Fusogenic Liposome-coated Porous Silicon Nanoparticles

Sponsor(s)

	National Science Foundation (NSF) https://www.nsf.gov/
	National Institutes of Health (NIH) https://www.nih.gov/

Partnerships

	Sanford Burnham Prebys Medical Discovery Institute https://sbpdiscovery.org/
	Huazhong University of Science and Technology https://english.hust.edu.cn/
	Massachusetts Institute of Technology (MIT) https://www.mit.edu/

Type of technology

Inorganic nanoparticles

Administration route

Subcutaneous, Intramuscular, Intravenous, Topical (Rectal), Transmucosal, Transdermal

Development state and regulatory approval

C-FNP (Chemosensitizing siRNA)

Pre-clinical

Not provided

Description

Fusogenic liposome-coated porous silicon nanoparticles represent a biodegradable drug delivery system for oligonucleotides, capable of bypassing endocytic pathways to facilitate direct cytoplasmic delivery. This system enables the release of the API from the nanoparticle core into the cell cytoplasm. The fusogenic liposome layer, which surrounds the silicon-based core, is functionalized with polyethylene glycol (PEG) to enhance stability and circulation time

Technology highlight

1. The API is encapsulated within the porous structure of the fusogenic liposomal nanoparticle. 2. The API is released into the cytoplasm through the shedding of the nanoparticle's fusogenic lipid coating, facilitating direct intracellular delivery. 3. The nanoparticle exhibits a high API loading capacity, allowing for the efficient delivery of therapeutic agents. 4. The formulation demonstrates low cytotoxicity, ensuring biocompatibility for in vitro and in vivo applications. 5. The fusogenic nanoparticles achieve 90–100% transfection efficiency due to enhanced cellular uptake through membrane fusion and endosomal escape.

Technology main components

a) Silicon-containing core b) Porous surface that is chemically linked to the core c) Plurality of cargo molecules that are physically associated with silicon-containing core material d) Metal silicate e) Fusogenic liposome coating with silicon-containing core material

Information on the raw materials sourcing, availability and anticipated price

The raw materials are obtained from Alza Corporation and Nova Pharmaceuticals.

Delivery device(s)

No delivery device

API desired features

Nucleic acids

Oligonucleotides such as small interfering RNA (siRNA) therapeutics are targeted for this fugogenic nanoparticle drug delivery system. Some examples of siRNA peptides that are targeted against Irf5 gene and Rev3l gene.

Proteins

Fusogenic porous silicon nanoparticles have the capacity to securely deliver proteins to the targeted site, but the choice of protein for this delivery system is not disclosed.

Additional solubility data

Not provided

Additional stability data

Not provided

API loading: Maximum drug quantity to be loaded

75-90 wt%

API co-administration

1 single API :

LogP

Not provided

Scale-up prospects

Not provided

Tentative equipment list for manufacturing

1. Teflon Etch 2. Square Wave Generator 3. Sonicator Bath 4. Liposome Extruder

Manufacturing

Fusogenic Nanoparticle Manufacturing in Small Scale includes: Etch a silicon wafer in HF using square wave currents (50 mA/cm² for 0.6 s, 400 mA/cm² for 0.36 s) for 500 cycles. Detach the porous layer (3.7 mA/cm², 250 s), rinse with ethanol and water, and store silicon chips in RNase-free water. Sonicate chips (35 kHz, 12 h). Prepare DMPC, DSPE-PEG, and DOTAP lipid film, hydrate with siRNA-loaded calcium-coated nanoparticles, and extrude 20× through a polycarbonate membrane. Conjugate targeting peptides (1 mg/mL, 20 min), purify via 30 kDa filtration, and store at -80°C.

Specific analytical instrument required for characterization of formulation

1. Dynamic Light Scattering (DLS) 2. Zeta Potential Analyzer 3. Transmission Electron Microscopy (TEM) 4. Confocal Laser Scanning Microscopy (CLSM) 5. Fourier Transform Infrared Spectroscopy (FTIR) 6. Ultraviolet-Visible Spectroscopy (UV-Vis) 7. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) 8. Differential Scanning Calorimetry (DSC) 9. Nanoparticle Tracking Analysis (NTA) 10. High-Performance Liquid Chromatography (HPLC)

Proprietary excipients used

No proprietary excipient used

Novel excipients or existing excipients at a concentration above Inactive Ingredients Database (IID) for the specified route of administration

No novel excipient or existing excipient used

Residual solvents used

No residual solvent used

Other features of the technology

• Biodegradable
• Drug-eluting

Release properties

Preclinical studies show that the fusogenic lipid-coated nanoparticles have very little initial burst release rate. It also proves that up to 75% of the API is released into the cells at 168 hours and complete release after 336 hours, i.e., 14 days.

Injectability

Fusogenic porous silicon nanoparticles can be administered via intravitreal, intravenous, and subcutaneous injection.

Safety

Not provided

Stability

Product Stability: Details regarding the formulation's stability are not disclosed. Nanoparticle Stability: The stability of the nanoparticles can be modified by adjusting their size to optimize the API payload.

Storage conditions and cold-chain related features

Not provided

Therapeutic area(s)

Use case(s)

Not provided

Use of technology

Description

Fusogenic liposome-coated porous silicon nanoparticles

Brief description

The disclosure describes a fusogenic liposome-coated porous silicon nanoparticles for high loading efficiency of anionic payloads (small molecules, dyes, nucleic acids), and for non-endocytic delivery of hydrophilic and lipophilic payloads by membrane fusion. The liposome coating can be further modified with targeting peptides or antibodies via covalent binding chemistry between the ligands and functionalized poly(ethylene glycol). The surface moieties can be transferred to the cellular membrane surface by fusogenic uptake. The composition of the disclosure can be applied in the treatment of diseases by delivering entrapped/encapsulated payloads.

Representative patent

US10702474B2

Category

Formulation

Patent holder

University of California San Diego UCSD

Exclusivity

Not provided

Expiration date

July 8, 2038

Status

Anticipated expiration

Publications

<p><span style="color: rgb(27, 27, 27);">Kim, B., &amp; Sailor, M. J. (2019). Synthesis, Functionalization, and Characterization of Fusogenic Porous Silicon Nanoparticles for Oligonucleotide Delivery.&nbsp;</span><em style="color: rgb(27, 27, 27);">Journal of visualized experiments : JoVE</em><span style="color: rgb(27, 27, 27);">, (146), 10.3791/59440. </span><a href="https://doi.org/10.3791/59440" rel="noopener noreferrer" target="_blank" style="color: rgb(27, 27, 27);">https://doi.org/10.3791/59440</a></p>

With the advent of gene therapy, the development of an effective in vivo nucleotide-payload delivery system has become of parallel import. Fusogenic porous silicon nanoparticles (F-pSiNPs) have recently demonstrated high in vivo gene silencing efficacy due to its high oligonucleotide loading capacity and unique cellular uptake pathway that avoids endocytosis. The synthesis of F-pSiNPs is a multi-step process that includes: (1) loading and sealing of oligonucleotide payloads in the silicon pores; (2) simultaneous coating and sizing of fusogenic lipids around the porous silicon cores; and (3) conjugation of targeting peptides and washing to remove excess oligonucleotide, silicon debris, and peptide. The particle’s size uniformity is characterized by dynamic light scattering, and its core-shell structure may be verified by transmission electron microscopy. The fusogenic uptake is validated by loading a lipophilic dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), into the fusogenic lipid bilayer and treating it to cells in vitro to observe for plasma membrane staining versus endocytic localizations. The targeting and in vivo gene silencing efficacies were previously quantified in a mouse model of Staphylococcus aureus pneumonia, in which the targeting peptide is expected to help the F-pSiNPs to home to the site of infection. Beyond its application in S. aureus infection, the F-pSiNP system may be used to deliver any oligonucleotide for gene therapy of a wide range of diseases, including viral infections, cancer, and autoimmune diseases.

<p><span style="color: rgb(34, 34, 34);">Kim, B., Sun, S., Varner, J. A., Howell, S. B., Ruoslahti, E., &amp; Sailor, M. J. (2019). Securing the payload, finding the cell, and avoiding the endosome: peptide‐targeted, fusogenic porous silicon nanoparticles for delivery of siRNA.&nbsp;</span><em style="color: rgb(34, 34, 34);">Advanced Materials</em><span style="color: rgb(34, 34, 34);">,&nbsp;</span><em style="color: rgb(34, 34, 34);">31</em><span style="color: rgb(34, 34, 34);">(35), 1902952.</span><span style="color: rgb(118, 118, 118);">&nbsp;</span><a href="https://doi.org/10.1002/adma.201902952" rel="noopener noreferrer" target="_blank" style="color: var(--wiley-blue-color);">https://doi.org/10.1002/adma.201902952</a></p>

Despite the promise of ribonucleic acid interference therapeutics, the delivery of oligonucleotides selectively to diseased tissues in the body, and specifically to the cellular location in the tissues needed to provide optimal therapeutic outcome, remains a significant challenge. Here, key material properties and biological mechanisms for delivery of short interfering RNAs (siRNAs) to effectively silence target-specific cells in vivo are identified. Using porous silicon nanoparticles as the siRNA host, tumor-targeting peptides for selective tissue homing, and fusogenic lipid coatings to induce fusion with the plasma membrane, it is shown that the uptake mechanism can be engineered to be independent of common receptor-mediated endocytosis pathways. Two examples of the potential broad clinical applicability of this concept in a mouse xenograft model of ovarian cancer peritoneal carcinomatosis are provided: silencing the Rev3l subunit of polymerase Pol ζ to impair DNA repair in combination with cisplatin; and reprogramming tumor-associated macrophages into a proinflammatory state.

Additional documents

• PhD Thesis of Fusogenic Porous Silicon Nanoparticles as a Platform Technology for Gene Therapy

Useful links

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