US009181319B2 (12) United States Patent (10) Patent No.: US 9,181,319 B2 Schrum et al. (45) Date of Patent: Nov. 10, 2015 (54) ENGINEERED NUCLEICACIDS AND 4,401,796 A 8, 1983 Itakura METHODS OF USE THEREOF 4411,657 A 10, 1983 Galindo 4.415,732 A 11/1983 Caruthers et al. 4,458,066 A 7/1984 Caruthers et al. (71) Applicant: Moderna Therapeutics, Inc., 4.474,569 A 10, 1984 Newkirk Cambridge, MA (US) 4,500,707 A 2/1985 Caruthers et al. 4,579,849 A 4, 1986 MacCoSS et al. (72) Inventors: Jason P. Schrum, Philadelphia, PA 4,588,585 A 5/1986 Market al. (US); Stephane Bancel, Cambridge, MA is, A 38. Athlet al. (US); Noubar B. Afeyan, Cambridge, 4,816,567- A 3/1989 Cabillyak ( eta. al. MA (US) 4,879,111 A 1 1/1989 Chong 4,957,735 A 9/1990 Huang (73) Assignee: Moderna Therapeutics, Inc., 4.959,314 A 9, 1990 Market al. Cambridge, MA (US) 4,973,679 A 11/1990 Caruthers et al. s 5,012,818 A 5/1991 Joishy (*) Notice: Subject to any disclaimer, the term of this 3. A &E El al. patent is extended or adjusted under 35 5,036,006 A 7, 1991 Sanford et al. U.S.C. 154(b) by 0 days. 5,047,524. A 9/1991 Andrus et al. 5,116,943 A 5/1992 Koths et al. (21) Appl. No.: 14/270,736 5,130,238 A 7, 1992 Malek et al. 5,132,418 A 7, 1992 Caruthers et al. 5,153,319 A 10, 1992 Caruthers et al. (22) Filed: May 6, 2014 5,168,038 A 12/1992 Tecott et al. 5,169,766 A 12/1992 Schuster et al. (65) Prior Publication Data 5,194,370 A 3/1993 Berninger et al. 5, 199441 A 4/1993 Hogle 5,262.530 A 11/1993 Andrus et al. Related U.S. Application Data (Continued) (62) Division of application No. 13/204,609, filed on Aug. 5, 2011, now Pat. No. 8822,663. FOREIGN PATENT DOCUMENTS (60) Provisional application No. 61/401,052, filed on Aug. CA 2376634 A1 12/2000 6, 2010. CA 2473135 C 6/2003 s CA 2795695 A1 10, 2011 EP O194809 B1 3, 1986 (51) Int. Cl. EP 0204401 A1 12, 1986 C7H 2L/04 (2006.01) EP O366400 B1 10, 1989 CI2N 15/63 (2006.01) EP O366400 A2 5, 1990 C07K I4/62 (2006.01) EP O427073 A2 5, 1991 A6 IK3I/395 (2006.01) E SER SEC A6 IK3I/7088 (2006.01) A 6LX3L/75 (2006.01) (Continued) A6 IK38/3 (2006.01) A6 IK38/9 (2006.01) OTHER PUBLICATIONS A6 IK38/28 (2006.01) AOIK 67/OO (2006.01) Attwood, T.K. The babel of bioinformatics. Science 290:471-473, (52) U.S. Cl. 2OOO.* CPC ............... C07K 14/62 (2013.01); A61 K3I/395 (Continued) (2013.01); A61K3I/7088 (2013.01); A61 K 3I/7115 (2013.01); A61K 38/13 (2013.01); A6 IK38/193 (2013.01); A61 K38/28 (2013.01) Primary Examiner Quang Nguyen (58) Field of Classification Search (74) Attorney, Agent, or Firm — DT Ward, PC; Donna T. CPC ............... C07K 14/62: A61Sig K31/7088: Ski A61 K Ward, Jennifer F. Bryan USPC ..................................... 536/23.51; 435/320.1 See application file for complete search history. (57) ABSTRACT (56) References Cited Provided are compositions and methods for delivering bio U.S. PATENT DOCUMENTS logical moieties Such as modified nucleic acids into cells to modulate protein expression. Such compositions and meth 2,008,526 A 7/1935 Wrappler et al. ods include the use of modified messenger RNAs, and are 35 52: A & 3. E. stal useful to treat or prevent diseases, disorders or conditions, or 37 66,907 A 10, 1973 MN, to improve a subjects heath or wellbeing. 3,906,092 A 9, 1975 Hilleman et al. 4,373,071 A 2f1983 Itakura 4,399.216 A 8, 1983 Axel et al. 9 Claims, 9 Drawing Sheets US 9,181,319 B2 Page 2 (56) References Cited 6,190,315 2, 2001 Kost et al. 6,210,931 4, 2001 Feldstein et al. U.S. PATENT DOCUMENTS 6,214,804 4, 2001 Felgner et al. 6,217,912 4, 2001 Park et al. 5,273,525 12, 1993 Hofmann 6,228,640 5/2001 Cezayirli et al. 5,298.422 3, 1994 Schwartz et al. 6,234,990 5/2001 Rowe et al. 5,332,671 T. 1994 Ferrara et al. 6,235,883 5/2001 Jakobovits et al. 5,399.491 3, 1995 Kacian et al. 6,239,116 5/2001 Krieg et al. 5,409,818 4, 1995 Davey et al. 6,251,665 6, 2001 Cezayirli et al. 5.426,180 6, 1995 Kool 6.255,076 T/2001 Widner et al. 5.437,990 8, 1995 Burg et al. 6,258,558 T/2001 Szostak et al. 5,457,041 10, 1995 Ginaven et al. 6,261,584 T/2001 Peery et al. 5,466,586 11, 1995 Davey et al. 6,265,387 T/2001 Wolff et al. 5,484.401 1, 1996 Rodriguez et al. 6,265,389 T/2001 Burke 5,514,545 5, 1996 Eberwine 6,267.987 T/2001 Park et al. 5,527.288 6, 1996 Gross et al. 6,291, 170 9, 2001 Van Gelder et al. 5,545,522 8, 1996 Van Gelder et al. 6,300,484 10, 2001 Duhl 5,554,517 9, 1996 Davey et al. 6,303,378 10, 2001 Bridenbaugh et al. 5,580,859 12, 1996 Felgner et al. 6,303,573 10, 2001 Ruoslahti et al. 5,588,960 12, 1996 Edwards et al. 6,322,967 11, 2001 Parkin 5,589,466 12, 1996 Felgner et al. 6,326,174 12, 2001 Joyce et al. 5,663,153 9, 1997 Hutherson et al. 6,334,856 1, 2002 Allen et al. 5,665,545 9, 1997 Malek et al. 6,355.245 3, 2002 Evans et al. 5,672.491 9, 1997 Khosla et al. 6,368,801 4, 2002 Faruqi 5,674.267 10, 1997 Mir et al. 6,376,248 4, 2002 Hawley-Nelson et al. 5,677,124 10, 1997 DuBois et al. 6,395.253 5/2002 Levy et al. 5,679,512 10, 1997 Laney et al. 6,399,061 6, 2002 Anderson et al. 5,693,622 12, 1997 Wolff et al. 6,406,705 6, 2002 Davis et al. 5,693,761 12, 1997 Queen et al. 6.410,276 6, 2002 Burg et al. 5,697,901 12, 1997 Eriksson 64 13,942 T/2002 Felgner et al. 5,700,642 12, 1997 Monforte et al. 6,433,155 8, 2002 Umansky et al. 5,702,384 12, 1997 Umeyama et al. 6,440,096 8, 2002 Lastovich et al. 5,703,055 12, 1997 Felgner et al. 6,455,043 9, 2002 Grillo-Lopez 5,712,127 1, 1998 Malek et al. 6,491,657 12, 2002 Rowe et al. 5,716,785 2, 1998 Van Gelder et al. 6,500,419 12, 2002 Hone et al. 5,736,137 4, 1998 Anderson et al. 6,500.919 12, 2002 Adema et al. 5,756.264 5, 1998 Schwartz et al. 6,514,498 22003 Antonsson et al. 5,759, 179 6, 1998 Balbierz 6,514,948 2, 2003 Raz et al. 5,766,903 6, 1998 Sarnow et al. 6,517,869 2, 2003 Park et al. 5,773,244 6, 1998 Ares, Jr. et al. 6,520,949 2, 2003 St. Germain 5,776.456 7, 1998 Anderson et al. 6,525, 183 2, 2003 Vinayak et al. 5,789,554 8, 1998 Leung et al. 6,527,216 3, 2003 Eagelman et al. 5,807,707 9, 1998 Andrews et al. 6,528,262 3, 2003 Giladet al. 5,824,307 10, 1998 Johnson 6,534.312 3, 2003 Shiver et al. 5,824,497 10, 1998 Andrews et al. 6,552,006 4/2003 Raz et al. 5,840,299 11, 1998 Bendig et al. 6,555,525 4/2003 Burke 5,843,439 12, 1998 Anderson et al. 6,565,572 5/2003 Chappius 5,848,996 12, 1998 Eldor 6,572.857 6, 2003 Casimiro et al. 5,849,546 12, 1998 Sousa et al. 6,586,524 T/2003 Sagara 5,851,829 12, 1998 Marasco et al. 6,589,940 T/2003 Raz et al. 5,861,501 1, 1999 Benseler et al. 6,610,044 8, 2003 Mathiesen 5,869,230 2, 1999 Sukhatime 6,610,661 8, 2003 Carson et al. 5,889,136 3, 1999 Scaringe et al. 6,613,026 9, 2003 Palasis et al. 5,891,636 4, 1999 Van Gelder et al. 6,617, 106 9, 2003 Benner 5,914,269 6, 1999 Bennett et al. 6,623.457 9, 2003 Rosenberg 5,955,310 9, 1999 Widner et al. 6,652,886 11/2003 Ahn et al. 5,958,688 9, 1999 Eberwine et al. 6,653,468 11/2003 GuZaev et al. 5,962,271 10, 1999 Chenchik et al. 6,664,066 12, 2003 Parks 5,962,272 10, 1999 Chenchik et al. 6,670,178 12, 2003 Selden et al. 5,965,720 10, 1999 Gryaznov et al. 6,676,938 1, 2004 Teti et al. 5,965,726 10, 1999 Pavlakis et al. 6,696,038 2, 2004 Mahato et al. 5,980,887 11, 1999 Isner et al.