BMS-202

The programmed death-ligand 1 (PD-L1) protein plays a key role in immune responses. Scutellarin (SCU), as a flavonoid, has a variety of bioactivities. In this study, the human PD-L1 was obtained by expression and purification, and the interaction mechanisms between PD-L1 and SCU were revealed through multi-spectroscopy and computer simulation. Fluorescence data indicated that the quenching of PD-L1 by SCU was mainly static quenching, and primarily driven by hydrogen bonding and van der Waals forces. The binding constant (K_a) was decreased from 2.05 ± 0.55 × 10⁴ L·mol^-1 to 0.28 ± 0.08 × 10⁴ L·mol^-1 with increasing temperature. Meanwhile, the changes in the microenvironment of PD-L1 were revealed by the synchronous and the 3D fluorescence data. In addition, the melting temperature of PD-L1 increased by 1.67 °C after binding with SCU. Moreover, the circular dichroism data showed that SCU changed the secondary structure of PD-L1 by increasing α-helix content and decreasing β-sheet content. Furthermore, the binding modes between SCU and PD-L1 and the key residues involved in the interaction were revealed by molecular docking and molecular dynamics. These findings supported SCU as an alternative ICB therapeutic strategy and provided evidence for computer-based drug design strategies.

We have previously identified a latent interaction mechanism between non-small cell lung cancer cells (NSCLCC) and their associated macrophages (TAM) mediated by mutual paracrine activation of the HMGB1/RAGE/NF-κB signaling. Activation of this mechanism results in TAM stimulation and PD-L1 upregulation in the NSCLCC. In the present work, we found that free DOX at a low concentration that does not cause DNA damage could activate the HMGB1/RAGE/NF-κB/PD-L1 pathway byinducing oxidative stress. It was thus proposed that a combination of low-dose DOX and a PD-L1 blocker delivered in the NSCLC tumor would achieve synergistic TAM stimulation and thereby synergetic anti-tumor potency. To prove this idea, DOX and BMS-202 (a PD-L1 blocker) were loaded to black phosphorus (BP) nanoparticles after dosage titration to yield the BMS-202/DOX@BP composites that rapidly disintegrated and released drug cargo upon mild photothermal heating at 40 °C. In vitro experiments then demonstrated that low-dose DOX and BMS-202 delivered via BMS-202/DOX@BP under mild photothermia displayed enhanced tumor cell toxicity with a potent synergism only in the presence of TAM. This enhanced synergism was due to an anti-tumor M1-like TAM phenotype that was synergistically induced by low dose DOX plus BMS-202 only in the presence of the tumor cells, indicating the damaged tumor cells to be the cardinal contributor to the M1-like TAM stimulation. In vivo, BMS-202/DOX@BP under mild photothermia exhibited targeted delivery to NSCLC graft tumors in mice and synergistic anti-tumor efficacy of delivered DOX and BMS-202. In conclusion, low-dose DOX in combination with a PD-L1 blocker is an effective strategy to turn TAM against their host tumor cells exploiting the HMGB1/RAGE/NF-κB/PD-L1 pathway. The synergetic actions involved highlight the value of TAM and the significance of modulating tumor cell-TAM cross-talk in tumor therapy. Photothermia-responsive BP provides an efficient platform to translate this strategy into targeted, efficacious tumor therapy.