrospective pooled analysis" 47 : 1312-1318, 2016

202 Suzuki E, "Efficacy of SMTP-7, a small-molecule anti-inflammatory thrombolytic, in embolic stroke in monkeys" 6 : e00448-, 2018

203 Si Z, "Effects of thrombolysis within 6 hours on acute cerebral infarction in an improved rat embolic middle cerebral artery occlusion model for ischaemic stroke" 23 : 2468-2474, 2019

204 Lapchak PA, "Effects of the spin trap agent disodium-[tert-butylimino)methyl]benzene-1,3-disulfonate N-oxide (generic NXY-059)on intracerebral hemorrhage in a rabbit large clot embolic stroke model: combination studies with tissue plasminogen activator" 33 : 1665-1670, 2002

205 Emberson J, "Effect of treatment delay, age, and stroke severity on the effects of intravenous thrombolysis with alteplase for acute ischaemic stroke: a meta-analysis of individual patient data from randomised trials" 384 : 1929-1935, 2014

206 Loskutoff DJ, "Effect of thrombin on the fibrinolytic activity of cultured bovine endothelial cells" 64 : 329-332, 1979

207 Chen S, "Early abrogation of gelatinase activity extends the time window for tPA thrombolysis after embolic focal cerebral ischemia in mice" 5 : 2018

208 Schroeter M, "Dynamic changes of magnetic resonance imaging abnormalities in relation to inflammation and glial responses after photothrombotic cerebral infarction in the rat brain" 101 : 114-122, 2001

209 Barta G, "Dyed fibrin plate assay of fibrinolysis" 44 : 233-240, 1966

210 Ishrat T, "Dose-response, therapeutic time-window and tPA-combinatorial efficacy of compound 21: a randomized, blinded preclinical trial in a rat model of thromboembolic stroke" 39 : 1635-1647, 2019

211 Furlan AJ, "Dose escalation of desmoteplase for acute ischemic stroke(DEDAS) : evidence of safety and efficacy 3 to 9hours after stroke onset" 37 : 1227-1231, 2006

212 Culp WC, "Dodecafluoropentane emulsion decreases infarct volume in a rabbit ischemic stroke model" 23 : 116-121, 2012

213 Harrison TC, "Displacement of sensory maps and disorganization of motor cortex after targeted stroke in mice" 44 : 2300-2306, 2013

214 Kim DE, "Direct thrombus imaging as a means to control the variability of mouse embolic infarct models : the role of optical molecular imaging" 42 : 3566-3573, 2011

215 Kim JY, "Direct imaging of cerebral thromboemboli using computed tomography and fibrin-targeted gold nanoparticles" 5 : 1098-1114, 2015

216 Zimmerman M, "Direct fluorescent assay of urokinase and plasminogen activators of normal and malignant cells : kinetics and inhibitor profiles" 75 : 750-753, 1978

217 Prasad S, "Development of an in vitro model to study clot lysis activity of thrombolytic drugs" 4 : 14-, 2006

218 Kuroiwa T, "Development of a rat model of photothrombotic ischemia and infarction within the caudoputamen" 40 : 248-253, 2009

219 Bonnard T, "Development and validation of a high throughput whole blood thrombolysis plate assay" 7 : 2346-, 2017

220 Ilich A, "Development and application of global assays of hyper-and hypofibrinolysis" 4 : 46-53, 2020

221 von Kummer R, "Desmoteplase 3 to 9 hours after major artery occlusion stroke : the DIAS-4 trial(efficacy and safety study of desmoteplase to treat acute ischemic stroke)" 47 : 2880-2887, 2016

222 Kazemali M, "Design of a novel chimeric tissue plasminogen activator with favorable vampire bat plasminogen activator properties" 67 : 82-86, 2014

223 Zhang L, "Demonstration of therapeutic window of cerebrolysin in embolic stroke: a prospective, randomized, blinded, and placebo-controlled study" 12 : 628-635, 2017

224 Sladojevic N, "Decreased thromboembolic stroke but not atherosclerosis or vascular remodelling in mice with ROCK2-deficient platelets" 113 : 1307-1317, 2017

225 Zhang N, "Cyclic RGD functionalized liposomes encapsulating urokinase for thrombolysis" 70 : 227-236, 2018

226 Wang X, "Crystal structure of the catalytic domain of human plasmin complexed with streptokinase" 281 : 1662-1665, 1998

227 Yakovlev S, "Conversion of fibrinogen to fibrin : mechanism of exposure of tPA-and plasminogen-binding sites" 39 : 15730-15741, 2000

228 Xu Z, "Conservation of critical functional domains in murine plasminogen activator inhibitor-1" 279 : 17914-17920, 2004

229 Bär FW, "Comparison of saruplase and alteplase in acute myocardial infarction. SESAM Study Group. The Study in Europe with Saruplase and Alteplase in Myocardial Infarction" 79 : 727-732, 1997

230 Faraji H, "Comparison of expression optimization of new derivative of staphylokinase (SAK-2RGDTTI) with the rSAK" 35 : e2819-, 2019

231 Xu XQ, "Comparative study of the relative signal intensity on DWI, FLAIR, and T2 images in identifying the onset time of stroke in an embolic canine model" 35 : 1059-1065, 2014

232 Tiebosch IA, "Combined treatment with recombinant tissue plasminogen activator and dexamethasone phosphate-containing liposomes improves neurological outcome and restricts lesion progression after embolic stroke in rats" 123 : 65-74, 2012

233 Kim DE, "Combined near-infrared fluorescent imaging and micro-computed tomography for directly visualizing cerebral thromboemboli" 2016

234 Armstrong PW, "Collaborative angiographic patency trial of recombinant staphylokinase(CAPTORS)" 139 : 820-823, 2000

235 Armstrong PW, "Collaborative angiographic patency trial of recombinant staphylokinase(CAPTORS II)" 146 : 484-488, 2003

236 Patchett SE, "Clot lysis by gastric juice : an in vitro study" 30 : 1704-1707, 1989

237 Pennica D, "Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli" 301 : 214-221, 1983

238 Pang PT, "Cleavage of proBDNF by tPA/plasmin is essential for longterm hippocampal plasticity" 306 : 487-491, 2004

239 Stewart RJ, "Characterization of the interactions of plasminogen and tissue and vampire bat plasminogen activators with fibrinogen, fibrin, and the complex of d-Dimer noncovalently linked to fragment E" 273 : 18292-18299, 1998

240 Jiao J, "Characterization of a recombinant chimeric plasminogen activator with enhanced fibrin binding" 1546 : 399-405, 2001

241 Hilger T, "Characterization of a novel chronic photothrombotic ring stroke model in rats by magnetic resonance imaging, biochemical imaging, and histology" 24 : 789-797, 2004

242 Ansar S, "Characterization of a new model of thromboembolic stroke in C57black/6J mice" 5 : 526-533, 2014

243 Bakhit C, "Cellular catabolism of recombinant tissue-type plasminogen activator. Identification and characterization of a novel high affinity uptake system on rat hepatocytes" 262 : 8716-8720, 1987

244 Gibson PR, "Cell associated urokinase activity and colonic epithelial cells in health and disease" 32 : 191-195, 1991

245 Moon SK, "Both compensation and recovery of skilled reaching following small photothrombotic stroke to motor cortex in the rat" 218 : 145-153, 2009

246 Kleinschnitz C, "Blocking of platelets or intrinsic coagulation pathway-driven thrombosis does not prevent cerebral infarctions induced by photothrombosis" 39 : 1262-1268, 2008

247 Lucore CL, "Biochemical determinants of clearance of tissue-type plasminogen activator from the circulation" 77 : 906-914, 1988

248 de Munk GA, "Binding of tissue-type plasminogen activator to lysine, lysine analogues, and fibrin fragments" 28 : 7318-7325, 1989

249 van Zonneveld AJ, "Autonomous functions of structural domains on human tissue-type plasminogen activator" 83 : 4670-4674, 1986

250 Zhang L, "Atorvastatin extends the therapeutic window for tPA to 6 h after the onset of embolic stroke in rats" 29 : 1816-1824, 2009

251 Menon BK, "Association of clinical, imaging, and thrombus characteristics with recanalization of visible intracranial occlusion in patients with acute ischemic stroke" 320 : 1017-1026, 2018

252 Provenzale JM, "Assessment of the patient with hyperacute stroke : imaging and therapy" 229 : 347-359, 2003

253 Mair G, "Arterial obstruction on computed tomographic or magnetic resonance angiography and response to intravenous thrombolytics in ischemic stroke" 48 : 353-360, 2017

254 Asahi M, "Antiactin-targeted immunoliposomes ameliorate tissue plasminogen activator-induced hemorrhage after focal embolic stroke" 23 : 895-899, 2003

255 Andersen OM, "Analysis of a two-domain binding site for the urokinase-type plasminogen activator-plasminogen activator inhibitor-1 complex in low-density-lipoprotein-receptor-related protein" 357 : 289-296, 2001

256 Wang Z, "An in vitro assay for sonothrombolysis based on the spectrophotometric measurement of clot thickness" 36 : 681-698, 2017

257 Kudo M, "An animal model of cerebral infarction. Homologous blood clot emboli in rats" 13 : 505-508, 1982

258 Helenius MA, "Amplification of urokinase gene in prostate cancer" 61 : 5340-5344, 2001

259 Maheshwari N, "Amino-terminal fusion of epidermal growth factor 4, 5, 6 domains of human thrombomodulin on streptokinase confers anti-reocclusion characteristics along with plasmin-mediated clot specificity" 11 : e0150315-, 2016

260 Sherry S, "Activity of plasmin and streptokinase-activator on substituted arginine and lysine esters" 16 : 18-31, 1966

261 Kotb E, "Activity assessment of microbial fibrinolytic enzymes" 97 : 6647-6665, 2013

262 Aguiar de Sousa D, "Access to and delivery of acute ischaemic stroke treatments : a survey of national scientific societies and stroke experts in 44 European countries" 4 : 13-28, 2019

263 Somerville DA, "A technique for demonstrating fibrinolysis by cutaneous bacteria" 25 : 740-741, 1972

264 Jespersen J, "A study of the fibrin plate assay of fibrinolytic agents. Optimal conditions, reproducibility and precision" 13 : 301-315, 1983

265 Hawkey CM, "A standard clot method for the assay of plasminogen activators, anti-activators, and plasmin" 17 : 175-181, 1964

266 Niinobe M, "A sensitive colorimetric assay for various proteases using naphthyl ester derivatives as substrates" 87 : 779-783, 1980

267 Kim HS, "A rat model of photothrombotic capsular infarct with a marked motor deficit: a behavioral, histologic, and microPET study" 34 : 683-689, 2014

268 Zhang RL, "A rat model of focal embolic cerebral ischemia" 766 : 83-92, 1997

269 Gidron E, "A rapid screening test for reduced fibrinolytic activity of plasma : streptokinase activated lysis time" 31 : 54-57, 1978

270 Moroz LA, "A rapid and sensitive 125I-fibrin solid-phase fibrinolytic assay for plasmin" 46 : 543-553, 1975

271 Roche PC, "A rapid and highly sensitive solid-phase radioassay for plasminogen activators" 31 : 269-277, 1983

272 Parsons M, "A randomized trial of tenecteplase versus alteplase for acute ischemic stroke" 366 : 1099-1107, 2012

273 Longstaff C, "A proposed reference method for plasminogen activators that enables calculation of enzyme activities in SI units" 2 : 1416-1421, 2004

274 Pevsner PH, "A photothrombotic model of small early ischemic infarcts in the rat brain with histologic and MRI correlation" 45 : 227-233, 2001

275 Fatemeh Davami, "A novel variant of t-PA resistant to plasminogen activator inhibitor-1; expression in CHO cells based on In Silico experiments" 생화학분자생물학회 44 (44): 34-39, 2011

276 Chen Y, "A novel mouse model of thromboembolic stroke" 256 : 203-211, 2015

277 Szemraj J, "A new recombinant thrombolytic and antithrombotic agent with higher fibrin affinity: a staphylokinase variant. I. In vitro study" 3 : 2156-2165, 2005

278 Zhang Z, "A new rat model of thrombotic focal cerebral ischemia" 17 : 123-135, 1997

279 Goldenberg NA, "A new global assay of coagulation and fibrinolysis" 116 : 345-356, 2005

280 Smith AA, "A new euglobulin clot lysis assay for global fibrinolysis" 112 : 329-337, 2003

281 Sun YY, "A murine photothrombotic stroke model with an increased fibrin content and improved responses to tPA-lytic treatment" 4 : 1222-1231, 2020

282 Shimamura N, "A model of rat embolic cerebral infarction with a quantifiable, autologous arterial blood clot" 4 : 564-570, 2013

283 Howell M, "A method for assessing clot lysis" 17 : 310-312, 1964

284 Mander P, "A low molecular weight chymotrypsin-like novel fibrinolytic enzyme from Streptomyces sp. CS624" 46 : 1449-1455, 2011

285 Thomas GR, "A long-half-life and fibrin-specific form of tissue plasminogen activator in rabbit models of embolic stroke and peripheral bleeding" 25 : 2072-2078, 1994

286 He S, "A laboratory method for determination of overall haemostatic potential in plasma. I. Method design and preliminary results" 96 : 145-156, 1999

287 Millar WT, "A highly sensitive solid phase assay for plasminogen activators using 125I-fibrinogen" 13 : 389-395, 1978

288 Kulseth MA, "A highly sensitive chromogenic microplate assay for quantification of rat and human plasminogen" 210 : 314-317, 1993

289 Kim PY, "A high affinity interaction of plasminogen with fibrin is not essential for efficient activation by tissue plasminogen activator" 287 : 4652-4661, 2012

290 Keyt BA, "A faster-acting and more potent form of tissue plasminogen activator" 91 : 3670-3674, 1994

291 Wu SC, "A fast-acting, modular-structured staphylokinase fusion with kringle-1 from human plasminogen as the fibrin-targeting domain offers improved clot lysis efficacy" 278 : 18199-18206, 2003

292 Salazar AM, "A comparative analysis of the clotting and fibrinolytic activities of the snake venom(Bothrops atrox)from different geographical areas in Venezuela" 120 : 95-104, 2007

293 김혜진, "A Novel Thrombolytic and Anticoagulant Serine Protease from Polychaeta, Diopatra sugokai" 한국미생물·생명공학회 28 (28): 275-283, 2018