USOO9266901 B2 (12) United States Patent (10) Patent No.: US 9.266,901 B2 Pietras et al. (45) Date of Patent: Feb. 23, 2016 (54) COMPOSITIONS AND METHODS FOR (58) Field of Classification Search TREATING CANCER CPC ..................................................... CO7D 493/04 USPC .......................................................... 549/299 (71) Applicant: The Regents of the University of See application file for complete search history. California, Oakland, CA (US) (56) References Cited (72) Inventors: Richard J. Pietras. Sherman Oaks, CA (US); Michael E. Jung, Los Angeles, U.S. PATENT DOCUMENTS CA (US); Diana C. Marquez-Garban, 6,890,946 B2 5/2005 Nakshatri et al. Los Angeles, CA (US); Gang Deng, Los 7.312,242 B2 12/2007 Crooks et al. Angeles, CA (US) 7,678,904 B2 * 3/2010 Crooks ................ CO7D 307.77 544,153 (73) Assignee: The Regents of The University of 8,124,652 B2 2/2012 Crooks et al. 2002fOO58077 A1 5, 2002 Shen et al. Califronia, Oakland, CA (US) 2004/0229936 A1 11/2004 Hsieh et al. (*) Notice: Subject to any disclaimer, the term of this 2010, OOO997O A1 1/2010 Johansen et al. patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. FOREIGN PATENT DOCUMENTS WO WO-2005/007103 A2 1, 2005 (21) Appl. No.: 14/505,288 OTHER PUBLICATIONS (22) Filed: Oct. 2, 2014 Nasim Bioorganic & Medicinal Chemistry Letters 18 (2008) 3870 (65) Prior Publication Data 3873.* Neukirch, Bioorganic & Medicinal Chemistry (2003), 11(7), 1503 US 2015/OO31725A1 Jan. 29, 2015 151O.* Hwang et al., “Synthesis and anti-viral activity of a series of Related U.S. Application Data sesquiterpene lactones and analogues in the Subgenomic HCV replicon system.” Bioorg Med. Chemistry, 13:83-91 (2006). (63) Continuation of application No. PCT/US2013/ Neelakantan et al., "Aminoparthenolides as novel anti-leukemic 035.010, filed on Apr. 2, 2013. agents: Discovery of the NF-kappaB inhibitor, DMAPT (LC-1).” (60) Provisional application No. 61/619.289, filed on Apr. Bioorg. Med. Chem. Letters. 19:4346-4349 (2009). 2, 2012. * cited by examiner (51) Int. Cl. C07D 307/00 (2006.01) Primary Examiner — Nizal Chandrakumar CO7D 49.3/04 (2006.01) (74) Attorney, Agent, or Firm — Mintz, Levin, Cohn, Ferris, A6 IK 45/06 (2006.01) Glovsky and Popeo, P.C. A6 IK3I/365 (2006.01) A6 IK3I/443 (2006.01) (57) ABSTRACT A6 IK3 L/4525 (2006.01) Provided herein, inter alia, are compositions and methods (52) U.S. Cl. useful for treating hyperproliferative diseases, including can CPC ............ C07D493/04 (2013.01); A61K3I/365 cer and non-malignant hyperproliferative diseases. (2013.01); A61 K3I/443 (2013.01); A61 K 3 1/4525 (2013.01); A61K 45/06 (2013.01) 3 Claims, 16 Drawing Sheets U.S. Patent Feb. 23, 2016 Sheet 1 of 16 US 9.266,901 B2 Fig. 1 10 9 - 8 SS 7 s O 6 cy S2 5 Q 4 i. On 3 - 2 1 O CON E2 Fig. 2 15252 O O 5 O p P3 P1 P3 ADH-CD33- ADH-CD 133 U.S. Patent Feb. 23, 2016 Sheet 2 of 16 US 9.266,901 B2 Fig. 3 100 5O O 10 20 50 PTL (micromolar) Fig. 4 1 OO 90 80 70 60 50 40 30 2O 10 Bulk CSC U.S. Patent Feb. 23, 2016 Sheet 3 of 16 US 9.266,901 B2 2 1 2 3 4. 5 6 8 7 8 .x-pass to9 --3028 11 xercissist:12 8 8 t U.S. Patent Feb. 23, 2016 Sheet 4 of 16 US 9.266,901 B2 Fig. 5 (cont'd) 30 2 3 100 4 : 5 6 : 8. 7 8 9 s O .xx. 33.303 10 38.338 Mira &issatist:12 U.S. Patent Feb. 23, 2016 Sheet 5 of 16 US 9.266,901 B2 Fig. 5 (cont'd) 2: WM wim 2:32 win 3282 3 8 . aw 203 4. x-E-- 3:3: 5 0. 53.98 6 88 ww.338 7 wi:2 8 w28 9 exei E2310 W. 2311 wratisatist: 2 U.S. Patent Feb. 23, 2016 Sheet 6 of 16 US 9.266,901 B2 Fig. 5 (cont'd) 4. 2 O O O LM O. M. LV 10 M OOM isa. 120 ar 8x. .3 : 48 5 100 as: rx -x-Ja26; ; --- 28 SSs - & ::...&.823 Yr 8-2 ------------------kissata ; ise 108 80 88 2 66 a. - 4 S. a 20 26 8 23 : & O2 2 --&-23 recisolatin U.S. Patent Feb. 23, 2016 Sheet 7 of 16 US 9.266,901 B2 28 S 28 202 23 Cispati + 5 uy Cispiatin 81 22 283 Cisplatin + S v Cispiatin 8 Fig. 7 p-NFkBp65 NFkBp65 U.S. Patent Feb. 23, 2016 Sheet 10 of 16 US 9.266,901 B2 Fig. F. :rest Cr-fig2. feath:8::ciay U.S. Patent Feb. 23, 2016 Sheet 11 of 16 US 9.266,901 B2 Fig. 12 inflammatory Cytokines, Growth Factors, CD23,CE)3CD28, LPS, Fas, etc. IL-1R K.E. S. &3;&de Cascais Caspase poly. Apoptosis ibiguitination eer Proteasone degradation Activated NFkB N Parthenoides inhibit plays a Central role Se:S36 NFkB activation and in regulating Active NF-kB binding to DNA malignant Se:2's progression Serai Cytokines, Chesnakiries, Nucleus raisciption Transcriptior Factors. T Adaesion votecuies KB site Actite raise Pigtairs, ix3-c. U.S. Patent Feb. 23, 2016 Sheet 12 of 16 US 9.266,901 B2 Fig. 12 cont'd t sixx c Sass :: Ssss 8:::::::: Fig. 3 6 & 3. S. : p-NFkBp65 U.S. Patent Feb. 23, 2016 Sheet 13 of 16 US 9.266,901 B2 Fig. 14 8 34 CO, TMR HER NFkB Reporter Fig. 15 4 1. 1. Drug Dose (micromolar) U.S. Patent Feb. 23, 2016 Sheet 14 of 16 US 9.266,901 B2 Fig. 6 Xs. 1 a ;3 {".g. 888 fi: 8; 8.3: U.S. Patent Feb. 23, 2016 Sheet 15 of 16 US 9.266,901 B2 Fig. 17 f 5 3. CO TR I-IFkB Reporter Fig. 18 ; 8: 88. k . - A - 203+TZ : -- TZ--2O3 Vh 8: 0.1 1.0 10 50 Drug Dose (micromolar) U.S. Patent Feb. 23, 2016 Sheet 16 of 16 US 9.266,901 B2 Fig. 19 -------------------------------------------------- US 9,266,901 B2 1. 2 COMPOSITIONS AND METHODS FOR either TSC1 or TSC2 and in hamartomas of TSC patients. TREATING CANCER TSC can occur in association with pulmonary lymphangiolei omyomatosis (LAM), a progressive and often fatal interstitial CROSS-REFERENCES TO RELATED lung disease that occurs largely in women and is character APPLICATIONS ized by proliferation of abnormal smooth muscle cells. TSC2 null Smooth muscle cells in angiomyolipomas (AML) are This application is a continuation application of interna very similar to those of pulmonary LAM, and data Suggest tional PCT application No. PCT/US2013/035.010, filed Apr. LAM may be the result of benign cell metastases. Rapamycin 2, 2013, which claims the benefit of U.S. Provisional Patent and related drugs (everolimus) which bind and inhibit Application No. 61/619.289, filed Apr. 2, 2012, which are 10 mTORC1 have clinical activity for therapy of TSC, including incorporated herein by reference in their entirety and for all renal AML and pulmonary LAM (72). However, rapamycin purposes. does not elicit complete TSC regression in most cases, and STATEMENT AS TO RIGHTS TO INVENTIONS termination of therapy oft leads to lesion re-growth. New MADE UNDER FEDERALLY SPONSORED 15 therapeutic approaches to control the growth of TSC-related RESEARCH AND DEVELOPMENT proliferative diseases are urgently needed. The poor prognosis of advanced non-Small cell lung cancer This invention was made with Government support under (NSCLC) is due, in part, to emergence of tumor resistance to DAMD17-03-1-0381, awarded by the U.S. Army, Medical chemotherapy (1). Recent data indicate that human tumors Research and Materiel Command. The Government has cer contain a small subset of cancer stem cells (CSC) responsible tain rights in the invention. for drug resistance and tumor maintenance (2-5). If Such minute subsets of CSC drive tumor formation and drug resis BACKGROUND tance, therapies targeting the bulk tumor mass but not CSC will fail. Since it is hypothesized that CSC are responsible for Lung Cancer is the leading cause of cancer death in women 25 tumor regeneration after chemo-therapy (6), we performed and men in the U.S. It is estimated that there will be more than preliminary studies to determine if drug treatment could 230,000 patients diagnosed with lung cancer this year, with enrich for CSCs. Treatment of human lung tumor NSCLC cell more than 160,000 deaths, and non-small cell lung cancer lines (A549, H23) with increasing doses of cisplatin resulted (NSCLC) will account for more than three quarters of these in selection of drug-resistant cells (DRC)(6). Using Fluores cases (1). Survival rates of NSCLC, the predominant type of 30 cence Activated Cell Sorting (FACS), DRC were selected for lung cancer are unacceptably low, and new approaches to expression of cell surface CD133+ and cytosolic ALDH1 by treat and prevent this disease are urgently needed. Feverfew Aldefluor assay (7.8), yielding cell subsets amounting to extracts and parthenolide show anticancer activity in NSCLC 2%-5% of bulk tumor cell populations of H23 and A549, in vitro, while parthenolides modified to enhance solubility respectively. Furthermore, CSC-like cells grew as tumor also show significant anticancer activity in vivo in NSCLC 35 spheres, maintained self-renewal capacity and differentiated, models. This anticancer effect may be associated with the with differentiated progenitors losing CD133 expression and ability of these substances to inhibit the nuclear transcription acquiring drug sensitivity.