Identification, synthesis and structural confirmation of process-related impurities in proparacaine hydrochloride.

Proparacaine hydrochloride is an ester-type local anesthetic agent that is extensively used in ophthalmic operations. The process-related impurities of proparacaine hydrochloride were investigated, and seven impurities were detected in the reaction solution of the last step at the level of 0.03-1.08 % by a newly developed HPLC method. Based on the synthetic process and the results of LC-HRMS, the structures of five impurities were proposed as 3-amino-4-propoxybenzoic acid (Imp-A), ethyl 3-amino-4-propoxybenzoate (Imp-B), 2-(ethylamino)ethyl 3-amino-4-propoxybenzoate hydrochloride (Imp-C), 2-(diethylamino)ethyl 3-nitro-4-propoxybenzoate hydrochloride (Imp-F), and 3-nitro-4-propoxybenzoic acid (Imp-G). And the structures were confirmed by synthesis, followed by varieties of spectral and chromatographic analyses. The structures of two impurities with almost same molecular weight in LC-HRMS were elucidated as 2-(diethylamino)ethyl 3-(ethylamino)-4-propoxy-benzoate hydrochloride (Imp-D) and 2-(diethylamino)ethyl 3-formamido-4-propoxybenzoate hydrochloride (Imp-E) by NMR and IR. An HPLC-based method was developed, and validation study demonstrated that the approach was precise, accurate, and sensitive. The impurities information of proparacaine hydrochloride can be used for the quality control of intermediate, raw material drug and its commercial products.

Proparacaine complexation with beta-cyclodextrin and p-sulfonic acid calix[6]arene, as evaluated by varied (1)H-NMR approaches.

This study focused on the use of NMR techniques as a tool for the investigation of complex formation between proparacaine and cyclodextrins (CDs) or p-sulfonic acid calix[6]arene. The pH dependence of the complexation of proparacaine with beta-CD and p-sulfonic acid calix[6]arene was studied and binding constants were determined by (1)H NMR spectroscopy [diffusion-ordered spectroscopy (DOSY)] for the charged and uncharged forms of the local anesthetic in beta-CD and p-sulfonic acid calix[6]arene. The stoichiometries of the complexes was determined and rotating frame Overhauser enhancement spectroscopy (ROESY) 1D experiments revealed details of the molecular insertion of proparacaine into the beta-CD and p-sulfonic acid calix[6]arene cavities. The results unambiguously demonstrate that pH is an important factor for the development of supramolecular architectures based on beta-CD and p-sulfonic acid calix[6]arene as the host molecules. Such host-guest complexes were investigated in view of their potential use as new therapeutic formulations, designed to increase the bioavailability and/or to decrease the systemic toxicity of proparacaine in anesthesia procedures.

Topology of a ternary complex (proparacaine-beta-cyclodextrin-liposome) by STD NMR.

The topologies of proparacaine (PPC) in beta-cyclodextrin (beta-CD), PPC in egg phosphatidylcholine (EPC) liposomes and PPC in beta-CD in EPC were investigated using NMR experiments (1D ROESY and saturation transfer difference (STD)). This is the first description of the STD technique applied to PPC-EPC-beta-CD system, revealing that not only PPC was imbedded in EPC bilayer, but beta-CD was also interacting with liposome vesicles. These results are novel and were rationalized as the spontaneous formation of a ternary complex with some beta-CD molecules bound to external liposome vesicles surfaces.