Pharmacophore hybridisation and nanoscale assembly to discover self-delivering lysosomotropic new-chemical entities for cancer therapy

Integration of the unique advantages of the fields of drug discovery and drug delivery is invaluable for the advancement of drug development. Here we propose a self-delivering one-component new-chemical-entity nanomedicine (ONN) strategy to improve cancer therapy through incorporation of the self-assembly principle into drug design. A lysosomotropic detergent (MSDH) and an autophagy inhibitor (Lys05) are hybridised to develop bisaminoquinoline derivatives that can intrinsically form nanoassemblies. The selected BAQ12 and BAQ13 ONNs are highly effective in inducing lysosomal disruption, lysosomal dysfunction and autophagy blockade and exhibit 30-fold higher antiproliferative activity than hydroxychloroquine used in clinical trials. These single-drug nanoparticles demonstrate excellent pharmacokinetic and toxicological profiles and dramatic antitumour efficacy in vivo. In addition, they are able to encapsulate and deliver additional drugs to tumour sites and are thus promising agents for autophagy inhibition-based combination therapy. Given their transdisciplinary advantages, these BAQ ONNs have enormous potential to improve cancer therapy.

In vivo toxicity studies. The toxicity of BAQ NPs was investigated on female FVB/N mice via iv injection.
Mice were administrated with various concentrations (10 mg kg -1 , 20 mg kg -1 or 40 mg kg -1 ) of Lys05, Liposomes@Lys05, BAQ12 NPs, and BAQ13 NPs every two days. The status of mice was monitored every day and their body weight was recorded every two days. Blood samples were collected and sent to the UCD Comparative Pathology Laboratory for tests of complete blood count (CBC) and serum chemistry.
In vivo pharmacokinetic study. The jugular vein of female Sprague-Dawley rats (200-250g) was implanted with a catheter for drug injection and blood collection (Harland, Indianapolis, IN, USA). Rats (n = 3) were injected with free DiD, BAQ12 NPs@DiD (10:1, mass ratio) and BAQ13 NPs@DiD (10:1, mass ratio), respectively, which contained an equivalent dose of DiD (0.5 mg kg -1 ). Blood samples were collected at the indicated time points and then were centrifuged to obtain the plasma. The plasma was diluted with DMSO (1:100), and the fluorescence intensity (λEx=595 nm, λEm=665 nm) was measured by a microplate reader (SpectraMax M2).
In/ex vivo biodistribution. Nude mice bearing the HT29 tumours were subjected to iv administration of BAQ13 NPs@DiD (10:1, mass ratio) at a dose of 1.0 mg kg -1 DiD. In vivo imaging studies were performed at the corresponding time point. Organs (brain, heart, lung, liver, spleen, kidney, intestines, and muscle) and tumours were collected from mice for ex vivo imaging. Biodistribution of BAQ13 NPs@ NAPA+DiD (10/2.5/1.0 mg kg -1 , iv) was studied on NRG mice bearing PCSC tumours. Both in vivo and ex vivo imaging studies were performed as above.
Supplementary Table 4 Primers and sequences used in RT-PCR analysis.