Table 2.
Characterization of heparin samples that combine different chemical modifications. Shadowed in gray are the formulations selected for in vivo assays.
| Sample | Mw (Da) | SO3−/Disaccharide | IC50 (µg/mL ± SD) | IC90 (µg/mL ± SD) | aPTT (IU/mg) | In Vitro Toxicity 1 (% ± SD) | Hemolysis 1 (% ± SD) |
|---|---|---|---|---|---|---|---|
| MMWH_1 | 10,497 | n.d. | 22.7 ± 2.4 | 178.1 ± 28.4 | 110 | 1.1 ± 3.4 | 0.1 ± 0.2 |
| MMWH_2 | 11,000 | 1.9 | 33.7 ± 21.8 | 374.8 ± 189.3 | 130 | 0.0 ± 6.0 | 0.5 ± 0.2 |
| 2-O-desulfated MMWH_1 | 8771 | 1.6 | 80.7 ± 5.0 | 270.3 ± 42.5 | 38 | 10.0 ± 3.0 | 0.3 ± 0.4 |
| 2-O-desulfated MMWH_2 | 8776 | 1.5 | 91.9 ± 9.0 | 351.9 ± 43.4 | 23 | 0.0 ± 2.3 | 2.0 ± 2.0 |
| 2-O-desulfated MMWH_3 | 10,198 | 1.5 | 59.1 ± 16.5 | 487.9 ± 161.3 | 53 | 17.9 ± 2.1 | 0.5 ± 0.2 |
| 2-O-desulfated MMWH_4 | 11,190 | 1.4 | 68.0 ± 14.5 | 404.8 ± 119.4 | 33 | 7.5 ± 1.8 | 0.0 ± 0.0 |
| 2-O-desulfated glycol-split MMWH_1 | 7388 | 1.8 | 79.6 ± 5.4 | 893.1 ± 321.0 | 6 | 7.3 ± 4.5 | 0.4 ± 0.1 |
| 2-O-desulfated glycol-split MMWH_2 | 7037 | 1.5 | 84.2 ± 13.4 | 303.4 ± 46.2 | 5 | 3.6 ± 5.4 | 0.0 ± 0.0 |
| ULMWH | 4270 | 2.2 | 49.3 ± 6.0 | 236.3 ± 54.9 | 6 | 6.8 ± 9.2 | 0.2 ± 0.1 |
| 2-O-desulfated ULMWH_1 | 4150 | 1.6 | 140.9 ± 15.9 | 322.5 ± 27.5 | ud. | 20.0 ± 16.7 | 0.0 ± 0.0 |
| 2-O-desulfated ULMWH_2 | 4450 | 1.4 | 129.2 ± 13.1 | 262.5 ± 16.1 | ud. | 37.0 ± 9.9 | 0.2 ± 0.0 |
| 2-O-desulfated glycol-split ULMWH_1 | 4024 | 1.7 | 104.4 ± 6.0 | 192.4 ± 16.2 | ud. | 2.5 ± 2.3 | 0.0 ± 0.2 |
| 2-O-desulfated glycol-split ULMWH_2 | 3800 | 1.6 | 130.3 ± 15.5 | 200.5 ± 111.6 | ud. | 11.5 ± 8.0 | 0.0 ± 0.1 |
| UH, commercial | 13,000 | 1.9–2.0 | 9.4 ± 4.4 | 135.5 ± 12.8 | 197 | 0.0 ± 6.0 | 0.0 ± 0.0 |
1 Reported in vitro toxicity in human umbilical vein endothelial cell (HUVEC) culture (% of cell death) and hemolysis data (% of lysed RBCs) have been calculated with 2 mg heparin/mL. The numbers after two compounds with the same name indicate that those samples are replicates, made in order to check the reproducibility of the procedure and tested separately to corroborate if there were any differences in antimalarial or anticoagulant activity. n.d.: not determined, ud.: undetected.