IP Library › Granted Patent US 12,478,588
Granted Patent B2
US 12,478,588 · App. 17/058,925 · Granted Nov 25, 2025

Nanoparticle for protein delivery

Inventors: Siyang Zheng (State College, PA); Cheng Gong (Malden, MA)
Assignee: The Penn State Research Foundation
A61K9/5123A61K9/5176A61K9/5192A61K38/02
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,478,588
App. No.
17/058,925
Granted
Nov 25, 2025
Kind
B2
Abstract

The present invention relates to compositions and methods for delivery of therapeutic agent. In certain aspects, the invention comprises a metal-organic framework nanoparticle encapsulating a protein. In some aspects, the metal-organic framework nanoparticle encapsulating a protein is coated with an extracellular vesicle membrane.

Claims (21)

1 . A nanoparticle comprising:

an outer surface comprising an extracellular vesicle membrane (EVM) removed from an extracellular vesicle (EV) of a cancer cell of a type of cancer such that the EVM substantially lacks tumorigenic constituents of the EV from which the EVM is derived, the EVM being between 20 nanometers (nm) and 200 nm in size;

an inner core comprising a metal-organic framework element including at least one metal ion and at least one organic molecule linker compound, the metal-organic framework element formed via mixing (i) a first aqueous solution comprising the at least one organic molecule linker compound and an agent, wherein the at least one organic molecule linker includes 2-methylimidazole and the agent includes a protein or a nucleic acid, with (ii) a second aqueous solution comprising zinc, wherein a ratio of moles of zinc to moles of the at least one organic molecule linker compound is between 0.01 and 0.1 so that the metal-organic framework element encapsulates the protein or the nucleic acid so that the inner core is positionable within the EVM having the size of between 20 nm and 200 nm,

wherein the EVM of the outer surface coating enveloping the metal-organic framework element therein so that the nanoparticle is deliverable to a cancer cell of the type of cancer, the metal-organic framework element facilitating intracellular transduction of the the protein or the nucleic acid to the cancer cell via a pH sensitive release of the the protein or the nucleic acid complex into the cancer cell.

2 . The nanoparticle of claim 1 , wherein the at metal-organic framework element encapsulates the protein.

3 . The nanoparticle of claim 2 , wherein the cancer cell of the type of cancer from which the EV is derived is a tumor cell of the type of cancer.

4 . The nanoparticle of claim 3 , wherein the tumor cell is from supernatants of cultured tumor cells or a tumor of a patient.

5 . The nanoparticle of claim 3 , wherein the EVM entirely lacks tumorigenic contents constituents of the EV from which the EVM is derived.

6 . A method of targeted delivery of a protein to a cell, the method comprising administering to a subject a composition comprising a nanoparticle of claim 2 .

7 . The nanoparticle of claim 1 , wherein the nanoparticle comprises a loading capacity of about 20% to 41% for a bovine serum albumin (BSA) concentration range of 1-2 mg/mL.

8 . The nanoparticle of claim 2 , wherein the nanoparticle comprises a loading capacity of about 20% to 41% for a bovine serum albumin (BSA) concentration range of 1-2 mg/mL.

9 . The nanoparticle of claim 1 , wherein the EVM entirely lacks tumorigenic constituents of the EV from which the EVM is derived.

10 . The nanoparticle of claim 1 , wherein the EVM substantially lacks tumorigenic constituents of the EV from which the EVM is derived such that the EVM lacks between 70% and 100% of the tumorigenic constituents of the EV.

11 . The nanoparticle of claim 2 , wherein the EVM substantially lacks tumorigenic constituents of the EV from which the EVM is derived such that the EVM lacks between 95% and 100% of the tumorigenic constituents of the EV.

12 . The nanoparticle of claim 1 , wherein the at least one organic molecule linker includes functional groups selected from the group consisting of a carboxylate, a phosphonate, an amine, an azide, a cyanide, a squaryl, an imidazole, and mixtures thereof.

13 . The nanoparticle of claim 12 , wherein the EVM encapsulates the metal-organic framework element via mixing of a solution having the EVM and the metal-organic framework at a ratio that ranges between 10:1 to 1:1 on a mass basis via ultrasonication to induce self-assembly of the EVM to the metal-organic framework element.

14 . The nanoparticle of claim 13 , wherein the metal-organic framework element encapsulates the protein.

15 . The nanoparticle of claim 14 , wherein the EVM has a diameter of between 20 nanometers (nm) and 100 nm.

16 . The nanoparticle of claim 1 , wherein the EVM has a diameter of between 20 nanometers (nm) and 100 nm.

17 . The nanoparticle of claim 1 , wherein the EVM substantially lacks tumorigenic constituents of the EV from which the EVM is derived such that the EVM lacks between 97% and 100% of the tumorigenic contents of the EV from which the EVM is derived.

18 . The nanoparticle of claim 1 , wherein the EVM substantially lacks tumorigenic constituents of the EV from which the EVM is derived such that the EVM lacks tumorigenic contents of the EV from which the EVM is derived and also lacks an entirety of proteins of the EV from which the EVM is derived and/or also lacks an entirety of nucleic acids of the EV from which the EVM is derived.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 29, 2021
From: ZHENG, SIYANG; GONG, CHENG
To: THE PENN STATE RESEARCH FOUNDATION
Reel/Frame 056098/0703 →
CONFIRMATORY LICENSE Recorded Dec 15, 2020
From: PENNSYLVANIA STATE UNIVERSITY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 054760/0150 →
Continuity (2)
Provisional Application 62676341 · May 25, 2018
Related Publication 20210212955A1 · Jul 15, 2021
References Cited (97)
US 4683202A · Mullis · 1987 [cited by applicant]
US 4708871A · Geysen · 1987 [cited by applicant]
US 4946778A · Ladner et al. · 1990 [cited by applicant]
US 5023243A · Tullis · 1991 [cited by applicant]
US 5190931A · Inouye · 1993 [cited by applicant]
US 5928906A · Koster et al. · 1999 [cited by applicant]
US 6103489A · Arakaki et al. · 2000 [cited by applicant]
US 6506559B1 · Fire et al. · 2003 [cited by applicant]
US 20130337066A1 · Zhang et al. · 2013 [cited by applicant]
WO WO2016000032A1 · 2016 [cited by examiner]
WO 2017078609A1 · 2017 [cited by applicant]
Rani, Sweta, and Thomas Ritter. “The exosome—A naturally secreted nanoparticle and its application to wound healing.” Advanced materials 28.27 (2016): 5542-5552 (Year: 2016). [cited by examiner]
Li et al. ACS Nano 2017, 11, 7006-7018. (Year: 2017). [cited by examiner]
Deria et al. J. Am. Chem. Soc. 2016, 138, 14449-14457. (Year: 2016). [cited by examiner]
Franzen et al. The Journal of Urology, 195, 1331-1339, May 2016. (Year: 2016). [cited by examiner]
Armstrong et al. ACS Nano 2017, 11, 69-83. (Year: 2017). [cited by examiner]
Coumans et al. Circulation Research. 2017;120:1632-1648. (Year: 2017). [cited by examiner]
Tiwari et al. Scientific reports 7.1 (2017): 12598. (Year: 2017). [cited by examiner]
Ren et al. Chem. Commun., 2014, 50, 1000-1002. (Year: 2014). [cited by examiner]
Dineen, K. The Jackson Laboratory. https://www.jax.org/news-and-insights/jax-blog/2016/august/women-in-science-remembering-henrietta-lacks, Aug. 1, 2016. (Year: 2016). [cited by examiner]
International Search Report and Written Opinion for International Application No. PCT/US2019/034119 dated Sep. 23, 2019. [cited by applicant]
Illes, B., Hirschle, P., Barnert, S., Cauda, V., Wuttke, S., & Engelke, H. (2017). Exosome-Coated Metal-Organic Framework Nanoparticles: An Efficient Drug Delivery Platform. Chemistry of Materials, 29(19), 8042-8046. do… [cited by applicant]
Agrwal, Sudhir, and John Goodchild. “Oligodeoxynucleoside methylphosphonates: synthesis and enzymic degradation.” Tetrahedron letters 28.31 (1987): 3539-3542. [cited by applicant]
Andaloussi, Samir EL, et al. “Extracellular vesicles: biology and emerging therapeutic opportunities.” Nature reviews Drug discovery 12.5 (2013): 347-357. [cited by applicant]
Banerjee, Rahul, et al. “High-throughput synthesis of zeolitic imidazolate frameworks and application to CO2 capture.” Science 319.5865 (2008): 939-943. [cited by applicant]
Barth, Andreas. “Infrared spectroscopy of proteins.” Biochimica et Biophysica Acta (BBA)—Bioenergetics 1767.9 (2007): 1073-1101. [cited by applicant]
Brodin, Jeffrey D., et al. “DNA-mediated cellular delivery of functional enzymes.” Journal of the American Chemical Society 137.47 (2015): 14838-14841. [cited by applicant]
Butcher, Neville J., Gysell M. Mortimer, and Rodney F. Minchin. “Unravelling the stealth effect.” Nature nanotechnology 11.4 (2016): 310-311. [cited by applicant]
Christian, Rosemarie B., et al. “Simplified methods for construction, assessment and rapid screening of peptide libraries in bacteriophage.” Journal of molecular biology 227.3 (1992): 711-718. [cited by applicant]
Chu, Ted C., et al. “Aptamer: toxin conjugates that specifically target prostate tumor cells.” Cancer research 66.12 (2006): 5989-5992. [cited by applicant]
Cwirla, Steven E., et al. “Peptides on phage: a vast library of peptides for identifying ligands.” Proceedings of the National Academy of Sciences 87.16 (1990): 6378-6382. [cited by applicant]
D'Astolfo, Diego S., et al. “Efficient intracellular delivery of native proteins.” Cell 161.3 (2015): 674-690. [cited by applicant]
Devlin, James J., Lucy C. Panganiban, and Patricia E. Devlin. “Random peptide libraries: a source of specific protein binding molecules.” Science 249.4967 (1990): 404-406. [cited by applicant]
Eckstein, Fritz, and Gerald Gish. “Phosphorothioates in molecular biology.” Trends in biochemical sciences 14.3 (1989): 97-100. [cited by applicant]
Eide, David J. “Zinc transporters and the cellular trafficking of zinc.” Biochimica et Biophysica Acta (BBA)—Molecular Cell Research 1763.7 (2006): 711-722. [cited by applicant]
Ellington, Andrew D., et al. “Targeting PSMA on tumor cells using novel aptamer-gelonin conjugates.” (2005): 1455-1455. Abstract 6183. [cited by applicant]
Escola et al., 1998; Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. Escola JM, Kleijmeer MJ, Stoorvogel W, Griffith JM, Y… [cited by applicant]
Furukawa, H., and K. E. Cordova. “O, Keeffe, M.; Yaghi, OM The chemistry and applications of metal-organic frameworks.” Science 341.6149 (2013): 1230444. [cited by applicant]
Gay, Laurie J., and Brunhilde Felding-Habermann. “Contribution of platelets to tumour metastasis.” Nature Reviews Cancer 11.2 (2011): 123-134. [cited by applicant]
Ghosh, Partha, et al. “Intracellular delivery of a membrane-impermeable enzyme in active form using functionalized gold nanoparticles.” Journal of the American Chemical Society 132.8 (2010): 2642-2645. [cited by applicant]
Gilleron, Jerome, et al. “Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape.” Nature biotechnology 31.7 (2013): 638-646. [cited by applicant]
Gu, Zhen, et al. “Tailoring nanocarriers for intracellular protein delivery.” Chemical Society Reviews 40.7 (2011): 3638-3655. [cited by applicant]
Howarth, Ashlee J., et al. “Chemical, thermal and mechanical stabilities of metal- organic frameworks.” Nature Reviews Materials 1.3 (2016): 1-15. [cited by applicant]
Hu, Che-Ming J., et al. “Nanoparticle biointerfacing by platelet membrane cloaking.” Nature 526.7571 (2015): 118-121. [cited by applicant]
Huang, Xiao-Chun, et al. “Ligand-directed strategy for zeolite-type metal-organic frameworks: zinc (II) imidazolates with unusual zeolitic topologies.” Angewandte Chemie International Edition 45.10 (2006): 1557-1559. [cited by applicant]
Iversen, Tore-Geir, Tore Skotland, and Kirsten Sandvig. “Endocytosis and intracellular transport of nanoparticles: Present knowledge and need for future studies.” Nano today 6.2 (2011): 176-185. [cited by applicant]
Jaiswal, Siddhartha, et al. “CD47 is upregulated on circulating hematopoietic stem cells and leukemia cells to avoid phagocytosis.” Cell 138.2 (2009): 271-285. [cited by applicant]
Jean, Steve, et al. “Starvation-induced MTMR 13 and RAB 21 activity regulates VAMP 8 to promote autophagosome-lysosome fusion.” EMBO reports 16.3 (2015): 297-311. [cited by applicant]
Jensen, Samuel A., et al. “Spherical nucleic acid nanoparticle conjugates as an RNAi-based therapy for glioblastoma.” Science translational medicine 5.209 (2013): 209ra152-209ra152. [cited by applicant]
Kamerkar, Sushrut, et al. “Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer.” Nature 546.7659 (2017): 498-503. [cited by applicant]
Kim, Mi-Young, et al. “Tumor self-seeding by circulating cancer cells.” Cell 139.7 (2009): 1315-1326. [cited by applicant]
Koivusalo, Mirkka, et al. “Endocytic trafficking of sphingomyelin depends on its acyl chain length.” Molecular biology of the cell 18.12 (2007): 5113-5123. [cited by applicant]
Kowal, J., et al. (2016). Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of the National Academy of Sciences, 113(8), E968-E977. doi: … [cited by applicant]
Lässer, Cecilia, et al. “Human saliva, plasma and breast milk exosomes contain RNA: uptake by macrophages.” Journal of translational medicine 9.1 (2011):9, 1-8. [cited by applicant]
Leader, Benjamin, Quentin J. Baca, and David E. Golan. “Protein therapeutics: a summary and pharmacological classification.” Nature reviews Drug discovery 7.1 (2008): 21-39. [cited by applicant]
Liang, Kang, et al. “Biomimetic mineralization of metal-organic frameworks as protective coatings for biomacromolecules.” Nature communications 6.1 (2015): 1-8. [cited by applicant]
Liu, Xiaojuan, et al. “CD47 blockade triggers T cell-mediated destruction of immunogenic tumors.” Nature medicine 21.10 (2015): 1209-1215. [cited by applicant]
Lu, Yue, Wujin Sun, and Zhen Gu. “Stimuli-responsive nanomaterials for therapeutic protein delivery.” Journal of controlled release 194 (2014): 1-19. [cited by applicant]
Macia, E., et al. “T. 110 Kirchhausen.” Dev. Cell 10 (2006): 839-850. [cited by applicant]
Mitragotri, Samir, Paul A. Burke, and Robert Langer. “Overcoming the challenges in administering biopharmaceuticals: formulation and delivery strategies.” Nature reviews Drug discovery 13.9 (2014): 655-672. [cited by applicant]
Monopoli, Marco P., et al. “Biomolecular coronas provide the biological identity of nanosized materials.” Nature nanotechnology 7.12 (2012): 779-786. [cited by applicant]
Moody, Harold M., et al. “Regiospecific inhibition of DNA duplication by antisense phosphate-methylated oligodeoxynucleotides.” Nucleic acids research 17.12 (1989): 4769-4782. [cited by applicant]
Moreira, Carla, et al. “Improving chitosan-mediated gene transfer by the introduction of intracellular buffering moieties into the chitosan backbone.” Acta biomaterialia 5.8 (2009): 2995-3006. [cited by applicant]
Nischan, Nicole, et al. “Covalent attachment of cyclic TAT peptides to GFP results in protein delivery into live cells with immediate bioavailability.” Angewandte Chemie International Edition 54.6 (2015): 1950-1953. [cited by applicant]
Ojida, Akio, et al. “Effective Disruption of Phosphoprotein—Protein Surface Interaction Using Zn (II) Dipicolylamine-Based Artificial Receptors via Two-Point Interaction.” Journal of the American Chemical Society 128.6 … [cited by applicant]
Pack, Daniel W., David Putnam, and Robert Langer. “Design of imidazole-containing endosomolytic biopolymers for gene delivery.” Biotechnology and Bioengineering 67.2 (2000): 217-223. [cited by applicant]
Pakulska, Malgosia M., Shane Miersch, and Molly S. Shoichet. “Designer protein delivery: From natural to engineered affinity-controlled release systems.” Science 351, No. 6279 (2016). [cited by applicant]
Pan, Yichang, et al. “Rapid synthesis of zeolitic imidazolate framework-8 (ZIF-8) nanocrystals in an aqueous system.” Chemical Communications 47.7 (2011): 2071-2073. [cited by applicant]
Parodi, Alessandro, et al. “Synthetic nanoparticles functionalized with biomimetic leukocyte membranes possess cell-like functions.” Nature nanotechnology 8.1 (2013): 61-68. [cited by applicant]
Parolini, Isabella, et al. “Microenvironmental pH is a key factor for exosome traffic in tumor cells.” Journal of Biological Chemistry 284.49 (2009): 34211-34222. [cited by applicant]
Raposo, Graca, et al. “B lymphocytes secrete antigen-presenting vesicles.” Journal of Experimental Medicine 183.3 (1996): 1161-1172. [cited by applicant]
Reedijk, M., et al. “Tyr721 regulates specific binding of the CSF-1 receptor kinase insert to PI 3′-kinase SH2 domains: a model for SH2-mediated receptor-target interactions.” The EMBO Journal 11.4 (1992): 1365-1372. [cited by applicant]
Roberge, Jacques Y., Xenia Beebe, and Samuel J. Danishefsky. “A strategy for a convergent synthesis of N-linked glycopeptides on a solid support.” Science 269.5221 (1995): 202-204. [cited by applicant]
Schöttler, Susanne, et al. “Protein adsorption is required for stealth effect of poly (ethylene glycol)-and poly (phosphoester)-coated nanocarriers.” Nature nanotechnology 11.4 (2016): 372-377. [cited by applicant]
Schwarze, Steven R., et al. “In vivo protein transduction: delivery of a biologically active protein into the mouse.” Science 285.5433 (1999): 1569-1572. [cited by applicant]
Smith, Douglas M., Jakub K. Simon, and James R. Baker Jr. “Applications of nanotechnology for immunology.” Nature Reviews Immunology 13.8 (2013): 592-605. [cited by applicant]
Sorkin, Alexander, and Mark Von Zastrow. “Signal transduction and endocytosis: close encounters of many kinds.” Nature reviews Molecular cell biology 3.8 (2002): 600-614. [cited by applicant]
Stec, Wojciech J., et al. “Synthesis and absolute configuration of P-chiral O-isopropyl oligonucleotide triesters.” Tetrahedron letters 26.18 (1985): 2191-2194. [cited by applicant]
Sun, Wujin, et al. “Self-assembled DNA nanoclews for the efficient delivery of CRISPR-Cas9 for genome editing.” Angewandte Chemie 127.41 (2015): 12197-12201. [cited by applicant]
Tenzer, Stefan, et al. “Rapid formation of plasma protein corona critically affects nanoparticle pathophysiology.” Nature nanotechnology 8.10 (2013): 772-781. [cited by applicant]
Ulysse, Luckner, Juanita Cubillos, and Jean Chmielewski. “Photoregulation of cyclic peptide conformation.” Journal of the American Chemical Society 117.32 (1995): 8466-8467. [cited by applicant]
Van Niel et al., 2003 Intestinal epithelial exosomes carry MHC class II/peptides able to inform the immune system in mice. Van Niel G, Mallegol J, Bevilacqua C, Candalh C, Brugière S, Tomaskovic-Crook E, Heath JK, Cerf-… [cited by applicant]
Varkouhi, Amir K., et al. “Endosomal escape pathways for delivery of biologicals.” Journal of Controlled Release 151.3 (2011): 220-228. [cited by applicant]
Veenendaal, Liesbeth M., et al. “In vitro and in vivo studies of a VEGF121/rGelonin chimeric fusion toxin targeting the neovasculature of solid tumors.” Proceedings of the National Academy of Sciences 99.12 (2002): 7866… [cited by applicant]
Wan, Yuan, et al. “Rapid magnetic isolation of extracellular vesicles via lipid-based nanoprobes.” Nature biomedical engineering 1.4 (2017): 1-11. [cited by applicant]
Wang, Ming, et al. “Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles.” Proceedings of the National Academy of Sciences 113.11 (2016): 2868-2873. [cited by applicant]
Wilhelm, Stefan, et al. “Analysis of nanoparticle delivery to tumours.” Nature reviews materials 1.5 (2016): 1-12. [cited by applicant]
Wozniak, Ann L., et al. “Intracellular proton conductance of the hepatitis C virus p7 protein and its contribution to infectious virus production.” PLoS pathogens 6.9 (2010): e1001087. [cited by applicant]
Wubbolts et al., 2003 Proteomic and biochemical analyses of human B cell-derived exosomes. Potential implications for their function and multivesicular body formation. Wubbolts R, Leckie RS, Veenhuizen PT, Schwarzmann G… [cited by applicant]
Xiang, Jian, et al. “Antigen-loaded upconversion nanoparticles for dendritic cell stimulation, tracking, and vaccination in dendritic cell-based immunotherapy.” ACS nano 9.6 (2015): 6401-6411. [cited by applicant]
Yang, Jian, et al. “Zr-based MOFs shielded with phospholipid bilayers: improved biostability and cell uptake for biological applications.” Chemistry of Materials 29.10 (2017): 4580-4589. [cited by applicant]
Mn, Hao, et al. “Non-viral vectors for gene-based therapy.” Nature Reviews Genetics 15.8 (2014): 541-555. [cited by applicant]
Mn, Hao, et al. “Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo.” Nature biotechnology 34.3 (2016): 328-333. [cited by applicant]
Yu, Mikyung, et al. “Nanotechnology for protein delivery: Overview and perspectives.” Journal of controlled release 240 (2016): 24-37. [cited by applicant]
Zhao, Muxun, et al. “Clickable protein nanocapsules for targeted delivery of recombinant p53 protein.” Journal of the American chemical society 136.43 (2014): 15319-15325. [cited by applicant]
Zhou, Li, et al. “DNA-mediated Construction of Hollow Upconversion Nanoparticles for Protein Harvesting and Near-Infrared Light Triggered Release.” Advanced Materials 26.15 (2014): 2424-2430. [cited by applicant]
Xi et al.; “Drug loading techniques for exosome-based drug delivery systems”; Pharmazie 76: 61-67 (2021); dated Dec. 23, 2020; 7 pages. [cited by applicant]