# US 12,178,907 B2 · Binary lipid bilayer-containing vesicles comprising embedded cytotoxic agents and methods of making and using the same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,178,907 B2 - Document ID: US12178907B2 - Status: Granted - Kind: B2 - Application number: 17/259,499 - Filed: 2019-07-11 - Published: 2021-08-05 - Granted: 2024-12-31 - Earliest claimed: 2018-07-12 - Assignees: The United States of America, as represented by the Secretary, Department of Health and Human Services - Applicants: The USA, as represented by the Secretary, Department of Health and Human Services - Inventors: Anu Puri; Mathias Viard - Web page: https://patentgrep.com/patent/US12178907B2 ## CPC - A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) - A61P 35/00: Antineoplastic agents - A61K 31/4745: condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines (yohimbine derivatives, vinblastine A61K31/475; ergoline derivatives A61K31/48) - A61K 41/0071: {PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines} - A61K 47/543: {Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine} - A61K 47/60: the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol - 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 Embodiments of vesicles comprising embedded cytotoxic agents are disclosed, as well as methods of making and using the vesicles. Pharmaceutical compositions including the vesicles also are disclosed. The vesicles include a binary lipid bilayer surrounding a cavity, wherein the vesicle binary lipid bilayer includes (i) a non-bilayer forming lipid (or combination of non-bilayer forming lipids) and a PEGylated lipid; and (i) a cytotoxic agent embedded within the vesicle wall. ## Claims ### Claim 1 A vesicle, comprising: a binary lipid bilayer comprising an alkyne-containing phospholipid and a PEGylated lipid; and a cytotoxic agent embedded in the binary lipid bilayer; wherein the binary lipid bilayer is free of, or does not comprise, a lipid other than the alkyne-containing phospholipid or the PEGylated lipid. ### Claim 2 (depends on 1) The vesicle of claim 1, wherein the binary lipid bilayer comprises greater than 6 mol % to 20 mol % of the PEGylated lipid. ### Claim 3 (depends on 1) The vesicle of claim 1, wherein the binary lipid bilayer comprises 10 mol % to 20 mol % of the PEGylated lipid. ### Claim 4 (depends on 1) The vesicle of claim 1, wherein the alkyne-containing phospholipid and the PEGylated lipid, taken together, and the cytotoxic agent are present at a ratio of 1:0.05, total lipid:cytotoxic agent. ### Claim 5 (depends on 1) The vesicle of claim 1, wherein the alkyne-containing phospholipid is an alkyne-containing phosphocholine lipid or a mixture of the alkyne-containing phosphocholine lipid and an alkyne-containing phosphoethanolamine lipid. ### Claim 6 (depends on 5) The vesicle of claim 5, wherein the alkyne-containing phosphocholine lipid is 1,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine (DC8,9PC) and wherein the alkyne-containing phosphoethanolamine lipid is 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphoethanolamine (DC8,9PE). ### Claim 7 (depends on 1) The vesicle of claim 1, wherein the PEGylated lipid is a PEGylated 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) lipid comprising a PEG group having a molecular weight ranging from 500 Da to 5000 Da. ### Claim 8 (depends on 1) The vesicle of claim 1, wherein the PEGylated lipid is a PEGylated DSPE lipid comprising a PEG group having a molecular weight ranging from 1000 Da to 3000 Da. ### Claim 9 (depends on 1) The vesicle of claim 1, wherein the PEGylated lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy (polyethylene glycol)-2000 (DSPE-PEG2000). ### Claim 10 (depends on 1) The vesicle of claim 1, wherein the cytotoxic agent is a tetrapyrrollic compound, a camptothecin compound, paclitaxel, daunorubicin, methotrexate, vincristine, etoposide, sorafenib, erlotinib, imatinib, or any combination thereof. ### Claim 11 (depends on 10) The vesicle of claim 10, wherein the tetrapyrrollic compound is 2-[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a (HPPH), (17S,18S)-18-(2-carboxyethyl)-20-(carboxymethyl)-12-ethenyl-7-ethyl-3,8,13,17-tetramethyl-17,18,22,23-tetrahydroporphyrin-2-carboxylic acid (Ce6), (3S,4S)-9-Ethenyl-14-ethyl-21-(methoxycarbonyl)-4,8,13,18-tetramethyl-20-oxo-3-phorbinepropanoic acid; 3,3′,3″,3″-(2,3-dihydroporphyrin-5,10,15,20-tetrayl)tetraphenol, 3-[(23S,24R)-14-ethenyl-5-(3-methoxy-3-oxopropyl)-22,23-bis(methoxycarbonyl)-4,10,15,24-tetramethyl-25,26,27,28-tetraazahexacyclo[16.6.1.13,6.18,11.113,16.019,24]octacosa-1,3,5,7,9,11 (27),12,14,16,18 (25), 19,21-dodecaen-9-yl]propanoic acid, or any combination thereof. ### Claim 12 (depends on 10) The vesicle of claim 10, wherein the camptothecin compound is camptothecin, silatecan 7-t-butyldimethylsilyl-10-hydroxycamptothecin, 7-ethyl-10-hydroxy-20(S)-camptothecin, topotecan, irinotecan, 9-nitro-camptothecin, lurtotecan, exatecan, gimatecan, or karenitecin. ### Claim 13 (depends on 1) The vesicle of claim 1, comprising a binary lipid bilayer consisting of an alkyne-containing lipid, a PEGylated lipid and a cytotoxic agent embedded within the binary lipid bilayer. ### Claim 14 A vesicle, comprising: a binary lipid bilayer comprising (i) DSPE-PEG2000 and (ii) DC8,9PC, or a combination of DC8,9PC and DC8,9PE; and HPPH, Ce6, and/or camptothecin embedded in the binary lipid bilayer; and wherein the binary lipid bilayer is free of, or does not comprise, a lipid other than the DC8,9PC, the DC8,9PE, and the DSPE-PEG2000. ### Claim 15 (depends on 1) A method, comprising: providing a vesicle according to claim 1; and irradiating the vesicle with targeted application of light having a selected wavelength in the near-infrared range and a selected intensity for an effective period of time to activate at least a portion of the cytotoxic agent. ### Claim 16 (depends on 15) The method of claim 15, wherein irradiating the vesicle with targeted application of light comprises irradiating the vesicle with a laser that produces light having a wavelength of 650-670 nm. ### Claim 17 (depends on 15) The method of claim 15, wherein the selected intensity is from 1 mW to 500 mW and/or the effective period of time is at least 30 seconds. ### Claim 18 (depends on 15) The method of claim 15, further comprising: identifying a subject as having a condition that may be treated with the cytotoxic agent; administering the vesicle to the subject; and wherein the targeted application of light is directed at a targeted portion of the subject. ### Claim 19 (depends on 18) The method of claim 18, wherein the subject has a tumor and the targeted portion of the subject includes an area proximate a location of the tumor and wherein administering the vesicle to the subject comprises intravenously injecting an amount of the vesicle effective to induce tumor size regression. ### Claim 20 (depends on 18) The method of claim 18, wherein irradiating is performed 4-6 hours after administering the vesicle to the subject. ### Claim 21 (depends on 18) The method of claim 18, wherein administering the vesicle to the subject comprises administering a pharmaceutical composition comprising the vesicle to the subject. ### Claim 22 (depends on 18) The method of claim 18, wherein the targeted application of light occurs by (i) externally applying the light to the targeted portion of the subject for the effective period of time, thereby transcutaneously applying the light to the tumor or (ii) internally applying the light to the targeted portion of the subject for the effective period of time using an endoscope or a fiber optic catheter. ### Claim 23 (depends on 1) A method for impairing growth of a tumor in a subject, comprising: administering to the subject a therapeutically effective amount of a vesicle according to claim 1; and irradiating the vesicle by targeted application of light having a selected wavelength in the near-infrared range and a selected intensity to a target area of the subject proximate a location of the tumor for an effective period of time to activate at least a portion of the cytotoxic agent to promote reactive oxygen species formation, thereby impairing growth of the tumor. ### Claim 24 (depends on 23) The method of claim 23, wherein the effective period of time is at least 30 seconds and irradiating is performed 4-6 hours after administering the vesicle to the subject. ### Claim 25 (depends on 23) The method of claim 23, wherein administering the vesicle to the subject comprises administering an amount of the vesicle effective to induce tumor size regression and/or wherein the vesicle is administered via intravenous injection. ### Claim 26 (depends on 23) The method of claim 23, wherein administering the vesicle to the subject comprises administering a pharmaceutical composition comprising the vesicle to the subject. ### Claim 27 (depends on 23) The method of claim 23, wherein irradiating the vesicle by targeted application of light comprises externally or internally applying the light to the targeted portion of the subject for the effective period of time using an endoscope or a fiber optic catheter.