20mg CTD 2.4

2.4 2.4... 5 2.4.1... 5 2.4.2... 7 2.4.3... 10 2.4.4... 13 2.4.5... 18 2.4.6... 20 3

2.4 2.4 2.4.1 JNS002 ( HSPC) N-(Carbonyl-methoxypolyethylene glycol 2000)-1,2- distearoyl-sn-glycero-3-phosphoethanolamine sodium salt (Sodium MPEG-2000-Carbonyl-DSPE MPEG-DSPE) 2.4.1-1 L- ph 2.4.1-1 STEALTH 2.4.1-2 MPEG-DSPE Methoxypolyethylene Glycol MPEG STEALTH 150 nm 2.4.1-1 MPEG-DSPE 2.4.1-1 20 mg mg/vial 20 Sodium MPEG-2000-Carbonyl-DSPE (MPEG-DSPE) 31.9 HSPC 95.8 31.9 20 940 L- 15.5 ph ph 15 5

2.4 MPEG-DSPE 2.4.1-2 (JNS002) STEALTH STEALTH SPL 20 SPL SPL MPEG-DSPE HSPC SPL MPEG-DSPE Doxil-1 Doxil-2 L- JNS002 Doxil-3 Doxil-2 Doxil-1 Doxil-2 JNS002 2.4.1-2 2.4.1-2 Doxil-1 Doxil-2 JNS002 mg/ml Doxil-1 Doxil-2 JNS002 Doxil-3 2.00 MPEG-DSPE 3.19 HSPC 9.58 3.19 2 94 NA NA NA L- 1.55 ph 6.5 NA 6

2.4 2.4.2 C26 HEY A2780 ( ) (A2780/AD) Doxil-2 PC- 3 SPL SPL SPL 2.4.2-1 JNS002 ( ) A2780/AD 10 mg/kg 4.2.1.1.3 a 1 3 9 mg/kg 4.2.1.3.1 SPL 5 ml/kg a 1 3 mg/kg 4.2.1.3.2, SPL 2 ml/kg a 1 3 mg/kg 4.2.1.3.2 SPL 2 ml/kg 2 mg/ml SPL a in vitro 4.2.1.3.4 a SPL SPL 1.5, 7.5, 4.2.1.3.5 15 ml/kg a SPL 1 ml/kg SPL, 0.25, 0.5, 1.0 ml/min a GLP 4.2.1.3.6 7

2.4 2.4.2-2 JNS002 Doxil-2 ( ) C26 1.5 3.0 6.0 mg/kg 1 2 4.2.1.1.1 Doxil-2 6 9 mg/kg HEY 4.2.1.1.2 6 9 mg/kg PC-3 Doxil-2 6 9 mg/kg 4.2.1.1.4 6 9 mg/kg A2780 5 mg/kg/day, 7 3 20 mg/kg/day 4.2.1.4.1 3 4 ( ) 1 Balb/c 1 12 mg/kg 2 9 mg/kg C26 1.5 6.0 mg/kg 1 2 6.0 mg/kg 1 3 1 1 HEY Doxil-2 6 mg/kg 9 mg/kg 1 Doxil- 2 6 mg/kg 9 mg/kg 6 mg/kg 9 mg/kg Doxil-2 A2780 A2780/AD A2780/AD P PC-3 Doxil-2 Doxil-2 9 mg/kg 9 mg/kg Doxil-2 9 mg/kg 64 19 18 9 mg/kg 20 19 Doxil-2 2 1 3 9 mg/kg FOB 9 mg/kg 8

2.4 1 3 mg/kg 1 SPL 1 3 mg/kg SPL 1 3 mg/kg SPL PR RR QRS QT QTc 1 3 mg/kg 1 ph 1 3 mg/kg SPL ph SPL SPL 9 1 ml/ SPL 7 0.25 0.5 1.0 ml/ 3 SPL SPL 2 H 1 H 2 SPL SPL SPL 3 A2780/CDDP Gallo 4.2.1.4.1 4 SPL 9

2.4 2.4.3 2.4.3-1 14 C- STEALTH JNS002 14 C-JNS002 Doxil-1 Doxil-2 JNS002 JNS002 STEALTH STEALTH 2.4.3-1 JNS002 0.3 1.0 mg/kg 4.2.2.2.1 Doxil-1 4.0, 8.0, 12.0 mg/kg 4.2.2.2.2 Doxil-2 4.0, 8.0, 12.0 mg/kg Doxil-250/Tris (JNS002) 0.3 mg/kg Doxil-250/Histidine (JNS002) 0.3 mg/kg 4.2.2.2.3 JNS002 1.0 mg/kg 1.0 mg/kg 4.2.2.2.4 JNS002 1.0 mg/kg 4.2.2.2.5 JNS002 1.0 mg/kg 1.0 mg/kg 4.2.2.2.6 Doxil-1 1.5 mg/kg 4.2.2.2.7 Doxil-2 1.5 mg/kg JNS002 0.25, 1.0 mg/kg 1.0 mg/kg 4.2.2.2.8 JNS002 6.0 mg/kg 6.0 mg/kg 4.2.2.3.1 Doxil-2 0.9 mg/kg 0.9 mg/kg 4.2.2.3.2 JNS002 1.0 mg/kg 1.0 mg/kg 4.2.2.3.3 14 C-JNS002 1.0 mg/kg 14 C- 1.0 mg/kg 4.2.2.3.4 JNS002 0.25, 1.0 mg/kg 1.0 mg/kg 4.2.2.3.5 JNS002 0.75, 1.0 mg/kg 4.2.2.3.6 JNS002 1.0 mg/kg 1.0 mg/kg 4.2.2.4.1 JNS002 0.25, 1.0 mg/kg 1.0 mg/kg 4.2.2.4.2 JNS002 0.75, 1.0 mg/kg 4.2.2.4.3 JNS002 1.0 mg/kg 1.0 mg/kg 4.2.2.5.1 14 C-JNS002 1.0 mg/kg 14 C- 1.0 mg/kg 4.2.2.5.2 10

2.4 1 1 HPLC-FLD FLD HPLC-FLD 0.005 0.01 µg/ml 0.005 µg/ml FLD 0.05 µg/ml 0.06 µg/ml 0.11 µg/ml 0.22 µg/ml response factor 2 TR 1.4 14 C- 14 C-JNS002 2 Doxil-2 V z V z 2 3 100% 96 STEALTH STEALTH MPEG-DSPE % LPC % CL V z CL 1.0 mg/kg nm V z 0.3 mg/kg nm nm Doxil-1 Doxil-2 Doxil-2 Doxil- 1 CL V z t 1/2 MRT 2 3 Doxil-2 100 Doxil-1 80 96 STEALTH Doxil-2 Doxil-1 Doxil-250/ Doxil-250/Histidine Histidine 3 1 13 72 11

2.4 V ss V z t 1/2 MRT CL 3 C26 AUC 1.8 2.4 3.2 3.8 7.0 PC-3 Doxil-2 Doxil-2 % of dose 1 1.3 2.5 3.0 24 10 3.0 80 STEALTH V z t 1/2 14 C-JNS002 14 C- 3 1 13 1) 4 2) 3 1 13 BLQ CL BLQ 5 72 3% 48 72 72 8 0 24 3) 14 C-JNS002 14 C- 12

2.4 2.4.4 Doxil-2 JNS002 2.4.4-1 ASA GLP JNS002 Doxil-2 2.4.4-1 mg/kg a) 0 4 8 12 4.2.3.1.8 a) 0 1.5 1.8 2.1 4.2.3.1.11 0 0.25 1.0 1.5 13 1 /3 1.0 4.2.3.2.3 SPL d) 10 1 /3 b) c) 0 0.25 0.75 1.0 1.0 4.2.3.2.7 SPL e) 0 0.1 0.5 1.0 0.2 0.4 4.2.3.5.2.1 SPL e) 0 0.5 1.5 2.5 4.2.3.5.2.2 JNS002 SPL 4.2.3.6.1 0.1 1.0mL JNS002 SPL 4.2.3.6.2 0.1 1.0mL 0.16 0.8 ASA 5 4.2.3.7.1.1 ASA PCA 5 0.8 ASA 4.2.3.7.1.2 5 a) Doxil-2 b) 6 9 12 15 6-15 c) 6 9 12 15 18 d) JNS002 1.5 mg/kg e) JNS002 1.0 mg/kg 1 Doxil-2 Doxil-2 4 8 12 mg/kg 12 mg/kg 4/10 8 mg/kg 1/10 8 mg/kg Doxil-2 1.5 1.8 2.1 mg/kg 28 2.1 mg/kg 1/3 2.1 mg/kg 2.1 mg/kg 13

2.4 2 JNS002 0.25 1.0 1.5 mg/kg 3 1 13 SPL 1.0 mg/kg SPL 1.5 mg/kg 8 22 7/15 1/15 1.0 mg/kg 1/15 36 2/15 66 68 1.0 mg/kg 0.25 mg/kg 1.0 mg/kg JNS002 JNS002 JNS002 JNS002 0.25 mg/kg JNS002 0.25 0.75 1.0 mg/kg 3 1 10 SPL 1.0 mg/kg SPL 0.75 mg/kg 0.75 mg/kg 4 JNS002 1/6 2/6 JNS002 JNS002 JNS002 JNS002 0.25 mg/kg 3 JNS002 JNS002 4 JNS002 JNS002 5 1 JNS002 0.1 0.5 1.0 mg/kg 6 3 1 4 SPL 0.2 0.4 mg/kg 6 15 1 1 14

2.4 SPL 0.1 mg/kg 0.5 mg/kg 1.0 mg/kg 2 JNS002 0.5 1.5 2.5 mg/kg 6 3 1 5 1.5 2.5 mg/kg 1/5 3/5 0.5 mg/kg 1.5 2.5 mg/kg JNS002 6 JNS002 0.1 1.0 ml JNS002 2.0 mg/ml 2.0 SPL JNS002 SPL JNS002 SPL JNS002 7 1 ASA PCA 2 JNS002 JNS002 JNS002 JNS002 JNS002 1.5 3 a) SPL SPL in vitro in vivo SPL SPL 2.4.4-2 15

2.4 2.4.4-2 SPL 13 1 /3 JNS002 1.5 mg/kg 4.2.3.2.3 10 1 /3 JNS002 1.0 mg/kg 4.2.3.2.7 410 273 137 41.0 27.2 13.7 in vitro µg/plate a) 4.2.3.7.7.3.1 L5178Y 41.0 20.5 10.3 5.13 2.56 TK TK +/- in vitro 1.28 µg/ml a) 4.2.3.7.7.3.2 CHO in vitro 41.0 30.8 20.5 10.3 µg/ml a) 4.2.3.7.7.3.3 2.05 4.10 8.20 mg/kg a) 4.2.3.7.7.3.4 b) JNS002 1.0 mg/kg 4.2.3.5.2.1 0.1 1.0mL 4.2.3.6.1 0.1 1.0mL 4.2.3.6.2 JNS002 0.8 mg/kg ASA 4.2.3.7.1.2 JNS002 5 mg/kg c) a) b) 6 9 12 15 c) SPL JNS002 JNS002 SPL SPL SPL SPL b) SPL SPL MPEG-DSPE HSPC MPEG-DSPE MPEG-DSPE 1000 mg/kg 2 100 mg/kg/ 1 420 42 HSPC HSPC 672 mg/kg 2 33.6 mg/kg/ 1 90 5 1 HSPC 10% HSPC 16

2.4 1 10% 300 mg/kg 1 200 13 886 mg/kg/ 1975 mg/kg/ 8 1 a) Doxil-2 Doxil-2 b) JNS002 3 1 13 3 1 10 JNS002 JNS002 c) JNS002 JNS002 JNS002 1.5 d) JNS002 JNS002 JNS002 JNS002 e) JNS002 17

2.4 f) JNS002 JNS002 SPL JNS002 4) 5) JNS002 6) JNS002 g) JNS002 7) 2 JNS002 JNS002 JNS002 JNS002 8) 9) JNS002 JNS002 2.4.5 2.4.5.1 1) Balb/c 1 12 mg/kg 2 9 mg/kg C26 1.5 6.0 mg/kg 1 2 6.0 mg/kg 1 3 18

2.4 2) HEY A2780/AD A2780 Doxil-2 3) PC-3 Doxil-2 4) SPL SPL 5) SPL PR RR QRS QT QTc 6) SPL 7) SPL SPL 1 2 SPL 1) STEALTH STEALTH 96 STEALTH 2) 3 1 13 3) STEALTH STEALTH 4) 5) 14 C-JNS002 14 C- 14 C-JNS002 14 C- 6) 0 24 48 72 19

2.4 7) 14 C-JNS002 14 C- 1) 8 mg/kg 2.1 2.1 mg/kg 2) 3) 4) 5) 6) SPL SPL STEALTH 2.4.6 1) Samuels BL, Vogelzang NJ, Ruane M, Simon MA. Continuous venous infusion of doxorubicin in advanced sarcomas. Cancer Treat Rep. 1987; 71: 971-2. 2) Arnold RD, Slack JE, Straubinger RM. Quantification of doxorubicin and metabolites in rat plasma and small volume tissue samples by liquid chromatography/electrospray tandem mass spectroscopy. J Chromatgr B. 2004; 808: 141-52. 3) Tavoloni N, Guarino AM. Disposition and metabolism of adriamycin in the rat. Pharmacology. 1980; 21: 244-55. 4) Maeda N, Takeuchi Y, Takada M, Sadzuka Y, Namba Y, Oku N. Anti-neovascular therapy by use of tumor neovasculature-targeted long-circulating liposome. J Control Release. 2004;100:41-52. 20

2.4 5) Cornali E, Zietz C, Benelli R, Weninger W, Masiello L, Breier G, et al. Vascular endothelial growth factor regulates angiogenesis and vascular permeability in Kaposi's sarcoma. Am J Pathol. 1996; 149: 1851-69. 6) Amantea M, Newman MS, Sullivan TM, Forrest A, Working PK. Relationship of dose intensity to the induction of palmar-plantar erythrodysesthia by pegylated liposomal doxorubicin in dogs. Hum Exp Toxicol. 1999;18:17-26. 7) Kato M, Makino S, Kimura H, Ota T, Furuhashi T, Nagamura Y. Sperm motion analysis in rats treated with adriamycin and its applicability to male reproductive toxicity studies. J Toxicol Sci. 2001;26:51-9. 8) Rudolph R, Stein RS, Pattillo RA. Skin ulcers due to adriamycin. Cancer. 1976;38:1087-94. 9) Garnick MB, Weiss GR, Steele GD Jr, Israel M, Schade D, Sack MJ, et al. Clinical evaluation of longterm, continuous-infusion doxorubicin. Cancer Treat Rep. 1983;67:133-42. 21