(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2007/0082861 A1
`(43) Pub. Date:
`Apr. 12, 2007
`Matsuo et al.
`
`US 20070O82861A1
`
`(54)
`
`(75)
`
`(73)
`
`(21)
`(22)
`(86)
`
`ENANUCLEC ACID DRUGS MODIFYING
`SPLICING IN MIRNA PRECURSOR
`
`Inventors: Masafumi Matsuo, Hyogo (JP);
`Yasuhiro Takeshima, Hyogo (JP);
`Makoto Koizumi, Tokyo (JP)
`Correspondence Address:
`OBLON, SPIVAK, MCCLELLAND, MAIER &
`NEUSTADT, P.C.
`194O DUKE STREET
`ALEXANDRIA, VA 22314 (US)
`Assignees: Masafumi Matsuo, Hyogo (JP); Yasu
`hiro Takeshima, Hyogo (JP); Nonpro.
`Org. Trans. Res. Org. of Duch. Musc.
`Dys., Hyogo (JP)
`
`Appl. No.:
`
`10/536,258
`
`PCT Filed:
`
`Nov. 21, 2003
`
`PCT No.:
`
`PCT/UP03/14915
`
`S 371(c)(1),
`(2), (4) Date: Dec. 13, 2005
`
`(30)
`
`Foreign Application Priority Data
`
`Nov. 25, 2002 (JP)...................................... 2002-340857
`Jul. 31, 2003 (JP)...................................... 2003-204381
`
`Publication Classification
`
`(51) Int. Cl.
`(2006.01)
`A6IR 48/00
`(2006.01)
`C7H 9/048
`(2006.01)
`C7H 2L/02
`(52) U.S. Cl. ......................... 514/44; 536/23.1; 536/27.1;
`536/281
`
`(57)
`
`ABSTRACT
`
`Oligonucleotides having a nucleotide sequence complemen
`tary to nucleotide numbers such as 2571-2607, 2578-2592,
`2571-2592, 2573-2592, 2578-2596, 2578-2601 or 2575
`2592 of the dystrophin cDNA (Gene Bank accession No.
`NM 004006.1) and therapeutic agents for muscular dystro
`phy comprising Such oligonucleotides.
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 1 of 22
`
`US 2007/0082861 A1
`
`FIG. 1
`
`Untreated AO1
`
`S-O i SO
`
`
`
`C.
`
`Band B
`{- Band A
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 2 of 22
`
`US 2007/0082861 A1
`
`FIG. 2
`
`Untreated AO1 AO14 AO15 AO16 AO17 AO18 AO19 AO24 AO25
`
`
`
`C- Band B
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 3 of 22
`
`US 2007/0082861 A1
`
`FIG 3
`
`Untreated A018
`
`
`
`A050
`
`AO51
`
`A052
`
`A053
`
`A054
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 4 of 22
`
`US 2007/0082861 A1
`
`FIG. 4
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 5 of 22
`
`US 2007/0082861 A1
`
`FIG. 5
`
`º pueg ->
`
`
`
`
`
`
`
`
`
`180V U80V 810V 810V 110V 910W W 190V 950WGGOW pº??eer nun
`
`
`
`
`
`
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 6 of 22
`
`US 2007/0082861 A1
`
`FIG. 6
`
`Untreated AO33 AO85 AO86 AO87
`
`
`
`-O- Band B
`
`- Band A
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 7 of 22
`
`US 2007/0082861 A1
`
`FIG. 7
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 8 of 22
`
`US 2007/0082861 A1
`
`FIG. 8
`
`Marker Untreated AO27 AO48
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 9 of 22
`
`US 2007/0082861 A1
`
`FIG. 9
`
`Marker . Untreated AO2 hAON8 AO27 hAON4 AO28 hAON6
`
`
`
`-- Band. A
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 10 of 22
`FIG 10
`
`US 2007/0082861 A1
`
`
`
`3s3s S
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 11 of 22
`
`US 2007/0082861 A1
`
`FIG. 11
`
`
`
`5gs3N
`
`C
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 12 of 22
`
`US 2007/0082861 A1
`
`FIG. 12
`
`
`
`2ES2 3
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 13 of 22
`
`US 2007/0082861 A1
`
`FIG. 13
`
`
`
`S&
`
`E
`
`

`

`Patent Application Publication Apr. 12
`
`2007 Sheet 14 Of 22
`
`US 2007/0082861 A1
`
`FIG. 14
`
`
`
`GOV/
`WOW
`
`COV (–)
`
`§§ §§
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 15 of 22
`
`US 2007/0082861 A1
`
`FIG. 15
`
`Marker
`
`(-)
`
`AO8
`
`AO9
`
`AO 10
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 16 of 22
`
`US 2007/0082861 A1
`
`FIG. 16
`
`Marker
`
`
`
`(-)
`
`AO37
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 17 of 22
`
`US 2007/0082861 A1
`
`FIG. 17
`
`
`
`C
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 18 of 22
`
`US 2007/0082861 A1
`
`F.G. 18
`
`Os
`t
`
`
`
`S
`
`N
`
`s
`
`3
`
`s
`
`S3
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 19 of 22
`
`US 2007/0082861 A1
`
`FIG. 19
`
`(-)
`
`AO95
`
`96
`
`97
`
`98
`
`
`
`1H- Band B
`
`-H Band A
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 20 of 22
`
`US 2007/0082861 A1
`
`FIG. 20
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 21 of 22
`FIG 21
`
`US 2007/0082861 A1
`
`O
`
`f
`O
`s
`
`
`
`

`

`Patent Application Publication Apr. 12, 2007 Sheet 22 of 22
`
`US 2007/0082861 A1
`
`FIG. 22
`
`
`
`s
`
`s
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`ENANUCLEC ACID DRUGS MODIFYING
`SPLICING IN MIRNA PRECURSOR
`
`TECHNICAL FIELD
`0001. The present invention relates to ENA nucleic acid
`pharmaceuticals capable of modifying splicing of mRNA
`precursors. More specifically, the present invention relates to
`antisense oligonucleotide compounds to splicing enhancer
`sequences within exon 19, 41, 45, 46, 44, 50, 55, 51 or 53
`of the dystrophin gene, as well as therapeutic agents for
`muscular dystrophy comprising the compounds.
`BACKGROUND ART
`0002 Muscular dystrophy, which is a genetic muscular
`disease, is roughly classified into Duchenne muscular dys
`trophy (DMD) and Becker muscular dystrophy (BMD).
`DMD is the most frequently occurring genetic muscular
`disease and occurs at a ratio of 1 per 3,500 male births. DMD
`patients show symptoms of weakening of muscles in their
`childhood; thereafter, muscular atrophy progresses consis
`tently and results in death at the age of around 20. Currently,
`there is no effective therapeutic for DMD. Development of
`therapeutics is strongly demanded by DMD patients
`throughout the world. BMD in many cases occurs in adult
`hood and most of the patients are capable of normal Survival
`though slight weakening of muscles is observed. Mutations
`of deletions in the dystrophin gene have been identified in 2/3
`of DMD and BMD cases. The progress of clinical symptoms
`in DMD or BMD patients is predictable depending on
`whether Such deletions disrupt the translational reading
`frame of mRNA or maintain that reading frame (Monaco A.
`P. et al., Genomics 1988: 2:90-95). Although molecular
`biological understanding of DMD has been thus deepened,
`no effective method for treating DMD has been established
`yet.
`0003) When DMD patients have a frame shift mutation,
`dystrophin protein disappears completely from patients
`skeletal muscles. On the other hand, dystrophin protein is
`produced from in-frame mRNA in BMD patient-derived
`muscle tissues, though the protein is incomplete. As a
`method for treating DMD, there is known a method in which
`an out-frame mutation (the reading frame of amino acids is
`shifted) is converted to an in-frame mutation (the reading
`frame is maintained) by modifying dystrophin mRNA (Mat
`suo M., Brain Dev 1996; 18:167-172). Recently, it has been
`reported that the mdx mouse synthesized a deletion-contain
`ing dystrophin as a result of induction of exon skipping with
`an oligonucleotide complementary to the splicing consensus
`sequence of the dystrophin gene (Wilton S. D. et al.,
`Neuromusc Disord 1999: 9:330-338; Mann C. J. et al., Proc
`Natl Acad Sci USA 2001: 98:42-47). In these studies, exon
`skipping is induced using as a target the splicing consensus
`sequence located on the border between two exons.
`0004. It is asserted that splicing is regulated by splicing
`enhancer sequences (SESs). In fact, it has been demon
`strated that by disrupting the SES within exon 19 of the
`dystrophin gene with an antisense oligonucleotide comple
`mentary thereto, complete skipping of exon 19 occurs in
`normal lymphoblastoid cells (Takeshima Y. et al., J. Clin
`Invest 1995: 95:515-520; Pramono Z. A. et al., Biochem
`Biophys Res Commun 1996: 226:445-449).
`0005. It has been also reported that by introducing an
`oligonucleotide complementary to the SES within exon 19
`
`of the dystrophin gene to thereby induce exon skipping, a
`deletion-containing dystrophin was successfully produced in
`muscular cells derived from DMD patients carrying exon 20
`deletion (Takeshima Y. et al., Brain & Development 2001:
`23:788-790; Japanese Unexamined Patent Publication No.
`H11-140930; Japanese Unexamined Patent Publication No.
`2002-10790). This indicates that repairing of the reading
`frame shift by inducing exon 19 skipping with an antisense
`oligonucleotide complementary to the SES within exon 19
`of the dystrophin gene results in production of a dystrophin
`protein whose function is partially restored; and thus it is
`possible to change DMD to BMD. If it is possible to convert
`DMD, a severe myoatrophy, to slight BMD, prolonging
`patients’ lives can be expected.
`0006. At present, oligonucleotide analogues having
`stable and excellent antisense activity are being developed
`(Japanese Unexamined Patent Publication No. 2000
`297097).
`0007. It is an object of the present invention to provide
`therapeutics with broader applicable range and higher effi
`cacy, by improving antisense oligonucleotides to the SES
`within exon 19, 41, 45, 46, 44, 50, 55, 51 or 53 of the
`dystrophin gene.
`
`DISCLOSURE OF THE INVENTION
`0008. As a result of extensive and intensive researches
`toward the achievement of the above-described object, the
`present inventors have succeeded in designing and synthe
`sizing those nucleotide sequences and antisense oligonucle
`otide compounds which have higher exon skipping effect on
`exon 19, 41, 45, 46, 44, 50, 55, 51 or 53 of the dystrophin
`gene. Thus, the present invention has been achieved.
`0009. The present invention may be summarized as fol
`lows.
`1. An oligonucleotide having the nucleotide
`0010)
`sequence as shown in any one of SEQ ID NOS: 2-6,
`10-22, 30-78, 87 or 88 in the SEQUENCE LISTING, or
`a pharmacologically acceptable salt thereof.
`0011) 2) The oligonucleotide of 1 above or a pharma
`cologically acceptable salt thereof, wherein at least one of
`the Sugars and/or the phosphates constituting the oligo
`nucleotide is modified.
`0012
`3) The oligonucleotide of 2 above or a pharma
`cologically acceptable salt thereof, wherein the Sugar
`constituting the oligonucleotide is D-ribofuranose and the
`modification of the sugar is modification of the hydroxyl
`group at position 2' of D-ribofuranose.
`0013
`4. The oligonucleotide of 3 above or a pharma
`cologically acceptable salt thereof, wherein the modifi
`cation of the sugar is 2'-O-alkylation and/or 2'-O,4'-C-
`alkylenation of the D-ribofuranose.
`0014
`5) The oligonucleotide of 2 above or a pharma
`cologically acceptable salt thereof, wherein the modifi
`cation of the phosphate is thioation of the phosphate
`group.
`6 A compound represented by the following gen
`00.15
`eral formula (I) or a pharmacologically acceptable salt
`thereof:
`
`BT—BM BE
`
`(I)
`
`

`

`US 2007/0082861 A1
`
`Apr. 12, 2007
`
`2
`
`[0016] where BT is a group represented by any one of the
`following (1a) to (1k):
`
`70*,
`*O-Bt-,
`’O'BC'Bt',
`*O-Bg-Bc-Bt-,
`*O-Ba-Bg-Bc-Bh
`, O-Bg-Ba—Bg-Bc-Bt-,
`*O-Bt—Bg-Ba-Bg-Bc-Btg
`, O-Bc-Bt—Bg-Ba-Bg-Bc-Bt—,
`, O-Bc—Bc-Bt-Bg-Ba-Bg-Bc-Bt-, or
`7O-Bg—Bc-Bc-Bt-Bg-Ba—Bg-Bc-Bt-;
`
`
`
`
`
`( a)
`( b)
`( c)
`( d)
`( e)
`(1f)
`( g)
`( h)
`(lj)
`( k)
`
`
`
`[0017] where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); BC is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U1) or (T2):
`
`(GI)
`
`o
`
`N
`
`|
`
`NH
`/l\
`
`NH2
`
`N
`</
`N
`
`O
`
`X
`
`Y
`
`-continued
`
`NHZ
`
`
`
`NH2
`
`\
`
`I
`
`N
`kN
`
`O
`
`O
`
`X
`
`Y
`
`0
`
`(G2)
`
`NH2
`
`
`
`Al
`
`(
`
`)
`
`O
`
`NH
`
`(A2)
`
`(C1)
`
`(C2)
`
`(U1)
`
`NH;
`
`\
`
`| J
`
`/
`N
`
`N
`
`</
`
`N
`
`O
`
`X
`
`Y
`
`' x
`
`N
`
`O
`
`O
`
`X
`
`Y
`
`Sarepta Exhibit 1051, Page 25 of 175
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`7. The compound according to claim 6 which is
`0025
`selected from the group consisting of the following com
`pounds (i) to (vi), or a pharmacologically acceptable salt
`thereof:
`0026 (i) a compound where Br is a group represented by
`(1k) and B is a group represented by (2h).
`0027 (ii) a compound where Br is a group represented by
`(1a) and B is a group represented by (2a),
`0028 (iii) a compound where Br is a group represented
`by (1a) and B is a group represented by (2h).
`0029 (iv) a compound where B is a group represented
`by (1e) and B is a group represented by (2a),
`0030 (v) a compound where Br is a group represented by
`(1k) and B is a group represented by (2a).
`0031 (vi) a compound where B is a group represented
`by (1a) and B is a group represented by (2f), and
`0032 (vii) a compound where Br is a group represented
`by (1a) and B is a group represented by (2d).
`0033
`8. The compound of 6 above which is selected
`from the group consisting of the following compounds
`(I1) to (IT), or a pharmacologically acceptable salt
`thereof:
`
`-continued
`
`(T2)
`
`
`
`0018 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`f
`o--on
`
`f
`
`(X1)
`
`(X2)
`
`0.019 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0020 B is a group represented by the following formula
`(2):
`(2)
`-Bg-Ba-Bt-Bc-Bt-Bg-Bc-Bt-Bg-Bg-Bc-Ba-Bt-Bc-Bt-
`0021 where Bg, Ba, Bt and Bc are as defined above:
`0022 B is a group represented by any one of the
`following (2a) to (2h):
`
`0023 where Bg, Ba, Bt and Bc are as defined above:
`0024 provided that at least one of the nucleosides con
`stituting the compound represented by formula (I) has
`2'-O,4'-C-alkylene group.
`
`0034 where Bg is a group represented by the following
`formula (G1), Ba is a group represented by the follow
`ing formula (A1); Bc is a group represented by the
`following formula (C1); Bt is a group represented by
`the following formula (U 1); Bg is a group represented
`by formula (G2); Ba is a group represented by formula
`(A2); Bc * is a group represented by formula (C2); and
`Bt is a group represented by formula (T2):
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`O
`
`(G1a)
`
`-continued
`
`(X2)
`
`NH als
`
`N
`
`NH2
`
`N (
`
`N
`
`O
`
`X OR
`
`NH2
`
`(A1)
`
`N (1. 2
`
`N
`
`N
`
`O
`
`X OR
`
`NH2
`
`NN
`
`1s.
`
`O
`
`X OR
`
`O
`
`NH 1s.
`
`O
`
`X OR
`
`(C1a)
`
`(U18)
`
`0035 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`(X1)
`
`0036) and R' is individually and independently an alkyl
`group with 1-6 carbon atoms.)
`0037
`9. The compound of 8 above where X in for
`mulas (G1), (A1), (C1) and (U1") is a group repre
`sented by formula (X2) and X in formulas (G2), (A2),
`(C2) and (T2) is a group represented by formula (X 1), or
`a pharmacologically acceptable salt thereof.
`0038)
`10 The compound of 8 above where X in all the
`formulas (G1), (A1), (C1), (U1), (G2), (A2), (C2) and
`(T2) is a group represented by formula (X2), or a phar
`macologically acceptable salt thereof.
`0039)
`11. The compound of 8 above which is repre
`sented by any one of the following formulas (Il-a), (I2-a),
`(I3-a), (I4-a), (I5-a), (I6-a), (I7-a). (I8-a) and (I9-a), or a
`pharmacologically acceptable salt thereof:
`
`(I5-a)
`
`(I7-a)
`
`(I8-a)
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`-continued
`
`(I9-a)
`
`0040 where Bg is a group represented by formula
`(G1"): Ba is a group represented by formula (A1); Bc
`is a group represented by formula (C1); Bt is a group
`represented by formula (U 1); Bg is a group repre
`sented by formula (G2); Ba is a group represented by
`formula (A2); Bc * is a group represented by formula
`(C2); Bt is a group represented by formula (T2); and in
`individual formulas, at least one of Bg, Ba, Bc, Bt,
`Bg, Ba, Bc and Bt has a group represented by
`formula (X2) as X and all of Bt 3, Bg", Bar, Ber) and Btri have
`a group represented by (X 1) as X.
`0041)
`12. The compound of any one of 6 to 11 above
`where Y in formulas (G1), (A1), (C1) and (U 1) is a
`methoxy group and Z in formulas (G2), (A2), (C2) and
`(T2) is an ethylene group, or a pharmacologically accept
`able salt thereof.
`0.042
`13 A compound represented by the following
`general formula (I) or a pharmacologically acceptable
`salt thereof:
`
`(I)
`BT, 1 BMI BBI
`0043 where B is a group represented by any one of the
`following (1a') to (1 o'):
`
`
`
`0044 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`(G1)
`
`(G2)
`
`(A1)
`
`(A2)
`
`(C1)
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`-continued
`
`NH2
`
`O
`
`NH 1s.
`
`O
`
`X Y
`
`O
`
`
`
`(C2)
`
`(U1)
`
`(T2)
`
`0045 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`o--on
`
`(X1)
`
`(X2)
`
`0046 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0047 B is a group represented by the following for
`mula (1"):
`-Ba-Ba-Ba-Bc-Bt-Bg-Ba-
`(1)
`0048 where Bg, Ba, Bt and Bc are as defined above:
`0049 B is a group represented by any one of the
`following (12a") to (121"):
`
`—CH2CH2OH,
`-Bg-CH2CH2OH,
`-Bg-Bc-CHCH-OH,
`-Bg-Bc-Ba-CH2CH2OH,
`-Bg-Bc-Ba-Ba-CH2CH2OH,
`
`-Bg-Bc-Ba-Ba-Ba-CH2CH2OH,
`-Bg-Bc-Ba-Ba-Ba-Bt-CH2CH2OH,
`
`-Bg-Bc-Ba-Ba-Ba-Bt-Bt-CHCH-OH,
`-Bg-Bc-Ba-Ba-Ba-Bt-Bt-Bt-CH2CH2OH,
`-Bg-Bc-Ba-Ba-Ba-Bt-Bt-Bt-Bg-CH2CH2OH,
`
`(12a")
`(12b')
`(12c')
`(12d')
`(12e")
`
`(12f)
`(12g)
`
`(12h")
`(12i)
`(12")
`
`-Bg-Bc-Ba-Ba-Ba-Bt-Bt-Bt-Bg-Bc-CHCH-OH, or
`(12k)
`-Bg-Bc-Ba-Ba-Ba-Bt-Bt-Bt-Bg-Bc-Bt-CH2CH2OH,
`(121')
`0050 where Bg, Ba, Bt and Bc are as defined above:
`0051 provided that at least one of the nucleosides con
`stituting the compound represented by formula (I) has
`2'-O,4'-C-alkylene group.
`0052
`14 A compound represented by the following
`general formula (II) or a pharmacologically acceptable
`salt thereof:
`
`(II)
`BT2 BM2 BB2
`0053 where Br is a group represented by any one of the
`following (2a") to (2):
`(2a")
`HO-
`(2b')
`HO-Bg-,
`(2c')
`HO-Ba-Bg-,
`(2d')
`HO-Ba-Ba-Bg-,
`(2e")
`HO-Ba-Ba-Ba-Bg-,
`(2f)
`HO-Bc-Ba-Ba-Ba-Bg-,
`(2g)
`HO-Bg-Bc-Ba-Ba-Ba-Bg-,
`(2h")
`HO-Bt-Bg-Bc-Ba-Ba-Ba-Bg-, or
`(2)
`HO-Bg-Bt-Bg-Bc-Ba-Ba-Ba-Bg-
`0054 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`

`

`US 2007/0O82861 A1
`
`7
`
`Apr. 12, 2007
`
`(G1)
`
`(G2)
`
`(A1)
`
`(A2)
`
`(C1)
`
`(C2)
`
`(U1)
`
`-continued
`
`NH2
`
`O
`
`NH 1s.
`
`O
`
`X Y
`
`O
`
`(T2)
`
`
`
`
`
`0055 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`o=l-oil
`
`O s=h-oh
`
`(X1)
`
`(X2)
`
`0056 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`
`NH2
`
`n
`
`4.
`
`N
`
`{ N
`
`O
`
`X Y
`
`NH2
`
`n
`
`2 N
`
`N
`
`{
`
`
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0057 B is a group represented by the following for
`mula (2):
`
`0.058 where Bg, Ba, Bt and Bc are as defined above:
`0059 B is a group represented by any one of the
`following (22a") to (22i"):
`—CHCH-OH,
`-Ba-CH2CH2OH,
`
`(22a")
`(22b')
`
`-Ba-Ba-CH2CH2OH,
`-Ba-Ba-Ba-CHCH-OH,
`-Ba-Ba-Ba-Ba-CH2CH2OH,
`
`(22c')
`(22d')
`(22e")
`
`-Ba-Ba-Ba-Ba-Bc-CH2CH2OH,
`(22f)
`-Ba-Ba-Ba-Ba-Bc-Bt-CHCH-OH,
`(22g)
`-Ba-Ba-Ba-Ba-Bc-Bt-Bg-CH2CH2OH, or
`(22h')
`-Ba-Ba-Ba-Ba-Bc-Bt-Bg-Ba-CH2CH2OH
`(221)
`0060 where Bg, Ba, Bt and Bc are as defined above:
`0061 provided that at least one of the nucleosides con
`stituting the compound represented by formula (II) has
`2'-O,4'-C-alkylene group.
`0062
`15 A compound represented by the following
`general formula (III) or a pharmacologically acceptable
`salt thereof:
`
`(III)
`BT3 BM3 BB3
`0063 where Br is a group represented by any one of the
`following (3a') to (3c'):
`
`HO-
`
`(3a)
`
`(3b')
`HO-Bc-, or
`(3c")
`HO-Bg-Bc-
`0064 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`(G1)
`
`N
`
`N
`
`O
`
`NH
`
`2
`N
`
`NH2
`
`
`
`
`
`-continued
`
`(G2)
`
`(A1)
`
`(A2)
`
`(C1)
`
`(C2)
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`-continued
`
`(U1)
`
`0066 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0067 B is a group represented by the following for
`mula (3'):
`-Bc-Bg-Bc-Bt-Bg-Bc-Bc-Bc-Ba-Ba-
`(3')
`0068 where Bg, Ba, Bt and Bc are as described above:)
`0069 B is a group represented by any one of the
`following (32a") to (32i"):
`
`
`
`(T2)
`
`0065 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`(X1)
`
`(X2)
`
`o--on
`
`s=-ol
`
`0070 where Bg, Ba, Bt and Bc are as described above:
`0071 provided that at least one of the nucleosides con
`stituting the compound represented by formula (III) has
`2'-O,4'-C-alkylene group.
`0072
`16 A compound represented by the following
`general formula (IV) or a pharmacologically acceptable
`salt thereof:
`
`(IV)
`BT, BMA BB4
`0073 where Bra is a group represented by any one of the
`following (4a') to (4m'):
`HO-
`
`(4a)
`
`HO-Ba-,
`
`HO-Ba-Ba-,
`HO-Bc-Ba-Ba-,
`
`(4b')
`
`(4c")
`(4d")
`
`(4e")
`HO-Ba-Bc-Ba-Ba-,
`(4f)
`HO-Bg-Ba-Bc-Ba-Ba-,
`(4g)
`HO-Bt-Bg-Ba-Bc-Ba-Ba-,
`(4h")
`HO-Bc-Bt-Bg-Ba-Bc-Ba-Ba-,
`(4)
`HO-Bt-Bc-Bt-Bg-Ba-Bc-Ba-Ba-,
`(4k)
`HO-Bt-Bt-Bc-Bt-Bg-Ba-Bc-Ba-Ba-,
`(41')
`HO-Bg-Bt-Bt-Bc-Bt-Bg-Ba-Bc-Ba-Ba-, or
`(4m")
`HO-Bt-Bg-Bt-Bt-Bc-Bt-Bg-Ba-Bc-Ba-Ba-
`0074 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`

`

`US 2007/0O82861 A1
`
`10
`
`Apr. 12, 2007
`
`(G1)
`
`(G2)
`
`(A1)
`
`(A2)
`
`(C1)
`
`-continued
`
`NH2
`
`
`
`
`
`O
`
`O
`
`X Y
`
`NH 1s.
`
`O
`
`(C2)
`
`(U1)
`
`(T2)
`
`0075 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`(X1)
`
`(X2)
`
`s=-ol
`
`0076 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`
`NH2
`
`n
`
`4.
`
`N
`
`{ N
`
`O
`
`X Y
`
`NH2
`
`n
`
`) 2 N
`
`N
`
`( .
`
`
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0.077
`B.M is a group represented by the following for
`mula (4):
`(4)
`-Bc-Ba-Bg-Bt-Bt-Bt-Bg-
`0078 where Bg, Ba, Bt and Bc are as described above:
`0079 B is a group represented by any one of the
`following (42a") to (421"):
`
`-continued
`
`0080 where Bg, Ba, Bt and Bc are as described above;
`0081 provided that at least one of the nucleosides con
`stituting the compound represented by formula (IV) has
`2'-O,4'-C-alkylene group.
`0082)
`17. A compound represented by the following
`general formula (V) or a pharmacologically acceptable
`salt thereof:
`
`(V)
`BT's BM's BB's
`0.083 where Bris is a group represented by any one of the
`following (5a') to (5g):
`
`0084 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`O
`
`(G1)
`
`
`
`
`
`(G2)
`
`(A1)
`
`(A2)
`
`(C1)
`
`(C2)
`
`NH
`
`N
`/
`
`{ll NH2
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`-continued
`
`(U1)
`
`(T2)
`
`
`
`0090 where Bg, Ba, Bt and Bc are as described above:
`0091 provided that at least one of the nucleosides con
`stituting the compound represented by formula (V) has
`2'-O,4'-C-alkylene group.
`0092]
`18 A compound represented by the following
`general formula (VI') or a pharmacologically acceptable
`salt thereof:
`
`(VI)
`BT6 BM6 BB6
`0093 where Br is a group represented by any one of the
`following (6a') to (6r'):
`
`0085 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`(X1)
`
`(X2)
`
`f
`o--on
`
`f
`s--on
`
`0.086 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0087 BMs is a group represented by the following for
`mula (5'):
`(5')
`-Bc-Bt-Bt-Bt-Bt-Ba-Bg-Bt-Bt-Bg-Bc-Bt-Bg-Bc-
`0088 where Bg, Ba, Bt and Bc are as described above;
`0089 Bs is a group represented by any one of the
`following (52a") to (52i):
`—CH2CH2OH,
`-Bt-CHCH-OH,
`-Bt-Bc-CHCH-OH,
`
`(52a")
`(52b')
`(52c')
`
`0094 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`(G1)
`
`O
`
`NH
`
`2
`N
`
`NH2
`
`N
`
`N
`
`O
`
`

`

`US 2007/0O82861 A1
`
`13
`
`Apr. 12, 2007
`
`-continued
`
`-continued
`
`(G2)
`
`O
`
`(U1)
`
`NH 1s.
`
`O
`
`X Y
`
`O
`
`
`
`(T2)
`
`(A1)
`
`(A2)
`
`0095 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`f
`o--on
`
`(C1)
`
`SEP-OH
`
`(C2)
`
`(X1)
`
`(X2)
`
`0096 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0097 B is a group represented by the following for
`mula (6'):
`-Bt-Bt-Bt-Bt-Bc-Bc-
`(6')
`0.098 where Bg, Ba, Bt and Bc are as described above:
`
`
`
`
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`0099 B is a group represented by any one of the
`following (62a") to (62m"):
`
`-continued
`
`(G2)
`
`(A1)
`
`(A2)
`
`(C1)
`
`(C2)
`
`0100 where Bg, Ba, Bt and Bc are as described above;
`0101 provided that at least one of the nucleosides con
`stituting the compound represented by formula (VI) has
`2'-O,4'-C-alkylene group.
`compound represented by the following
`01.02
`19 A
`d
`d by the followi
`general formula (VII) or a pharmacologically acceptable
`salt thereof:
`
`(VII)
`BT, BM7 BB7
`0103 where B, is a group represented by any one of the
`following (7a) to (7f.):
`
`0104 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`(G1)
`
`O
`
`NH
`
`2
`N
`
`NH2
`
`N
`
`N
`
`O
`
`
`
`
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`-continued
`
`(U1)
`
`20) The compound of any one of 13 to 19
`0.111
`above which is selected from the group consisting of the
`following compounds (i') to (xiii"), or a pharmacologically
`acceptable salt thereof:
`0112 (i') a compound represented by the following for
`mula (i"):
`
`0113 (ii) a compound represented by the following for
`mula (ii"):
`
`
`
`(T2)
`
`0114 (iii') a compound represented by the following
`formula (iii"):
`
`0115 (iv) a compound represented by the following
`formula (iv):
`
`0116 (v) a compound represented by the following for
`mula (v):
`
`(vi') a compound represented by the following
`0.117
`formula (vi"):
`
`0105 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`f
`o--on
`
`f
`
`(X1)
`
`0118 (vii') a compound represented by the following
`formula (vii"):
`
`0119 (viii) a compound represented by the following
`formula (viii"):
`
`(X2)
`
`0120 (ix) a compound represented by the following
`formula (ix):
`
`0106 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms;
`0107 B, is a group represented by the following for
`mula (7"):
`(7")
`-Bc-Bt-Bg-Bc-Bt-Bt-Bc-Bc-Bt-Bc-Bc-Ba-Ba-Bc-Bc-
`0108 where Bg, Ba, Bt and Bc are as described above;
`0109 B, is a group represented by the following (72a"):
`–CHCH-OH
`(72a")
`0110 provided that at least one of the nucleosides con
`stituting the compound represented by formula (VII") has
`2'-O,4'-C-alkylene group.
`
`0121 (x) a compound represented by the following for
`mula (x):
`
`(Xi') a compound represented by the following
`0.122
`formula (xi"):
`
`0123 (xii') a compound represented by the following
`formula (xii"):
`
`(xiii") a compound represented by the following for
`mula (xiii):
`
`0.124 where Bg is a group represented by the following
`formula (G1) or (G2); Ba is a group represented by the
`
`

`

`US 2007/0O82861 A1
`
`16
`
`Apr. 12, 2007
`
`following formula (A1) or (A2); Bc is a group represented
`by the following formula (C1) or (C2); and Bt is a group
`represented by the following formula (U 1) or (T2):
`
`-continued
`
`NH2
`
`O
`
`(G1)
`
`{ll
`
`NH
`
`4.
`
`N
`
`N
`
`O
`
`X Y
`
`N
`
`{ N
`
`O
`
`X Y
`
`O
`
`NH2
`
`n
`
`%
`
`NH2
`
`N
`
`( .
`
`n
`
`) 2 N
`
`
`
`
`
`
`
`O
`
`l
`
`N
`
`O
`
`O
`
`X Y
`
`(G2)
`
`(A1)
`
`(C2)
`
`(U1)
`
`(T2)
`
`0.125 where X is individually and independently a group
`represented by the following formula (X 1) or (X2):
`
`(A2)
`
`OR – OH
`
`(C1)
`
`SF -OH
`
`(X1)
`
`(X2)
`
`0.126 Y is individually and independently a hydrogen
`atom, a hydroxyl group or an alkoxy group with 1-6
`carbon atoms; and Z is individually and independently a
`single bond or an alkylene group with 1-5 carbon atoms.
`
`

`

`US 2007/0O82861 A1
`
`17
`
`21 The compound of any one of 13 to 20
`0127
`above which is represented by any one of the following
`compounds (I'l) to (I'20), or a pharmacologically accept
`able salt thereof:
`
`Apr. 12, 2007
`
`(G1)
`
`(I2)
`
`ORI
`
`NH2
`
`(A1)
`
`N
`
`n N
`
`{l N
`
`4.
`
`ORI
`
`NH
`
`NN
`
`(C1a)
`
`(U18)
`
`(G2)
`
`CHCH-OH
`
`CHCH-OH
`
`CHCH-OH
`HO
`
`CHCH-OH
`
`CHCH-OH
`
`2 C
`
`HCH-OH
`
`HCH-OH
`HO
`B
`ck-CHCH-OH
`B
`HO
`B
`Bt:
`
`HO
`Bc
`
`Bek 8-Bitsi-Biki-Biki-Biki-Ba-Bg-Bt-Bt
`
`CHCH-OH
`HO
`
`18)
`
`19)
`
`
`
`HO
`'20)
`(
`Bc
`0128 where Bg is a group represented by the following
`formula (G1"): Ba is a group represented by the follow
`ing formula (A1); Bc is a group represented by the
`following formula (C1); Bt is a group represented by
`the following formula (U 1); Bg is a group represented
`by the following formula (G2); Ba is a group repre
`sented by the following formula (A2); Bc * is a group
`represented by the following formula (C2); and Bt is a
`group represented by the following formula (T2):
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`22 The compound of 21 above where X in
`0131
`formulas (G1), (A1), (C1) and (U1") is a group repre
`sented by formula (X2) and X in formulas (G2), (A2),
`(C2) and (T2) is a group represented by formula (X 1), or
`a pharmacologically acceptable salt thereof.
`0132) 23. The compound of 21 above where X in all
`the formulas (G1"), (A1), (C1"), (U1), (G2), (A2), (C2)
`and (T2) is a group represented by formula (X2), or a
`pharmacologically acceptable salt thereof.
`0.133
`24 The compound of 21 above which is repre
`sented by any one of the following formulas (I'1-a) to
`(I'20-b), or a pharmacologically acceptable salt thereof:
`
`
`
`
`
`
`
`-continued
`
`NH2
`
`HC
`
`NH2
`
`NN
`
`1s,
`
`
`
`
`
`
`
`(A2)
`
`(C2)
`
`(T2)
`
`0129 where X is individually and independently a group
`represented by the following formula (X1) or (X2); R is
`individually and independently an alkyl group with 1-6
`carbon atoms;
`0130 and Z is individually and independently a single
`bond or an alkylene group with 1-5 carbon atoms:
`
`(X1)
`
`(X2)
`
`

`

`US 2007/0O82861 A1
`
`Apr. 12, 2007
`
`-continued
`
`(I'10-a)
`
`(I'13-a)
`-Bt-Bit-Bt
`HO-Bd. --Bc * :-Bt-Bt. -Bt is
`Bita-Ba-Bar-Bt-Bit--Bck --Bc * :-Bt. -Bar --Bck -
`
`(I.14-a)
`
`(I16-a)
`
`Bass-Bar-Bck-Bck -CH2CH2OH
`
`individual formulas, at least one of Bg, Ba, Bc, Bt
`Bg, Ba, Bc and Bt has a group represented by
`formula (X2) as X and all of st"). Bg7. Barr J. Beri and Et's have
`a group represented by (X 1) as X.
`0135), 25. The compound of any one of 13 to 24
`above where Y in formulas (G1), (A1), (C1) and (U 1) is
`a methoxy group and Z in formulas (G2), (A2), (C2) and
`(T2) is an ethylene group, or a pharmacologically accept
`able salt thereof.
`0.136
`(26 A compound represented by the following
`general formula (I") or a pharmacologically acceptable
`salt thereof:
`
`(I")
`BT1-BM. 1 BB,
`0137 where B is a group represented by any one of the
`following (1a") to (1m"):
`(1a")
`HO-
`(1b")
`HO-Bt-,
`(1c")
`HO-Bt-Bt-,
`(1d")
`HO-Bt-Bt-Bt-,
`(1e")
`HO-Ba-Bt-Bt-Bt-,
`(1f")
`HO-Bt-Ba-Bt-Bt-Bt-,
`(1g")
`HO-Bg-B