Synthesis and In Vitro Activity of Novel Melphalan Analogs in Hematological Malignancy Cells.

Anastazja Poczta,Piotr Krzeczyński,Agnieszka Marczak,Arkadiusz Gajek,Joanna Tobiasz,Aneta Rogalska

doi:10.3390/ijms23031760

Abstract

Despite the continuous developments in pharmacology and the high therapeutic effect of new treatment options for patients with hematological malignancies, these diseases remain a major health issue. Our study aimed to synthesize, analyze in silico, and determine the biological properties of new melphalan derivatives. We obtained three methyl esters of melphalan having in their structures amidine moieties substituted with thiomorpholine (EM–T–MEL), indoline (EM–I–MEL), or 4-(4-morpholinyl) piperidine (EM–MORPIP–MEL). These have not yet been described in the literature. The in vitro anticancer properties of the analogs were determined against THP1, HL60, and RPMI8226 cells. Melphalan derivatives were evaluated for cytotoxicity (resazurin viability assay), genotoxicity (alkaline comet assay), and their ability to induce apoptosis (Hoechst33342/propidium iodide double staining method; phosphatidylserine translocation; and caspase 3/7, 8, and 9 activity measurements). Changes in mitochondrial membrane potential were examined using the specific fluorescence probe JC–1 (5,5′,6,6′-tetrachloro-1,1′,3,3′–tetraethylbenzimidazol carbocyanine). The EM–T–MEL derivative had the highest biological activity, showing higher cytotoxic and genotoxic properties than the parent drug. Moreover, it showed a high ability to induce apoptosis in the tested cancer cells. This compound also had a beneficial effect in peripheral blood mononuclear cells (PBMC). In conclusion, we verified and confirmed the hypothesis that chemical modifications of the melphalan structure improved its anticancer properties. The conducted study allowed the selection of the compound with the highest biological activity and provided a basis for chemical structure-biological activity analyses.

Highlights

Introduction published maps and institutional affilBlood cancers are currently among the most serious medical problems
Samples of melphalan derivatives used in biological research were obtained by threestage chemical synthesis from commercially available substrates (Figure 1)
The first step in the synthesis involved the reaction of commercial melphalan (MEL)

Summary

Introduction

Introduction published maps and institutional affilBlood cancers are currently among the most serious medical problems. Recent data have shown that one in eight cancer cases worldwide come from blood cells, bone marrow, or the lymphatic system. Malignant hematopoiesis, such as leukemia and lymphomas, is divided into more than 100 subtypes with varying survival rates [1]. Multiple myeloma (MM) is characterized by the infiltration of monoclonal plasma cells into the bone marrow, which secretes monoclonal immunoglobulin that is found in the urine and/or blood. Treatment of MM involves a combination of drugs with distinct mechanisms of action, including immunomodulating drugs, proteasome inhibitors, monoclonal antibodies, alkylating agents, and histone deacetylase inhibitors Despite these numerous modern therapies, high doses of melphalan followed by autologous stem cell transplantation remain key in iations

Methods

Results

Discussion

Conclusion