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1 20mg CTD 2.4

2

3

4

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

6 2.4 MPEG-DSPE (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 JNS Doxil-1 Doxil-2 JNS002 mg/ml Doxil-1 Doxil-2 JNS002 Doxil MPEG-DSPE 3.19 HSPC NA NA NA L ph 6.5 NA 6

7 C26 HEY A2780 ( ) (A2780/AD) Doxil-2 PC- 3 SPL SPL SPL JNS002 ( ) A2780/AD 10 mg/kg a mg/kg SPL 5 ml/kg a 1 3 mg/kg , SPL 2 ml/kg a 1 3 mg/kg SPL 2 ml/kg 2 mg/ml SPL a in vitro a SPL SPL 1.5, 7.5, ml/kg a SPL 1 ml/kg SPL, 0.25, 0.5, 1.0 ml/min a GLP

8 JNS002 Doxil-2 ( ) C mg/kg Doxil mg/kg HEY mg/kg PC-3 Doxil mg/kg mg/kg A mg/kg/day, mg/kg/day ( ) 1 Balb/c 1 12 mg/kg 2 9 mg/kg C mg/kg mg/kg 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 mg/kg Doxil mg/kg FOB 9 mg/kg 8

9 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 ml/ 3 SPL SPL 2 H 1 H 2 SPL SPL SPL 3 A2780/CDDP Gallo SPL 9

10 C- STEALTH JNS C-JNS002 Doxil-1 Doxil-2 JNS002 JNS002 STEALTH STEALTH JNS mg/kg Doxil-1 4.0, 8.0, 12.0 mg/kg 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 JNS mg/kg 1.0 mg/kg JNS mg/kg JNS mg/kg 1.0 mg/kg Doxil mg/kg Doxil mg/kg JNS , 1.0 mg/kg 1.0 mg/kg JNS mg/kg 6.0 mg/kg Doxil mg/kg 0.9 mg/kg JNS mg/kg 1.0 mg/kg C-JNS mg/kg 14 C- 1.0 mg/kg JNS , 1.0 mg/kg 1.0 mg/kg JNS , 1.0 mg/kg JNS mg/kg 1.0 mg/kg JNS , 1.0 mg/kg 1.0 mg/kg JNS , 1.0 mg/kg JNS mg/kg 1.0 mg/kg C-JNS mg/kg 14 C- 1.0 mg/kg

11 HPLC-FLD FLD HPLC-FLD µg/ml µg/ml FLD 0.05 µg/ml 0.06 µg/ml 0.11 µg/ml 0.22 µg/ml response factor 2 TR C- 14 C-JNS002 2 Doxil-2 V z V z % 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 Doxil STEALTH Doxil-2 Doxil-1 Doxil-250/ Doxil-250/Histidine Histidine

12 2.4 V ss V z t 1/2 MRT CL 3 C26 AUC PC-3 Doxil-2 Doxil-2 % of dose STEALTH V z t 1/2 14 C-JNS C ) 4 2) BLQ CL BLQ % ) 14 C-JNS C- 12

13 Doxil-2 JNS ASA GLP JNS002 Doxil mg/kg a) a) / SPL d) 10 1 /3 b) c) SPL e) SPL e) JNS002 SPL mL JNS002 SPL mL ASA ASA PCA ASA a) Doxil-2 b) c) d) JNS mg/kg e) JNS mg/kg 1 Doxil-2 Doxil mg/kg 12 mg/kg 4/10 8 mg/kg 1/10 8 mg/kg Doxil mg/kg mg/kg 1/3 2.1 mg/kg 2.1 mg/kg 13

14 2.4 2 JNS mg/kg SPL 1.0 mg/kg SPL 1.5 mg/kg /15 1/ mg/kg 1/ / mg/kg 0.25 mg/kg 1.0 mg/kg JNS002 JNS002 JNS002 JNS mg/kg JNS mg/kg SPL 1.0 mg/kg SPL 0.75 mg/kg 0.75 mg/kg 4 JNS002 1/6 2/6 JNS002 JNS002 JNS002 JNS mg/kg 3 JNS002 JNS002 4 JNS002 JNS JNS mg/kg SPL mg/kg

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

16 SPL 13 1 /3 JNS mg/kg /3 JNS mg/kg in vitro µg/plate a) L5178Y TK TK +/- in vitro 1.28 µg/ml a) CHO in vitro µg/ml a) mg/kg a) b) JNS mg/kg mL mL JNS mg/kg ASA JNS002 5 mg/kg c) a) b) c) SPL JNS002 JNS002 SPL SPL SPL SPL b) SPL SPL MPEG-DSPE HSPC MPEG-DSPE MPEG-DSPE 1000 mg/kg mg/kg/ HSPC HSPC 672 mg/kg mg/kg/ HSPC 10% HSPC 16

17 % 300 mg/kg mg/kg/ 1975 mg/kg/ 8 1 a) Doxil-2 Doxil-2 b) JNS JNS002 JNS002 c) JNS002 JNS002 JNS d) JNS002 JNS002 JNS002 JNS002 e) JNS002 17

18 2.4 f) JNS002 JNS002 SPL JNS002 4) 5) JNS002 6) JNS002 g) JNS002 7) 2 JNS002 JNS002 JNS002 JNS002 8) 9) JNS002 JNS ) Balb/c 1 12 mg/kg 2 9 mg/kg C mg/kg mg/kg

19 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) ) STEALTH STEALTH 4) 5) 14 C-JNS C- 14 C-JNS C- 6)

20 2.4 7) 14 C-JNS C- 1) 8 mg/kg mg/kg 2) 3) 4) 5) 6) SPL SPL STEALTH ) Samuels BL, Vogelzang NJ, Ruane M, Simon MA. Continuous venous infusion of doxorubicin in advanced sarcomas. Cancer Treat Rep. 1987; 71: ) 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: ) Tavoloni N, Guarino AM. Disposition and metabolism of adriamycin in the rat. Pharmacology. 1980; 21: ) 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:

21 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: ) 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: ) 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: ) Rudolph R, Stein RS, Pattillo RA. Skin ulcers due to adriamycin. Cancer. 1976;38: ) 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:

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2 94.3 91.3 5.1 7.5 0.0 0.0 0.1 0.5 0.6 0.1 0.1 0.4 21.4% 15.8% 14.8% 15.0% 16.0% 16.5% 0.5% 16.1% 15.2% 16.9% 15.7% 17.1% 18.6% 0.4% 21.4% 15.8% 14.8 15 7 8,000 15 4 1 0 5 15 4 2 15 10 1 15 4 1 6 11 4,500 3,500 16 26 35 27 34 16 2 19 16 2 24 16 3 15 1 2 94.3 91.3 5.1 7.5 0.0 0.0 0.1 0.5 0.6 0.1 0.1 0.4 21.4% 15.8% 14.8% 15.0% 16.0% 16.5% 0.5% 16.1%

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