A Novel Method to Synthesize Phosphocreatine and Phosphocreatine Prodrugs

摘要

Background: Adenosine triphosphate (ATP) is the energy currency of the body; it takespart in various and indispensable metabolic processes for the maintenance of cell homeostasis, degradingto its hydrolysis product, adenosine diphosphate (ADP). Efficient ways to restore ATP aretherefore necessary in the cells. When the cell lacks energy due to ischemic conditions or highATP demand, phosphocreatine gives its phosphate group to ADP that converts to ATP, in a reactioncatalyzed by the enzyme creatine kinase. For this reason, phosphocreatine is utilized as apharmacological treatment in human diseases that involve a failure of the cellular energy, most notablyin coronary artery disease.Objective: Commercially available phosphocreatine is currently synthesized using different methods,each of one characterized by a rather low yield of the final product, probably due to the lowreactivity of the guanylating reagent. The aim of this work is to overcome the drawbacks of thesynthetic methods currently employed, devising a novel synthetic route to obtain phosphocreatineand phosphocreatine prodrugs in higher yields and purity.Method: To obtain a higher yield of the final product and a lower number of sub-products, thismethod utilizes a new guanylating agent characterized by high reactivity, endowed with a protectinggroup t-Boc on one of the two nitrogen atoms of the guanidinic function and a protected phosphateon the other one; that compound is then conjugated with an opportune secondary amine. Theobtained product is cleaved first with acidic conditions to obtain the phosphocreatine prodrug(phosphocreatine ethyl ester) and then with an enzymatic method to obtain the phosphocreatine.Result: Have been obtained in good yield and purity as demonstrated by HPLC and mass spectrometryanalysis.Conclusion: This novel synthetic route permits to obtain the phosphocreatine molecule in higheryield and purity compared to the methods currently employed with a combination of chemical andenzymatic methods.