IP Library Granted Patent US 12,357,580
Granted Patent B2
US 12,357,580 · App. 18/744,370 · Granted Jul 15, 2025

Lipid nanoparticle compositions for delivery of mRNA and long nucleic acids

Inventors: Daniel J. Siegwart (Dallas, TX); Qiang Cheng (Dallas, TX)
Assignee: The Board of Regents of The University of Texas System
A61K9/5123A61K48/0033C12N9/22C12N15/11C12N15/111C12N15/113C12N15/88C12N2310/11C12N2310/14C12N2310/20C12N2320/32
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Quick Facts
Patent No.
US 12,357,580
App. No.
18/744,370
Granted
Jul 15, 2025
Kind
B2
Abstract

In some aspects, the present disclosure provides compositions of lipid nanoparticles useful for the delivery of large RNAs including mRNAs. These compositions may include a cationic ionizable lipid, a phospholipid, a PEGylated lipid, and a steroid including using less of a cationic ionizable lipid than compositions with shorter nucleic acids. These compositions may be used to treat a disease or disorder for which the delivery of an mRNA is therapeutically effective.

Claims (64)

1. A method for delivering a messenger ribonucleic acid (mRNA) into a cell, the method comprising contacting said cell with a lipid composition encapsulating said mRNA, wherein the lipid composition comprises:

a cationic ionizable lipid at a molar percentage from about 5 to about 30;

a phospholipid at a molar percentage from about 10 to about 45;

a steroid or steroid derivative at a molar percentage from about 15 to about 50; and

a polymer-conjugated lipid at a molar percentage from about 1 to about 6,

wherein the molar percentage is determined based on the total mols of lipids present in the lipid composition;

thereby delivering said mRNA into said cell,

wherein the cationic ionizable lipid is a compound having the structure of Formula (I):

Core-(Repeating Unit) n -Terminating Group  (I),

or a pharmaceutically acceptable salt thereof,

wherein the compound of Formula (I) or the pharmaceutically acceptable salt thereof is a dendron or dendrimer, wherein:

the core is linked to one or more repeating units, wherein:

the core corresponds to the structure of Formula (IV):

wherein, in Formula (IV):

X 3 is selected from —NR 6 —, —O—, substituted or unsubstituted alkylaminodiyl (C≤8) , substituted or unsubstituted alkoxydiyl (C≤8) , substituted or unsubstituted arenediyl (C≤8) , substituted or unsubstituted heteroarenediyl (C≤8) , and substituted or unsubstituted heterocycloalkanediyl (C≤8) ,

wherein R 6 is hydrogen, unsubstituted alkyl (C≤8) , or substituted alkyl (C≤8) ;

R 3 and R 4 are each independently selected from amino, hydroxy, mercapto, substituted or unsubstituted alkylamino (C≤12) , and substituted or unsubstituted dialkylamino (C≤12) ; and

c and d are each independently 1, 2, 3, 4, 5, or 6;

the repeating unit comprises a degradable diacyl group and optionally a linker; wherein:

the degradable diacyl group has the formula:

wherein, in Formula (VII):

A 1 and A 2 are each independently —O— or —NR a —,

wherein R a is hydrogen or substituted or unsubstituted alkyl (C≤6) ;

Y 3 is selected from substituted or unsubstituted alkanediyl (C≤12) , substituted or unsubstituted alkenediyl (C≤12) , substituted or unsubstituted arenediyl (C≤12) , and a group of the formula:

wherein:

X 3A and X 4 are each independently selected from substituted or unsubstituted alkanediyl (C≤12) , substituted or unsubstituted alkenediyl (C≤12) , and substituted or unsubstituted arenediyl (C≤12) ; and

Y 5 is selected from a covalent bond, substituted or unsubstituted alkanediyl (C≤12) , substituted or unsubstituted alkenediyl (C≤12) , and substituted or unsubstituted arenediyl (C≤12) ; and

R 9 is substituted or unsubstituted alkyl (C≤8) ; and

the linker group has the formula:

wherein, in Formula (VI):

Y 1 is selected from substituted or unsubstituted alkanediyl (C≤12) , substituted or unsubstituted alkenediyl (C≤12) , and substituted or unsubstituted arenediyl (C≤12) ; and

wherein when the repeating unit comprises a linker group, then the linker group is attached to the degradable diacyl group on both the nitrogen and the sulfur atoms of the linker group, wherein the first group in the repeating unit is the degradable diacyl group, wherein for each linker group, the next group comprises two degradable diacyl groups attached to the nitrogen atom of the linker group; and

wherein n is 1, 2, 3, 4, 5, or 6; and

the terminating group has the formula:

wherein, in Formula (VIII):

Y 4 is unsubstituted alkanediyl (C≤18) or alkanediyl (C≤18) substituted with one or more substituents independently selected from —OH, —F, —Cl, —Br, —I, —SH, —OCH 3 , —OCH 2 CH 3 , —SCH 3 , and —OC(O)CH 3 ;

R 10 is selected from hydrogen, carboxy, hydroxy, aryl (C≤12) , alkylamino (C≤12) , dialkylamino (C≤12) , N-heterocycloalkyl (C≤12) , —C(O)N(R 11 )-alkanediyl (C≤6) -heterocycloalkyl (C≤12) , —C(O)-alkyl-amino (C≤12) , —C(O)-dialkylamino (C≤12) , and —C(O)—N-heterocyclo-alkyl (C≤12) , wherein:

R 11 is hydrogen or substituted or unsubstituted alkyl(C≤6); and

wherein the final degradable diacyl in the chain of repeating unit(s) is attached to the terminating group.

2. The method of claim 1 , wherein, prior to said contacting, said cell exhibits an aberrant expression or activity of the protein encoded by said mRNA.

3. The method of claim 2 , wherein said aberrant expression or activity of said protein comprises the expression of a non-functional variant of said protein.

4. The method of claim 3 , wherein said aberrant expression or activity of said protein is associated with a genetic disease or disorder.

5. The method of claim 3 , wherein said mRNA is expressed in said cell, upon said contacting, to produce a functional variant of said protein.

6. The method of claim 3 , wherein the expression of said mRNA in said cell increases the amount of a functional variant of said protein as compared to the amount of said functional variant of said protein generated in absence of said contacting.

7. The method of claim 1 , wherein said contacting is in vivo.

8. The method of claim 1 , wherein said cell is in a tissue or organ of a subject.

9. The method of claim 8 , wherein said tissue or organ is a functionally compromised tissue or organ.

10. The method of claim 1 , wherein said contacting comprises administering to said subject said lipid composition assembled with said mRNA.

11. The method of claim 1 , further comprising repeating said contacting.

12. The method of claim 1 , wherein said contacting comprises contacting a plurality of cells that comprises said cell.

13. The method of claim 12 , wherein said mRNA is expressed in at least 40% of said plurality of cells, upon said contacting, to produce a functional variant of the protein encoded by said mRNA.

14. The method of claim 1 , wherein the lipid composition comprises said mRNA and said cationic ionizable lipid at a weight ratio from about 1:1 to about 1:100.

15. The method of claim 1 , wherein said phospholipid is a zwitterionic phospholipid.

16. The method of claim 1 , wherein said phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).

17. The method of claim 1 , wherein the lipid composition comprises said polymer-conjugated lipid at a molar percentage from about 1 to about 4, wherein the molar percentage is determined based on the total mols of lipids present in the lipid composition.

18. The method of claim 17 , wherein said polymer-conjugated lipid is a polyethylene glycol (PEG)-conjugated lipid.

19. The method of claim 1 , wherein said steroid or said steroid derivative comprises a cholesterol moiety.

20. The method of claim 1 , wherein, in the compound of Formula (I), or the pharmaceutically acceptable salt thereof, the core corresponds to a structure selected from the group consisting of:

21. The method of claim 1 , wherein, in the terminating group of Formula (VIII), Y 4 is alkanediyl (C≤18) ; and R 10 is hydrogen.

22. The method of claim 1 , wherein the compound of Formula (I), or the pharmaceutically acceptable salt thereof, is of a generation selected from 0, 1, 2, and 3.

23. The method of claim 1 , wherein the lipid composition comprises said cationic ionizable lipid at a molar percentage from about 5 to about 25, wherein the molar percentage is determined based on the total mols of lipids present in the lipid composition.

24. The method of claim 1 , wherein the lipid composition comprises said cationic ionizable lipid at a molar percentage from about 5 to about 20, wherein the molar percentage is determined based on the total mols of lipids present in the lipid composition.

25. The method of claim 1 , wherein the lipid composition comprises a lipid nanoparticle.

26. The method of claim 1 , wherein the composition is formulated for administration: orally, intraadiposally, intraarterially, intraarticularly, intracranially, intradermally, intralesionally, intramuscularly, intranasally, intraocularly, intrapericardially, intraperitoneally, intrapleurally, intraprostatically, intrarectally, intrathecally, intratracheally, intratumorally, intraumbilically, intravaginally, intravenously, intravesicularlly, intravitreally, liposomally, locally, mucosally, parenterally, rectally, subconjunctival, subcutaneously, sublingually, topically, transbuccally, transdermally, vaginally, in crèmes, via a catheter, via a lavage, via continuous infusion, via infusion, via inhalation, via injection, via local delivery, or via localized perfusion.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 16, 2024
From: CHENG, QIANG; SIEGWART, DANIEL J.
To: THE BOARD OF REGENTS OF THE UNIVERSITY OF TEXAS SYSTEM
Reel/Frame 068592/0279 →
Continuity (15)
Continuation In Part 18590737 · Feb 28, 2024
Continuation 18534974 · Dec 11, 2023
Continuation 18529992 · Dec 5, 2023
Continuation 18186105 · Mar 17, 2023
Continuation 17929704 · Sep 4, 2022
Continuation 17711911 · Apr 1, 2022
Continuation 17572615 · Jan 10, 2022
Continuation 17473863 · Sep 13, 2021
Continuation 17191895 · Mar 4, 2021
Continuation In Part 17124462 · Dec 16, 2020
Continuation PCTUS2019049565 · Sep 4, 2019
Continuation PCTUS2019037904 · Jun 19, 2019
Provisional Application 62726741 · Sep 4, 2018
Provisional Application 62687010 · Jun 19, 2018
Related Publication 20240325315A1 · Oct 3, 2024
References Cited (246)
US 5820873A · Choi et al. · 1998 [cited by applicant]
US 7314956B2 · Li et al. · 2008 [cited by applicant]
US 7404969B2 · Chen et al. · 2008 [cited by applicant]
US 8017804B2 · Keil et al. · 2011 [cited by applicant]
US 8058069B2 · Yaworski et al. · 2011 [cited by applicant]
US 8450298B2 · Mahon et al. · 2013 [cited by applicant]
US 9326939B2 · Paulson et al. · 2016 [cited by applicant]
US 9562086B2 · Upton · 2017 [cited by applicant]
US 11229609B2 · Cheng et al. · 2022 [cited by applicant]
US 11247968B2 · Siegwart et al. · 2022 [cited by applicant]
US 11304911B2 · Cheng et al. · 2022 [cited by applicant]
US 11510880B2 · Cheng et al. · 2022 [cited by applicant]
US 11858884B2 · Siegwart et al. · 2024 [cited by applicant]
US 20040236015A1 · Kozlowski et al. · 2004 [cited by applicant]
US 20050008689A1 · Semple et al. · 2005 [cited by applicant]
US 20060008910A1 · MacLachlan et al. · 2006 [cited by applicant]
US 20070298006A1 · Tomalia et al. · 2007 [cited by applicant]
US 20080242626A1 · Zugates et al. · 2008 [cited by applicant]
US 20090221684A1 · Grinstaff et al. · 2009 [cited by applicant]
US 20100048888A1 · Chen et al. · 2010 [cited by applicant]
US 20100178267A1 · Puerta et al. · 2010 [cited by applicant]
US 20100196277A1 · DeSimone et al. · 2010 [cited by applicant]
US 20100303884A1 · Upton et al. · 2010 [cited by applicant]
US 20110009641A1 · Anderson et al. · 2011 [cited by applicant]
US 20110038941A1 · Lee et al. · 2011 [cited by applicant]
US 20110165223A1 · Sgouros et al. · 2011 [cited by applicant]
US 20130171241A1 · Geall · 2013 [cited by applicant]
US 20130195967A1 · Guild et al. · 2013 [cited by applicant]
US 20140186332A1 · Ezrin et al. · 2014 [cited by applicant]
US 20140206753A1 · Guild et al. · 2014 [cited by applicant]
US 20140371293A1 · Brown et al. · 2014 [cited by applicant]
US 20150110859A1 · Heartlein et al. · 2015 [cited by applicant]
US 20150118288A1 · Lee · 2015 [cited by applicant]
US 20150272886A1 · Chen et al. · 2015 [cited by applicant]
US 20150297749A1 · Hahn et al. · 2015 [cited by applicant]
US 20160081944A1 · Lee · 2016 [cited by applicant]
US 20160158354A1 · DeRosa et al. · 2016 [cited by applicant]
US 20160220681A1 · Siegwart et al. · 2016 [cited by applicant]
US 20160317647A1 · Ciaramella et al. · 2016 [cited by applicant]
US 20170121279A1 · Siegwart et al. · 2017 [cited by applicant]
US 20170240501A1 · DeRosa et al. · 2017 [cited by applicant]
US 20170326254A1 · Chen et al. · 2017 [cited by applicant]
CN 101591428 · 2011 [cited by applicant]
CN 103999853 · 2014 [cited by applicant]
EP 3315125 · 2018 [cited by applicant]
EP 3757570 · 2020 [cited by applicant]
EP 3810148 · 2021 [cited by applicant]
EP 3950003 · 2022 [cited by applicant]
JP 2014103108 · 2014 [cited by applicant]
JP 2014529328 · 2014 [cited by applicant]
JP 2015519346 · 2015 [cited by applicant]
JP 2019515016 · 2019 [cited by applicant]
KR 20110090661 · 2011 [cited by applicant]
WO WO2006138380 · 2006 [cited by applicant]
WO WO2010053329 · 2010 [cited by applicant]
WO WO2010129687 · 2010 [cited by applicant]
WO WO2010141069 · 2010 [cited by applicant]
WO WO2012090223 · 2012 [cited by applicant]
WO WO2012170930 · 2012 [cited by applicant]
WO WO2012170952 · 2012 [cited by applicant]
WO WO2013177415 · 2013 [cited by applicant]
WO WO2013177419 · 2013 [cited by applicant]
WO WO2014026283 · 2014 [cited by applicant]
WO WO2014105985 · 2014 [cited by applicant]
WO WO2014106208 · 2014 [cited by applicant]
WO WO2014144196 · 2014 [cited by applicant]
WO WO2015089462 · 2015 [cited by applicant]
WO WO2015148247 · 2015 [cited by applicant]
WO WO2015191693 · 2015 [cited by applicant]
WO WO2016010840 · 2016 [cited by applicant]
WO WO2016094342 · 2016 [cited by applicant]
WO WO2016118697 · 2016 [cited by applicant]
WO WO2016118725 · 2016 [cited by applicant]
WO WO2017048789 · 2017 [cited by applicant]
WO WO2017053713 · 2017 [cited by applicant]
WO WO2017173054 · 2017 [cited by applicant]
WO WO2017180917 · 2017 [cited by applicant]
WO WO2017201091 · 2017 [cited by applicant]
WO WO2017201350 · 2017 [cited by applicant]
WO WO2018029586 · 2018 [cited by applicant]
WO WO2018078053 · 2018 [cited by applicant]
WO WO2019246203 · 2019 [cited by applicant]
WO WO2020051220 · 2020 [cited by applicant]
WO WO2020051223 · 2020 [cited by applicant]
Adams et al., “Trial design and rationale for APOLLO, a Phase 3, placebo-controlled study of patisiran in patients with hereditary ATTR amyloidosis with polyneuropathy,” [cited by applicant]
Akinc et al., “A combinatorial library of lipid-like materials for delivery of RNAi therapeutics,” [cited by applicant]
Amoasii et al., “Gene editing restores dystrophin expression in a canine model of Duchenne muscular dystrophy,” [cited by applicant]
Bartsch et al., “Massive and selective delivery of lipid-coated cationic lipoplexes of oligonucleotides targeted in vivo to hepatic endothelial cells,” [cited by applicant]
Blasco et al., “Simple and rapid in vivo generation of chromosomal rearrangements using CRISPR/Cas9 technology,” [cited by applicant]
Bosman et al., “About dendrimers: Structure, physical properties, and applications,” [cited by applicant]
Boyerinas et al., “The role of let-7 in cell differentiation and cancer,” [cited by applicant]
Bryantsev et al., “pKa calculations of aliphatic amines, diamines, and aminoamides via density functional theory with a Poisson-Boltzmann continuum solvent model,” J. Phys. Chem. A., 111:4422-4430, 2007. [cited by applicant]
Carlmark et al., “New methodologies in the construction of dendritic materials,” [cited by applicant]
Chatani et al., “Facile and Efficient Synthesis of Dendrimers and One-Pot Preparation of Dendritic-Linear Polymer Conjugates via a Single Chemistry: Utilization of Kinetically Selective Thiol-Michael Addition Reactions,” [cited by applicant]
Cheng and Lee, “The role of helper lipids in lipid nanoparticles (LNPs) designed for oligonucleotide delivery,” [cited by applicant]
Cheng et al., “Dendrimer-Based Lipid Nanoparticles Deliver Therapeutic FAH mRNA to Normalize Liver Function and Extend Survival in a Mouse Model of Hepatorenal Tyrosinemia Type I,” [cited by applicant]
Cheng et al., “Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR-Cas gene editing,” [cited by applicant]
Chew et al., “A multifunctional AAV-CRISPR-Cas9 and its host response,” [cited by applicant]
Coelho et al., “Safety and efficacy of RNAi therapy for transthyretin amyloidosis,” [cited by applicant]
Cong et al., “Multiplex genome engineering using CRISPR/Cas systems,” [cited by applicant]
Cui et al., “Correlation of the cytotoxic effects of cationic lipids with the headgroups,” [cited by applicant]
Dahlman et al., “In vivo endothelial siRNA delivery using polymeric nanoparticles with low molecular weight,” [cited by applicant]
Derosa et al., “Therapeutic efficacy in a hemophilia B model using a biosynthetic mRNA liver depot system,” [cited by applicant]
Dong et al., “Lipopeptide nanoparticles for potent and selective siRNA delivery in rodents and nonhuman primates,” [cited by applicant]
Dong et al., “Poly(glycoamidoamine) Brushes Formulated Nanomaterials for Systemic siRNA and mRNA Delivery in Vivo,” [cited by applicant]
Doudna & Charpentier, “Genome editing. The new frontier of genome engineering with CRISPR-Cas9,” [cited by applicant]
Duncan and Izzo, “Dendrimer biocompatibility and toxicity,” [cited by applicant]
Ex parte Rolf Bergmann, Maria Lundqvist, Stig Mannberg, Bjorn Lundgren, and Robert Shimizu, Board of Patent Appeals and Interferences, Appeal 2011-013450, Feb. 1, 2012. [cited by applicant]
Extended European Search Report issued in European Application No. 16847193.6, mailed Feb. 19, 2019. [cited by applicant]
Extended European Search Report issued in European Application No. 19857774.4, mailed Jun. 7, 2022. [cited by applicant]
Extended European Search Report issued in European Application No. 19858575.4, mailed Sep. 12, 2022. [cited by applicant]
Extended European Search Report issued in European Application No. 19822888.4, mailed May 6, 2022. [cited by applicant]
Extended European Search Report issued in European Application No. 17800042.8, mailed Dec. 19, 2019. [cited by applicant]
Fenton et al., “Bioinspired Alkenyl Amino Alcohol Ionizable Lipid Materials for Highly Potent In Vivo mRNA Delivery,” [cited by applicant]
Fenton et al., “Synthesis and biological evaluation of ionizable lipid materials for the in vivo delivery of messenger RNA to B lymphocytes,” [cited by applicant]
Finn et al., “A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing,” [cited by applicant]
Franc and Kakkar, ““Click” methodologies: efficient, simple and greener routes to design dendrimers,” [cited by applicant]
Gilleron et al., “Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape,” [cited by applicant]
Gillies and Fréchet, “Designing macromolecules for therapeutic applications: Polyester dendrimer-poly(ethylene oxide) ”bow-tie“ hybrids with tunable molecular weight and architecture,” [cited by applicant]
Grayson and Fréchet, “Convergent dendrons and dendrimers: From synthesis to applications,” [cited by applicant]
Grompe et al., “Pharmacological correction of neonatal lethal hepatic dysfunction in a murine model of hereditary tyrosinaemia type I,” [cited by applicant]
Gustafson et al., “Nanoparticle Uptake: The Phagocyte Problem,” [cited by applicant]
Hafez et al., “On the mechanism whereby cationic lipids promote intracellular delivery of polynucleic acids,” [cited by applicant]
Hajj & Whitehead, “Tools for translation: non-viral materials for therapeutic mRNA delivery,” [cited by applicant]
Hao et al., “Rapid Synthesis of a Lipocationic Polyester Library via Ring-Opening Polymerization of Functional Valerolactones for Efficacious siRNA Delivery,” [cited by applicant]
Harvie et al., “Characterization of lipid DNA interactions. I. Destabilization of bound lipids and DNA dissociation,” [cited by applicant]
Hendel et al., “Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells,” [cited by applicant]
Hoyle et al., “Thiol-click chemistry: a multifaceted toolbox for small molecule and polymer synthesis,” [cited by applicant]
Jarzbińska et al., “A Single Methylene Group in Oligoalkylamine-Based Cationic Polymers and Lipids Promotes Enhanced mRNA Delivery,” [cited by applicant]
Jayaraman et al., “Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo,” [cited by applicant]
Jiang et al., “A non-viral CRISPR/Cas9 delivery system for therapeutically targeting HBV DNA and pcsk9 in vivo,” [cited by applicant]
Jinek et al., “A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity,” [cited by applicant]
Kaczmarek et al., “Polymer-Lipid Nanoparticles for Systemic Delivery of mRNA to the Lungs,” [cited by applicant]
Kanasty et al., “Delivery materials for siRNA therapeutics,” [cited by applicant]
Kang et al., “Tat-conjugated PAMAM dendrimers as delivery agents for antisense and siRNA oligonucleotides,” [cited by applicant]
Kauffman et al., “Optimization of Lipid Nanoparticle Formulations for mRNA Delivery in Vivo with Fractional Factorial and Definitive Screening Designs,” [cited by applicant]
Khan et al., “Ionizable amphiphilic dendrimer-based nanomaterials with alkyl-chain-substituted amines for tunable siRNA delivery to the liver endothelium in vivo,” [cited by applicant]
Killops et al., “Robust, efficient, and orthogonal synthesis of dendrimers via thiol-ene “click” chemistry,” [cited by applicant]
Kormann et al., “Expression of therapeutic proteins after delivery of chemically modified mRNA in mice,” [cited by applicant]
Lee et al., “Designing dendrimers for biological applications,” [cited by applicant]
Leung et al., “Lipid nanoparticles containing siRNA synthesized by microfluidic mixing exhibit an electron-dense nanostructured core,” [cited by applicant]
Li and Szoka, “Lipid-based nanoparticles for nucleic acid delivery,” [cited by applicant]
Li et al., “A biomimetic lipid library for gene delivery through thiol-yne click chemistry,” [cited by applicant]
Li et al., “An Orthogonal Array Optimization of Lipid-like Nanoparticles for mRNA Delivery in Vivo,” [cited by applicant]
Li et al., “Effects of local structural transformation of lipid-like compounds on delivery of messenger RNA,” [cited by applicant]
Love et al., “Lipid-like materials for low-dose, in vivo gene silencing,” [cited by applicant]
Lowe, “mRNA vaccines; what happens,” located at https://www.science.org/content/blog-post/mrna-vaccines-what-happens, accessed Jan. 15, 2022, originally published Jan. 21, 2021. [cited by applicant]
Lowe, “RNA vaccines and their lipids,” In the Pipeline, https://blogs.sciencemag.org/pipeline/archives/2021/01/11/rna-vaccines-and-their-lipids, accessed Aug. 19, 2021, originally published Jan. 11, 2021. [cited by applicant]
Lowe, “What mRNA is good for, and what it maybe isn't,” https://www.science.org/content/blog-post/what-mrna-good-and-what-it-maybe-isn-t accessed Jan. 15, 2022, originally published Jun. 29, 2021. [cited by applicant]
Lu et al., “Toxicity of cationic lipids and cationic polymers in gene delivery,” [cited by applicant]
Ma et al., “Facile synthesis of polyester dendrimers from sequential click coupling of asymmetrical monomers,” [cited by applicant]
Maddalo et al., “In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system,” [cited by applicant]
Mali et al., “RNA-guided human genome engineering via Cas9,” [cited by applicant]
Marshall et al., “Cationic lipid structure and formulation considerations for optimal gene transfection of the lung,” [cited by applicant]
Miller et al., “Non-viral CRISPR/Cas gene editing in vitro and in vivo enabled by synthetic nanoparticle co-delivery of Vas9 mRNA and sgRNA,” [cited by applicant]
Murat and Grest, “Molecular dynamics study of dendrimer molecules in solvents of varying quality,” [cited by applicant]
Nelson et al., “Balancing cationic and hydrophobic content of PEGylated siRNA polyplexes enhances endosome escape, stability, blood circulation time, and bioactivity in vivo,” ACS Nano, 7:8870-8880, 2013. [cited by applicant]
Notice of Allowance issued in U.S. Appl. No. 15/265,064, mailed Oct. 1, 2021. [cited by applicant]
Notice of Allowance issued in U.S. Appl. No. 17/191,895, mailed Sep. 1, 2021. [cited by applicant]
Office Action issued in British Application No. 2104769.1, mailed Jun. 1, 2021. [cited by applicant]
Office Action issued in British Application No. 2104777.4, mailed Jun. 1, 2021. [cited by applicant]
Office Action issued in British Application No. GB 2219700.8, mailed Feb. 3, 2023. [cited by applicant]
Office Action issued in British Application No. GB2100678.8, mailed Jun. 11, 2021. [cited by applicant]
Office Action issued in British Application No. GB2104777.4, mailed Mar. 2, 2022. [cited by applicant]
Office Action issued in British Application No. GB2111272.7, mailed Mar. 2, 2022. [cited by applicant]
Office Action issued in Chinese Application No. 201980070622.2, mailed Nov. 28, 2023, and English translation thereof. [cited by applicant]
Office Action issued in Japanese Application No. 2018-513463, mailed Sep. 7, 2020. [cited by applicant]
Office Action issued in Japanese Application No. 2021-536681, mailed Nov. 9, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 15/265,064, mailed Aug. 3, 2020. [cited by applicant]
Office Action issued in U.S. Appl. No. 15/597,063, mailed Apr. 22, 2020. [cited by applicant]
Office Action issued in U.S. Appl. No. 15/597,063, mailed Aug. 22, 2019. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Apr. 22, 2021. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Apr. 6, 2023. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Jul. 12, 2021. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Jun. 22, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Jan. 25, 2024. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Nov. 15, 2021. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/124,462, mailed Sep. 20, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/191,895, mailed Jun. 16, 2021. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/191,975, mailed Aug. 25, 2021. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/191,975, mailed Dec. 14, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/191,975, mailed Feb. 11, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/191,975, mailed Jul. 26, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/191,975, mailed Nov. 1, 2021. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/572,615, mailed Mar. 24, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/572,615, mailed May 5, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/839,699, mailed Sep. 6, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 17/929,704, mailed Oct. 28, 2022. [cited by applicant]
Office Action issued in U.S. Appl. No. 18/534,974, mailed Feb. 16, 2024. [cited by applicant]
Pankowicz et al., “Reprogramming metabolic pathways in vivo with CRISPR/Cas9 genome editing to treat hereditary tyrosinaemia,” [cited by applicant]
Pardi et al., “Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes,” [cited by applicant]
Patel et al., “Boosting Intracellular Delivery of Lipid Nanoparticle-Encapsulated mRNA,” [cited by applicant]
PCT International Preliminary Report on Patentability issued in International Application No. PCT/US2019/049552, mailed Mar. 18, 2021. [cited by applicant]
PCT International Preliminary Report on Patentability issued in International Application No. PCT/US2019/049565, mailed Mar. 18, 2021. [cited by applicant]
PCT International Search Report and Written Opinion issued in International Application No. PCT/US2019/037904, mailed Oct. 2, 2019. [cited by applicant]
PCT International Search Report and Written Opinion issued in International Application No. PCT/US2016/051648, mailed Feb. 7, 2017. [cited by applicant]
PCT International Search Report and Written Opinion issued in International Application No. PCT/US2019/049552, mailed Dec. 30, 2019. [cited by applicant]
PCT International Search Report and Written Opinion issued in International Application No. PCT/US2019/049565, mailed Feb. 7, 2020. [cited by applicant]
PCT International Search Report and Written Opinion issued in International Application No. PCT/US2017/032967, mailed Aug. 11, 2017. [cited by applicant]
Percec et al., “Self-assembly of Janus dendrimers into uniform dendrimersomes and other complex architectures,” [cited by applicant]
Petsch et al., “Protective efficacy of in vitro synthesized, specific mRNA vaccines against influenza A virus infection,” [cited by applicant]
Ramaswamy et al., “Systemic delivery of factor IX messenger RNA for protein replacement therapy,” [cited by applicant]
Regnaud, “Design and synthesis of dendrimers by combination of ‘click’ chemistry and A3-coupling,” Thesis, McGill University, pp. 1-67, 2013. [cited by applicant]
Richner et al., “Modified mRNA Vaccines Protect against Zika Virus Infection,” [cited by applicant]
Sahay et al., “Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling,” [cited by applicant]
Sahin et al., “mRNA-based therapeutics—developing a new class of drugs,” [cited by applicant]
Sander & Joung, “CRISPR-Cas systems for editing, regulating and targeting genomes,” Nat. Biotechnol., 32:347-355, 2014. [cited by applicant]
Schaffert et al., “Solid-phase synthesis of sequence-defined T-, i-, and U-shape polymers for pDNA and siRNA delivery,” [cited by applicant]
Semple et al., “Rational design of cationic lipids for siRNA delivery,” [cited by applicant]
Shobaki et al., “Mixing lipids to manipulate the ionization status of lipid nanoparticles for specific tissue targeting,” [cited by applicant]
Siegwart et al., “Combinatorial synthesis of chemically diverse core-shell nanoparticles for intracellular delivery,” [cited by applicant]
Staahl et al., “Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes,” [cited by applicant]
Stiriba et al., “Dendritic polymers in biomedical applications: From potential to clinical use in diagnostics and therapy,” [cited by applicant]
Sun et al., “Self-assembled DNA nanoclews for the efficient delivery of CRISPR-Cas9 for genome editing,” [cited by applicant]
Sundaram et al., “Reversibly switchable polymer with cationic/zwitterionic/anionic behavior through synergistic protonation and deprotonation,” [cited by applicant]
Tabebordbar et al., “In vivo gene editing in dystrophic mouse muscle and muscle stem cells,” Science, 351:407-411, 2016. [cited by applicant]
Taratula et al., “Surface-engineered targeted PPI dendrimer for efficient intracellular and intratumoral siRNA delivery,” [cited by applicant]
Tousignant et al., “Comprehensive analysis of the acute toxicity systemic administration of cationic lipid:plasmid DNA complexes in mice,” [cited by applicant]
Uchida et al., “Modulated protonation of side chain aminoethylene repeats in N-substituted polyaspartamides promotes mRNA transfection,” [cited by applicant]
Wang et al., “Cas9-mediated allelic exchange repairs compound heterozygous recessive mutations in mice,” Nat. Biotechnol., 36(9):839-842, 2018. [cited by applicant]
Wang et al., “CRISPR/Cas9-Based Genome Editing for Disease Modeling and Therapy: Challenges and Opportunities for Nonviral Delivery,” [cited by applicant]
Whitehead et al., “Degradable lipid nanoparticles with predictable in vivo siRNA delivery activity,” [cited by applicant]
Wilhelm et al., “Analysis of nanoparticle delivery to tumours,” Nat. Rev. Mater., 1:16014, 2016. [cited by applicant]
Wittrup et al., “Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown,” [cited by applicant]
Wood, “Traumatic brain injury induces transmissible tau pathology,” [cited by applicant]
Wu et al., “Dendrimers in medicine: Therapeutic concepts and pharmaceutical challenges,” [cited by applicant]
Wu et al., “Efficiency and fidelity in a click-chemistry route to triazole dendrimers by the copper(I)-catalyzed ligation of azides and alkynes,” [cited by applicant]
Wu et al., “RNAi therapies: drugging the undruggable,” [cited by applicant]
Xu et al., “Fluorescent water-soluble perylenediimide-cored cationic dendrimers: synthesis, optical properties, and cell uptake,” [cited by applicant]
Xue et al., “CRISPR-mediated direct mutation of cancer genes in the mouse liver,” Nature, 514:380-384, 2014. [cited by applicant]
Yan et al., “Functional polyesters enable selective siRNA delivery to lung cancer over matched normal cells,” [cited by applicant]
Yan et al., “Systemic mRNA Delivery to the Lungs by Functional Polyester-based Carriers,” [cited by applicant]
Yin et al., “Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype,” [cited by applicant]
Yin et al., “Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing,” Nat. Biotechnol., 35:1179-1187, 2017. [cited by applicant]
Yin et al., “Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo,” [cited by applicant]
Yu et al., “An amphiphilic dendrimer for effective delivery of small interfering RNA and gene silencing in vitro and in vivo,” [cited by applicant]
Yung et al., “Lipid nanoparticles composed of quaternary amine-tertiary amine cationic lipid combination (QTsome) for therapeutic delivery of antimiR-21 for lung cancer,” [cited by applicant]
Zelphati and Szoca, “Intracellular distribution and mechanism of delivery of oligonucleotides mediated by cationic lipids,” [cited by applicant]
Zhang et al., “Biodegradable amino-ester nanomaterials for Cas9 mRNA delivery in vitro and in vivo,” [cited by applicant]
Zhang et al., “Knockdown of Anillin Actin Binding Protein Blocks Cytokinesis in Hepatocytes and Reduces Liver Tumor Development in Mice Without Affecting Regeneration,” [cited by applicant]
Zhang et al., “The Polyploid State Plays a Tumor-Suppressive Role in the Liver,” [cited by applicant]
Zhou et al., “Balancing biocompatibility, internalization and pharmacokinetics of polycations/siRNA by structuring the weak negative charged ternary complexes with hyaluronic acid,” [cited by applicant]
Zhou et al., “Modular degradable dendrimers enable small RNAs to extend survival in an aggressive liver cancer model,” [cited by applicant]
Zhou et al., “Modular degradable dendrimers enable small RNAs to extend survival in an aggressive liver cancer model,” Supporting Information, [cited by applicant]
Zhou et al., “PAMAM dendrimers for efficient siRNA delivery and potent gene silencing,” [cited by applicant]
Zuris et al., “Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo,” Nat. Biotechnol., 33:73-80, 2015. [cited by applicant]