# US 12,213,994 B2 · Compositions and methods for treating pain with extracellular vesicles > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,213,994 B2 - Document ID: US12213994B2 - Status: Granted - Kind: B2 - Application number: 18/063,227 - Filed: 2022-12-08 - Published: 2023-09-07 - Granted: 2025-02-04 - Earliest claimed: 2021-12-08 - Assignees: Zeo ScientifiX, Inc. - Applicants: Organicell Regenerative Medicine, Inc. - Inventors: Maria Ines Mitrani; Michael Bellio; Albert Mitrani - Web page: https://patentgrep.com/patent/US12213994B2 ## CPC - A61K 9/5176: {Compounds of unknown constitution, e.g. material from plants or animals (oils, fats, waxes, shellac A61K9/5123)} - A61K 35/14: Blood; Artificial blood (perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42) - A61K 9/0014: {Skin, i.e. galenical aspects of topical compositions (non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00)} - 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 9/127: Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant - A61K 35/16: Blood plasma; Blood serum (umbilical cord blood A61K35/51) - A61K 35/19: Platelets; Megacaryocytes - A61K 47/6425: {the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell} - A61P 19/02: for joint disorders, e.g. arthritis, arthrosis - A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract Described herein are cell-free therapeutic compositions derived from blood or plasma and uses thereof for the treatment of selected diseases and disorders. ## Claims ### Claim 1 A method of treating a disease or disorder in a subject in need thereof, the method comprising: administering to the subject having the disease or disorder a therapeutically effective amount of a composition comprising: blood-derived nanoparticles, the blood-derived nanoparticles having a diameter less than 200 nm, the blood-derived nanoparticles derived from substantially platelet-free and cell-free blood, and a concentration of blood-derived nanoparticles in the composition being at least 1×108 nanoparticles/ml. ### Claim 2 (depends on 1) The method of claim 1, wherein the composition is made by a process comprising: (a) obtaining a volume of blood; (b) centrifuging the volume of blood at 2,000×g for at least 10 minutes (min) to obtain a first supernatant; (c) centrifuging the first supernatant at 100,000×g for at least 90 min to obtain a pellet comprising the nanoparticles; (d) isolating and resuspending the pellet in a volume of resuspension fluid to obtain a resuspended volume; and (e) filtering at least a portion of the resuspended volume using a 0.22 uM filter. ### Claim 3 (depends on 1) The method of claim 1, wherein the composition is made by a process comprising: (a) obtaining a volume of blood; (b) centrifuging the volume of blood at 2,000×g for at least 10 minutes (min) to obtain a first supernatant; (c) centrifuging the first supernatant at 2,000×g for at least 5 min to obtain a second supernatant; (d) centrifuging the second supernatant at 100,000×g for at least 90 min to obtain a pellet comprising the nanoparticles; (e) isolating and resuspending the pellet in a volume of resuspension fluid to obtain a resuspended volume; and (f) filtering at least a portion of the resuspended volume using a 0.22 uM filter. ### Claim 4 (depends on 1) The method of claim 1, wherein the disease or disorder is joint pain. ### Claim 5 (depends on 4) The method of claim 4, wherein the joint pain is derived from arthritis. ### Claim 6 (depends on 5) The method of claim 5, wherein the arthritis is osteoarthritis or rheumatoid arthritis. ### Claim 7 (depends on 4) The method of claim 4, wherein the joint is selected from the group consisting of hand, hip, shoulder, and knee. ### Claim 8 (depends on 1) The method of claim 1, wherein the nanoparticles are extracellular vesicles (EVs). ### Claim 9 (depends on 8) The method of claim 8, wherein the extracellular vesicles are autologous to the subject. ### Claim 10 (depends on 8) The method of claim 8, wherein the extracellular vesicles are heterologous to the subject. ### Claim 11 (depends on 8) The method of claim 8, wherein the extracellular vesicles comprise exosomes. ### Claim 12 (depends on 8) The method of claim 8, wherein the extracellular vesicles comprise any of the surface-bound proteins CD41a, CD9, CD63, or CD81. ### Claim 13 (depends on 12) The method of claim 12, wherein the concentration of EVs comprising CD41a is at least 107 EVs per mL. ### Claim 14 (depends on 12) The method of claim 12, wherein the concentration of EVs comprising CD41a ranges from 109 to 1013 EVs per mL. ### Claim 15 (depends on 1) The method of claim 1, wherein a protein concentration is at least 0.02 mg/mL. ### Claim 16 (depends on 1) The method of claim 1, wherein a protein concentration ranges from 0.02 to 10 mg/mL. ### Claim 17 (depends on 16) The method of claim 16, wherein the protein concentration is about 0.045 mg/mL, 2.25 mg/mL, or 4.5 mg/mL. ### Claim 18 (depends on 1) The method of claim 1, wherein the diameter of the nanoparticles ranges from 20 nm to 200 nm. ### Claim 19 (depends on 1) The method of claim 1, wherein the diameter of the nanoparticles is, on average, 60 nm. ### Claim 20 (depends on 1) The method of claim 1, wherein the concentration of the nanoparticles is at least 109 particles/mL. ### Claim 21 (depends on 20) The method of claim 20, wherein the concentration of the nanoparticles is at least 1.0×1011 particles/mL. ### Claim 22 (depends on 1) The method of claim 1, wherein the concentration of nanoparticles ranges from 1.0×108 particles/mL to 1.0×1013 particles/mL. ### Claim 23 (depends on 1) The method of claim 1, wherein the concentration of nanoparticles is about 3.0×109, 1.5×1011, or 3.0×1011 particles per mL. ### Claim 24 (depends on 1) The method of claim 1, wherein the wherein the composition is administered by intravenous or intraarticular injection. ### Claim 25 (depends on 1) The method of claim 1, wherein the subject is administered at least one dose of the composition. ### Claim 26 (depends on 25) The method of claim 25, wherein the subject is administered two doses of the composition. ### Claim 27 (depends on 1) The method of claim 1, wherein the nanoparticles are derived from the blood of the subject. ### Claim 28 (depends on 24) The method of claim 24, wherein the volume of the composition administered by intravenous injection is at least 100 mL. ### Claim 29 (depends on 24) The method of claim 24, wherein the volume of the composition administered by intraarticular injection is about 1 mL or 2 mL. ### Claim 30 (depends on 24) The method of claim 24, wherein the volume of the composition administered by intraarticular injection is at least 1 mL.