US 20150284724A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0284724 A1 COLLARD et al. (43) Pub. Date: Oct. 8, 2015 (54) TREATMENT OF SIRTUIN (SIRT) RELATED (60) Provisional application No. 61/330,427, filed on May DISEASES BY INHIBITION OF NATURAL 3, 2010, provisional application No. 61/415,891, filed ANTISENSE TRANSCRIPT TO A SIRTUN on Nov. 22, 2010, provisional application No. 61/412, (SIRT) 066, filed on Nov. 10, 2010, provisional application No. 61/409,136, filed on Nov. 2, 2010. (71) Applicant: CuRNA, Inc., Miami, FL (US) Publication Classification (72) Inventors: Joseph COLLARD, Delray Beach, FL (US): Olga Khorkova Sherman, (51) Int. Cl. Tequesta, FL (US); Carlos Coito, West CI2N IS/II3 (2006.01) Palm Beach, FL (US); Belinda De Leon, (52) U.S. Cl. San Francisco, CA (US) CPC ........ CI2N 15/1137 (2013.01); C12N 2310/11 (2013.01) (21) Appl. No.: 14/741,164 (57) ABSTRACT (22) Filed: Jun. 16, 2015 The present invention relates to antisense oligonucleotide that modulate the expression of and/or function of a Sirtuin Related U.S. ApplicationO O Data RidSIRT), in particular,p a Sirtuin by Rs. targeting E. inventionnatural antisense also ria poly o (62) Division of application No. 13/695,801, filed on Nov. the identification of these antisense oligonucleotides and their 2, 2012, now Pat. No. 9,089,588, filed as application use in treating diseases and disorders associated with the No. PCT/US2011/021052 on Jan. 13, 2011. expression of Sirtuins (SIRT)s. Patent Application Publication Oct. 8, 2015 Sheet 1 of 14 US 2015/0284724 A1 F}, i Foid tiererica in SiRi expression after treatnerit with siris to Sas c. : 3 3 38 s s 3. S. a S 6 8 S s s: 3 $ 3. 3. S 3 in FC Fesix difference in sistas expressics after treatment with risis. 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In another embodiment, an oligonucleotide targets a DISEASES BY INHIBITION OF NATURAL natural antisense sequence of a Sirtuin (SIRT) polynucle ANTISENSE TRANSCRIPT TO A SIRTUN otide, for example, nucleotides set forth in SEQID NO: 9 to (SIRT) 23, 141 to 143, and any variants, alleles, homology, mutants, derivatives, fragments and complementary sequences FIELD OF THE INVENTION thereto. Examples of antisense oligonucleotides useful in practicing the methods of the present invention are set forth as 0001. The present application claims the priority of U.S. SEQID NOS: 24 to 127. provisional patent application No. 61/330,427 filed May 3, 0007 Another embodiment provides a method of modu 2010, U.S. provisional patent application No. 61/409,136 lating function and/or expression of a Sirtuin (SIRT) poly filed Nov 2, 2010, U.S. provisional patent application No. nucleotide in patient cells or tissues in Vivo or in vitro com 61/412,066 filed Nov. 10, 2010 and U.S. provisional patent prising contacting said cells or tissues with an antisense application No. 61/415,891 filed Nov. 22, 2010. oligonucleotide 5 to 30 nucleotides in length wherein said 0002 Embodiments of the invention comprise oligonucle oligonucleotide has at least 50% sequence identity to a otides modulating expression and/or function of a Sirtuin reverse complement of an antisense of the Sirtuin (SIRT) (SIRT) and associated molecules. polynucleotide; thereby modulating function and/or expres sion of the Sirtuin (SIRT) polynucleotide in patient cells or BACKGROUND tissues is vivo or in vitro. 0008 Another embodiment provides a method of modu 0003 DNA-RNA and RNA-RNA hybridization are lating function and/or expression of a Sirtuin (SIRT) poly important to many aspects of nucleic acid function including nucleotide in patient cells or tissues in Vivo or in vitro com DNA replication, transcription, and translation. Hybridiza prising contacting said cells or tissues with an antisense tion is also central to a variety of technologies that either oligonucleotide 5 to 30 nucleotides in length wherein said detect a particular nucleic acid or alter its expression. Anti oligonucleotide has at least 50% sequence identity to an anti sense nucleotides, for example, disrupt gene expression by sense oligonucleotide to a Sirtuin (SIRT) antisense poly hybridizing to target RNA, thereby interfering with RNA nucleotide; thereby modulating function and/or expression of splicing, transcription, translation, and replication. Antisense the Sirtuin (SIRT) polynucleotide in patient cells or tissues in DNA has the added feature that DNA-RNA hybrids serve as vivo or in vitro. a substrate for digestion by ribonuclease H, an activity that is 0009. In one embodiment, a composition comprises one or present in most cell types. Antisense molecules can be deliv more antisense oligonucleotides which bind to sense and/or ered into cells, as is the case for oligodeoxynucleotides antisense Sirtuin (SIRT) polynucleotides. (ODNs), or they can be expressed from endogenous genes as 0010. In another embodiment, the oligonucleotides com RNA molecules. The FDA recently approved an antisense prise one or more modified or substituted nucleotides. drug, VITRAVENETM (for treatment of cytomegalovirus 0011. In another embodiment, the oligonucleotides com retinitis), reflecting that antisense has therapeutic utility. prise one or more modified bonds. 0012. In yet another embodiment, the modified nucle SUMMARY otides comprise modified bases comprising phosphorothio 0004. In one embodiment, the invention provides methods ate, methylphosphonate, peptide nucleic acids, 2'-O-methyl, for inhibiting the action of a natural antisense transcript by flouro- or carbon, methylene or other locked nucleic acid using antisense oligonucleotide(s) targeted to any region of (LNA) molecules.