Studies on the Hemorrhagic Sweet Clover Disease Iv. The Isolation and Crystallization of the Hemorrhagic Agent

Abstract

1. The hemorrahgic agent is spoiled sweet clover hay (Melilotus alaba) has been isolatd in a pure state, m. p. 288‐289°, from experimentally produced spoiled hays as well as form hyas that killed cattle in argicultural practice. 2. This substance has the empirical formula C12H12O6. It is optically inactive. There are two acidic hydroxyls in the molecule. The acidity of the pure substance falls between that of the phenols and the carboxylic acids. A crystalline dimethyl other C12H12O4(OCH2)2 with a melting point fo 168‐170° (physiologically inactive) has been prepared by methylation with diazomethane. 3. In the pure state the substance has a low soulubility in the ordinary organic solvents. It is insoluble in acid media. Basic solvenets. and dilute alkali effect solution radily (salt formation). 4. The substance could not be characterized or identified on the basis of its behavior towards the usual identification reagents. Its occurrence in nature has not previosly been reproted. 5. 1.5 mg. of hte crystalline hemorrhagic agent cause approximately the same reduciton in the prothrombin level or activity of the plasma of standardized susceptible reabbits in 40 hours as 50 gm. of the standard spoiled hay sample. 6. Spoiled sweet cover hays produced experimentally from Melilotus alba and those relized in agricultural parctice contain approximately 0.003 per cent of the hemorrhagic agent on the dry substance basis. The over‐all yield of the subsance in a fractionation scheme involving sixteen steps approximates 86 per cent of the quantity present.

A facile and practical p-Toluenesulfonic acid catalyzed route to dicoumarols containing an Aroyl group

“…1 This compound is used for the prevention and treatment of thrombosis. Furthermore, dicoumarol derivatives exhibit bioactivity as inhibitor of reductases.…”

Vitamin K and Bone: Past, Present, and Future

“…This is derived from the observation that the incidence of osteoporotic fracture is relatively lower in regions of the country where dietary intake of natto is high. (23) There are also some suggestions that the anti-osteoporotic effects of MK 4 are independent of g-carboxylation.…”

“…(24) In a recent meta-analysis of 13 small randomized controlled trials with data on bone loss, the authors concluded that supplementation with either MK 4 or K 1 has a positive effect on BMD. Further analysis, including only the trials that provided both BMD and fracture data, concluded that supplementation with 45 mg/d of menatetrenone was associated with increased BMD and reduced fracture incidence.…”

“…(2,3) Almost simultaneously, dicoumarol was identified as the causative agent of a hemorrhagic disease in cattle fed spoiled sweet clover hay, and its derivative, warfarin, was established as a potent anticoagulant. (4) The mechanism of action of vitamin K remained unknown until 1974 when the modified amino acid, gcarboxyglutamic acid (Gla), was identified in normal prothrombin. (5,6) The formation of Gla occurs through a unique vitamin K-dependent post-translational carboxylation of specific peptide-bound glutamate residues in a process that is inhibited by warfarin.…”

Place of Dabigatran in Contemporary Pharmacotherapy

“…The hemorrhagic factor isolated from sweet clover was first described in an article published in 1940. 2 One year later, the identity of this hemorrhagic agent was characterized, and the first anticoagulant, dicumarol, was discovered. 3 The first branded form of warfarin, developed by the Wisconsin Alumni Research Foundation, was approved by the FDA on June 8, 1954.…”