Fig. 4: Reversal of thrombin inhibition.
From: Development of supramolecular anticoagulants with on-demand reversibility
a, Schematic representation of antidote addition and reversal of inhibition. b, Chemical structures of adenine and diaminopurine forming hydrogen bonds with thymine. c, Fluorogenic assay data showing the reversal of thrombin inhibition by the addition of different concentrations of antidote after 30 min of inhibition. Data are presented as mean ± s.d., with n = 3 replicates. d, Fibrinogen assay data showing the reversal of thrombin inhibition by the addition of antidote (1 equiv.) after 30 min of inhibition. Data are presented as mean ± s.d., with n = 3 replicates. e, CAT of A8–E1 with and without antidote. Data are presented as mean values, with n = 3 replicates. f, Average fibrin intensity and average thrombus volume for control-treated animals (n = 7), argatroban-treated animals (n = 4; 2 mg per kg (body weight) bolus followed by an infusion of 12 mg per kg (body weight)), A8–E1-treated animals (n = 3; 5 mg per kg (body weight) bolus) and A8–E1 + AD2-treated animals (n = 3; 5 mg per kg (body weight) + 5 molar equiv. antidote). Statistical significance between multiple treatment groups was analyzed using an ordinary one-way ANOVA with Tukey’s multiple comparisons testing and a single pooled variance. For multiple comparisons, the mean of each column was compared to the mean of every other column. Data are presented as mean ± s.e.m., where ‘n’ equals the number of independent experiments performed. The P values for the average thrombus volume data are *P = 0.0412 (control versus A8–E1), P = 0.7759 (NS; control versus A8–E1 + AD2), ***P = 0.0001 (control versus argatroban), **P = 0.0021 (A8–E1 versus A8–E1 + AD2), P = 0.1208 (NS; A8–E1 versus argatroban) and ****P < 0.0001 (A8–E1 + AD2 versus argatroban). The P values for the average fibrin intensity data are P = 0.4545 (NS; control versus A8–E1 + AD2), P = 0.7333 (NS; A8–E1 versus argatroban) and ****P < 0.0001 (all other comparisons). g, Exemplar image of a thrombus 15 min after needle injury without inhibitor (left), with A8–E1 (center) and with A8–E1 + AD2 (right). Platelets are shown in red, fibrin is shown in green, and collagen is shown in white; scale bars, 10 μm.
