US 201201498.43A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0149843 A1 Chien et al. (43) Pub. Date: Jun. 14, 2012 (54) NANOFIBERS AND MORPHOLOGY Publication Classification SHIFTING MICELLES (51) Int. Cl. (75) Inventors: Miao-Ping Chien, San Diego, CA C08F 29/2 (2006.01) (US); Nathan C. Gianneschi, San A 6LX 9/70 (2006.01) Diego, CA (US) CI2O I/68 (2006.01) CO8F 299/00 (2006.01) (73) Assignee: THE REGENTS OF THE BOSD 7/4 (2006.01) UNIVERSITY OF A6IR 8/02 (2006.01) CALIFORNLA, Oakland, CA (US) CI2P 19/34 (2006.01) B82Y 4O/OO (2011.01) (21) Appl. No.: 13/363,645 B82Y5/00 (2011.01) (22) Filed: Feb. 1, 2012 (52) U.S. Cl. ......... 525/54.1; 424/401; 424/443; 435/6.1: 435/91.1 : 525/54.2; 427/216:977/788; 977/915; Related U.S. Application Data 977/890; 977/906 (63) Continuation of application No. PCT/US2010/ 044321, filed on Aug. 3, 2010. (57) ABSTRACT (60) Provisional application No. 61/230.924, filed on Aug. The invention discloses novel morphology shifting micelles 3, 2009, provisional application No. 61/316,325, filed and amphiphilic coated metal nanofibers. Methods of using on Mar. 22, 2010. and making the same are also disclosed. Assetsy whe Yxxx x: wa toxia 8A: is:tex : ona ... 8:kyi-xxx8 tapex 88-se: PEG - DNA-5-o- s ot-on-o-8.as -o-on-oh'ss taxi & 8 8-1 cataceoA-8 NA as S-PEG-38AGASKSACrestsgescGA-3 DNA as 8-cacAccoAgios CEO: C(-3 Patent Application Publication Jun. 14, 2012 Sheet 1 of 27 US 2012/0149843 A1 $peritai ixeie *:::s 8x : DNA-5-o-o: * DNA is 8-CGCAccoA-3 DNA a 5-PEG-GSAGAGAGACTGGGGCGA-3 DNA is 5'-cacAccoAGOC CCC-3 Patent Application Publication Jun. 14, 2012 Sheet 2 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 3 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 4 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 5 of 27 US 2012/0149843 A1 0xxx 8-888&Aaxx xxxico88088.xxxarcicacea. www.irs. 8-888-888-38A3Kxxx88088. Yxx, is 8.8x3xxxxx3:38 iyciropric :8:8::::::: Patent Application Publication Jun. 14, 2012 Sheet 6 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 7 of 27 US 2012/0149843 A1 i. § Patent Application Publication Jun. 14, 2012 Sheet 8 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 9 of 27 US 2012/0149843 A1 x figg Patent Application Publication Jun. 14, 2012 Sheet 10 of 27 US 2012/0149843 A1 Fig. 8 Patent Application Publication Jun. 14, 2012 Sheet 11 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 12 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 13 of 27 US 2012/0149843 A1 xt.8six Nicy Pli sixtixx xxix. g &* x8:88:8::::::::388& 3 * x8::::::::::::::::::::::: Patent Application Publication Jun. 14, 2012 Sheet 14 of 27 US 2012/0149843 A1 Fig. if S exity re-extixie keptigiite 8. 8::::::::::::: proteases . ... *:::8. x: 8:8-8 aaaaaaa-Xax 8::::::::::: phos. Kinase pratase * ATP x: x:ix8 six: Patent Application Publication Jun. 14, 2012 Sheet 15 of 27 US 2012/0149843 A1 i Patent Application Publication Jun. 14, 2012 Sheet 16 of 27 US 2012/0149843 A1 Fig. 3. Patent Application Publication Jun. 14, 2012 Sheet 17 of 27 US 2012/0149843 A1 Fig. { 1. 3. s s 88 38 : Tirse ars + xpitatase * 3: y * Kike 3 recialysis gy 2 8xxixiysis Patent Application Publication Jun. 14, 2012 Sheet 18 of 27 US 2012/0149843 A1 sexy Patent Application Publication Jun. 14, 2012 Sheet 19 of 27 US 2012/0149843 A1 Fig. *:::::: 4x :- .. •x s: Nikayae & X s & ck six &3. (guitari & : -: : s x as & is 38 ft 8 &:8: 8:388:::::::::::::: 8888:888: Fig. 8 --x"R" Patent Application Publication Jun. 14, 2012 Sheet 20 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 21 of 27 US 2012/0149843 A1 Patent Application Publication Jun. 14, 2012 Sheet 22 of 27 US 2012/0149843 A1 X TT Assay Patent Application Publication Jun. 14, 2012 Sheet 23 of 27 US 2012/0149843 A1 ig. 22A is go is: 8: . .x : o.3s to is x : xi x i: it wittees ig. 228 is grox s 0.85 ..is is s 8: ine wintes Patent Application Publication Jun. 14, 2012 Sheet 24 of 27 US 2012/0149843 A1 Fig. 23. : : : :x:8 : : : : *:38: 8+ Corp. : : I 8:888 : 3 8 : &g: .338g x838g 88: 8:88:8 Fig. 23B : & W Fiters Fibers + Corp S:38:88. : 3. : 3. ..: i: Kickey &rarr restine Patent Application Publication Jun. 14, 2012 Sheet 25 of 27 US 2012/0149843 A1 Fig. 3 8 &:88 sk:xxx& 8:::::::::::::::: -w Disra N s :::::::::::::::: : &ga:::::::::::::::::::::: N ::::::: Patent Application Publication Jun. 14, 2012 Sheet 26 of 27 US 2012/0149843 A1 : : 88: 8. : ::igi: s 8: 0.8 8 -88.88, 8-3-8:::::::::::::wa & &so.6 . i. 8 8:38.88:::::::::::: 8 *:::::::::::::::::::::: .gs: & "is fire in Patent Application Publication Jun. 14, 2012 Sheet 27 of 27 US 2012/0149843 A1 &xtix. 888xy 88 AA a 88: too 8- + xixxyge it ki O. ni. *:: * Kiky:8: 2 * 0.0 inv { { 0.1 ! . 5 to 15 8. P. s r. sson at Tine (min. {xy 88xy:8-2 sixy-8- US 2012/0149843 A1 Jun. 14, 2012 NANOFIBERS AND MORPHOLOGY BRIEF DESCRIPTION OF THE DRAWINGS SHIFTING MICELLES 0009 FIG. 1 depicts DNA-programmed lipid phase tran CROSS-REFERENCES TO RELATED sitions. DNA-programmed lipid assembles to form spherical APPLICATIONS lamellar vesicles capable of Switching phase to form Small 0001. This application is a continuation of PCT/US2010/ spherical micelles in a fully reversible fashion via DNA 044321, filed Aug. 3, 2010, which claims the benefit of U.S. hybridization (+DNA) and strand invasion (+DNA) cycles. Application Nos. 61/230.924, filed Aug. 3, 2009 and 61/316, Sequences: DNA (SEQID NO:1); DNA (SEQID NO:2). 325, filed Mar. 22, 2010, each of which is incorporated by 0010 FIGS. 2A-2F depict characterization of unilamellar reference in its entirety and for all purposes. vesicle structures formed from the self-assembly of DNA lipid in aqueous solution. FIGS. 2A-2B: Cryogenic-TEM REFERENCE TO A “SEQUENCE LISTING. A (cryo-TEM) images showing unilamellar bilayer morphol TABLE, ORACOMPUTER PROGRAM LISTING ogy of the vesicles at different scales of magnification. The APPENDIX SUBMITTED AS ANASCII TEXT scales for FIGS. 2A-2B are provided in the figures. FIGS. FILE 2C-2D: SEM of vesicles at scales depicted in the figures. FIG. 0002. The Sequence Listing written in file 88654 2E: AFM depiction of vesicle structures dry, on a mica sur 830139 ST25.TXT, created on Jan. 25, 2012, 10,939 bytes, face. FIG.2F: AFM data showing the flattened height profile machine format IBM-PC, MS-Windows operating system, is of the vesicle structures provided in FIG. 2E. hereby incorporated by reference. (0011 FIGS. 3A-3C depict DNA-directed vesicle to micelle phase transition. FIG.3A: FIG. 3A depicts cartoon of BACKGROUND OF THE INVENTION phase transition reaction and results monitored by DLS. “D,” 0003 Various non-informational, non-programmable refers to Hydrodynamic Diameter). FIG. 3B: Spherical nanoparticles have been known in the art, Such as those dis micelles monitored by TEM (negative stain). FIG. 3C: closed in Zhang, et al., Science 272:1777-1779, 1996; LaRue Vesicles monitors by TEM (negative stain). Experimental et al., Macromolecules 39:309-314, 2006: Ishihara et al., conditions: Tris (pH 7.4, 50 mM), MgCl, (50 mM), DNA Chem. Eur: J. 13:4560-4570, 2007; Kim et al., Angew: Chem., lipid (1 uM), DNA (2 uM), room temperature. Int. Ed 46:5779-5782, 2007: Li et al., Macromolecules (0012 FIGS. 4A-4H depict specificity and reversibility of 41:6605-6607, 2008; Roy et al., Chem. Commun. 2106-2108, DNA-programmed phase shifting. Scale bar-1 lum. Two dif 2009; and Fernyhough et al., Soft Matter 5:1674-1682, 2009. ferent DNA-lipids were 3'-labeled with two different dyes: There is a need in the art for micelles that are capable of DNA-Fluorescein and DNA-Rhodamine. All fluorescence changing morphology in a predictable or programmable way. images are red/green channel merges. FIG. 4H: FIG. 4H Provided herein are solutions to these and other problems in depicts a cartoon reaction scheme having reacting species the art. corresponding to each of the images set forth in FIGS. 4A-4F, as indicated by proximity of the reacting species to FIGS. BRIEF SUMMARY OF THE INVENTION 4A-4F. Legend for FIG. 4H: Green fluorescence: gray; red 0004 Provided herein, inter alia, are novel micelles fluorescence: crosshatched; yellow fluorescence: stippled. capable of changing morphology (e.g. shape) in a controlled FIG. 4A: Representative bright field image of labeled DNA manner upon application of the appropriate stimulus as well lipid assemblies. FIG. 4B: DNA-Rhodamine hybridizes to as amphiphilic coated metal nanofibers. The micelles may be DNAs resulting in observed (i.e., green) fluorescence only. used in a variety of chemical, pharmaceutical and electronic FIG. 4C: DNA (complementary to DNAs) causes mixing of applications as described herein. the two Surfactants to give a colocalized signal (i.e., yellow 0005. In one aspect, a micelle is provided that includes a signal) from both dyes. FIG. 4D: DNA-Fluorescein hybrid plurality of aggregated brush copolymers having a hydro izes to DNA to generate micelles resulting in observed (i.e., philic portion and a hydrophobic portion: red) fluorescence only.