TRICYCLO-DNA ANTISENSE OLIGONUCLEOTIDES, COMPOSITIONS, AND METHODS FOR THE TREATMENT OF DISEASE

• US 20170260524A1
• Filed: 02/09/2017
• Published: 09/14/2017
• Est. Priority Date: 04/10/2009
• Status: Active Grant

• Current Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH), Centre National De La Recherche Scientifique, Institut de Myologie, Sorbonne Universite
• Original Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH), Centre National De La Recherche Scientifique, Institut de Myologie, Universitã¤T Bern, University Pierre et Marie Curie
• Inventors: SCHUMPERLI, Daniel, LEUMANN, Christian, FURLING, Denis, GARCIA, Luis, VOIT, Thomas
• Granted Patent: US 10,465,191 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 31/711 Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/712 Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

Provided are tricyclo-DNA (tc-DNA) AON and methods employing tc-DNA AON for modifying splicing events that occur during pre-mRNA processing. Tricyclo-DNA (tc-DNA) AON are described that may be used to facilitate exon skipping or to mask intronic silencer sequences and/or terminal stein-loop sequences during pre-mRNA processing and to target RNase-mediated destruction of processed mRNA. Tc-DNA AON described herein may be used in methods for the treatment of Duchenne Muscular Dystrophy by skipping a mutated exon 23 or exon 51 within a dystrophin gene to restore functionality of a dystrophin protein; in methods for the treatment of Spinal Muscular Atrophy by masking an intronic silencing sequence and/or a terminal stem-loop sequence within an SMN2 gene to yield modified functional SMN2 protein, including an amino acid sequence encoded by exon 7, which is capable of at least partially complementing a non-functional SMN1 protein; and in methods for the treatment of Steinert'"'"'s Myotonic Dystrophy by targeting the destruction of a mutated DM1 mRNA comprising 3′-terminal CUG repeats.

Claims

1-65. -65. (canceled)

66. A method for the delivery of an antisense oligonucleotide (AON) into a human patient comprising administering to the patient a tricyclo-DNA (tc-DNA) antisense oligonucleotide (AON), wherein the tc-DNA AON has a length of 10 to 18 nucleotides.

67. The method of claim 66, wherein the tc-DNA AON has a length of 12 to 16 nucleotides

68. The method of claim 66, wherein the tc-DNA AON has a length of 13 to 15 nucleotides

69. The method of claim 66, comprising intramuscular (i.m.), intravenous (i.v.), intraperitoneal (i.p.), intra-arterial, sub-cutaneous or transdermic injection of the tc-DNA AON.

70. The method of claim 66, comprising administering 1mg/kg to 100 mg/kg body weight/day to the patient.

71. The method of claim 66, comprising injecting the tc-DNA AON directly into the muscle of the patient.

72. The method of claim 66, wherein 8-16 nucleotides of said tc-DNA AON are complementary to a dystrophin pre-mRNA intronic splice donor site, wherein 2-8 nucleotides of said tc-DNA AON are complementary to a dystrophin pre-mRNA exonic region, and wherein said exonic region is contiguous with and 3′

to said intronic splice donor site.

73. The method of claim 66, wherein the tc-DNA AON comprises the nucleotide sequence 5′

-AACCTCGGCTTACCT-3′

(SEQ ID NO;

     1).

74. The method of claim 66 wherein said tc-DNA AON comprises the nucleotide sequence 5′

-AGAAATGCCATCTTC-3′

(SEQ ID NO;

     2).

75. The method of claim 66 wherein said tc-DNA AON comprises the nucleotide sequence 5′

-AAATGCCATCTTCCT-3′

(SEQ ID NO;

     3).

76. The method of claim 66 wherein said tc-DNA AON comprises the nucleotide sequence 5′

-TGCCATCTTCCTTGA-3′

(SEQ ID NO;

     4).

77. The method of claim 66, wherein the tc-DNA AON is complementary to an SMN2 pre-mRNA intronic silencer sequence (ISS).

78. The tc-DNA AON of claim 77, wherein said intronic silencer sequence is ISS-N1.

79. The tc-DNA AON of claim 78, wherein said tc-DNA AON comprises the nucleotide sequence 5′

-CUUUCAUAAUGCUGG-3′

(SEQ ID NO;

     5).

80. The method of claim 66, wherein the tc-DNA AON is complementary to an SMN2 pre-mRNA terminal stem-loop (TSL).

81. The method of claim 80, wherein the terminal stem-loop is TSL2.

82. The method of claim 81, wherein the tc-DNA AON comprises the nucleotide sequence 5′

-UUAAUUUAAGGAA-3′

(SEQ ID NO;

     6).

83. The method of claim 66, wherein the tc-DNA AON is complementary to a mutated DM1 mRNA comprising one or more 3′

CUG amplification(s).

84. The method of claim 83, wherein the tc-DNA AON comprises the nucleotide sequence 5′

-CAGCAGCAGCAGCAG-3′

(SEQ ID NO;

     8) or 5′

-CAGCAGCAGCAGCAGCAGCAG-3′

(SEQ ID NO;

     9).

85. The method of claim 66, wherein the patient has Duchene Muscular Dystrophy (DMD), Spinal Muscular Atrophy, or Steinert'"'"'s Myotonic Dystrophy.