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. 2025 Feb 4;17(2):191.
doi: 10.3390/pharmaceutics17020191.

Exploratory Study on Nanoparticle Co-Delivery of Temozolomide and Ligustilide for Enhanced Brain Tumor Therapy

Gang Ke  1   2 , Mingxia Zhang  1 , Pengyi Hu  1 , Jing Zhang  1 , Abid Naeem  3 , Lianfang Wang  1 , Huixin Xu  1 , Yu Liu  1 , Ming Cao  1 , Qin Zheng  1
Affiliations

Exploratory Study on Nanoparticle Co-Delivery of Temozolomide and Ligustilide for Enhanced Brain Tumor Therapy

Gang Ke et al. Pharmaceutics. .

Abstract

Background: Temozolomide (TMZ) is the first-line therapy for glioblastoma (GBM), but its clinical efficacy is limited by its short half-life, poor brain targeting, adverse side effects, and the development of drug resistance. Ligustilide (LIG) has been shown to enhance blood-brain barrier permeability and reduce P-glycoprotein activity, thereby potentiating the synergistic effect of TMZ against GBM. Methods: The dual-drug-loaded nanoparticles encapsulating both TMZ and LIG (TMZ/LIG-NPs) were prepared using Poly (d,l-lactic-co-glycolide)-monomethoxy poly (ethylene glycol) (PLGA-mPEG). The physicochemical properties of the NPs, including particle size and zeta potential, were characterized. Cellular uptake of NPs was evaluated using flow cytometry and fluorescence staining. The pharmacokinetic profile and cytotoxicity of TMZ/LIG-NPs were compared to those of free TMZ and a mixture of TMZ and LIG in rat and glioma cells, respectively. Results: The mean particle size of TMZ/LIG-NPs was 117.6 ± 0.7 nm, with a zeta potential of -26.5 ± 0.4 mV. Cellular uptake of NPs was significantly higher than that of free drug in U251 cells. Encapsulation of TMZ in NPs significantly increased its half-life by 1.62-fold compared to free TMZ and significantly improved its pharmacokinetic profile. Moreover, the storage stability of the TMZ/LIG-NPs solution was extended to one month. The toxicity of TMZ/LIG-NPs to glioma cells C6 and U251 was markedly enhanced compared to the mixture of TMZ and LIG. Conclusions: The development of TMZ/LIG-NPs using PLGA-mPEG effectively enhanced the stability and efficacy of both TMZ and LIG. This dual drug-loaded nanoparticle system represents a promising strategy for glioblastoma therapy.

Keywords: dual drug-loaded nanoparticles; glioblastoma multiforme; ligustilide; temozolomide.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Chemical structure of (A) temozolomide and (B) ligustilide.
Figure 2
Figure 2
Preparation of TMZ/LIG-NPs.
Figure 3
Figure 3
Synergistic inhibitory effects and Q values of TMZ and LIG on U251 cells. Cell inhibition of TMZ (A) and LIG (B). (C) Q values of TMZ (40 µg/mL) with LIG (5–30 µg/mL). (D) Q values of combination groups for TMZ and LIG. Values are the mean ± standard deviation (n = 6).
Figure 4
Figure 4
Characteristics of TMZ/LIG-NPs. (A) Particle-size distribution by DLS. (B) SEM. Scale bar = 500 nm. (C) TEM. Scale bar = 100 nm.
Figure 5
Figure 5
Stability upon dilution and storage and the matrix effect. (A) Mean particle size, PDI, and zeta potential after dilution. (B) Mean particle size, PDI, and zeta potential after storage (50-fold dilution). (C) Mean particle size, PDI, and zeta potential to test matrix effect. Values are the mean ± standard deviation (n = 3), *** p < 0.001., **** p < 0.0001.
Figure 6
Figure 6
In vitro release of TMZ and LIG. (A) TMZ release. (B) LIG release. Values are the mean ± standard deviation (n = 3).
Figure 7
Figure 7
Pharmacokinetics and brain distribution. (A) Concentration-time profiles of TMZ in blood after injection with TMZ, CTL, TMZ/LIG-NPs via the tail vein. Values are the mean ± standard deviation (n = 6). (B) Concentration of TMZ in brain at 3 h after post-dose of TMZ, CTL, or TMZ/LIG- NPs. Values are the mean ± standard deviation (n = 3), * p < 0.05, *** p < 0.001.
Figure 8
Figure 8
Uptake of free Cou-6 and Cou-6-NPs in U251 cells. (A) Time-dependent mean fluorescence intensity according to flow cytometry. (B) Representative FITC spectra. (C) Representative fluorescence-microscope images after 4-h incubation. Cell nuclei were stained with DAPI. Scale bar = 100 μm. Values are the mean ± standard deviation (n = 3). Differs from free Cou-6 group: **** p < 0.0001.
Figure 9
Figure 9
Inhibition by CTL and TMZ/LIG-NPs on U251 cells (A) and C6 cells (B). Values are the mean ± standard deviation (n = 3). **** p < 0.0001.
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References

    1. Ostrom Q.T., Patil N., Cioffi G., Waite K., Kruchko C., Barnholtz-Sloan J.S. CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2013–2017. Neuro-Oncology. 2020;22((Suppl. S1)):iv1–iv96. doi: 10.1093/neuonc/noaa200. - DOI - PMC - PubMed
    1. Stupp R., Hegi M.E., Mason W.P., van den Bent M.J., Taphoorn M.J.B., Janzer R.C., Ludwin S.K., Allgeier A., Fisher B., Belanger K., et al. Effects of Radiotherapy with Concomitant and Adjuvant Temozolomide versus Radiotherapy Alone on Survival in Glioblastoma in a Randomised Phase III Study: 5-Year Analysis of the EORTC-NCIC Trial. Lancet Oncol. 2009;10:459–466. doi: 10.1016/S1470-2045(09)70025-7. - DOI - PubMed
    1. Arko L., Katsyv I., Park G.E., Luan W.P., Park J.K. Experimental Approaches for the Treatment of Malignant Gliomas. Pharmacol. Ther. 2010;128:1–36. doi: 10.1016/j.pharmthera.2010.04.015. - DOI - PMC - PubMed
    1. Louis D.N., Perry A., Wesseling P., Brat D.J., Cree I.A., Figarella-Branger D., Hawkins C., Ng H.K., Pfister S.M., Reifenberger G., et al. The 2021 WHO Classification of Tumors of the Central Nervous System: A Summary. Neuro-Oncology. 2021;23:1231–1251. doi: 10.1093/neuonc/noab106. - DOI - PMC - PubMed
    1. Lee S.Y. Temozolomide Resistance in Glioblastoma Multiforme. Genes Dis. 2016;3:198–210. doi: 10.1016/j.gendis.2016.04.007. - DOI - PMC - PubMed

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