US 20190022235A1 ( 19) United States (12 ) Patent Application Publication (10 ) Pub. No. : US 2019 /0022235 A1 Durfee et al. (43 ) Pub . Date : Jan . 24 , 2019 ( 54 ) OSTEOTROPIC NANOPARTICLES FOR Publication Classification PREVENTION OR TREATMENT OF BONE (51 ) Int. Cl. METASTASES A61K 47 /54 (2006 .01 ) A61K 47 /69 (2006 . 01) ( 71 ) Applicants : Paul N . Durfee, Albuquerque , NM A61K 47 / 10 ( 2006 .01 ) (US ) ; Charles Jeffrey Brinker , A61K 9 / 16 (2006 . 01 ) Albuquerque , NM (US ); Yu - shen Lin , A61P 35 /00 (2006 .01 ) Seattle , WA ( US ) ; Hon Leong , London ( 52 ) U .S . CI. (CA ) CPC . .. A61K 47 /548 ( 2017 . 08 ) ; A61K 47 /6923 (72 ) Inventors: Paul N . Durfee , Albuquerque , NM ( 2017 . 08 ) ; B82Y 5 / 00 ( 2013 .01 ) ; A61K 9 / 16 (US ) ; Charles Jeffrey Brinker , ( 2013 .01 ) ; A61P 35 / 00 ( 2018 .01 ) ; A61K 47/ 10 Albuquerque, NM (US ) ; Yu - shen Lin , ( 2013. 01 ) Seattle , WA (US ) ; Hon Leong , London (57 ) ABSTRACT (CA ) The present disclosure is directed to protocells or nanopar ticles, which are optionally coated with a lipid bilayer, which (21 ) Appl. No. : 16 /068 , 235 can be used for targeting bone tissue for the delivery of bioactive agents useful in the treatment and /or diagnosis of ( 22 ) PCT Filed : Jan . 6 , 2017 bone cancer , often metastatic bone cancer which often occurs secondary to a primary cancer such as prostate ( 86 ) PCT No .: PCT/ US2017 /012583 cancer, breast cancer, lung cancer and ovarian cancer, among $ 371 ( C ) ( 1 ) , numerous others . These protocells or nanoparticles target ( 2 ) Date : Jul. 5 , 2018 bone cancer especially metastatic bone cancer with bioactive agents including anticancer agents and /or diagnostic agents for purposes of treating , diagnosing and / or monitoring the Related U . S . Application Data therapy of the bone cancer . Osteotropic protocells or nano (60 ) Provisional application No .62 / 395 ,196 , filed on Sep . particles , pharmaceutical compositions comprising a popu 15, 2016 , provisional application No . 62 / 276 , 388 , lation of osteotropic protocells or nanoparticles and methods filed on Jan . 8 , 2016 , provisional application No. of diagnosing , treating and / or monitoring therapy of bone 62 / 276 ,297 , filed on Jan . 8 , 2016 . cancer are representative aspects . P 1 . TILLL Patent Application Publication Jan . 24 , 2019 Sheet 1 of 28 US 2019 /0022235 A1 WHI Fig. 11 WMWA Fig. 1B Wwwwwww Fig. 16 Patent Application Publication Jan . 24 , 2019 Sheet 2 of 28 US 2019 /0022235 A1 Fig.2 Therapeutic Payload WK MMMM WWWWMWMNWAM SUMMMM Deliveryof MSNPS(esultsinspecificdelivery oftherapeuticto WWW osteotrophic reducemetastatic . 6 ????????????????????????????i: WWWWWWWWWWWWWWWWWW Metastases UWEN X 1913 . WAS ????????????????? W . Prostatecancerpatientwith ???? UmSdNSWpeoked 34 w * * yumSdNSW Patent Application Publication Jan . 24 , 2019 Sheet 3 of 28 US 2019 /0022235 A1 Cy" ge o ALENDRONATE COOH ( BISPHOSPHONATE ) HOJA OD Fig. 31 Fig. 3B RIS WOW MSNP P2 - A ( ALENDRONATE CONJUGATED ) WM ESTRE WEWE MMMMMMMWMMMMM MUL MSNP P2- COOH (BARE CARBOXYLIC ACID, NEGATIVE CONTROL) Fig. 4B Patent Application Publication Jan . 24 , 2019 Sheet 4 of 28 US 2019 /0022235 A1 TON : Fig. 5A 21 Fig . 5B NOT dronate -FITC NOTdronate -FITO Fig. 50 Alendronate- FITC Alendronate - Firo Fig. 50 Patent Application Publication Jan . 24 , 2019 Sheet 5 of 28 US 2019 / 0022235 A1 Alendronste -ETC NOTdronate - FITC Wwww ?! w soogoo Fig. 58 . 77777777777777777777777777777 . Fig. 57 Patent Application Publication Jan . 24 , 2019 Sheet 6 of 28 US 2019 /0022235 A1 Fig.60" P2-C Fig.6F . P2-A . Fig.6B 68.Fig . www. Fig.61 NEGATIVECONTROL(NOMSNP) 19-621 Patent Application Publication Jan . 24 , 2019 Sheet 7 of 28 US 2019 /0022235 A1 GATED TO RHODAMINE INTENSITY(RFI) riviriwwrrrrrrrrrrrrrwis: LONGANGLESCATTER(SIZE) SM www * * * * * * * * * * * * * LONG ANGLE SCA irriters LONG ANGLE SCATTER ( SIZE ) - Fig. 7 Patent Application Publication Jan . 24 , 2019 Sheet 8 of 28 US 2019 /0022235 A1 AR Fig. 8A Fig. 8B Patent Application Publication Jan . 24 , 2019 Sheet 9 of 28 US 2019 /0022235 A1 NIIN . ZIS] SUH+7 6??? 1 16Bey ????????????????????????????????????????????? ???? RFL RHODAMINE 02 ! ! . ! ! ! ! ! ! ! . 9.4-1?*INTINI 97 . 02 .III1'1 .. SDH9 . JZIS 6?i GI SUNOH w . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . : . OL . Wia . § la serie de la SEMANA RFI 0 RHODAMINE 0+ > LLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLLL 00008 00002 00009 00009 00001 00002 00002 00001 w * * * SOHO * DZIS V6bey * * * 00000000 RFI RHODAMINE Patent Application Publication Jan . 24 , 2019 Sheet 10 of 28 US 2019 /0022235 A1 MERGE Fig. 104 NUCLEATED CELLS LUIDDLE .: Fig. 10B * * * * * WWW LECTIN (VESSELS ) Fig. 100 MSNPS Fig. 100 Patent Application Publication Jan . 24 , 2019 Sheet 11 of 28 US 2019 /0022235 A1 KIDNEY BONE . LIVER VENOUS SPLEEN ARTERIAL BLOOD / BLOOD CELL PLASMA PANCREAS PLASMA - GI TRACT TRACT OTHER - - LUNG - - - - BLOOD PLASMA FLOW LYMPH FLOW Fig. 11 TV Q - L : :BLOOD CELLS : KDEG Kres l ov, L PLASMAPLASMA (1 -0 ) -4 KDEG MSNPA MSNPHYDRÓXYAPATITE INTERSTITIAL SPACE INTRACELLULAR SPACE Fig .12 Patent Application Publication Jan . 24 , 2019 Sheet 12 of 28 US 2019 /0022235 A1 W WWWWWWWWWWWWWWWWWWWWWWW HA ) VIA Y SCHILE COOH -modified MSNP Fig. 134 M Wh Wh XOVE PS . Bisphosphonate. -modified MSNP Fig . 13B Patent Application Publication Jan . 24 , 2019 Sheet 13 of 28 US 2019 /0022235 A1 Hydrodynamic Size Measurements 2007 . Z-Average(d.nm) . COOHBisphophonate-MSNP MSNP Fig . 14 HO ) $ PLOH | a=o 0 0 15 Patent Application Publication Jan . 24 , 2019 Sheet 14 of 28 US 2019 /0022235 A1 Fig.16A ONH Nt . AzidoaceticacidNHSester PROTEINSBY-PRODUCTACTIVEESTER INH)2(laoW9 Alendronate PRIMARYAMINE N=NEN H2N- Omoo?H 0-ACYLISOUREACROSSLINKEDISOUREA EDC •Heterobifunctionalcrosslinkerchemistry(EDC) .CTA NF +OH CARBOXYLICACID •Carboxylicacidfunctionalizedcore(PD2) Dibenzocyclooctyne-Nhydroxysuccinimidylester(DBCO) CoreMSNP 3-(TRIETHOXYSILYL)PROPYLSUCCINICANHYDRIDE .ClickChemistry Patent Application Publication Jan . 24 , 2019 Sheet 15 of 28 US 2019 /0022235 A1 PANY SS AR Fig . 16B Fig. 166 PD2 WWW With Control a a20 b Fig. 17A Patent Application Publication Jan . 24 , 2019 Sheet 16 of 28 US 2019 /0022235 A1 Hydrodynamic Size Measurements 200 Z-Average(d.nm) COOH-BisphophonateMSNP MSNP Fig. 17B Patent Application Publication Jan . 24 , 2019 Sheet 17 of 28 US 2019 /0022235 A1 WWW?K? 7 WWWWWWWWWWWWWWWWWWWWWWW COOH -modified MSNP Fig. 170 Bisphosphonate -modified aSNP Fig . 170 Patent Application Publication Jan . 24 , 2019 Sheet 18 of 28 US 2019 /0022235 A1 PD42 All WW WA WASANII OSNO Control A B Fig. 17 Patent Application Publication Jan . 24 , 2019 Sheet 19 of 28 US 2019 /0022235 A1 PD2-Alendronate 180.Pig PD2-COOHcontrol 881041 Hydroxyapatite(HA) control Fig.181 Patent Application Publication Jan . 24 , 2019 Sheet 20 of 28 US 2019 /0022235 A1 Fig.19 . CombinedMW=507.42g/mol til. PEGsilaneMW550-5000g/mol rits TRIMETHOXYSILYL)PROPYLJAMMONIO}PROPANE-1SULFONATE Op-OH OH :OH HOP-OH Alendronate MW=329.485g/mol H?N 3-{[DIMETHYL( 1 179 •Zwitterioniccoating OCH3 OCH3 •Stericbarrier •PEG H2C0-Si (3-Glycidyloxypropyl)trimethoxysilane Patent Application Publication Jan . 24 , 2019 Sheet 21 of 28 US 2019 /0022235 A1 ES WIZAZ. TER BE A292 . LINK W WIN Fig. 20A Patent Application Publication Jan . 24 , 2019 Sheet 22 of 28 US 2019 /0022235 A1 bonne 20B.Fig . 900 OPEREBORNOPOSODE19392 W Patent Application Publication Jan . 24 , 2019 Sheet 23 of 28 US 2019 /0022235 A1 WRXXXXXXXX 10–12nmporesizebyTEM Fig.200 500 Patent Application Publication Jan . 24 , 2019 Sheet 24 of 28 US 2019 /0022235 A1 PD47 (alendronate /zwitter ) Fig. 211 PD48 (zwitter ) Fig. 21B Patent Application Publication Jan . 24 , 2019 Sheet 25 of 28 US 2019 /0022235 A1 PD42 Control PD2 HA Fig. 224 PD42 - DBCO modified IN WWXXN FISHER PEMKE KIWA KUWA AMES D WARN RETE SETAUTOM 9 Fig . 228 Patent Application Publication Jan . 24 , 2019 Sheet 26 of 28 US 2019 /0022235 A1 PD47 Tibia Pig. 234 PD47 Liver 2 . Fig. 238 PD47 Heart Fig. 236 Patent Application Publication Jan . 24 , 2019 Sheet 27 of 28 US 2019 /0022235 A1 Fig. 248 WWW Fig. 240 Fig . 24B Patent Application Publication Jan . 24 , 2019 Sheet 28 of 28 US 2019 /0022235 A1 WWW Fig . 248 Fig. 240 Fig. 24F Fig. 246 US 2019 / 0022235 A1 Jan . 24 , 2019 OSTEOTROPIC NANOPARTICLES FOR tissues outside of bone . By concentrating drug within the PREVENTION OR TREATMENT OF BONE bony structures, where the vast majority of prostate cancer METASTASES metastases are found , greater therapeutic effect is achieved . These osteotropic nanoparticles are composed of mesopo CROSS -REFERENCE TO RELATED rous silica that can contain various payload types within the APPLICATIONS pores and where the surface of the nanoparticle and / or the protocell is decorated ( generally, through chemical conju [ 0001 ] This application claims the benefit of the filing date gation ) with therapeutic agents , such as bisphosphonates . of U . S . application Ser . No. 62 /276 ,388 , filed on Jan . 8 , Bisphosphonates are widely used to inhibit bone loss / for 2016 , U . S . application Ser. No . 62 / 276 ,297 , filed Jan . 8 , mation and covalently bind to hydroxyapatite (Russell et al. , 2016 , and U . S . application Ser . No . 62 /395 , 196 , filed on 2007 ) , the mineralized calcium - based component of bone , Sep . 15 , 2016 , the disclosures of which are incorporated by and therefore bone targeting using bisphosphonate -deco reference herein . rated nanoparticles is a logical goal. After nanoparticle