A novel enediyne‐integrated antibody–drug conjugate shows promising antitumor efficacy against CD30⁺ lymphomas

Abstract

CD30 is a 120‐kDa type I transmembrane glycoprotein belonging to the tumor necrosis factor receptor superfamily. Overexpression of CD30 has been reported in Hodgkin's lymphoma (HL) and anaplastic large‐cell lymphoma (ALCL). CD30‐targeted treatment with antibody–drug conjugates (ADCs) can lead to promising clinical benefit. Lidamycin (LDM), consisting of an apoprotein LDP and an active enediyne chromophore AE, is a member of the enediyne antibiotic family and one of the most potent antitumor agents. AE and LDP can be dissociated and reconstituted under certain conditions in vitro. LDM is an ideal payload for the preparation of ADCs. In this study, we show the generation, production, and antitumor activity of anti‐CD30‐LDM, a novel ADC which consists of the intact anti‐CD30 antibody and LDM. First, the anti‐CD30‐LDP fusion protein was constructed and expressed in CHO/dhFr− cells. Anti‐CD30‐LDP showed specific and high‐affinity binding to CD30 and could be internalized into target cells. It also exhibited excellent tumor‐targeting capability in vivo. Next, anti‐CD30‐LDM was prepared by assembling the enediyne molecule AE to the fusion protein anti‐CD30‐LDP. Anti‐CD30‐LDM was highly cytotoxic to HL and ALCL cell lines, with IC50 values of 5–50 pm. It can also induce cell apoptosis and G2/M cell cycle arrest. In the Karpas299 xenograft model, the tumor growth was inhibited by 87.76% in mice treated with anti‐CD30‐LDM and with no discernible adverse effects. Taken together, anti‐CD30‐LDM shows attractive tumor‐targeting capability and antitumor efficacy both in vitro and in vivo and could be a promising candidate for the treatment of CD30+ lymphomas.

Preliminary biological evaluation of 123I-labelled anti-CD30-LDM in CD30-positive lymphomas murine models

“…Therefore, we speculated that anti-CD30-LDM might have the potential to overcome resistance induced by BV. We had reported anti-CD30-LDM either alone or combination with crizotinib show excellent antitumor activities in CD30þ/ALK þ anaplastic large cell lymphoma both in vitro and in vivo [13,16]. It indicated that anti-CD30-LDM could be a promising candidate for the treatment of CD30þ lymphomas.…”

“…The composition of the ADC was finally confirmed by reverse-phase HPLC. For more detailed information, see literature [13].…”

“…We have previously described a novel ADC, anti-CD30-LDM [13], which is comprised of two molecules of LDM linked to an anti-CD30 monoclonal antibody (mAb) via noncleavable linkers. Thus, the LDM is delivered to the CD30positive cancer cells by the mAb, causes DNA damage by inducing double-strand breaks, and results in cell cycle arrest and apoptosis [14,15].…”

“…Thus, the LDM is delivered to the CD30positive cancer cells by the mAb, causes DNA damage by inducing double-strand breaks, and results in cell cycle arrest and apoptosis [14,15]. This ADC displays specific affinity and extremely potent cytotoxicity against CD30-overexpressing tumour cells in vitro, with IC 50 (half-maximal inhibitory concentration) values ranging from 0.54 Â 10 À11 to 3.74 Â 10 À11 M. In a Karpas299 xenograft model, tumour growth was inhibited by 87.76% in mice treated with 0.7 mg/kg anti-CD30-LDM [13]. The combination of anti-CD30-LDM with crizotinib exhibits a synergistic inhibitory antitumor effect in ALCL [16].…”

<p>Synergistic Treatment of Tumor by Targeted Biotherapy and Chemotherapy via Site-Specific Anchoring of Aptamers on DNA Nanotubes</p>

“…Samples were collected by centrifugation (14,000×g, 10 min, 4°C) and quantified by a BCA kit (Bradford Protein Assays, Thermo Fisher Scientific, Germany). 24 Subsequently, the samples (20 µg) were separated on a sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE; 12%, 120 V, 1 h) and transferred onto a polyvinylidene fluoride (PVDF) membrane via wet transfer (200 mA, 1 h). After being blocked with nonfat milk at room temperature (5%, in PBST (0.1% of Tween 20 in PBS); 3 h, 100 rpm), the membrane was incubated with primary antibody (1:1000; PBST; overnight, 4°C).…”