US00566293OA United States Patent (19) 11 Patent Number: 5,662.930 Ahl et al. I45 Date of Patent: *Sep. 2, 1997 54 REDUCTION OF LPOSOME-INDUCED 5,030,453 7/1991 Lenk et al............................... 424/450 ADVERSE PHYSOLOGICAL REACTIONS 5,041278 8/1991 Janoff et al. .. ... 424/1.1 5,077.056 12/1991 Bally et al. ... ... 424/450 75 Inventors: Patrick L. Ahl, Princeton; Suresh K. 5,213,804 5/1993 Martin et al. ........................... 424/450 Bhatia, Plainsboro; Sharma R. Minchey, Monmouth Junction, all of FOREIGN PATENT DOCUMENTS N.J.; Andrew S. Janoff, Yardley, Pa. O 213 523 3/1987 European Pat. Off. ......... A61K 9/50 0312 212 4/1989 European Pat. Off. ..... G01N 33/531 73) Assignee: The Liposome Company, Inc., 61-088887 5/1986 Japan ............... ... A61K 3/127 Princeton, N.J. 85/00968 3/1985 WIPO .............................. A61K 9/22 86/00238 1/1986 WTPO ............................ B01D 13/00 86/01102 2/1986 WIPO .............................. A61K 9/60 * Notice: The term of this patent shall not extend 37/00043 1/1987 WEPO .............................. A61K 9/00 beyond the expiration date of Pat. No. 87/02219 4/1987 WIPO ............................ A01N 25/28 5,614,214. 88/06443 9/1988 WIPO .............................. A61K 9/66 88/09165 5/1989 WIPO .............................. A61K 9/50 90/14105 11/1990 WIPO ............................ A61K 43/00 21 Appl. No.: 433,665 92/05773 4/1992 WIPO ............................ A61K 9/27 22 Filed: May 4, 1995 OTHER PUBLICATIONS Related U.S. Application Data Bangham, et al., “Diffusion of Univalent Ions across the 60) Division of Ser. No. 247,053, May 20, 1994, which is a Lamellae of Swollen Phospholipids”. J. Mol. Biol., continuation-in-part of Ser. No. 207,651, Mar. 7, 1994, 13:238-252 (1965). abandoned, which is a continuation-in-part of Ser. No. Deamer and Uster, "Liposome Preparation: Methods and 65,928, May 21, 1993, abandoned. Materials.” in: Liposomes. Marcel Dekker, Inc., New York 51 Int. Cl. ... A61K9/127; A61 K9/133 (1983) 27-51. 52 U.S. Cl. ... 424/450; 424/1.21 58) Field of Search ................................... 424/450, 1.21; (List continued on next page.) 436/829 Primary Examiner-Gollamudi S. Kishore Attorney, Agent, or Firm-Kenneth B. Rubin 56) References Cited 57 ABSTRACT U.S. PATENT DOCUMENTS Provided herein is a method of administering a liposome 3,577,446 5/1971 Rakhit .................................... 260/403 3,993,754 11/1976 Rahman et al. ... 424/177 composition to an animal, the method involving adminster 4,145,410 3/1979 Sears ......................................... 424/19 ing to the animal a liposome composition containing an 4.224,179 9/1980 Schneider ........ 252/316 adverse physiological reaction-reducing effective amount of 4229,360 10/1980 Schneider et al. .......... ... 260/403 a liposome which has, in addition to a bioactive agent, a lipid 4,235,871 11/1980 Papahadjopoulos et al. ............ 424/19 bilayer containing a lipid and a surface agent-modified 4,522,803 6/1985 Lenk et al. ............................... 424/1.1 molecule. An adverse physiological reaction which may be 4,588,578 5/1986 Fountain et al. ......................... 424/1.1 experienced by the animal upon administration of a lipo 4,721,612 1/1988 Janoff et al. ............................. 424/1.1 some composition is reduced by way of the presence of the 4,861,580 8/1989 Janoff et al. ............................. 424/1.1 surface agent-modified molecule in the liposome's lipid 4,880,635 11/1989 Janoff et al. ... ... 424/450 bilayer. 4,975,282 12/1990 Bally et al. ..... ... 424/450 5,008,050 4/1991 Cullis et al. ............................. 264/4.3 5,013,556 5/1991 Woodle et al. ......................... 424/450 17 Claims, 16 Drawing Sheets o ^nuacio 5,662.930 Page 2 OTHER PUBLICATIONS Mayer, et al., "Solute distributions and trapping efficiencies observed in freeze-thawed multimalar vesicles”, Biochim. Ivancev et al., “Effect of Intravenously Injected Iodinated Biophys. Acta, 1985, 817:193-196. Lipis Emulsions on the Liver", Acta Radiologica, 30, Papahadjopoulos, et al., “Phospholipid Model Membrane, I. (1989), 291-298. Structural Characteristics of hydrated Liquid Crystals". Bio chim. Biophys. Acta, 1968, 135:624-638. Ivancev, et al., "Experimental Investigation of a New Iodi Park, et al., "Some negatively charged phospholipid deriva nated Lipid Emulsion for Computed Tomography of the tives prolong the liposome circulation time in vivo” Bio Liver". Acta Radiologica 30 (1989), Fasc. 4, p407-413. chimica et Biophysica Acta, 1992, 1108(2), 257-60. 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E Nus? * d) E H s U.S. Patent Sep. 2, 1997 Sheet 13 of 16 5,662.930 Fig. 13 s R e Se O C O O O O C O CD s CO N up r N vis euSe u eSOO % U.S. Patent Sep. 2, 1997 Sheet 14 of 16 5,662.930 Fig. 14 7O.OO 60.00 50.00 40.00 : 30.00 20.00 1.O.OO O.OO E O - Sp e (s) (S) 5 V O up 2 g"O g9 (S) ; Co o O U.S. Patent Sep. 2, 1997 Sheet 15 of 16 5,662.930 Fig. 15 7O.OO 60.00 50.00 40.00 30.00 2O.OO O.OO O.OO O 2 (S) 5 to (S) C3 ar.9 o O U.S. Patent Sep. 2, 1997 Sheet 16 of 16 5,662,930 Fig. 16 (esopjo%9z)•Mºna6||6uu1ºg 06 09 5,662,930 1. 2 REDUCTION OF LPOSOME-ENDUCED lether (Woodie et al., U.S. Pat. No. 5,013,556 "Liposomes ADVERSE PHYSOLOGICAL REACTIONS with Enhanced Circulation Time", issued May 7, 1991, the contents of which are incorporated herein by reference). This application is a division of U.S. application Ser. No. 247,053, filed May 20, 1994, pending which is a 5 SUMMARY OF THE INVENTON continuation-on-part of U.S. application Ser. No. 207,651, This invention provides a method of administering a filed Mar. 7, 1994 and now abandoned, which in turn is a liposcome composition to an animal which comprises continuation-in-part of U.S. application Ser. No. 065.928, administering to the animal a liposome composition com filed May 21, 1993 and now abandoned. prising an adverse physiological reaction-reducing effective This invention relates to a method for the reduction of amount of a liposome comprising a bioactive agent and a liposome induced adverse physiological reactions when lipid bilayer having a lipid and a surface agent-modified liposome compositions are administered to animals, includ molecule which comprises a Surface-modifying agent and an ing mammals such as humans. These liposome compositions anchor. The liposome used in the method of this invention can be used to encapsulate bioactive agents. typically has a diameter of from about 200 nm to about 5000 Liposomes are completely closed lipid bilayer mem 15 nm, more preferably, from about 400 nm to about 5000 nm, branes containing an entrapped aqueous volume. The bilayer and most preferably, presently, from about 400 nm to about is composed of lipid molecules which comprise monolayers 1000 nm. Liposomes having such diameters are preferably having a hydrophobic region and a hydrophilic region. The interdigitation-fusion liposomes (IFVs), but can also be structure of the bilayer is such that the hydrophobic large unilamellar liposomes (LUVs) or multilamellar lipo (nonpolar) acyl chain region of the lipid monolayers orient somes (MLVs). The concentration of the surface agent toward the center of the bilayer while the hydrophilic (polar) modified molecule in the bilayer is typically at least about 2 lipid headgroups orient towards the aqueous phase. mole %, more preferably at least about 5 mole %, and most In a liposome-drug delivery system, a bioactive agent preferably at least about 10 mole %. Preferably, the adverse such as a drug is entrapped in the liposome and then physiological reaction-reducing effective amount of the administered to the patient to be treated (Rahman et al., U.S. 25 liposome is about 50 mg of the liposome per kg of the Pat.