# US 11,717,548 B2 · Synthetic oncolytic LNP-replicon RNA and uses for cancer immunotherapy > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,717,548 B2 - Document ID: US11717548B2 - Status: Granted - Kind: B2 - Application number: 16/739,407 - Filed: 2020-01-10 - Published: 2020-09-10 - Granted: 2023-08-08 - Earliest claimed: 2019-03-08 - Assignees: Massachusetts Institute of Technology; Ohio State Innovation Foundation - Applicants: Massachusetts Institute of Technology; Ohio State Innovation Foundation - Inventors: Darrell J. Irvine; Karl Dane Wittrup; Ron Weiss; Yingzhong Li; Noor Momin; Yizhou Dong - Web page: https://patentgrep.com/patent/US11717548B2 ## CPC - A61K 35/768: Oncolytic viruses not provided for in groups A61K35/761 - A61K35/766 - C12N 15/86: Viral vectors - A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)} - A61K 31/7105: Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links - A61P 35/00: Antineoplastic agents - C07K 14/5434: {IL-12} - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 15/88: using microencapsulation, e.g. using {amphiphile} liposome vesicle - C12N 2770/36121: Viruses as such, e.g. new isolates, mutants or their genomic sequences - C12N 2770/36132: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present disclosure relates to synthetic oncolytic viruses comprising a lipid nanoparticle comprising one or more types of lipid and a self-amplifying replicon RNA comprising a sequence that encodes an immunomodulatory molecule. ## Claims ### Claim 1 A synthetic oncolytic virus, comprising: (i) a lipid nanoparticle comprising N1,N3,N5-tris(3-(didodecylamino)propyl)benzene-1,3,5-tricarboxamide (TT3); and (ii) a self-amplifying replicon RNA comprising a sequence that encodes an interleukin (IL)-12 molecule; and wherein the IL-12 molecule is expressed by the self-amplifying replicon RNA. ### Claim 2 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle further comprises one or more types of lipid, where the one or more types of lipid comprises a cationic lipid. ### Claim 3 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle further comprises 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, and C14-PEG2000. ### Claim 4 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the self-amplifying replicon RNA is derived from an alphavirus or a hepatitis C virus. ### Claim 5 (depends on 4) The synthetic oncolytic virus of claim 4, wherein the alphavirus is Venezuela Equine Encephalitis virus, Semliki Forest virus, or Sindbis virus. ### Claim 6 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the sequence that encodes the IL-12 molecule is located in a subgenomic region of the self-amplifying replicon RNA. ### Claim 7 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the self-amplifying replicon RNA comprises a nucleotide sequence that has at least 90% sequence identity to the sequence set forth in SEQ ID NO: 1. ### Claim 8 (depends on 7) The synthetic oncolytic virus of claim 7, wherein the self-amplifying replicon RNA comprises a point mutation of G3936C and/or A4758G of SEQ ID NO: 1. ### Claim 9 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the self-amplifying replicon RNA further comprises a serum albumin coding sequence. ### Claim 10 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the self-amplifying replicon RNA further comprises a lumican coding sequence. ### Claim 11 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the IL-12 molecule is IL-12, an IL-12 subunit, or a mutant IL-12 molecule that retains the immunomodulatory function. ### Claim 12 (depends on 11) The synthetic oncolytic virus of claim 11, wherein the IL-12 molecule comprises IL-12α and/or IL-12β subunits. ### Claim 13 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle has a diameter of about 100-120 nm. ### Claim 14 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle has a zeta potential of about 3-6 mv. ### Claim 15 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle and the self-amplifying replicon RNA have a mass ratio of about 1:1. ### Claim 16 (depends on 1) A pharmaceutical composition, comprising the synthetic oncolytic virus of claim 1 and a pharmaceutically acceptable carrier. ### Claim 17 (depends on 16) The pharmaceutical composition of claim 16, wherein the pharmaceutical composition is formulated for intratumoral injection. ### Claim 18 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle further comprises 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol. ### Claim 19 (depends on 16) The pharmaceutical composition of claim 16, wherein the lipid nanoparticle further comprises 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol. ### Claim 20 (depends on 19) The pharmaceutical composition of claim 19, further comprising C14-PEG2000. ### Claim 21 A pharmaceutical composition, comprising a synthetic oncolytic virus and a pharmaceutically acceptable carrier; wherein the synthetic oncolytic virus comprises: (i) a lipid nanoparticle comprising TT3, 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and cholesterol; and (ii) a self-amplifying replicon RNA comprising a sequence that encodes an interleukin (IL)-12 molecule; wherein the self-amplifying replicon RNA comprises a nucleotide sequence that has at least 90% sequence identity to the sequence set forth in SEQ ID NO: 1; wherein the lipid nanoparticle is capable of triggering immunogenic cell death; wherein the IL-12 molecule is expressed by the self-amplifying replicon RNA; and wherein the pharmaceutical composition further comprises C14-PEG2000. ### Claim 22 (depends on 1) The synthetic oncolytic virus of claim 1, wherein the lipid nanoparticle is capable of triggering immunogenic cell death.