(19) TZZ Z__T (11) EP 2 726 601 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C07D 233/60 (2006.01) C07D 233/88 (2006.01) 26.12.2018 Bulletin 2018/52 C07D 495/04 (2006.01) C07D 277/36 (2006.01) C07D 277/587 (2006.01) C07D 277/64 (2006.01) (2006.01) (2010.01) (21) Application number: 12804411.2 C07D 213/50 C12N 15/873 A61K 31/05 (2006.01) A61K 31/137 (2006.01) A61K 31/277 (2006.01) A61K 31/352 (2006.01) (22) Date of filing: 13.04.2012 A61K 31/4745 (2006.01) A61K 31/498 (2006.01) A61K 35/14 (2015.01) A61K 35/28 (2015.01) A61K 35/51 (2015.01) A61K 35/54 (2015.01) C12N 5/071 (2010.01) C12N 5/075 (2010.01) (86) International application number: PCT/US2012/033672 (87) International publication number: WO 2013/002880 (03.01.2013 Gazette 2013/01) (54) COMPOSITIONS AND METHODS FOR ENHANCING BIOENERGETIC STATUS IN FEMALE GERM CELLS ZUSAMMENSETZUNGEN UND VERFAHREN ZUR VERBESSERUNG DES BIOENERGETISCHEN STATUS BEI WEIBLICHEN KEIMZELLEN COMPOSITIONS ET PROCÉDÉS POUR AMÉLIORER L’ÉTAT BIOÉNERGÉTIQUE DE CELLULES GERMINALES FEMELLES (84) Designated Contracting States: • SINCLAIR, David, A. AL AT BE BG CH CY CZ DE DK EE ES FI FR GB Chestnut Hill, MA 02467 (US) GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR (74) Representative: Reitstötter Kinzebach Patentanwälte (30) Priority: 29.06.2011 US 201161502840 P Sternwartstrasse 4 17.02.2012 US 201261600529 P 81679 München (DE) (43) Date of publication of application: (56) References cited: 07.05.2014 Bulletin 2014/19 WO-A2-01/30980 WO-A2-2006/105440 US-A1- 2006 010 508 US-A1- 2006 010 508 (60) Divisional application: US-A1- 2008 050 347 16194807.0 18203134.4 • Y.-T Huang ET AL: "Effects of luteolin and quercetin, inhibitors of tyrosine kinase, on cell (73) Proprietors: growth and metastasis-associated properties in • The General Hospital Corporation A431 cells overexpressing epidermal growth Boston, MA 02114 (US) factor receptor 1", , 1 November 1999 • President and Fellows of Harvard College (1999-11-01), XP055174027, Retrieved from the Cambridge, MA 02138 (US) Internet: URL:http://www.ncbi.nlm.nih.gov/pmc/articl (72) Inventors: es/PMC1571723/pdf/128-0702879a.pdf [retrieved • TILLY, Jonathan, Lee on 2015-03-05] Windham, NH 03087 (US) Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 726 601 B1 Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 2 726 601 B1 • ESTHERKELLENBERGER ET AL: "Flavonoids as • NATALIA IGOSHEVA ET AL: "Maternal inhibitors of human CD38", BIOORGANIC & Diet-Induced Obesity Alters Mitochondrial MEDICINAL CHEMISTRY LETTERS, vol. 21, no. Activity and Redox Status in Mouse Oocytes and 13, 14 May 2011 (2011-05-14), pages 3939-3942, Zygotes", PLOS ONE, vol. 5, no. 4, 9 April 2010 XP055164821, ISSN: 0960-894X, DOI: (2010-04-09), page e10074, XP055174031, DOI: 10.1016/j.bmcl.2011.05.022 10.1371/journal.pone.0010074 2 EP 2 726 601 B1 Description CROSS-REFERENCE TO RELATED SUBJECT MATTER 5 [0001] This application claims benefit under 35 U.S.C. § 119(e) of the U.S. provisional application Ser. Nos. 61/502,840, filed June 29, 2011, and U.S. provisional application Ser. No. 61/600,529, filed February 17, 2012. STATEMENT OF RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH 10 [0002] This work was supported in part by National Institutes of Health Grant No. NIH R37-AG012279. The government has certain rights to the invention. BACKGROUND OF THE INVENTION 15 [0003] Effects of luteolin and quercetin, inhibitors of tyrosine kinase, on cell growth and metastasis-associated prop- erties in A431 cells overexpressing epidermal growth factor receptor, are described in British Journal of Pharmacology 1999; 128: 999-1010. [0004] WO 2006/105540 A2 describes methods and means comprising nicotinamide riboside for modulating the sirtuin pathway.WO 01/30980 A2 relates to methods and compositions for enhancing developmental potential of oocytes and 20 zygotes. [0005] Igosheva et al., PLoS ONE 2010 5 (4): e10074 describe that maternal diet-induced obesity alters mitochondrial activity and redox status in mouse oocytes and zygotes. [0006] Since the early 1950s, clinical management of problems associated with ovarian insufficiency and failure, including infertility due to aging or insults, has been restricted by the belief that the pool of oocytes set forth at birth is 25 not amenable to replacement or renewal (Zuckerman, Recent Prog Horm Res 1951 6:63-108). In other words, any therapeutic intervention had to conform to manipulation of the existing stockpile of oocyte-containing follicles to produce a desired clinical outcome. In 2004, however, studies with mice challenged the idea of a fixed ovarian reserve of oocytes being endowed at birth (Johnson et al., Nature 2004 428:145-150). Based on results from several experimental ap- proaches, it was concluded that ovaries of adult female mammals retain rare germline or oogonial stem cells (OSCs) 30 that routinely produce new oocytes in a manner analogous to germline stem cell support of sperm production in the adult testis (Spradling, Nature 2004 428:133-134). Several years later, OSCs were successfully isolated from neonatal and adult mouse ovaries (Zou et al., Nat Cell Biol 2009 11:631-636; Pachiarotti et al., Differentiation 2010 79:159-170). Collectively, these investigations, along with several other reports from studies of mice (Johnson et al., Cell 2005 122:303-315; Wang et al., Cell Cycle 2010 9:339-349; Niikura et al., Aging 2010 2:999-1003) have conceptually validated 35 the use of OSCs as agents for transplantation and as targets for new therapies to modulate ovarian function and female fertility (Tilly et al., Biol Reprod 2009 80:2-12; Tilly et al., Mol Hum Reprod 2009; 15:393-398). In addition, the identificat ion of dormant OSCs in atrophic ovaries of aged mice, which spontaneously resume oocyte formation when exposed to a young adult ovarian environment, indicates that ovarian aging may be reversible (Niikura et al., Aging 2009 1:971-978; Massasa et al., Aging 2010; 2:1-2). The clinical utility of OSCs is now further confirmed by evidence shown herein that 40 a comparable population of oocyte-producing stem cells exists in, and can be purified from, ovaries of healthy repro- ductive-age women. [0007] Although these new studies indicate that oocyte numbers in adult ovaries are amenable to therapeutic expansion through OSC-based technology, ovarian aging and failure is determined by both a decline in oocyte number as well as a decline in the quality of the oocytes present in the ovaries. Hence, it is imperative to identify methods for improving 45 oocyte quality, especially in women of advancing maternal age. During the past few decades, because of cultural and social changes, women in the developed world have significantly delayed childbirth. For example, first birth rates for women 35-44 years of age in the United States have increased by more than 8-fold over the past 40 years (Ventura Vital Health Stat 2009 47:1-27; Matthews, NCHS Data Brief 2009 21:1-8). It is well known that pregnancy rates in women at 35 or more years of age are significantly lower, both naturally and with assisted reproduction. The decline in live birth 50 rate reflects a decline in response to ovarian stimulation by gonadotropin hormones (follicle-stimulating hormone or FSH, and luteinizing hormone or LH), reduced oocyte and embryo quality and pregnancy rates, and an increased incidence of miscarriages and fetal aneuploidy. In fact, aging-associated chromosomal and meiotic spindle abnormalities in eggs are considered the major factors responsible for the increased incidence of infertility, fetal loss (miscarriage) and con- ceptions resulting in birth defects - most notably trisomy 21 or Down syndrome - in women at advanced reproductive 55 ages (Henderson et al., Nature 1968 218:22-28; Hassold et al., Hum Genet 1985 70:11-17; Battaglia et al., Hum Reprod 1996 11:2217-2222; Hunt et al., Trends Genet 2008 24:86-93). Although the occurrence and consequences of aging- related aneuploidy in oocytes of humans and animal models have been extensively studied (Tarín et al., Biol Reprod 2001 65:141-150; Pan et al., Dev Biol 2008 316:397-407; Duncan et al., Biol Reprod 2009 81:768-776), approaches to 3 EP 2 726 601 B1 maintain fidelity of chromosome segregation during meiotic cell division with age have remained elusive. At present there is no known intervention to improve the pregnancy outcome of older female patients. In animal studies, chronic administration of pharmacologic doses of anti-oxidants during the juvenile period and throughout adult reproductive life has been reported to improve oocyte quality in aging female mice (Tarín et al., Mol Reprod Dev 2002 61:385-397). 5 However, this approach has significant long-term negative effects on ovarian and uterine function, leading to higher fetal death and resorptions as well as decreased litter frequency and size in treated animals (Tarín et al., Theriogenology 2002 57:1539-1550). Thus, clinical translation of chronic anti-oxidant therapy throughout reproductive life for maintaining or improving oocyte quality in aging females is impractical.