Apparent Cytotoxicity and Intrinsic Cytotoxicity of Lipid Nanomaterials Contained in a COVID-19 mRNA Vaccine

Authors

  • Gabriele Segalla Multichem R&D

DOI:

https://doi.org/10.56098/ijvtpr.v3i1.84

Keywords:

mRNA vaccine, LNP, lipid nanoparticles, ROS, reactive oxygen species, pKa, apparent pKa, intrinsic pKa

Abstract

The medicinal preparation called Comirnaty by Pfizer-BioNTech is an aqueous dispersion of lipid nanomaterials, intended to constitute, after thawing and dilution, the finished product for intramuscular injection. In the present study, we examine some evident chemical-physical criticalities of the preparation, particularly regarding the apparent and the intrinsic pKa (acid dissociation constant) of its main excipient, the ionizable cationic lipid ALC-0315. The very high value of its intrinsic pKa causes, after internalization and endosomal escape of LNPs, a sudden increase of its cationic charge concentration and consequently the formation of pro-inflammatory cytokines and ROS (reactive oxygen species), that can disrupt the mitochondrial membrane and release its content, cause RNA mistranslation, polymerization of proteins and DNA, DNA mutations, destruction of the nuclear membrane and consequent release of its content. Additionally, the apparently low pKa value (6.09) of ALC-0315 associated with other lipids in the LNP, is not suitable for intramuscular application. Its value is too low to enable a proper transfection of host cells, despite what is stated by EMA (European Medicines Agency) in its Assessment report dated 19 February 2021, in flagrant contradiction with the same bibliographic source therein cited. Furthermore, the exceptional penetrability, mobility, chemical reactivity and systemic accumulation of uncontrollable cationic lipid nanoparticles, with high cytotoxicity levels, shed in unpredictable biological locations, even far from the site of inoculation, are all factors that can lead to an unprecedented medical disaster. Meanwhile, further immediate studies and verifications are recommended, taking into consideration, in accordance with the precautionary principle, the immediate suspension of vaccinations with the COVID-19 mRNA- LNP-based vaccines.

References

Alabi, C., A., Love, K.T., Sahay, G., Anderson, D.G. (2013). Multiparametric approach for the evaluation of lipid nanoparticles for siRNA delivery. PNAS, 110 (32), 12881–12886. https://doi.org/10.1073/pnas.1306529110

Barone, F., De Angelis, I., Andreoli, C., Battistelli, C.L., Arcangeli, C., & Leter, G. (2017).Metodi in vitro e in silico per la valutazione del potenziale tossicologico dei nanomateriali[In vitroand in silicomethods for evaluating the toxicological potential of nanomaterials]. ENEA -Focus 3/2017 Energia, ambiente e innovazione. DOI 10.12910/EAI2017-045

Buschmann, M.D., Carrasco, M.J., Alishetty, S., Paige, M., Alameh, M.G., Weissman, D. (2021). Nanomaterial Delivery Systems for mRNA Vaccines. Vaccines, 9(1), 65. https://doi.org/10.3390/vaccines9010065

Buyens, K., De Smedt, S.C., Braeckmans, K., Demeester, J., Peeters, L., Van Grunsven, L.A., de Mollerat du Jeu, X., Sawant, R., Torchilin, V., Farkasova, K., Ogris, M., Sanders, N.N. (2012). Liposome based systems for systemic siRNA delivery: stability in blood sets the requirements for optimal carrier design. J. Control, 158, 362-370. https://doi.org/10.1016/j.jconrel.2011.10.009

Demple, B., Harrison, L. (1994). Repair of oxidative damage to DNA: enzymology and biology. Annu Rev Biochem. 63:915-48. doi: 10.1146/annurev.bi.63.070194.004411.

Draz, M.S., Fang, B.A., Zhang, P., Hu, Z., Gu, S., Weng, K.C., et al. (2014). Nanoparticle-mediated systemic delivery of siRNA for treatment of cancers and viral infections. Theranostics 4, 872. https://doi.org/10.7150/thno.9404

Fröhlich, E. (2012). The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles. Int J Nanomedicine. 7:5577-5591 https://doi.org/10.2147/IJN.S36111

Hassett, K., J., Benenato, K.E., Jacquinet, E., Lee, A., Woods, A., Yuzhakov, O., Himansu, S., Deterling, J., Geilich, B.M., Ketova, T., Mihai, C., Lynn, A., McFadyen, I., Moore, M.J., Senn, J.J., Stanton, M.G., Almarsson, Ö., Ciaramella, G., Brito, L.A. (2019). Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines. Molecular Therapy: Nucleic Acids Vol. 15, 1-11 https://doi.org/10.1016/j.omtn.2019.01.013

Hou, X., Zaks, T., Langer, R., Dong, Y. (2021). Lipid nanoparticles for mRNA delivery. Nat Rev Mater 6, 1078–1094. https://doi.org/10.1038/s41578-021-00358-0

Hu, Y.B., Dammer, E., Ren, R.J., Wang, G. (2015). The endosomal-lysosomal system: from acidification and cargo sorting to neurodegeneration. Transl Neurodegener 4, 18. https://doi.org/10.1186/s40035-015-0041-1

Imlay, J.A., Linn, S. (1988, June 3). DNA damage and oxygen radical toxicity. Science. 240(4857):1302-9. https://www.science.org/doi/10.1126/science.3287616

Jayaraman, M., Ansell, S.M., Mui, B.L., Tam, Y. K., Chen, J., Du, X., Butler, D., Eltepu, L., Matsuda, S., Narayanannair, J.K., Rajeev, K.G., Hafez, I.M., Akinc, A., Maier, M.A., Tracy, M.A., Cullis, P.R., Madden, T. D., Manoharan, M., Hope, M.J. (2012). Maximizing the Potency of siRNA Lipid Nanoparticles for Hepatic Gene Silencing In Vivo. Angew. Chem. Int. Ed., 51, 8529 –8533. https://doi.org/10.1002/anie.201203263

Kanasty, R.L, Whitehead, K.A, Vegas, A.J., Anderson, D.G. (2012). Action and Reaction: The Biological Response to siRNA and Its Delivery Vehicles. Molecular Therapy 20 (3), 513–524. https://doi.org/10.1038/mt.2011.294

Lee, J.M., Yoon, T.J., Cho, Y.S.. (2013). Recent developments in nanoparticle-based siRNA delivery for cancer therapy. BioMed Res. Int. 2013. https://doi.org/10.1155/2013/782041

Maki, H., Sekiguchi, M. (1992). MutT protein specifically hydrolyses a potent mutagenic substrate for DNA synthesis. Nature 355, 273–275. https://doi.org/10.1038/355273a0

Maruggi, G., Zhang, C., Li, J., Ulmer, J.B., Yu, D. (2019). mRNA as a Transformative Technology for Vaccine Development to Control Infectious Diseases. Molecular Therapy. https://doi.org/10.1016/j.ymthe.2019.01.020

Midoux, P., Pichon, C. (2015). Lipid-based mRNA vaccine delivery systems. Expert Review of Vaccines, 14 (2), 221-234. https://doi.org/10.1586/14760584.2015.986104

Nance, K.D., Meier, J.L. (2021). Modifications in an emergency: the role of n1-methylpseudouridine in COVID-19 vaccines. ACS Central Science, 7(5), 748–756. https://doi.org/10.1021/acscentsci.1c00197

Ndeupen, S., Qin, Z., Jacobsen, S., Bouteau, A., Estanbouli, H., Igyarto, B.Z. (2021). The mRNA-LNP platform’s lipid nanoparticle component used in preclinical vaccine studies is highly inflammatory. iScience 24, 103479. https://doi.org/10.1016/j.isci.2021.103479

Ozpolat, B., Sood, A.K., Lopez-Berestein, G. (2014). Liposomal siRNA nanocarriers for cancer therapy. Adv. Drug Deliv. Rev. 66, 110–116. https://doi.org/10.1016/j.addr.2013.12.008

Palmer, M., Bhakdi, S., Wodarg, W. (2022). Expertise on the genotoxic risks of the Pfizer COVID-19 vaccine. https://childrenshealthdefense.eu/wp-content/uploads/2022/07/att.-3-genotoxicity-mRNA-vaccines-scientific-report.pdf

Parry, P.I., Lefringhausen, A., Turni, C., Neil, C. J., Cosford, R., Hudson, N.J., Gillespie, J. (2023). ‘Spikeopathy’: COVID-19 Spike Protein Is Pathogenic, from Both Virus and Vaccine mRNA. Biomedicines, 11, 2287. https://doi.org/10.3390/biomedicines11082287

Patel, P., Ibrahim, N.M., Cheng, K. (2021). The Importance of Apparent pKa in the Development of Nanoparticles Encapsulating siRNA and mRNA. Trends in Pharmacological Sciences, 42(6), 448–460. https://doi.org/10.1016/j.tips.2021.03.002

Patel, S., Kim, J., Herrera, M., Mukherjee, A., Kabanov, A.V., Sahay, G. (2019). Brief update on endocytosis of nanomedicines. Advanced Drug Delivery Reviews, 144, 90-111. https://doi.org/10.1016%2Fj.addr.2019.08.004

Sahay G., Alakhova, D.Y., Kabanov, A.V. (2010) Endocytosis of nanomedicines. J Control Release, 145-182 https://doi.org/10.1016/j.jconrel.2010.01.036

Szebeni, J. (2014). Complement activation-related pseudoallergy: a stress reaction in blood triggered by nanomedicines and biologicals. Mol. Immunol. 61, 163–173. https://doi.org/10.1016/j.molimm.2014.06.038

Segalla, G. (2023). Chemical-physical criticality and toxicological potential of lipid nanomaterials contained in a COVID-19 mRNA vaccine. International Journal of Vaccine Theory, Practice, and Research, 3(1), 787–817. https://doi.org/10.56098/ijvtpr.v3i1.68

Tam, Y.Y.C., Chen, S., Cullis, P.R. (2013). Advances in lipid nanoparticles for siRNA delivery. Pharmaceutics 5 (3), 498-507. https://doi.org/10.3390%2Fpharmaceutics5030498

Tenchov, R., Bird, R., Curtze, A. E., Zhou, Q. (2021). Lipid Nanoparticles - From Liposomes to mRNA Vaccine Delivery, a Landscape of Research Diversity and Advancement, ACS Nano 2021 15 (11), 16982-17015, https://pubs.acs.org/doi/pdf/10.1021/acsnano.1c04996

Wan, C., Allen, T., Cullis, P. (2014). Lipid nanoparticle delivery systems for siRNA-based therapeutics. Drug Deliv Transl Res. 4 (1), 74-83. https://doi.org/10.1007/s13346-013-0161-z

Wick, P., Malek, A., Manser, P., Meili, D., Maeder-Althaus, X., Diener, L., Diener, P.A., Zisch, A., Krug, H.F., von Mandach, U. (2010). Barrier capacity of human placenta for nanosized materials. Environ. Health Perspect., 118, 432–436. https://doi.org/10.1289/ehp.0901200

Yu, Z., Li, Q., Wang, J., Yu, Y., Wang, Y., Zhou, Q. (2020). Reactive Oxygen Species-Related Nanoparticle Toxicity in the Biomedical Field - Nanoscale Res Lett 15, 115 https://doi.org/10.1186/s11671-020-03344-7

Yun, C.H., Bae, C.S., & Ahn, T. (2016). Cargo-Free Nanoparticles Containing Cationic Lipids Induce Reactive Oxygen Species and Cell Death in HepG2 Cells - Biol Pharm Bull. 39(8):1338-46 https://doi.org/10.1248/bpb.b16-00264

Zhang, J., Li, X., Huang, L. (2014). Non-viral nanocarriers for siRNA delivery in breast cancer. J. Control. Release. 190, pp. 440 - 450. https://doi.org/10.1016/j.jconrel.2014.05.037

Zhang, C., Ma, Y., Zhang, J., Kuo, J.C.T., Zhang, Z., Xie, H., Zhu, J., Liu, T. (2022). Modification of Lipid-Based Nanoparticles: An Efficient Delivery System for Nucleic Acid-Based Immunotherapy. Molecules 27(6), 1943. https://doi.org/10.3390/molecules27061943

Zhou, Y., Peng, Z., Seven, E.S., Leblanc, R.M. (2018). Crossing the blood-brain barrier with nanoparticles. J. Control Release, 270, 290–303. https://doi.org/10.1016/j.jconrel.2017.12.015

Downloads

Published

2023-10-16

How to Cite

Apparent Cytotoxicity and Intrinsic Cytotoxicity of Lipid Nanomaterials Contained in a COVID-19 mRNA Vaccine. (2023). International Journal of Vaccine Theory, Practice, and Research , 3(1), 957-972. https://doi.org/10.56098/ijvtpr.v3i1.84

Similar Articles

1-10 of 54

You may also start an advanced similarity search for this article.