Synthesis of Succinimidyl 4-(N-Maleimidomethyl)-Cyclohexane-1-Carboxylate (Smcc) As a Linker and Conjugating Trastuzumab to Maytansinoid Derivative (Dm1) Through Smcc As an Antibody-Drug Conjugate Publisher



Ahani M¹ ; Salarian M¹ ; Salehi M¹ ; Jalili N¹ ; Shafiee S¹ ; Taheri A¹ ; Farahmand L¹
Source: Pharmaceutical Chemistry Journal Published:2023

Abstract

HER2 is overexpressed in 20–25% of breast cancer cases and is responsible for more aggressive tumor behavior. Monoclonal antibodies such as trastuzumab can specifically bind to targeted antigen on the surface of cancer cells and have been approved for the treatment of HER2-positive breast cancer. In order to increase the efficiency in cancer therapy, antibody–drug conjugates (ADCs) can be used that combine the antigen-targeting specificity of monoclonal antibodies with the cytotoxicity of chemotherapeutic drugs. All three components of ADCs affect the efficacy and toxicity of the drug. In this study, an ADC was synthesized by conjugating trastuzumab to maytansinol. Succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) was synthesized in five different steps and compounds at each step were analyzed using infrared spectroscopy. The structure of the final product was confirmed using NMR. DM1 attached to SMCC in chloroform solvent and was purified by recrystallization. Antibody purification was performed by chromatography and bound to SMCC-DM1. SMCC was obtained with a 75.8% yield and melting point at 187–189°C. Conjugation of DM1 with SMCC occurred in acetonitrile. Antibodies with a concentration of 10 mg/mL and SMCC-DM1 with concentration of 20 mg/mL had the best results in terms of drug-bound antibody production. ADCs are a promising approach to the targeted therapy of cancer. One way to achieve safe and potent drugs is to optimize each component before synthesis, including the selection of the optimal specific antibody against the targeted antigen, the conjugation method, and the chemical properties of the linker. © 2024, Springer Science+Business Media, LLC, part of Springer Nature.