title: Antithrombotic activity of HY023016, a novel Dabigatran prodrug evaluated in animal thrombosis models.
abstract: BACKGROUND
  Thrombin is a multifunctional trypsin-like serine protease that plays key roles in coagulation and thrombogenesis. HY023016, a novel Dabigatran prodrug, is an oral direct thrombin inhibitor. The purpose of this study was to compare the anti-thrombotic activities and haemorrhagic effects of HY023016 with Dabigatran etexilate and tetramethylpyrazine in several animal thrombosis models.
  
  
  METHODS
  To investigate drug exposure, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used to determine the pharmacokinetic profile of HY023016. After single intragastric administrations of HY023016, Dabigatran etexilate or tetramethylpyrazine, the anti-thrombotic activities were evaluated through rabbit jugular vein thrombosis model, rat inferior vena cava thrombosis model, ex vivo rabbit platelet aggregation assay, in vivo rabbit coagulation assay, and direct thrombin binding assay. Meanwhile, we evaluated the effect of HY023016 on expression of tissue factor (TF) by RT-PCR. Rabbit cuticle bleeding assay and mouse tail bleeding assay were applied to evaluate the effects of HY023016 on haemorrhage.
  
  
  RESULTS
  Pharmacokinetic parameters indicated that HY023016 can convert to Dabigatran and tetramethylpyrazine. Our studies showed that HY023016 was able to significantly inhibit thrombus formation in a dose-dependent manner in rabbit and rat models (P<0.05). Similarly, it was able to dose-dependently inhibit thrombin- or ADP-induced platelet aggregation, prolonging the activated partial thromboplastin time (APTT) and prothrombin time (PT), inhibiting the activity of thrombin and inhibiting thrombin- or ADP-induced expression of TF (P<0.05 or 0.01). Dabigatran etexilate was also able to dose-dependently and significantly inhibit thrombus formation (P<0.01) but was unable to affect ADP-induced platelet aggregation and expression of TF. In contrast, tetramethylpyrazine could only exhibit mild antithrombotic activity compared with HY023016 and Dabigatran etexilate (P<0.05). HY023016 could prolong bleeding time (P<0.001), but the prolongations were significantly less than Dabigatran etexilate (P<0.05).
  
  
  CONCLUSIONS
  HY023016 showed thrombosis-inhibition activities comparable to those of Dabigatran etexilate, but better than those of tetramethylpyrazine. The attendant bleeding risk of HY023016 was lower than Dabigatran etexilate in rabbits and mice.
authors.affiliation: Department of Pharmacology, China Pharmaceutical University, Nanjing, Jiangsu, China.
authors.forename: Yun-Zhan
authors.initials: YZ
authors.lastname: Li
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Gong
authors.forename: Wen-Hui
authors.initials: WH
authors.lastname: Yang
authors.forename: Xin-Hui
authors.initials: XH
authors.lastname: Wang
authors.forename: Mei-Ling
authors.initials: ML
authors.lastname: Jiang
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhou
authors.forename: Xiao-Zhi
authors.initials: XZ
authors.lastname: Yang
authors.forename: Yun-Gen
authors.initials: YG
authors.lastname: Xu
authors.forename: Guang-Wei
authors.initials: GW
authors.lastname: He
mesh_terms: D000818:Animals; D000991:Antithrombins / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology*; D001562:Benzimidazoles / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology*; D000069604:Dabigatran; D004195:Disease Models, Animal; D051379:Mice; D011355:Prodrugs / Q000493:pharmacokinetics / Q000494:pharmacology*; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D011817:Rabbits; D051381:Rats; D013917:Thrombin / Q000378:metabolism; D013925:Thromboplastin / Q000096:biosynthesis / Q000235:genetics; D013927:Thrombosis / Q000097:blood / Q000188:drug therapy* / Q000378:metabolism*; D015091:beta-Alanine / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology
chemical_list: D000991:Antithrombins; D001562:Benzimidazoles; D011355:Prodrugs; D012333:RNA, Messenger; D015091:beta-Alanine; D013925:Thromboplastin; D013917:Thrombin; D000069604:Dabigatran