Computational and experimental therapeutic efficacy analysis of andrographolide phospholipid complex self-assembled
Atul Mourya1, Purva Pingle1, Chanti Katta Babu1
1Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research, Hyderabad, Telangana, India.
Background:
Neuroblastoma is one of the most common malignancies in childhood, accounts for approximately 7% of all malignancies. Andrographolide (AN) inhibits cancer cells progression via multiple pathways like cell cycle arrest, mitochondrial apoptosis, NF-κβ inhibition, and antiangiogenesis mechanism. Despite multiple advantages, application of AN is very limited due to its low aqueous solubility (6.39 ± 0.47 μg/mL), high lipophilicity (log P ∼ 2.632 ± 0.135), and reduced stability owing to pH sensitive lactone ring.
Objectives And Results:
In present investigation, a molecular complex of AN with soya-L-α-phosphatidyl choline (SPC) was synthesized as ANSPC and characterized by FT-IR and1H NMR spectroscopy. Spectral and molecular simulation techniques confirmed the intermolecular interactions between the 14-OH group of AN and the N+(CH3)3part of SPC. In addition, molecular dynamics (MD) simulation was used to determine the degree of interaction between various proteins such as TNF-α, caspase-3, and Bcl-2. Later, ANSPC complex was transformed in to self-assembled soft nanoparticles of size 201.8 ± 1.48 nm with PDI of 0.092 ± 0.004 and zeta potential of -21.7 ± 0.85 mV. The IC50 offree AN (8.319 μg/mL) and the self-assembled soft ANSPC nanoparticles (3.406 μg/mL ∼ 1.2 μg of AN) against Neuro2a cells was estimated with significant (P < 0.05) difference. Interestingly, the self-assembled soft ANSPC nanoparticles showed better endocytosis compared to free AN in Neuro2a cells. In-vitrobiological assays confirmed that self-assembled soft ANSPC nanoparticles induces apoptosis in Neuro2a cells by declining the MMP (Δψm) and increasing the ROS generation.
Conclusion:
Self-assembled soft ANSPC nanoparticles warrant further in-depth antitumor study in xenograft model of neuroblastoma to establish the anticancer potential.
Insights
This study developed self-assembled nanoparticles of Andrographolide (AN) with soya-L-α-phosphatidyl choline (SPC), named ANSPC. These ANSPC nanoparticles show enhanced anticancer effects against neuroblastoma cells, warranting further investigation.
Area of Science:
- Nanotechnology
- Pharmacology
- Biochemistry
Background:
- Neuroblastoma is a common childhood cancer.
- Andrographolide (AN) has anticancer properties but poor solubility and stability limit its use.
- Developing effective drug delivery systems for AN is crucial.
Purpose of the Study:
- To synthesize and characterize a molecular complex of AN with soya-L-α-phosphatidyl choline (SPC), named ANSPC.
- To formulate ANSPC into self-assembled soft nanoparticles.
- To evaluate the anticancer efficacy and cellular uptake of ANSPC nanoparticles against neuroblastoma cells.
Main Methods:
- Synthesis and characterization of ANSPC complex using FT-IR and 1H NMR.
- Molecular dynamics simulations to study interactions.
- Formation of self-assembled soft nanoparticles.
- In vitro evaluation of cytotoxicity, endocytosis, and apoptosis induction in Neuro2a cells.
Main Results:
- ANSPC complex formation was confirmed spectroscopically.
- Self-assembled ANSPC nanoparticles (201.8 nm) were successfully created.
- ANSPC nanoparticles exhibited significantly higher cytotoxicity (lower IC50) against Neuro2a cells compared to free AN.
- Enhanced cellular uptake and apoptosis induction (decreased MMP, increased ROS) were observed with ANSPC nanoparticles.
Conclusions:
- Self-assembled ANSPC nanoparticles represent a promising delivery system for Andrographolide.
- ANSPC nanoparticles demonstrate enhanced anticancer activity against neuroblastoma cells in vitro.
- Further studies in xenograft models are warranted to establish the in vivo anticancer potential.
