US 20150307441A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0307441 A1 Ogrodzinski et al. (43) Pub. Date: Oct. 29, 2015 (54) NEW COMPOUNDS AND USES THEREOF A 6LX3L/277 (2006.01) A63L/36 (2006.01) (71) Applicant: Biostatus Limited. Shepshed, A6II 45/06 (2006.01) Leicestershire (GB) (52) U.S. Cl. CPC ............. C07C 225/36 (2013.01); A61 K3I/136 (72) Inventors: Stefan Ogrodzinski, Shepshed, (2013.01); A61K 45/06 (2013.01); A61 K Leicestershire (GB); Paul Smith, 3I/277 (2013.01); G0IN 31/225 (2013.01) Shepshed, Leicestershire (GB); Stephanie McKeown, Shepshed, Leicestershire (GB); Laurence (57) ABSTRACT Patterson, Shepshed, Leicestershire (GB); Rachel Jane Errington, An anthraquinone compound of formula I (Such as the com Shepshed, Leicestershire (GB) pounds of formulae II to X) and processes for making the (73) Assignee: Biostatus Limited. Shepshed, same are provided. Pharmaceutical compositions for use in the treatment of cancer, optionally in combination with an Leicestershire (GB) agent capable of reducing the level of oxygenation of a (21) Appl. No.: 14/420,184 tumour, are also provided. Additionally, an option for com bination with chemotherapeutic and radiotherapeutic modali (22) PCT Filed: Aug. 7, 2013 ties to enhance overall tumour cell kill is provided. Methods for the detection of cellular hypoxia, both in vivo and in vitro, (86). PCT No.: PCT/GB2O13/O52106 are additionally provided. S371 (c)(1), (2) Date: Feb. 6, 2015 Formula I O (30) Foreign Application Priority Data X4 X e sas Aug. 8, 2012 (GB) ................................... 12141693 e as Publication Classification S. 1. X3 X2 (51) Int. Cl. O CD7C 225/36 (2006.01) GOIN3L/22 (2006.01) Patent Application Publication Oct. 29, 2015 Sheet 1 of 20 US 2015/0307441 A1 FGRE AQ4 x 4 days air OC100 x 4 days air Equivalent arrest of cells in G2-M as a function of drug dose in air Patent Application Publication Oct. 29, 2015 Sheet 2 of 20 US 2015/0307441 A1 {RE 12 n a M -------- f -- kat air VP-6 0.8 s 8 -är Jurkat air AQ4N E \ - E O.8 Ye -8-Jurkat air OCOO2 8 a -O-rkat 9 WP-6 S. -A - Bufkat 1% AC4N 5 O. 4. -- Jurkat 1%. OCT002 -C O -- GS. C.O OOO 1 OOOO drug dose (nM x 4 days) Patent Application Publication Oct. 29, 2015 Sheet 3 of 20 US 2015/0307441 A1 GRE 3 t 2 | C A549 air AQ4N is... no noxia Sy. 8 A549 air OCTOR i. 8 -A-A549 3% AC4N O 8 A-A549 3%, OC1002 O. 4. -- A549 1% O2 AC4N St. A549 1% OC1002 O 50 OO Drug dose (ny x 4 days) Patent Application Publication Oct. 29, 2015 Sheet 4 of 20 US 2015/0307441 A1 FR 3. Ai - OC. CO2 3% O2 - OCOO2 1% O2 + OC1002 8t; iivaert arras of ces 1% O2 - ACR4N 2, as a car situg cose ?er hypoxia Patent Application Publication Oct. 29, 2015 Sheet 5 of 20 US 2015/0307441 A1 FGRE 5(A) -- CH-21% OCOO2 -- Do 21% AC&N -- Do-23% OCTOO2 --DoHH23% AQ4N --O-O-O-H2 air OCTO02 -- Doh-2 air AQ4N W O 5 OO Drug conc. (nM) Patent Application Publication Oct. 29, 2015 Sheet 6 of 20 US 2015/0307441 A1 FGRESS) -- SU---4 16 OCOO2 -i-Su-DH-4 1% AC4N N. --S-OH-4.3%OCTOO2 N. - & - S -OH,-43% y AC4N -Q- SJ-3H-4 air OCI 10O2 -8- S-O-3-4 air i AGN .2 SO OO Drug conc. (nw) Patent Application Publication Oct. 29, 2015 Sheet 7 of 20 US 2015/0307441 A1 GRES SO 200 s {Q Air OCT12 A 3% oxygen OCTQ02 C 1% oxygen OCT1002 100 - a a Y- Linear (Air OC1002) s - - - - - - - - Sinear (3% oxygen OC1002) - 3% oxygen. A - - - - - - linear 1% oxygen OCT002) s - - ----- 5 ...--A non-Oxia 9 O 50 OO Drug dose (n\ x 4 days) Patent Application Publication Oct. 29, 2015 Sheet 8 of 20 US 2015/0307441 A1 FGRE Air Control OCOO x 4. days, a s??ando AQ4 x 4 days, 3. 3)) 8 800 Fluorescence intensity (ch. Fo s Ex 633 fam; Em >695 nm) Patent Application Publication Oct. 29, 2015 Sheet 9 of 20 US 2015/0307441 A1 GRE 3 25O 2OO O O 50 OO 150 200 250 Fluorescence intensity (ch. no.; Em-695 nm) Patent Application Publication Oct. 29, 2015 Sheet 10 of 20 US 2015/0307441 A1 FGRE 9A) far-red fluorescence trans ission Fluorescence intensity (ch, no. Ex633 nm, AEm >695 nm) Patent Application Publication Oct. 29, 2015 Sheet 11 of 20 US 2015/0307441 A1 FGRE ( 2. one vehicle only a8a Bicalutamide $2mg/kgiday) 18 5 14 3.2 10 2 A. s 8 O 12 4. 5 8 20 22 24 28 28 Day of Breatment Patent Application Publication Oct. 29, 2015 Sheet 12 of 20 US 2015/0307441 A1 FGE AY Patent Application Publication Oct. 29, 2015 Sheet 13 of 20 US 2015/0307441 A1 FGRE 2{A} ------------- arrrrrrrrrry------------- - r----------- a---------------------------------- A 7 .--Bicatutamide only --- -$o vehicle only --AQ4Nk - OCT1002 only only (50mg/kg) (50mg/kg) $ f 5 4. 3. 3. |- ---- --------------------- - ----- -------- ----- --- Q 5 10 5 2O 25 30 tine days} Patent Application Publication Oct. 29, 2015 Sheet 14 of 20 US 2015/0307441 A1 Figure 28 & OCT1002 only (50mg/kg) B -8-AQ4N only (50mg/kg) 8 is 30 8-Bicalutamide only SN s -m- Cs is 25 s s 20 15 2 e 10 5 O ic----- - - - - - ----------------------------------------------------------------- 0 2 4 6 8 to 12tine 1416 (day) 18 20 22 24 26 28 so Patent Application Publication Oct. 29, 2015 Sheet 15 of 20 US 2015/0307441 A1 FiGRE 3A) --Bicatutamide only ---------------- --&M vehicle only - - - - -8- - - - ACR4-N + 8icalutamide s & - OC1002 + Bicalutamide 4.5 3 Patent Application Publication Oct. 29, 2015 Sheet 16 of 20 US 2015/0307441 A1 FGURE 3(8) 7 : ------&AQ4N - ---------------Bicalutamide poly)------------ B 88:calutamide (exp} y = 0.5788e & CT1002 + Bicalutamide (lin) R = 09915 5 4. -$ $ 8 3 -' & y = 0.001.4x + 0.0728x + 0.58& 8 A. A R* x 0.9882 ... a8 - * - y a 0.0756x + 0.477 O i - - -------------------------- 5 O 5 2O 25 30 time (days) Patent Application Publication Oct. 29, 2015 Sheet 17 of 20 US 2015/0307441 A1 GRE a 1400- or-n Vehicle only re- vehicle - OCOO2 risk 1200- x- 3icalutamide ... " e8.o Sicautamide + OCOO ** S 1. wev" R 800 s s 600 - 400 & M we & S N N S S S - & 8 Tire days post treatinent Patent Application Publication Oct. 29, 2015 Sheet 18 of 20 US 2015/0307441 A1 FRE is (A) Vehicle + OCT1002 (day 7) (8) Bicalutamide + OCT1002 (day 7) Patent Application Publication Oct. 29, 2015 Sheet 19 of 20 US 2015/0307441 A1 FGE 5 - eiggs O 7 14. 2 days Patent Application Publication Oct. 29, 2015 Sheet 20 of 20 US 2015/0307441 A1 F&SR is e 8 se Treatment groups US 2015/0307441 A1 Oct. 29, 2015 NEW COMPOUNDS AND USES THEREOF distant tissues. The development of more malignant meta static tumours is often the precursor to a more significant CROSS REFERENCE TO RELATED disease-related morbidity and the death of the patient. APPLICATION 0005. An attractive approach is the use of a hypoxia acti 0001. The present application claims priority to Great vated prodrug that is non-toxic towards adequately oxygen Britain Patent Application No. 1214169.3, filed Aug. 8, 2012, ated cells found in Systemic tissues, but becomes activated or incorporated herein in its entirety. converted to a cytotoxic form under reduced oxygenation conditions. N-oxide derivatives of cytotoxic alkylaminoan TECHNICAL FIELD thraquinones provide anthraquinone pro-drugs that show almost no cytotoxicity. Importantly these prodrugs are 0002 The present invention relates to novel capable of being converted in vivo under the anaerobic/hy anthraquinone compounds and uses of the same, for example poxic conditions found within neoplastic tissue. Specificity in the treatment of cancer. for the tumour is ensured since systemic tissues, except for tumours, almost never experience oxygen levels low enough BACKGROUND to facilitate the production of the cytotoxic drug. 0003. The therapeutic advantage of an anticancer drug 0006. The anthraquinone N-oxide AQ4N (CAS#136470 depends primarily on the extent to which the agent shows 65-0) is a prodrug that is selectively bioreduced to AQ4, a selective activity for tumour cells and the limiting toxicity potent DNA topoisomerase II inhibitor, in hypoxic tumour towards non-target tissues. Frequently the poor quality of the cells. Previous publications have taught the fundamental vasculature within the growing tumour mass compromises properties and in-vitro I in-vivo characteristics of the prodrug the delivery of drugs, nutrients and oxygen. It is recognised AQ4N (for example, see U.S. Pat. No. 5,132.327). that tumours can have significantly lower median oxygen 0007. The invention seeks to address the need for levels (approximately 1% oxygen; pO2 7.5 mmHg) com improved cancer treatments by providing novel pared to normal tissues (~5.5% oxygen; 42 mmHg) (Sum anthraquinone compounds with a combination of preferable marised from data presented by Brown and Wilson, 2004). In pharmacological and hypoxia-sensing properties. addition, oxygenation levels can vary throughout the tumour due to intermittent opening and closing of tumour blood SUMMARY vessels; poor vascularisation, especially in the tumour core, contributes to oxygen levels often being below 0.1% oxygen 0008. The first aspect of the invention provides a com (1 mm Hg). Tumour cells experiencing varying degrees of pound of Formula I hypoxia, relative to normally perfused tissues, can compro mise treatment effectiveness and contribute to the malig Formula I nancy.