Neuroprotective molecules and methods of treating neurological disorders and inducing stress granules

• US 9,676,810 B2
• Filed: 07/08/2011
• Issued: 06/13/2017
• Est. Priority Date: 07/08/2010
• Status: Active Grant

• Current Assignee: Brigham and Women's Hospital Incorporated
• Original Assignee: Brigham and Women's Hospital Incorporated
• Inventors: Anderson, Paul, Ivanov, Pavel, Emara, Mohammed
• Primary Examiner(s): Babic, Christopher M
• Assistant Examiner(s): Aron, Kimberly A
• Application Number:
  US13/808,863
• Publication Number:
  US 20130197067A1
• Time in Patent Office: 2,167 Days
• Field of Search: None
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• 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 2310/18 acting by a non-sequence specific mechanism
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin

Abstract

Provided herein are neuroprotective molecules containing a sequence of 25-35 contiguous nucleotides that is at least 80% identical to a contiguous sequence between nucleotide 1 and nucleotide 50 of a mature human tRNA and at least four contiguous guanosine-containing nucleotides, where the sequence of 25-35 contiguous nucleotides contains a D-loop stem structure, the at least four contiguous guanosine-containing nucleotides are located at the 5′ end of the neuroprotective molecule, and the neuroprotective molecule contains at least one deoxyribonucleotide. Also provided are methods of inducing or increasing stress granule formation in a cell, decreasing protein translation in a cell, decreasing stress-induced cell death, or treating a neurological disorder associated with neuron death in a subject using at least one of these neuroprotective molecules.

Claims

1. A neuroprotective molecule consisting of:

(i) a sequence of 25-35 contiguous nucleotides that is identical to a contiguous sequence between nucleotide 1 and nucleotide 50 of a mature human transfer ribonucleic acid (tRNA) having a sequence selected from the group consisting of SEQ ID NOs;

32, 37, 40-46 and 48-55, wherein a first nucleotide of the 25-35 contiguous nucleotides corresponds to nucleotide position 1 of the mature human tRNA sequence;

at least part of the sequence of 25-35 contiguous nucleotides forms a D-loop stem structure; and

at least part of the sequence of 25-35 contiguous nucleotides binds to YB-1;

(ii) optionally a 5′

-monophosphate, one or more modified nucleotides, and/or one or more modifications in a phosphate backbone in the nucleotide(s) in (i) of the neuroprotective molecule; and

(ii) optionally a 5′

-protective group and/or a 3-′

protective group;

wherein at least four contiguous nucleotides in (i) of the neuroprotective molecule at the 5′

end are guanosine deoxyribonucleotides;

the neuroprotective molecule binds YB-1; and

the neuroprotective molecule is capable of decreasing protein translation in a rabbit reticulocyte assay.

2. The neuroprotective molecule of claim 1, wherein the neuroprotective molecule comprises a 5′

-monophosphate, one or more modified nucleotides, and/or one or more modifications in a phosphate backbone of the nucleotide(s) in (i) and (ii) of the neuroprotective molecule.

3. The neuroprotective molecule of claim 2, wherein the at least one modified nucleotide contains a modified base or a modified sugar.

4. The neuroprotective molecule of claim 1, wherein the neuroprotective molecule comprises a 5′

- and/or a 3′

-protective group.

5. The neuroprotective molecule of claim 1, wherein the neuroprotective molecule has a total length of between 39 to 60 nucleotides.

6. A pharmaceutical composition comprising at least one neuroprotective molecule of claim 1.

7. A neuroprotective molecule consisting of:

(i) a sequence of 25-35 contiguous nucleotides that is identical to a contiguous sequence between nucleotide 1 and nucleotide 50 of a mature human tRNA^Ala, wherein a first nucleotide of the 25-35 contiguous nucleotides corresponds to nucleotide position 1 of the mature human tRNA^Ala sequence;

at least part of the sequence of 25-35 contiguous nucleotides forms a D-loop stem structure; and

at least part of the sequence of 25-35 contiguous nucleotides binds to YB-1;

(ii) optionally a 5′

-monophosphate, one or more modified nucleotides, and/or one or more modifications in a phosphate backbone in the nucleotide(s) in (i) of the neuroprotective molecule; and

(ii) optionally a 5′

-protective group and/or a 3′

-protective group;

wherein at least four contiguous nucleotides in (i) of the neuroprotective molecule at the 5′

end are guanosine deoxyribonucleotides;

the neuroprotective molecule binds YB-1; and

the neuroprotective molecule is capable of decreasing protein translation in a rabbit reticulocyte assay.

8. The neuroprotective molecule of claim 7, wherein the neuroprotective molecule comprises a 5′

-monophosphate, one or more modified nucleotides, and/or one or more modifications in a phosphate backbone of the nucleotide(s) in (i) and (ii) of the neuroprotective molecule.

9. The neuroprotective molecule of claim 8, wherein the at least one modified nucleotide contains a modified base or a modified sugar.

10. The neuroprotective molecule of claim 7, wherein the neuroprotective molecule comprises a 5′

- and/or a 3′

-protective group.

11. The neuroprotective molecule of claim 7, wherein the neuroprotective molecule has a total length of between 39 to 60 nucleotides.

12. A pharmaceutical composition comprising at least one neuroprotective molecule of claim 7.