HITTA DENNAUS UNUNI 20180021259A1 IN UN IN THE COUNT ( 19) United States (12 ) Patent Application Publication ( 10) Pub . No. : US 2018 / 0021259 A1 HELLER et al. (43 ) Pub . Date : Jan . 25 , 2018 ( 54 ) DYE - STABILIZED NANOPARTICLES AND A61K 31/ 4184 (2006 .01 ) METHODS OF THEIR MANUFACTURE AND A61K 31 /4155 (2006 .01 ) THERAPEUTIC USE A61K 31 / 343 ( 2006 .01 ) A61K 31/ 404 ( 2006 .01 ) (71 ) Applicant: Memorial Sloan Kettering Cancer A61K 31 /4745 ( 2006 . 01 ) Center , New York , NY (US ) A61K 31 /517 ( 2006 . 01 ) A61K 47 / 22 ( 2006 .01 ) ( 72 ) Inventors: Daniel A . HELLER , New York , NY ( 52 ) U . S . CI. (US ) ; Yosef SHAMAY , New York , NY CPC .. .. .. A61K 9 / 1617 (2013 . 01 ) ; A61K 9 / 1694 (US ) ( 2013 .01 ) ; A61K 47 / 22 (2013 .01 ) ; A61K 31/ 44 ( 2013 .01 ) ; A61K 31 / 337 ( 2013 .01 ) ; A61K ( 73 ) Assignee : Memorial Sloan Kettering Cancer 31/ 565 (2013 . 01 ) ; A6IK 31/ 519 ( 2013 . 01 ) ; Center , New York , NY (US ) A61K 31 / 553 ( 2013 . 01 ) ; A61K 31/ 635 ( 2013 . 01 ) ; A61K 31/ 42 ( 2013 . 01 ) ; A61K ( 21 ) Appl. No. : 15 /549 , 985 31 /4166 (2013 .01 ) ; A61K 31/ 436 (2013 .01 ) ; A61K 31/ 4184 (2013 .01 ) ; A61K 31/ 4155 ( 22 ) PCT Filed : Feb . 9 , 2016 ( 2013 .01 ) ; A61K 31/ 343 ( 2013 .01 ) ; A61K 31/ 404 ( 2013 .01 ) ; A61K 31/4745 (2013 . 01 ) ; ( 86 ) PCT No. : PCT /US16 / 17153 A61K 31/ 517 ( 2013 .01 ) ; G06F 19 /706 $ 371 (c ) ( 1 ) , ( 2013 .01 ) ( 2 ) Date : Aug. 9 , 2017 ( 57 ) ABSTRACT Related U . S . Application Data Described herein are nanoparticles which are largely made of ( e . g ., 90 wt. % ) hydrophobic drugs and are stabilized by ( 60 ) Provisional application No . 62 / 114 , 507 , filed on Feb . water soluble dyes . The nanoparticles can range in size from 10 , 2015 . 30 nm to 150 nm and have highly negative surface charge ( e . g . , -55 mV ) . These nanoparticles are highly soluble in Publication Classification water , stable for days in PBS buffer and can be easily (51 ) Int . Cl. lyophilzed and reconstituted in water. Using quantitative A61K 9 / 16 ( 2006 .01 ) self - assembly prediction calculations , topochemical A61K 31 /44 ( 2006 . 01 ) molecular descriptors were identified and validated as highly A61K 31 / 337 ( 2006 .01 ) predictive indicators of nano -assembly , nanoparticle size , A61K 31 / 565 ( 2006 . 01 ) and drug loading . The resulting nanoparticles selectively A61K 31 /519 ( 2006 .01 ) targeted kinase inhibitors to caveolin - 1 - expressing human A61K 31 /553 ( 2006 .01 ) colon cancer and autochthonous liver cancer models to yield A61K 31/ 635 ( 2006 .01 ) striking therapeutic effects while avoiding PERK inhibition A61K 31 /42 ( 2006 .01 ) in healthy skin . The nanoparticles exhibited remarkable A61K 31 /4166 ( 2006 .01 ) anti -tumor efficacy in vitro and in vivo in models of hepato A61K 31 /436 ( 2006 . 01 ) cellular carcinoma. NANO -PRECIPITATION SORAFENB (80 % ) ORGANIC PHASE to w PROBE SONICATOR wwwww .3 07 wwwwwBaxt DYE- SORAFENIB NANOPARTICLE www . SALINE, SUCROSE 5 % CENTRIFUGATION ULTRA- SONICATION IR - 783/ 820 ( 14 % ) w hout 20 GO* WATER PHASE PELLET SEM X200 . 000 wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww Hzc CHI W HIC CHE Patent Application Publication Jan . 25 , 2018 Sheet 1 01of JT51 US 2018 / 0021259 A1 ONa CH3H3C 0 : 00 : 0 E E OOO WW taongitoNao. 1R820FIG.1C 1C.FIG wit FIG.1F CH3H3C OO 0 : 02 CH3 CHHICdi FIG.1B 1R806FIG.1B -CH3CHE ENTECH3C 1R780 FIG.1E NOCHEO CH3 : 09 : Q CHEHzC EHOJEHO NaOS v FIG.1AR783 FIG.1A 1R775 FIG.1D CHaHiC CHOCH : 00 : 0 Patent Application Publication Jan . 25 , 2018 Sheet 2 of 51 US 2018 / 0021259 A1 DYE-SORAFENIB NANOPARTICLE SEM.X200000 m 20> U ULTRA-SONICATION PROBESONICATOR Whil . www d wusuwwwwwwwwwwantin E PELLET ORGANICPHASE CENTRIFUGATION FIG.2 maurreau Sovies. WATERPHASE NANO-PRECIPITATION www ZI SORAFENIB(80%) och -SON HO R.783/620(14%) o CHOSH mmmmmmmmmmmmmmmmmmmmmmmmmmmmm CO* mmmmmmmmmmmmmmmmmmmmmmmmmmmmmmmmmm Patent Application Publication Jan . 25 , 2018 Sheet 3 of 51 US 2018 / 0021259 A1 167900 DirectMag:150000X AMTCameraSystem 61.50nm 001WU FIG.3B WU6779 X PrintMag:[email protected] 15:51367/312014 50.06nm 150K2mes, ! * TAI E E = * * CFFFE re* * PEEI * * r * * . TE with r * 00 = * SIZE(d.nm):%INTENSITY Errr 100 - = re:r. = - = - = .LX 4722 = re - SIZEDISTRIBUTIONBYINTENSITY re = - PEAK3:0.000 = - SIZE(d.nm) res . - - - PEAK1:1028 - - . - PEAK2: . - RH - - r.- - - - FIG.3A - - . - - - - r - - - r - - - ra - - - km - www .r wwwwwwww - - ' * - .r- - ' - FF ' Pal:0.154 - - INTERCEPT:0.975 - - FFF - - E - - TE - Z-AVERAGE(d.nm):9582 - - o. RESULTQUALITY:GOOD - o 77 fecer Patent Application Publication Jan . 25 , 2018 Sheet 4 of 51 US 2018 / 0021259 A1 1.5 1 CONCENTRATIONOFSEBINug/mi 100mm 11000-XPEIDEÁ69660=74 05 FIG.3E ??? ????????? ? omjo?? ABSORBANCE o aan 001 98USA%, wwwwe' YORUANOsoovnossomorenosokoos ous wwwwwwwwwwwwwwwwwwwwwwwwwwww 098 watch WAVELENGTH(nm) 008 FIG.3D MULTUM wwwwwwwwwwwwwwwww * AR * NO ovom o wwwwww 097 Luta 69806100wanang 0.240 02 007 ABS FIG.3C Patent Application Publication Jan . 25, 2018 Sheet 5 of 51 | US 2018/ 0021259 A1 REE- SEE R83 C ALII ) LLC) Engis ? LI RAREA MP II IP FIG . 4A Patent Application Publication Jan . 25 , 2018 Sheet 6 of 51 US 2018 / 0021259 A1 O R783- SFB O R783 gwodoo ???? MEANFLUORESCENCE TRRRRRRRRRRR menganggangmangangampangang que 313 RAW BMOM SB2 bend3 BAEC 13 264 FIGgggggggg . 4B 313 SB2 1R783 - SFB 313 IR783 -SFB FIG . 4C Patent Application Publication Jan . 25 , 2018 Sheet 7 of 51 US 2018 / 0021259 A1 NR -FLUORESCENCE (820mm GP. CANCER (520mm 5 TUMOR NORMAL TUMOR NORMAL * * OLX-9 o FIG . 5A WY NIRVIR - FLUORESCENCE E (820m GFP- CANCER (5200m * CANCER * 1R783- SF3 & CANCER X106 R/ 83- SF YTY NORMAL 42000W FIG . 5B GPP N IR VERGE R783RESEP- SF6 R783- SFB 1R783 FIG0000000 . 5C Patent Application Publication Jan . 25 , 2018 Sheet 8 of 51 US 2018 / 0021259 A1 R783-SFB30mgkgiv ws TERMINATION SORAFENB30mgkgPO DAY49 35 28 TREATMENT FIG.6A FIG.OB LI 21 0 UNTREATED rrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrfunn INOCULATION DAY DAY21 Patent Application Publication Jan . 25 , 2018 Sheet 9 of 51 US 2018 / 0021259 A1 UNTREATED SORAFENS SOmg kg PO 1R783. SFB 30mg kg iv wwwwwwwwwwwwww FIG . 7A 15. SORAFENB 30mg ko po 1R783 SB Zimg kgiv 0LX-9 299FIG . 7B LIVERWEIGHT(mg) TUMORVOLUME(TOT Suwalubunda wwwwwwwwwwwwwwww * * IIIIIII Www w # H on .5 RIX . SER E R783913 SORAFENBUV 33 UNTREATED Rabbabababababababababond WOWR723. SFB ! SORAFENIB UNTREATED FIG . 7C FIG . 7D N SCRAFENB R783 SFB A FIG . 7E Patent Application Publication Jan . 25 , 2018 Sheet 10 of 51 US 2018 /0021259 A1 NIR 1 MIN X5 DIC 24h X20 R783- SFB FIG . 8A DIC ( SB2 +313 CO- CULTURE ) DIC 1 MIN POST IRRADIATION 24h POST IRRADIATION WWWWWWWWWWWWWW WE R783SFB 373- GFP 373 GFP FIGzooooooo . 8B Patent Application Publication Jan . 25 , 2018 Sheet 11 of 51 US 2018 /0021259 A1 NODRUG NODRUG SORAFONB -SUNDING -ERLOTINIB .FORSKOLIN -CAMPTOTECIN -DMSO co-PACLITAXEL com-ENZALUTAMIDE -TRAMETNB -DMSO 900 ww 900 800 ey 700 800600700 WAVELENGTH(im 600 33333 10000333333 * * 500 w * * * 2 500 FIG.9B ABSORBANCE ABSORBANCE Y 900 900 KKOG www ww*0.1%aut -0.003%wt 800 -1%DNISO .40%DVSO Y 700 800700600 WAVELENGTH(mm WARR www 600 wife puyuyuwayuuuwwwww 500 N -- - 500 s ABSORBANCE ARADANACS mon PACLITAXELSORAFENB PRAMETINIBFULVESTRANT FIG.9A SUNTINGERLOTINB CAMPIOTEONTASELISB DRUGALONE INDOCYANINET + DRUG Patent Application Publication Jan . 25 , 2018 Sheet 12 of 51 US 2018 /0021259 A1 u * «•NOTRAVETINB -0SORAFENIB -RAPAMYCIN -*PACLITAXEL -WOW.AULESTRANT •*ENZALUTAMIDE centre amcare pe CE i schumannren omwon 42 DRUGINDOCYANINERATIO(mol) precum FIG.9D * * wo *** wowo una kunna woun in& medmetweemo en* 8 yuyuqoruyuyorpusuyurgau T ( SIZE PARTIOLE Rannnnnnnnnnnnkannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnkannnnnnnnnnnndununnnnnn PACUTAXEL FULVESTRANT FIG.IC SORAFENIB TRAMETNB Patent Application Publication Jan . 25 , 2018 Sheet 13 of 51 US 2018 /0021259 A1 wwwwwmmmmm ? ? ? ? ? ? ? ? ? ? u ? ? 30W ? ? 4 4 ? 0 ? 0 ? 0 ? ? 3/16VZN3000HddWosil1010WN ? ? ? ? ? ? ? u INVISVINTEXUINONGNSNLBMWX7301 ? ? ? ? ? SZOLIDAYONUNSEOA ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ???? ?? ?? ? ? wwwwww VOLOS Mdud wwwwwwwwwwwww OzidomATS9393941130NOSDIO01 GNIONIONSBN88081817NOSI01&NOI wwwwwwwww » » » » » » ? ? ? L > )s ( Bh _Splexd wwwwwwwwwwwwmmmmaawwwwwwwwwwwwwwwwwwwwww Patent Application Publication Jan . 25 , 2018 Sheet 14 of 51 US 2018 /0021259 A1 R2 - 0. 960 SoMazoBhis fuwwwwwwwwwwwwwwwwww 0 20 40 60 80 100 20 MAX DRUG DYE RATIO FIG . 10B 13 13700 £ 8 ? ? ??? ?? - wwwwwwwwwwwwwwwwwwww wonu ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? SpMaxd_Bhís 7CT510 SUOHO NYLONYNJDI $ mannahanannyian dodd SIM 1S wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww 0 090 019 00 02 01 00 01 02 02 067 097 082 ?? , } ?? FIG . 10C > . * TRAINING SET * VALIDATION SET PREDICTEDSIZE(d.mm . * # GATEWAYR4e . CXII go3D? 0 . # que porannnnnnngannnnnnnnnnnnnnnnnnnnnnnnndu wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww 60 nie80 100 120 140 09 01 00 91 091 SIZE BY NUMBER (MM OBSERVED SIZE (d . om FIG . 10D FIG . 10Exood Patent Application Publication Jan . 25 , 2018 Sheet 15 of 51 US 2018 /0021259 A1 TASEL ISB hat SORA FENB FIG.10H mame e BONOS HYDROGEN OF NUMBER Arnoon FENIBUSB2 many / / SORA.TASEL . Equity FIG.10G ????????? uuuuuuuuuuuuuu .www % 90 AIN ITSTAANHTV ?? TASELSB-R783 SoMAX4-4,88 FIG.10F SORA-ONBR783 SpMax&=743 Patent Application Publication Jan . 25 , 2018 Sheet 16 of 51 US 2018 /0021259 A1 SORAFENIB INPS TRAMETINBINPS R783 BAEC SK-136 C1116 WCF HL60 FIG . 11Anoon Patent Application Publication