USOO9468603B2 (12) United States Patent (10) Patent No.: US 9.468,603 B2 McChesney et al. (45) Date of Patent: Oct. 18, 2016 (54) NANOPARTICULATE COMPOSITIONS FOR A61 K3I/7076 (2013.01); A61K 45/06 TARGETED DELIVERY OF ACID LABILE, (2013.01); A61K 47/48061 (2013.01) LIPOPHILIC PRODRUGS OF CANCER (58) Field of Classification Search CHEMOTHERAPEUTICS AND THEIR CPC ........... A61K 47/48061; A61K 45/06; A61 K PREPARATION 9/0019; A61K 9/5123; C07D 305/14: CO7D 314/24: CO7D 407/12: C07D 487/04; C07D (71) Applicant: Arbor Therapeutics, LLC, Etta, MS 491/14: CO7D 493/06; C07D 493/08: C07D (US) 519700 See application file for complete search history. (72) Inventors: James D. McChesney, Etta, MS (US); Igor Nikoulin, San Diego, CA (US); (56) References Cited Steve J. Bannister, Tampa, FL (US); Douglas L. Rodenburg, Thaxton, MS U.S. PATENT DOCUMENTS (US) 4,762,720 A * 8/1988 Jizomoto ............. A61K9/1277 264/4.1 (73) Assignee: ARBOR THERAPEUTICS, LLC, 8.440,714 B2 * 5/2013 McChesney ......... CO7D 305,14 Etta, MS (US) 514,449 2004/0213837 A1* 10/2004 Mantripragada ...... A61K 9,127 424/450 (*) Notice: Subject to any disclaimer, the term of this 2011 0077291 A1* 3, 2011 Chen .................... A61K9/0019 patent is extended or adjusted under 35 514,449 U.S.C. 154(b) by 0 days. 2012/030861.6 A1* 12/2012 Liu ...................... A61K9/0019 424/400 (21) Appl. No.: 14/485,713 2012/0309819 A1 12/2012 McChesney 2015,0297749 A1* 10, 2015 Hahn ................. A61K 49, 1839 (22) Filed: Sep. 13, 2014 424, 1.37 (65) Prior Publication Data FOREIGN PATENT DOCUMENTS US 2015/OO79189 A1 Mar. 19, 2015 WO 2014, 159851 A1 10, 1994 WO WOOOf 45791 * 8/2OOO WO WO 2011050739 * 5/2011 Related U.S. Application Data OTHER PUBLICATIONS (60) Provisional application No. 61/877,521, filed on Sep. 13, 2013. Brown, Sarah, et al., Journal of Drug Delivery, vol. 2012 (Jan. 2012) pp. 1-7.* (51) Int. Cl. Arasto, Docetaxel, pp. 1-3, accessed Feb. 23, 2016.* A 6LX 9/27 (2006.01) Nikanjam et al., “Synthetic nano-LDL witith paclitaxel oleate as a A6 IK3I/337 (2006.01) targeted drug delivery vehicle for glioblastoma multiforme”, Jour A6 IK3I/365 (2006.01) nal of Controlled Release, Elsevier, Amsterdam, NL, vol. 124, No. 3, Nov. 16, 2007 pp. 163-171. A6 IK3I/366 (2006.01) Hackett Michael J. et al., “Fatty acids as therapeutic auxiliaries for A6 IK 3/427 (2006.01) oral and parenteral formulations'. Advanced Drug Delivery A6 IK 3/4745 (2006.01) Reviews, vol. 65, No. 10, Aug. 17, 2012, pp. 1331-1339. A6 IK 3/475 (2006.01) A6 IK3I/5685 (2006.01) (Continued) A 6LX 3/575 (2006.01) A6 IK3I/704 (2006.01) Primary Examiner — Brian-Yong Kwon A6 IK3I/7056 (2006.01) Assistant Examiner — Mark V Stevens A6 IK3I/7068 (2006.01) (74) Attorney, Agent, or Firm — Sam L. Nguyen; A 6LX 3L/7076 (2006.01) Hamilton, DeSanctis & Cha, LLP A6 IK 45/06 (2006.01) A6 IK 47/48 (2006.01) (57) ABSTRACT A6 IK 9/00 (2006.01) In one embodiment, the present application discloses Syn A 6LX 9/5 (2006.01) thetic LDL nanoparticles comprising mixtures of compo A 6LX 3L/357 (2006.01) nents selected from the group consisting of phospholipids, (52) U.S. Cl. triglycerides, cholesterol ester and free cholesterol: option CPC ........... A6IK 9/1275 (2013.01); A61K 9/0019 ally further comprising an agent selected from the group (2013.01); A61 K9/5123 (2013.01); A61 K consisting of natural antioxidants, ubiquinol and vitamin E, 31/337 (2013.01); A61K3I/357 (2013.01); and methods for preparing the synthetic nanoparticles. The A6 IK3I/365 (2013.01); A61K 3 1/366 disclosed synthetic LDL nanoparticles are capable of selec (2013.01); A61 K3I/427 (2013.01); A61 K tively delivering lipophilic drugs and prodrugs to cellular 31/475 (2013.01); A61K 31/4745 (2013.01); targets expressing LDL receptors after intra venous injec A6 IK3I/5685 (2013.01); A61K 3 1/575 tion. (2013.01); A61 K3I/704 (2013.01); A61 K 3 1/7056 (2013.01); A61 K3I/7068 (2013.01); 10 Claims, 45 Drawing Sheets US 9,468,603 B2 Page 2 (56) References Cited Lundberg, B., "Preparation of Drug-Carrier Emulsion Stabilized with Phosphatidylcholine-Surfactant Mixtures”. Journal of Pharma ceutical Sciences, vol. 83, No. 1, 1994, 72-75. OTHER PUBLICATIONS Norvaisas P. et al. The Role of Payload Hydrophobicity in Nanotherapeutic Pharmacokinetics. Journal of Pharmaceutical Sci Nikanjam et al., “Synthetic nano-low density lipoprotein as targeted ences 2014; 103(7): 2147-56. Fayad W. et al., Identification of Agents that Induce Apoptosis of drug delivery vehicle for glioblastoma multiforme”. International Multicellular Tumour Spheroids: Enrichment for Mitotic Inhibitors Journal of Pharmaceutics 328 (2007) 86-94. with Hydrophobic Properties. Chemical Biology & Drug Design Lundberg et al., “A lipophilic paclitaxel derivative incorporated in 2011; 78(4):547-57. a lipid emulsion for parenteral administration'. Journal of Con trolled Release 86 (2003) 93-100. * cited by examiner U.S. Patent Oct. 18, 2016 Sheet 1 of 45 US 9,468,603 B2 Figure 1: Particle size reaches the 55-60 nm plateau after 40 discrete passes. 160 140 120 Filtration 100 s 8O wintensity 60 xx Number 40 s - - - C s Y 10 s 20 i. s s S s 0 or&s O O.O 20.0 40.O 6O.O 80.0 100.0 Fig.1. Number of passes Figure 2: Particle size and stability of drug free formulation. Particle size and stability of drug-free formulation. --OO2.103.1 U.S. Patent Oct. 18, 2016 Sheet 2 of 45 US 9,468,603 B2 Figure 3: Cooling conditions allowed further decreasing of the particle size to 43 nm Filtration 4. asam intensity is 15.0 3 O. O : Volume ox Number Particle size and stability of drug-free firrituatics, -- 302,103.1 i ses (02,164.1 Fis. 4. Timc. hays 0.103.1-processed at , (). 103.1-processed at... U.S. Patent Oct. 18, 2016 Sheet 3 of 45 US 9,468,603 B2 Figure 5: Representative graph showing particle size reaches the plateau or resistance. Lt C.S. & 15, d 1. 500 100,0 Filtrati 40. (mama Intensity s 80.0 * , alume & O * Flumber O. Figure 6: Representative graph showing different particle size and Stability over time. s-------------------- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -------------------------------------------------- w S Particle size and stability of ART-2O7 containing formulation. 200 180 160 E 140 o n 12O u Cl .2 100 a k 8O h 60 sec 40 ----------------------------------------- 2O is . ................. -- O is assassssssssssssssssssssssssssssssss O 40 Time, Days U.S. Patent Oct. 18, 2016 Sheet 4 of 45 US 9,468,603 B2 Figure 7: Representative graph showing particle size at different temperatures. OOO2.108.1 10 15 Fig. 8. Time, Days U.S. Patent Oct. 18, 2016 Sheet S of 45 US 9,468,603 B2 Figure 9: Representative graph showing change in particle size over time and temperature. 12O 120 1OO - OO Filtration S 80 8O \sas intens ity U v.n 60 so “s Volume S C wasa Number a- s S 40 YAVM 40 C VN Yawass S S 2O a N--MVVVMVVVMAWWAV 20 O Rattraverrarrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrti. O Particle size and stability of ART-207 Containing formulation. --002.109.1 Fig. 10. Time, Days k &\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\ U.S. Patent Oct. 18, 2016 Sheet 6 of 45 US 9,468,603 B2 Figure T 1: Representative graph showing relative particle size over processing time and temperatures 002.111.1 co 6O Filtration as Ya R2 s N y \a Sas sasantens ity NY as A. O wVolume S. w Number s axSaxas x s s yS. C s 20 Salaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa-ra-as-as-a-s.S.------------- 20 S. s s s w O - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -. O s O 2O AO 60 Fig.11. Processing time, min Figure 12: Representative graph showing particle size and Stability over time -0-002111.1 Fig. 12. Time, Days U.S. Patent Oct. 18, 2016 Sheet 7 of 45 US 9,468,603 B2 Figure 13: Representative graph showing particle size over time and temperature 12O 1OO Filtration 8 O &N Interns ity 6 O x Volume Nsaw Number 4O Fig.13. Processing time, min w Figure 14: Representative graph showing particle size and stability of ART-207 over time. Particle size and stability of ART-2O7 containing formulation. 80 70 60 50 x 40 30 nue -- OO2.111.2 20 10 on US 9,468,603 B2 Figure 16: Representative graph showing particle size and Stability of ART-207 formulation &rlsrurrarvarvars rurrivers ravarvervs rurrivervs rurrive U.S. Patent Oct. 18, 2016 Sheet 9 of 45 US 9,468,603 B2 Figure 17: Representative graph showing particle size and processing times 6O Filtration \ssa Inten sity 40 xxx Volume asa Number C O 2O 40 Fig.17. Processing time, min wxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Figure 18: Representative graph showing particle size and stability of drug-free formulations Particle size and stability of drug-free formulations. ar.002.118.0 was OO2.118.00 Fig. 18. Time, Days S----------------------------------------------------------------------------------------------------------- U.S. Patent Oct. 18, 2016 Sheet 10 of 45 US 9,468,603 B2 Figure 19: Representative graph showing particle size over processing times and temperatures Filtration 30 &N Intensit y 6 O 6O wVolume asa Number 4O 40 2 O Fig.19. Processing time, min Figure 20: Representative graph showing particle size and stability over time Particle size and stability of ART-207 containing formulation. 2OO 150 100 -O-OO2.119.4 Fig. 20. Time, Days U.S. Patent Oct. 18, 2016 Sheet 11 of 45 US 9,468,603 B2 Figure 21: Representative graph showing stability of drug-free and ART-207 containing formulations Stability of drug-free and ART- containing Tormulations shipped to/from MTD site (- - -) and retained by Arbor Therapeutics O OO2-119-4 O02-119-3 -902-119-2 OO2-119-1 50 iss------ OQ2-118-00 O 5 10 15 2O 25 Fig.