Chalcogenide Variation In Organic Conjugated Molecules Based Nanoagents For Non-small Cell Lung Cancer Treatment

BackgroundLung cancer is the most common malignant tumor among adults,and non-small cell lung cancer accounts for about 80%.Recent popular therapies include chemotherapy,small molecule therapy,immunotherapy and a combination of these strategies.However,chemotherapy usually causes systemic side effects,while radiation therapy is limited by the cumulative radiation dose.In recent years,photoacoustic imaging(PAI)guided photothermal therapy(PTT)and photodynamic therapy(PDT)have received more and more attention.Compared with the disadvantages of biological tissue toxicity and non-degradability of inorganic nanomaterials,organic conjugated compound nanomaterials have the advantages of higher biocompatibility and biosecurity,higher photothermal conversion efficiency and stronger modification.The development of organic conjugated compound NIR nanomaterials and how to improve the photothermal properties of materials are still one of the current hot spots.Based on this idea,this article focuses on the molecular engineering of chalcogen-substituted organic conjugated near-infrared nanodiagnostic agents(PTA)to explore the relationship between the molecular structure of organic compounds and the photothermal or photodynamic properties,as well as the role of non-small cell lung cancer treatment.First,three kinds of bis-diketopyrrolopyrrole conjugated polymer are designed and synthesized,where only a single heteroatom of acceptor units changes from oxygen to sulfur to selenium,allowing for systematic investigation of the molecular structure-PTT property relationship.Second,thionated cis-/trans-isomer PDI-CS and PDI-TS were designed and prepared to investigate thionation engineering on therapeutic performance.ObjectiveTo construct a photothermal nanoparticles with high photothermal stability,good extinction coefficient and high photothermal conversion efficiency,and to explore its effect of non-small lung cancer cells as a photothermal preparation.Method1.The preparation and characterization of PTA:The structure and optical properties were investigated by UV-vis-NIR spectroscopy and 1H NMR.2.The preparation and characterization of PTA NPs:The nanoparticles were constructed using the coprecipitation method,and the particle size and appearance of the nanoparticles were detected3.Detection of photothermal properties:PTA NPs was tested for concentration dependence,power dependence,photothermal stability and photothermal conversion efficiency4.Detection of photodynamic properties:1,3-Diphenylisobenzofuran(DPBF)was used as the active oxygen generation indicator to detect the photodynamic performance of PTA under laser irradiation.5.In vitro research on non-small cell lung cancer cells(A549):1)Cell proliferation experiment:CCK-8 method was used to detect the toxic effects of different experimental groups on A549 cells with or without laser irradiation2)Cell fluorescence detection of anti-A549 cells in vitro:The cytotoxic effect of PTA NPs on A549 cells under laser irradiation was detected by calcein-AM/Propidium iodide double staining experiment.Annexin V-FITC/PI double staining was used to detect the toxic effect of PTA NPs on A549 cells under laser irradiation6.In vivo anti-small cell lung cancer research:1)Construction and grouping of non-small cell lung cancer tumor models:select right age nude mice and subcutaneous inoculation of A549 cells.When the tumor was about 100 mm3,it was randomly divided into four groups,five in each group2)Photoacoustic imaging experiment:PA imaging in nude mice was performed by MOST imaging system3)Anti-tumor effect evaluation:record the tumor volume and weight of each group,compare and evaluate the anti-tumor effect4)Biological safety assessment:testing the blood routine and important biochemical indicators of nude mice and staining the main organs of nude mice with hematoxylin and eosin(H&E)to evaluate the efficacy and biological safetyResult1.three kinds of bis-diketopyrrolopyrrole conjugated polymer(DPP-SO,DPP-SS,DPP-SSe)are designed and synthesized to study how the chalcogen atom(from oxygen to sulfur to selenium atom)engineering affects the therapeutic performance of cancer in vivo and in vitro1)Preparation and characterization of CPs:The absorption and extinction coefficient of these CPs can be facilely tuned by changing the heteroatoms of the acceptor units2)Preparation and characterization of CPNs:the three CPNs have a similar particle size of about 65-75 nm and are uniformly spherical3)Detection of photothermal properties:After calculation,their photothermal conversion efficiency is similar(74%-79%).Among them,DPP-SO showed the best mass extinction coefficient(66.5 1 L g-1 cm-1),which resulted in the best photothermal performance4)Anti-A549 cells in vitro:At the cellular level,the half-maximum inhibitory concentration(IC50)of DPP-SO NPs,DPP-SS and DPP-SSe NPs on A549 cells was 4.78,9.29 and 10.59 ?g mL-1,respectively.Cell flow cytometry detection of DPP-SO NPs showed that about 68%of A549 cell apoptosis,of which 60%is late apoptosis5)In vivo anti-non-small cell lung cancer research:DPP-SO NPs can be used as a contrast agent for photoacoustic imaging to monitor the accumulation of nanoparticles at the tumor site in real time.Finally,a subcutaneous A549 tumor-bearing nude mouse model was established for PTT detection of DPP-SO NPs2.Thionated cis-/trans-isomer PDI-CS and PDI-TS were designed and prepared to investigate thionation engineering on therapeutic performance1)Synthesis and characterization of PDI-CS and PDI-TS:1H,13C and 1H-1H NOESY 2D NMR of nuclear magnetic resonance verified the correct structure of the compound.2)Preparation and characterization of PDI-CS and PDI-TS NPs:while they have a similar particle size of about 50 nm3)Detection of photothermal/photodynamic properties:The position of sulfur atoms has little effect on the generation of active oxygen.In sharp contrast,the photothermal conversion efficiency of trans-PDI-TS NPs(58.4%)is significantly higher than that of cis-PDI-CS NPs(41.2%)4)Cell proliferation test:on A549 cells,660 nm laser irradiation for 5 min,the IC50 of PDI-TS NPs is 7.78 ?g mL-1,which is about half of PDI-CS NPs(13.23 ?g mL-1).PDI-TS NPs have better photothermal properties than PDI-CS NPs.Cell flow cytometry analysis showed that 60.68%of A549 cells died in the form of apoptosis,and most of them were in the state of late apoptosis5)In vivo anti-small cell lung cancer research:PDI-TS NPs can be used as a contrast agent for photoacoustic imaging to monitor the accumulation of nanoparticles in the tumor site in real time,so as to achieve precise and maximum ablation of cells by PDI-TS NPs Change.Under the guidance of photoacoustic imaging,PDI-TS NPs have obvious ablation effect and excellent biosecurity on A549 tumor-bearing nude miceConclusion1.Three kinds of bis-diketopyrrolopyrrole conjugated polymer(CPs:DPP-SO,DPP-SS and DPP-SSe)and their nanoparticles(CPNs)were successfully prepared.Experiments proved that changing the heteroatoms in a DPP unit from oxygen to sulfur to selenium can easily adjust the absorption spectrum of CPs and light-heat conversion efficiency of CPNs.Thus,this work provides a simple but novel strategy to manipulate the photothermal performance of CPs through changing the chalcogen atom at key positions2.Successfully prepared thio trans-isomer PDI-TS and cis-isomer PDI-CS and their nanoparticles(NPs).The experiment proves that photothermal performance of PDI-TS is better than PDI-CS,which is attributed to the fact that the strong ?-? and C…S interactions in nanoagents,resulting in a higher photothermal conversion efficiency of PDI-TS.This work provides simple but novel concept-to-proof of synergistically photothermal/photodynamic treatment of tumor through regioisomer-modulated engineering.