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(12) United States Patent (10) Patent No.: US 9,682,943 B2 Guedat Et Al

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(12) United States Patent (10) Patent No.: US 9,682,943 B2 Guedat Et Al USOO9682943B2 (12) United States Patent (10) Patent No.: US 9,682,943 B2 Guedat et al. (45) Date of Patent: Jun. 20, 2017 (54) BENZYLIDENEGUANIDINE DERIVATIVES (58) Field of Classification Search AND THERAPEUTIC USE FOR THE None TREATMENT OF PROTEIN MISFOLDING See application file for complete search history. DISEASES (56) References Cited (71) Applicants:Medical Research Council, Swindon (GB); InFlectis BioScience, Nantes U.S. PATENT DOCUMENTS (FR) 3,541,218 A 11/1970 Marshall et al. (72) Inventors: Philippe Guedat, Montenois (FR): Anne Bertolotti, Cambridge (GB) FOREIGN PATENT DOCUMENTS CA 958O18 11, 1974 (73) Assignees: MEDICAL RESEARCH COUNCIL, WO WO93,03714 3, 1993 Swindon (GB); INFLECTIS WO WO O2, 11715 2, 2002 BIOSCIENCE, Nantes (FR) (Continued) (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 OTHER PUBLICATIONS U.S.C. 154(b) by 0 days. Hughes, BMJ, vol. 324, Feb. 2002, pp. 466-469.* (21) Appl. No.: 14/760,350 (Continued) (22) PCT Filed: Jan. 10, 2014 Primary Examiner — Zinna Northington Davis (74) Attorney, Agent, or Firm — McDonnell Boehnen (86). PCT No.: PCT/EP2014/OSO422 Hulbert & Berghoff LLP S 371 (c)(1), (57) ABSTRACT (2) Date: Jul. 10, 2015 The present invention relates to a compound of formula (I), (87) PCT Pub. No.: WO2014/108520 or a tautomer and/or a pharmaceutically acceptable salt thereof, wherein: R is alkyl, Cl, F or Br; R is H or F. R. PCT Pub. Date: Jul. 17, 2014 is selected from H and alkyl; R is selected from H and C(O)R. Rs is H; or R and Rs are linked to form a (65) Prior Publication Data heterocyclic group which is optionally substituted with one US 2016/OO46589 A1 Feb. 18, 2016 or more Rio groups; R is selected from R7, OR, and NRsRo: R7, Rs and R are each independently selected from alkyl, (30) Foreign Application Priority Data cycloalkyl, aralkyl, cycloalkenyl, heterocyclyl and aryl, each of which is optionally substituted with one or more Rio Jan. 10, 2013 (GB) ................................... 1300.435.3 groups; each Ro is independently selected from halogen, OH, CN, NO, COO-alkyl, aralkyl, SO-alkyl, SO-aryl, (51) Int. Cl. COOH, CO-alkyl, CO-aryl, NH, NH-alkyl, N(alkyl), CF, CO7D 213/6 (2006.01) alkyl and alkoxy; X and Z are each independently CR, and CD7C 281/8 (2006.01) Y is selected from CR and N; and R is H or F: for use in A6 IK3I/55 (2006.01) treating a disorder associated with protein misfolding stress A6 IK 3/44 (2006.01) and in particular associated with accumulation of misfolded C07D 25.3/075 (2006.01) proteins. A6 IK3I/53 (2006.01) A6 IK 45/06 (2006.01) A6 IK3I/I65 (2006.01) (I) A6 IK 3L/26 (2006.01) R R3 CD7C 28/6 (2006.01) (52) U.S. Cl. f N N N1- - YR CPC ........ C07D 253/075 (2013.01); A61K 31/155 (2013.01); A61K 31/165 (2013.01); A61 K YN 2 N 3 1/216 (2013.01); A61 K3I/44 (2013.01); Nx R2 YR A6 IK3I/53 (2013.01); A61K 45/06 (2013.01); C07C 281/16 (2013.01); C07C 281/18 (2013.01); C07D 213/61 (2013.01) 13 Claims, 6 Drawing Sheets US 9,682,943 B2 Page 2 (56) References Cited FOREIGN PATENT DOCUMENTS WO WO 2005/031000 A2 * 4, 2005 OTHER PUBLICATIONS Jiang et al. “Synthesis and biological evaluation of novel 2-(2- arylmethylene)hydrazinyl-4-aminoquina Zoline derivatives as potent antitumor agents.' European Journal of medicinal Chemistry, vol. 54, Aug. 1, 2012, pp. 534-541. Tribouillard-Tanvier, et al., “Antihypertensive drug guanabenz is active in vio against both yeast and mammalian prions.” PLOS One, Public Library of Science, vol. 3., No. 4, Jan. 1, 2008, p. 2. Tsaytler et al. “Selective Inhibition of a Regulatory subunit of Protein Phosphatase 1 Restores Proteostasis,” Science, vol. 332, No. 6025, Apr. 1, 2011, pp. 91-94. International Search Report and Written Opinion mailed May 13, 2014 for International Application No. PCT/EP2014/050422 filed Jan. 10, 2014. * cited by examiner U.S. Patent Jun. 20, 2017 Sheet 1 of 6 US 9,682,943 B2 Example 1 8 09% 6 09% 40% 20% Concentration uM FIGURE 1 U.S. Patent Jun. 20, 2017 Sheet 2 of 6 US 9,682,943 B2 Tunicamycin Tunicamycin + Example 1 Time, h O 2 5 10 Time, h O 2 5 10 FIGURE 2 U.S. Patent Jun. 20, 2017 Sheet 3 of 6 US 9,682,943 B2 Guanabenz Example 1 FIGURE 3 U.S. Patent Jun. 20, 2017 Sheet 4 of 6 US 9,682,943 B2 Cell number ER-retained POS63del Example 1, 10puM FIGURE 4 U.S. Patent Jun. 20, 2017 Sheet S of 6 US 9,682,943 B2 % of initial UT CMP 4 UM Htt48O SOD1A4V FIGURE 5 U.S. Patent Jun. 20, 2017 Sheet 6 of 6 US 9,682,943 B2 Cell number P23H aggregates 4.5 - 4 3.5 3 2.5 15 Example 1, 10 uM FIGURE 6 US 9,682,943 B2 1. 2 BENZYLDENEGUANDINE DERVATIVES Guanabenz has also been shown to promote survival of AND THERAPEUTIC USE FOR THE HeLa cells exposed to otherwise cytotoxic ER-stress TREATMENT OF PROTEIN MISFOLDING induced by the N-glycosylation inhibitor tunicamycin, in a DISEASES dose-dependent manner (P. Tsaytler, H. P. Harding, D. Ron and A. Bertolotti, Science, 332, 1 Apr. 2011, 91-94). Quan titative assessment of cell viability revealed that guanabenz This application is a U.S. national phase application of doubled the number of cells surviving ER stress with a International Patent Application No. PCT/EP2014/050422 median effective concentration of ~0.4 uM. Neither the filed on Jan. 10, 2014, which claims the benefit of Great C2-adrenergic receptor agonist clonidine, nor the C2-adren Britain patent application no. 13.00435.3, filed Jan. 10, 2013. ergic receptor antagonist efaroxan protected cells from cyto The present invention relates to compounds that have 10 toxic ER stress and efaroxan did not interfere with gua potential therapeutic applications in treating disorders asso nabenz’s protective effect (P. Tsaytler, H. P. Harding, D. Ron ciated with protein misfolding stress and in particular with and A. Bertolotti, Science, 332, 1 Apr. 2011, 91-94). These an accumulation of misfolded proteins. In particular, the observations demonstrate that guanabenz rescues cells from invention provides compounds that are capable of exhibiting lethal ER stress by a mechanism independent of the 15 C2-adrenergic receptor. Guanabenz protects cells from oth a protective effect against cytotoxic endoplasmic reticulum erwise lethal accumulation of misfolded proteins by binding (ER) stress. to a regulatory subunit of protein phosphatase 1, PPP1R15A (GADD34), selectively disrupting the stress-induced BACKGROUND TO THE INVENTION dephosphorylation of the C. subunit of translation initiation factor 2 (eIF2C.). Guanabenz sets the translation rates in The compound 2-(2,6-dichlorobenzylidene)hydrazinecar stressed cells to a level manageable by available chaperones, boximidamide, also referred to as guanabenZ, is an alpha thereby restoring protein homeostasis. It was reported that agonist of the alpha-2 type that is used as an antihyperten Guanabenz does not bind to the constitutive PPP1R15B sive drug. (CReP) and therefore does not inhibit translation in non 25 stressed cells. (P. Tsaytler, H. P. Harding, D. Ron and A. Bertolotti, Science, 332, 1 Apr. 2011, 91-94). C Failure to maintain proteostasis in the ER by mounting an N NH2 adequate unfolded protein response (UPR) is recognized as n N1 s a contributing factor to many pathological conditions. Thus, 30 the molecules described here, which inhibit eIF2C. phos NH2 phatase to fine-tune protein synthesis, may be of therapeutic C benefit to a large number of diseases caused protein mis Guanabenz folding stress and in particular with an accumulation of misfolded proteins. Various derivatives of guanabenz have also been reported. 35 The present invention seeks to provide alternative com For example, U.S. Pat. No. 3,982,020 (Sandoz, Inc.) dis pounds based on a guanabenz core structure that have closes substituted benzylidene hydrazines and their use as potential therapeutic applications in treating disorders asso hypoglycemic-antihyperglycemic agents, anti-obesity ciated with protein misfolding stress and in particular with an accumulation of misfolded proteins. agents and anti-inflammatory agents. US 2004/0068017 40 (Bausch & Lomb Inc.) discloses substituted benzylidene hydrazines that are capable of increasing the activity of STATEMENT OF INVENTION gelatinase A in ocular cells. The molecules have applications A first aspect of the invention relates to a compound of in the treatment of primary open angle glaucoma. WO formula (I), or a pharmaceutically acceptable salt thereof, 2008/061647 (Acure Pharma AB) discloses the use of N-(2- 45 chloro-3,4-dimethoxybenzylideneamino)guanidine as a VEGFR inhibitor and its associated applications in the (I) treatment or prevention of undesired blood vessel formation R R3 during tumour growth and/or inflammatory conditions. WO N - - 2005/031000 (Acadia Pharmaceuticals, Inc.) discloses sub 50 f N N1 YR stituted benzylidene hydrazines and their use in treating acute pain and chronic neuropathic pain. Finally, EP YN 2 N 1908464 (CNRS) discloses guanabenz and chloroguanabenz Nx R2 YR and their use in the treatment of polyglutamine expansion associated diseases, including Huntington's disease. 55 wherein: More recently it has been reported that guanabenz has R is alkyl, Cl, F or Br; therapeutic potential in a number of other areas. Guanabenz, R is H or F: was recently noted to have anti-prion activity (D. Tribouil R is selected from H and alkyl: lard-Tanvier et al., 2008 PLoS One 3, e1981).
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