USO10722467B2 United States Patent ( 10 ) Patent No.: US 10,722,467 B2 Hayes et al. (45 ) Date of Patent : * Jul. 28 , 2020 ( 54 ) REMOTE LOADING OF SPARINGLY (58 ) Field of Classification Search WATER -SOLUBLE DRUGS INTO CPC A61K 9/127 ; A61K 9/1278 ; A61K 9/1271; LIPOSOMES A61K 9/1275 ; A61K 31/5377 ; (Continued ) ( 71 ) Applicant : ZONEONE PHARMA , INC . , San Francisco , CA (US ) (56 ) References Cited (72 ) Inventors : Mark E. Hayes , San Francisco , CA U.S. PATENT DOCUMENTS (US ) ; Charles O. Noble , San Francisco , 3,932,657 A 1/1976 Rahman CA (US ) ; Francis C. Szoka , Jr., San 4,016,290 A 4/1977 Rahman Francisco , CA (US ) ( Continued ) (73 ) Assignee : ZONEONE PHARMA , INC ., San FOREIGN PATENT DOCUMENTS Francisco , CA (US ) EP 2415470 A1 2/2012 ( * ) Notice : Subject to any disclaimer , the term of this WO WO 2000/009089 Al 2/2000 patent is extended or adjusted under 35 ( Continued ) U.S.C. 154 ( b ) by 0 days . OTHER PUBLICATIONS This patent is subject to a terminal dis claimer . Anderson , B. et al. , “ Enhanced active liposomal loading of a poorly soluble ionizable drug using supersaturated drug solutions. ” Journal ( 21) Appl. No .: 16 / 454,555 of Controlled Release , 162 , pp . 330-339 (2012 ) . (Continued ) ( 22 ) Filed : Jun . 27 , 2019 Primary Examiner Gollamudi S Kishore (65 ) Prior Publication Data (74 ) Attorney, Agent, or Firm Morgan , Lewis & US 2019/0314282 A1 Oct. 17 , 2019 Bockius LLP ; Jeffry S. Mann (57 ) ABSTRACT Related U.S. Application Data The present invention provides liposome compositions con taining sparingly soluble drugs that are used to treat life (63 ) Continuation of application No. 16 / 126,799, filed on threatening diseases . A preferred method of encapsulating a Sep. 10 , 2018 , now Pat. No. 10,507,182, which is a drug inside a liposome is by remote or active loading . ( Continued ) Remote loading of a drug into liposomes containing a transmembrane electrochemical gradient is initiated by co (51 ) Int. Cl. mixing a liposome suspension with a solution of drug, A61K 9/127 ( 2006.01 ) whereby the neutral form of the compound freely enters the A61K 31/4196 ( 2006.01 ) liposome and becomes electrostatically charged thereby ( Continued ) preventing the reverse transfer out of the liposome. There is (52 ) U.S. Ci. a continuous build - up of compound within the liposome CPC A61K 9/1278 ( 2013.01 ) ; A61K 9/0019 interior until the electrochemical gradient is dissipated or all ( 2013.01 ) ; A61K 9/1271 (2013.01 ) ; the drug is encapsulated in the liposome. However, this ( Continued ) (Continued ) Liposome Classification Multilamellar vesicles (MLV ) - Multiple bilayers Surunding uueous CON Unilamellar vesicles - Single bilayer surrounding Ous core Small uniluzycia vesicles (SUV ) + Diamom ***** Large unilanelas vesicles (LUVs ) . Dixeter > : US 10,722,467 B2 Page 2 process as described in the literature has been limited to 9,737,485 B2 * 8/2017 Hayes A61K 9/0019 10,004,759 B2 * 6/2018 Hayes A61K 9/1271 drugs that are freely soluble in aqueous solution or solubi 2001/0006643 A1 7/2001 Hope lized as a water - soluble complex . This invention describes 2001/0033860 A1 10/2001 Gwathmey compositions and methods for remote loading drugs with 2002/0102293 Al 8/2002 Sachse et al . low water solubility ( < 2 mg/ mL ). In the preferred embodi 2002/0168355 Al 11/2002 McDonald et al. 2004/0126886 Al 7/2004 Kan et al . ment the drug in the solubilizing agent is mixed with the 2005/0175684 Al 8/2005 Gwathmey liposomes in aqueous suspension so that the concentration of 2005/0208119 Al 9/2005 Takemoto solubilizing agent is lowered to below its capacity to com 2006/0039965 Al 2/2006 Bock et al. pletely solubilize the drug . This results in the drug precipi 2007/0014845 Al 1/2007 Zhang et al . tating but remote loading capability is retained . The process 2007/0110798 Al 5/2007 Drummond et al. 2007/0116753 Al 5/2007 Hong et al. is scalable and, in liposomes in which the lipid composition 2008/0107747 Al 5/2008 Roederer and remote loading agent are optimized , the resulting drug 2008/0213353 A1 9/2008 Barenholz et al. loaded liposomes are characterized by a high drug - to -lipid 2009/0028931 A1 1/2009 Wasan et al . ratios and prolonged drug retention when the liposome 2009/0092661 Al 4/2009 Huang et al. encapsulated drug is administered to a subject . 2009/0196918 Al 8/2009 Joguparthi et al. 2010/0311731 Al 12/2010 King , Jr. 2011/0178287 A1 7/2011 Glucksmann et al. 10 Claims, 18 Drawing Sheets 2012/0021042 A1 1/2012 Panzner et al . 2016/0324780 A1 11/2016 Hayes et al. 2017/0202776 A1 7/2017 Hayes et al . Related U.S. Application Data FOREIGN PATENT DOCUMENTS continuation of application No. 15 /429,113 , filed on WO WO 2011/092708 8/2011 Feb. 9 , 2017, now abandoned , which is a continuation WO WO 2012/118376 A1 9/2012 of application No. 15/ 216,416 , filed on Jul . 21 , 2016 , WO WO 2014/153192 9/2014 now Pat. No. 9,737,485 , which is a continuation of application No. 14 / 171,654 , filed on Feb. 3 , 2014 , OTHER PUBLICATIONS now abandoned . Berge, S. et al. , “ Pharmaceutical Salts .” Journal of Pharmaceutical Science , 66 , pp . 1-19 ( 1977 ) . (60 ) Provisional application No. 61/ 759,914 , filed on Feb. Brittenham , G.“ Iron - Chelating Therapy for transfusional Iron Over 1 , 2013 . load” , N Engl J Med 364 : 146-56 ( 2011) . Challa , R. et al ., “ Cyclodextrins in Drug Delivery : An Updated (51 ) Int. Cl. Review .” AAPS PharmSciTech , 6 ( 2 ), pp . E329 - E357 (2005 ) . A61K 31/496 ( 2006.01 ) Cullis et al ., “ Influence of pH gradients on the transbilayer transport A61K 38/07 ( 2006.01 ) of drugs, lipids, peptides and metal ions into large unilamellar A61K 31/5377 ( 2006.01 ) vesicles ” , Biochimica et Biophysica Acta , 1331 187-211 ( 1997 ) . A61K 9/00 ( 2006.01 ) Drummond D. et al ., “ Pharmacokinetics and in vivo drug release B01 ) 13/08 ( 2006.01) rates in liposomal nanocarrier development .” J. Pharm.Sci. , 97 ( 11) , A61K 9/19 ( 2006.01 ) pp . 4696-4740 ( 2008 ) . ( 52 ) U.S. CI. Fleming et al. , " Iron overload in human disease ” . N Engl J Med . CPC A61K 9/1275 ( 2013.01 ) ; A61K 31/4196 366 ( 4 ) : 348-59 ( 2012 ) . (2013.01 ) ; A6IK 31/496 ( 2013.01) ; A6IK Fritze et al ., “ Remote loading of doxorubicin into liposomes driven 31/5377 ( 2013.01) ; A61K 38/07 ( 2013.01) ; by a transmembrane phosphate gradient. ” Biochimica et Biophysica B01J 13/08 (2013.01 ) ; A61K 9/19 (2013.01 ) Acta 1758 , 1633-1640 (2006 ) . Guilmette et al. , “ Pharmacokinetics of the iron chelating agent (58 ) Field of Classification Search desferrioxamine as affected by liposome encapsulation : potential in CPC ...... A61K 9/0019 ; A61K 31/4196 ; A61K treatment of chronic hemosiderosis ” . Life Sci . 22( 4 ) :313-2 ( 1978 ) . 31/496 ; A61K 38/07 ; A61K 9/19 ; B01J Ihnat et al. , " Synthesis and solution properties of deferoxamine 13/08 amides ” , J Pharm Sci. 89 ( 12 ) : 1525-36 ( 2000 ) . See application file for complete search history . Lau et al. , “ Liposome - encapsulated desferrioxamine in experimen tal iron overload” . Br J Haematol . 47 ( 4 ): 505-18 ( 1981) . ( 56 ) References Cited Li, C. et al. , “ Novel sulfobutyl ether cyclodextrin gradient leads to U.S. PATENT DOCUMENTS highly active liposomal irinotecan formulation .” Journal of Phar macy and Pharmacology, 63 , pp . 765-773 ( 2011) . 4,310,506 A 1/1982 Baldeschwieler et al. Liu et al. , “ A Versatile Prodrug Approach for Liposomal Core 4,397,867 A 8/1983 Blake Loading of Water- Insoluble Camptothecin Anticancer Drugs .” Jour 5,043,166 A 8/1991 Barenholz nal of the American Chemical Society , vol . 124 , No. 6 ( 2002) . 5,192,549 A 3/1993 Barenholz et al. Loftsson , T. and Brewster , M., “ Pharmaceutical applications of 5,316,771 A 5/1994 Barenholz et al. cyclodextrins : basic science and product development. ” Journal of 5,616,341 A 4/1997 Mayer et al. Pharmacy and Pharmacology , 62 , pp . 1607-1621 (2010 ) . 5,756,069 A 5/1998 Torchilin et al. 5,800,833 A 9/1998 Hope et al . Postma et al. , “ Absorption and biodistribution of 111indium 5,827,532 A 10/1998 Mehlhorn labelled desferrioxamine (111In - DFO ) after subcutaneous injection 5,837,282 A 11/1998 Fenske et al. of 111 In -DFO liposomes " . J Control Release. 58 ( 1 ) :51-60 ( 1999 ) . 5,939,096 A 8/1999 Clerc et al. Rahman et al. , “ Intracellular plutonium removal by liposome 8,029,795 B2 10/2011 Gwathmey encapsulated chelating agent” . Science (Wash . D.C.) 180 :300-302 8,329,213 B2 12/2012 Hong et al . ( 1973 ) . 8,591,942 B2 11/2013 Javeri et al . Weinberg et al ., “ Iron withholding: a defense against disease ” . J 8,871,253 B2 10/2014 Li et al. Alzheimers Dis . 13 ( 4 ): 451-63 (2008 ) . US 10,722,467 B2 Page 3 ( 56 ) References Cited OTHER PUBLICATIONS Young et al. , “ Liposome entrapped desferrioxamine and iron trans porting ionophores : a new approach to iron chelation therapy” . Br J Haematol. 41 ( 3 ) : 357-63 (1979 ) . Zucker , D. et al. , “ Liposome drugs' loading efficiency : a working model based on loading conditions and drug's physicochemical properties .” Journal of Controlled Release , 139 , pp . 73-80 ( 2009 ). * cited by examiner U.S. Patent Jul. 28 ,9 2020 Sheet 1 of 18 US 10,722,467 B2 FIG . 1 Liposome Classification Multilamellar vesicles (MLV ) - Multiple bilayers surrounding aqueous core Unilamellar vesicles - Single bilayer surrounding Small unilamellar vesicles wen - Large unilamellar vesicles (LUV :) U.S. Patent Jul. 28 ,9 2020 Sheet 2 of 18 US 10,722,467 B2 FIG . 2 150 T 100 drug/lipidratio(gmolHSPC) 50 0 3 100 g drug /mol HSPC 200 g drug /mol HSPC U.S.