PIKFYVE antisense oligonucleotides

Abstract

The present disclosure relates to PIKFYVE antisense oligonucleotides (ASOs), pharmaceutical compositions containing the antisense oligonucleotides, and methods of treating, inhibiting, suppressing, and preventing neurological diseases using the same.

Description

[0001] Citation of Related Applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 483,517, filed on February 6, 2023. The entire disclosures of the above-referenced applications are incorporated herein by reference in their entireties.

[0003] Reference to a sequence listing submitted electronically

[0004] The contents of the electronically submitted Sequence Listing (Name: 3817_225PC01_SequenceListing_ST26.xml; Size: 5,488,433 bytes; Creation Date: February 6, 2024) are incorporated herein by reference in their entirety. Technical Field

[0005] The present disclosure relates to PIKFYVE antisense oligonucleotides (ASOs), pharmaceutical compositions containing them, and methods of using them to treat, inhibit, suppress, and prevent nervous system or neurodegenerative diseases. Background Art

[0006] Many neurodegenerative disorders in patients are difficult to treat effectively, particularly when the pathology of a particular patient's neurodegenerative disorder is not fully understood.

[0007] International Publication No. WO 2016 / 210372 discloses methods of treating neurodegenerative diseases by administering PIKFYVE inhibitors.

[0008] Many neurodegenerative disorders, such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), still require effective treatments. Summary of the Invention

[0009] The present disclosure relates to PIKFYVE antisense oligonucleotides (ASOs), pharmaceutical compositions containing them, and their use in treating neurodegenerative disorders.

[0010] In some aspects, the present disclosure provides a single-chain ASO that suppresses PIKFYVE expression, wherein the ASO has a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NO: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782); and unmodified versions of SEQ ID NO: 1646-2321. In some aspects, the present disclosure provides a single-chain ASO that suppresses PIKFYVE expression, wherein the ASO has a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0011] The nucleobase sequence of the ASO may comprise up to 30, 25, 24, 23, 22, 21 or 20 consecutive nucleobases of any of the following nucleobase sequences: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782); and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the nucleobase sequence of the ASO can comprise up to 30, 25, 24, 23, 22, 21, or 20 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1780 1882-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0012] The ASO may also be any of the following: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782); and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the ASO can also be any of the following: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0013] In some aspects, the present disclosure provides an oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782); and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the present disclosure provides an oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781;and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0014] The oligonucleotide may comprise up to 25, 24, 23, 22, 21 or 20 consecutive nucleobases of any of the following nucleobase sequences: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782); and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the oligonucleotide can comprise up to 25, 24, 23, 22, 21, or 20 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0015] In some aspects, at least one internucleoside linkage is a modified internucleoside linkage, and the modified internucleoside linkage can be a phosphorothioate internucleoside linkage or a phosphodiester internucleoside linkage. At least one nucleoside can also be a modified nucleobase.

[0016] In some aspects, at least one nucleoside of the ASO can be a modified sugar moiety, wherein the modified sugar moiety can be a bicyclic sugar moiety, or the modified sugar moiety can contain a 2'-O-methoxyethyl group. In certain aspects, the bicyclic sugar moiety contains a 4'-CH(R)-O-2' bridge, wherein the R groups are independently H, C 1-12 an alkyl group or a protecting group.

[0017] In some aspects, the ASO is a gapmer (e.g., a MOE gapmer), wherein the gap segment can consist of 8 to 12 linked deoxynucleosides, the 5' wing segment consists of 3 to 5 linked nucleosides, and the 3' wing segment consists of 3 to 5 linked nucleosides. In certain aspects, the gap segment can be located between the 5' wing segment and the 3' wing segment, wherein the nucleosides of each wing segment comprise a modified sugar moiety (e.g., a sugar moiety having a 2'-O-methoxyethyl group).

[0018] In some aspects, the oligonucleotide consists of 12 to 30 linked nucleosides and has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782); and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the oligonucleotide consists of 12 to 30 linked nucleosides and has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the following nucleobase sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781;and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0019] In some aspects, the present disclosure provides a pharmaceutical composition comprising a PIKFYVE ASO of the present disclosure and one or more pharmaceutically acceptable carriers, diluents and / or excipients. In some aspects, the pharmaceutical composition is suitable for parenteral administration, such as intracerebroventricular injection or intrathecal administration.

[0020] In some aspects, the present disclosure provides methods of inhibiting, suppressing, or preventing PIKFYVE expression in a patient (e.g., a patient with a neurological or neurodegenerative disease) by administering to the patient (e.g., by intracerebroventricular injection or intrathecal administration) a PIKFYVE ASO or pharmaceutical composition described herein (e.g., an effective amount thereof).

[0021] In some aspects, the present disclosure provides methods for treating a subject suffering from a neurological disease or a neurodegenerative disease by administering a therapeutically effective amount of a PIKFYVE ASO or pharmaceutical composition as described herein. In some aspects, the disease is amyotrophic lateral sclerosis (ALS) (e.g., C9orf72-related ALS). In some aspects, the disease is frontotemporal dementia (FTD), such as FTD with TDP-43 pathology or FTD with tau pathology. In some aspects, the disease is C9orf72-related FTD (C9-FTD). In some aspects, the disease is microtubule-associated protein tau (MAPT)-related FTD (MAPT-FTD), such as FTD with a V337M MAPT mutation.

[0022] In some aspects, the present disclosure provides methods of treating a subject suffering from a PIKFYVE disease or disorder by administering a therapeutically effective amount of a PIKFYVE ASO or pharmaceutical composition described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1A and Figure 1B Skipping of PIKFYVE exon 3 is predicted to generate a stop codon, leading to premature termination of PIKFYVE protein translation. "*" represents a stop codon. The sequence shown is taken from the Ensembl database (ENST00000264380).

[0024] Figure 2A and Figure 2B Skipping of PIKFYVE exon 5 is predicted to result in significant structural changes in the PIKFYVE protein. "*" represents a stop codon. The sequence shown is taken from the Ensembl database (ENST00000264380). DETAILED DESCRIPTION

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art. In the event of conflict, this document (including definitions) shall prevail. Although methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present disclosure, preferred methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated herein by reference in their entirety. The materials, methods and examples disclosed herein are illustrative only and are not intended to be limiting. The terms used herein are only for the purpose of describing specific aspects and are not intended to be limiting.

[0026] definition

[0027] As used herein, the terms "includes," "comprising," "have," "have," "may," "contain," "possibly," and variations thereof are open-ended transitional phrases, terms, or words that do not exclude the possibility of additional actions or structures.

[0028] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0029] This disclosure also contemplates other aspects "comprising," "consisting of," and "consisting essentially of" the aspects or elements presented herein, whether explicitly recited or not.

[0030] As used herein, an "unmodified version" of an ASO / sequence refers to an ASO / sequence that comprises only DNA or RNA nucleosides joined by phosphodiester linkages.

[0031] As used herein, "2'-deoxynucleoside" means a nucleoside comprising a 2'-H(H) furanosyl sugar moiety and a nucleobase as found in naturally occurring deoxyribonucleic acid (DNA). In some aspects, a 2'-deoxynucleoside may comprise a modified nucleobase and a furanosyl sugar moiety, or may comprise an RNA nucleobase (uracil) and a furanosyl sugar moiety.

[0032] As used herein, "2'-substituted nucleosides" means nucleosides comprising a 2'-substituted sugar moiety. As used herein, "2'-substituted" with respect to a sugar moiety means a sugar moiety comprising at least one 2'-substituent other than H or OH.

[0033] As used herein, "antisense molecule" means an oligomeric nucleic acid or oligomeric duplex capable of exerting at least one antisense activity.

[0034] The modifier "about" used in conjunction with a quantity is inclusive of the stated value and has the meaning dictated by the context (e.g., it includes at least the degree of error associated with the measurement of the particular quantity). The modifier "about" should also be considered to disclose a range defined by the absolute values ​​of the two endpoints. For example, the expression "about 2 to about 4" also discloses the range "2 to 4." The term "about" can refer to ±10% of the stated value. For example, "about 10%" can mean a range of 9% to 11%, and "about 1" can mean 0.9-1.1. Other meanings of "about" will be apparent from the context, such as rounding, so that, for example, "about 1" can also mean 0.5 to 1.4.

[0035] For the recitation of numerical ranges herein, each number therebetween is expressly contemplated with equal precision. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range of 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0036] As used herein, "bicyclic sugar" or "bicyclic sugar moiety" means a modified sugar moiety comprising two rings, wherein the second ring is formed by connecting a bridge of two atoms in the first ring, thereby forming a bicyclic structure. In some aspects, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In some aspects, the bicyclic sugar moiety does not include a furanosyl moiety. As used herein, "bicyclic nucleoside" or "BNA" means a nucleoside comprising a bicyclic sugar moiety.

[0037] As used herein, "chirally enriched population" means a plurality of molecules having the same molecular formula, wherein the number or percentage of molecules within the population containing a particular stereochemical configuration at a particular chiral center is greater than the number or percentage of molecules within the population that would be expected to contain the same particular stereochemical configuration at the same particular chiral center if the particular chiral center were stereorandom. A population of chirally enriched molecules having multiple chiral centers within each molecule can contain one or more stereorandom chiral centers. In some aspects, the molecule is a modified oligonucleotide. In some aspects, the molecule is a compound comprising a modified oligonucleotide.

[0038] As used herein, "complementary" with respect to an oligonucleotide means that when the nucleobase sequence of an oligonucleotide and another nucleic acid are arranged in opposite directions, at least 70% of the nucleobases of an oligonucleotide or one or more regions and the nucleobases of another nucleic acid or one or more regions thereof can form hydrogen bonds with each other. Complementary nucleobases mean nucleobases that can form hydrogen bonds with each other. Complementary nucleobase pairs include adenine (A) and thymine (T); adenine (A) and uracil (U); cytosine (C) and guanine (G); and 5-methylcytosine (mC) and guanine (G). Complementary oligonucleotides and / or nucleic acids do not necessarily have nucleobase complementarity at each nucleoside. On the contrary, some mismatches are tolerated. As used herein, "completely complementary" or "100% complementary" with respect to an oligonucleotide means that an oligonucleotide is complementary to another oligonucleotide or nucleic acid at each nucleoside of an oligonucleotide.

[0039] As used herein, " gapmer " means the oligonucleotide comprising the modification of the internal region with multiple nucleosides, and the nucleosides support the RNase H cleavage between the external region with one or more nucleosides, wherein the nucleosides comprising the internal region are chemically different from the nucleosides comprising the external region. The internal region can be referred to as "gap", and the external region can be referred to as "wing". Unless otherwise indicated, "gapmer" refers to a sugar motif. Unless otherwise indicated, the sugar moiety of the nucleosides in the gap of the gapmer is unmodified 2'-deoxyfuranosyl. Therefore, the term "MOE gapmer" indicates a gapmer with a sugar motif and a gap of a 2'-deoxynucleoside in two wings. Unless otherwise indicated, a MOE gapmer can include one or more modified internucleoside linkages and / or modified core bases, and such modifications do not necessarily follow a sugar-modified gapmer pattern. Tables 2 and 6 below provide exemplary MOE gapmers.

[0040] In some aspects, the oligonucleotide comprises one or more types of modified sugars and / or unmodified sugar moieties arranged in a defined pattern or sugar motif along the oligonucleotide or a region thereof. In some cases, such sugar motifs include, but are not limited to, any sugar modifications discussed herein.

[0041] In some respects, the oligonucleotide of modification comprises the region with gap polymer motif or is made up of the region with gap polymer motif, and this region is limited by two external regions or " wing " and central or inner region or " breach ".The three regions of gap polymer motif comprise " 5 ' wing ", " breach " and " 3 ' wing ", and they form the continuous sequence of nucleoside, and wherein at least some sugar moieties of the nucleoside of each wing are different from at least some sugar moieties of the nucleoside of breach. Specifically, the sugar moiety (most 3 ' nucleoside of 5 '-wing and most 5 ' nucleoside of 3 '-wing) of at least the nucleoside closest to breach of each wing is different from the sugar moiety of adjacent breach nucleoside, thereby defines the boundary (wing / breach junction) between wing and breach. In some respects, the sugar moiety in breach is identical to each other. In some respects, breach comprises one or more nucleoside, and described nucleoside has the sugar moiety of the sugar moiety of one or more other nucleoside that is different from breach. In some respects, the sugar motifs of two wings are identical to each other (symmetrical gap polymer). In some aspects, the sugar motif of the 5'-wing is different from the sugar motif of the 3'-wing (asymmetric gapmer).

[0042] In some aspects, the wings of the gapmer comprise 1-5 nucleosides. In some aspects, each nucleoside of each wing of the gapmer is a modified nucleoside.

[0043] In some aspects, the gap of the gapmer comprises 7-12 nucleosides (eg, 10 nucleosides). In some aspects, each nucleoside of the gap of the gapmer is an unmodified 2'-deoxynucleoside.

[0044] In some respects, the gap polymer is a deoxy gap polymer. In some respects, the nucleoside on the gap side of each wing / gap junction is an unmodified 2'-deoxynucleoside, and the nucleoside on the wing side of each wing / gap junction is a modified nucleoside. In some respects, each nucleoside of breach is an unmodified 2'-deoxynucleoside. In some respects, each nucleoside of each wing of the gap polymer is a modified nucleoside.

[0045] In some respects, the oligonucleotide of modification comprises the region with the sugar motif of complete modification or is made up of the region with the sugar motif of complete modification.In some respects, each nucleoside of the complete modified region of the oligonucleotide of modification comprises the sugar motif of modification.In some respects, each nucleoside of the oligonucleotide of whole modification comprises the sugar motif of modification.In some respects, the oligonucleotide of modification comprises the region with the sugar motif of complete modification or is made up of the region with the sugar motif of complete modification, wherein each nucleoside in the region of complete modification comprises the sugar motif of identical modification, is referred to as the sugar motif of unified modification in this article.In some respects, the oligonucleotide of modification comprises the oligonucleotide of unified modification.In some respects, each nucleoside of the oligonucleotide of unified modification comprises identical 2 '-modification.

[0046] As used herein, "inhibit" refers to the ability to substantially antagonize, arrest, prevent, contain, limit, slow, destroy, alter, eliminate, stop, or reverse the progression or severity of the activity of a particular agent (e.g., infectious agent) or disease.

[0047] As used herein, the term "internucleoside linkage" refers to a covalent linkage between adjacent nucleosides in an oligonucleotide. As used herein, "modified internucleoside linkage" means any internucleoside linkage other than a phosphodiester internucleoside linkage. A "phosphorothioate linkage" is a modified internucleoside linkage in which one of the non-bridging oxygen atoms of the phosphodiester internucleoside linkage is replaced by a sulfur atom.

[0048] In some aspects, the nucleosides of the modified oligonucleotide can be linked together using any internucleoside linkage. Two major categories of internucleoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include, but are not limited to, phosphates, phosphotriesters, methylphosphonates or other alkylphosphonates, phosphoramidates, phosphorothioates, and phosphorodithioates containing phosphodiester bonds (also known as unmodified or naturally occurring linkages). Representative non-phosphorus-containing internucleoside linking groups include, but are not limited to, methylenemethylimino (-CH2-N(CH3)-O-CH2), thiodiesters, thiocarbamates (-OC(=O)(NH)-S-); siloxanes (-O-SiH2-O-); and N,N'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-). Modified internucleoside linkages can be used to alter (usually increase) the nuclease resistance of oligonucleotides compared to naturally occurring phosphate linkages. Methods for preparing phosphorus-containing and non-phosphorus-containing internucleoside linkages are well known to those skilled in the art.

[0049] Representative internucleoside linkages with chiral centers include but are not limited to alkylphosphonates and phosphorothioates. The oligonucleotides comprising the modified internucleoside linkages with chiral centers can be prepared as modified oligonucleotide populations comprising stereorandom internucleoside linkages, or modified oligonucleotide populations comprising the phosphorothioate linkages of specific stereochemical configurations. In some aspects, the oligonucleotide populations of modification comprise phosphorothioate internucleoside linkages, wherein all phosphorothioate internucleoside linkages are stereorandom. Such modified oligonucleotides can be produced with a synthetic method that results in the random selection of the stereochemical configuration of each phosphorothioate linkage. Nevertheless, as is well known to those skilled in the art, each single phosphorothioate of each single oligonucleotide molecule has a definite stereoconfiguration. In some aspects, the oligonucleotide populations of modification are rich in modified oligonucleotides, and the oligonucleotides of the modification comprise one or more specific phosphorothioate internucleoside linkages that are in specific, independently selected stereochemical configurations. In some aspects, the specific configuration of specific phosphorothioate linkages is present in at least 65% of the molecules in the population. In some respects, the specific configuration of specific thiophosphate linkage is present in at least 70% molecule in the colony. In some respects, the specific configuration of specific thiophosphate linkage is present in at least 80% molecule in the colony. In some respects, the specific configuration of specific thiophosphate linkage is present in at least 90% molecule in the colony. In some respects, the specific configuration of specific thiophosphate linkage is present in at least 99% molecule in the colony. The chiral enriched population of such modified oligonucleotides can be produced using synthetic methods known in the art, such as those described in Oka et al., JACS 125, 8307 (2003); Wan et al., Nuc.Acid.Res. 42, 13456 (2014); Chapter 10 of Locked Nucleic Acid Aptamers in Nucleic Acid and Peptide Aptamers: Methods and Protocols v 535, Barciszewski et al., 2009, editor Gunter Mayer and; and WO 2017 / 015555. In some aspects, the oligonucleotide population of modification is enriched for oligonucleotides having at least one modification of the phosphorothioate in (Sp) configuration. In some aspects, the oligonucleotide population of modification is enriched for oligonucleotides having at least one modification of the phosphorothioate in (Rp) configuration.

[0050] As used herein, "MOE" means methoxyethyl. "2'-MOE" means the -OCH2CH2OCH3 group at the 2' position of the furanosyl ring.

[0051] "Nervous system disease" refers to any disease that causes electrical, biochemical or structural abnormalities in the brain, spine or neurons. For example, a nervous system disease can be a neurodegenerative disease. For example, a neurodegenerative disease can cause degeneration of motor neurons. A nervous system disease can be, for example, amyotrophic lateral sclerosis (ALS), Huntington's disease, Alzheimer's disease or frontotemporal dementia. Other examples of nervous system diseases include, but are not limited to, Parkinson's disease, multiple sclerosis, peripheral myopathy, Rasmussen's encephalitis, attention deficit hyperactivity disorder, autism, central pain syndrome, anxiety and / or depression.

[0052] Neurological diseases may be associated with abnormal endosomal trafficking. For example, endosomal pathways and endosomes are essential components of the recycling or degradation of membrane-bound proteins, the transport of Golgi-associated proteins, and the extracellular release of proteins in exosomes. These processes contribute to neurotransmission and drive the balance between recycling and degradation of synaptic vesicles or neurotransmitter receptors, for example.

[0053] Neurological diseases may be associated with abnormal lysosomal degradation. Alterations in lysosomal degradation may be present in neurological diseases such as neurodegenerative diseases. Imbalances in cathepsin production during aging and age-related diseases may have deleterious effects on central nervous system (CNS) neurons, and lysosomes may be sites for the unfolding and partial degradation of membrane proteins or their precursors, which are then expelled from the cell or released from dying cells and accumulate as pathological entities.

[0054] Healthcare professionals can diagnose a subject with a disorder related to motor neuron degeneration by evaluating one or more of these symptoms. To diagnose a neurological disorder, a physical exam may be followed by a thorough neurological examination. This examination may assess motor and sensory skills, nerve function, hearing and speech, vision, coordination and balance, mental status, and changes in mood or behavior. Non-limiting symptoms of illness associated with a neurological disorder may include weakness in the arms, legs, feet, or ankles; slurred speech; difficulty lifting the front of the foot and toes; weakness or clumsiness of the hands; muscle paralysis; muscle stiffness; involuntary jerking or writing movements (chorea); involuntary, persistent muscle contractions (dystonia); bradykinesia; loss of automatic movements; impaired posture and balance; lack of flexibility; tingling in parts of the body; electric shock sensations with head movement; twitching of the arms, shoulders, and tongue; difficulty swallowing; difficulty breathing; difficulty chewing; partial or complete loss of vision; double vision; slow or abnormal eye movements; tremors; unsteady gait; fatigue; memory loss; dizziness; difficulty thinking or concentrating; difficulty reading or writing; misunderstanding spatial relationships; disorientation; depression; anxiety; difficulty making decisions and judgments; loss of impulse control; difficulty planning and performing familiar tasks; aggression; irritability; social withdrawal; mood swings; confusion; changes in sleep habits; and changes in appetite.

[0055] Tests may be performed to rule out diseases and conditions that may have symptoms similar to those of a neurological disorder, to measure muscle involvement, and to assess neuronal degeneration. Non-limiting examples of tests are electromyography (EMG); nerve conduction velocity studies; laboratory tests of blood, urine, or other substances; magnetic resonance imaging (MRI); magnetic resonance spectroscopy; muscle or nerve biopsy; transcranial magnetic stimulation; genetic screening; X-rays; fluoroscopy; angiography; computed tomography (CT); positron emission tomography; cerebrospinal fluid analysis; intrathecal contrast-enhanced CT scan; electroencephalogram (EEG); electronystagmography; evoked responses; polysomnography; thermography; and ultrasound. A healthcare professional may also assess a patient's family history of diseases associated with motor neuron degeneration and make a diagnosis based in part on a family history of neurological diseases. A healthcare professional may diagnose a disease associated with a neurological disorder after the subject presents with one or more symptoms.

[0056] Neurodegenerative diseases result in the progressive destruction of neurons, thereby affecting neuronal signaling. For example, neurodegeneration can be amyotrophic lateral sclerosis, Alzheimer's disease, Huntington's disease, Friedreich's ataxia, Lewy body disease, Parkinson's disease, spinal muscular atrophy, primary lateral sclerosis, progressive muscular atrophy, progressive bulbar palsy, and pseudobulbar palsy.

[0057] Diseases associated with motor neuron degeneration may result in the progressive destruction of motor neurons, which interferes with the signals they send to muscles, leading to muscle weakness and atrophy. In healthy individuals, upper motor neurons transmit signals from the brain to lower motor neurons in the brainstem and spinal cord, which then carry the signals to muscles to produce voluntary muscle movement. Destruction of both upper and lower motor neurons can affect activities such as breathing, speaking, swallowing, and walking, and over time, these functions may be lost. Examples of motor neuron diseases include, but are not limited to, amyotrophic lateral sclerosis, primary lateral sclerosis, progressive muscular atrophy, progressive bulbar palsy, and pseudobulbar palsy.

[0058] Neuronal hyperexcitation can occur when receptors for the excitatory neurotransmitter glutamate (glutamate receptors), such as NMDA receptors and AMPA receptors, are overactivated by excess glutamate or other compounds or neurotransmitters that act on glutamate receptors. Excitotoxicity can be caused by neuronal hyperexcitation. Excitotoxicity is a pathological process in which nerve cells are damaged or die due to overstimulation. Excessive stimulation can cause high levels of calcium ions (Ca 2+ ) enters the cell. 2+ The influx of phospholipases into cells activates a variety of enzymes, including phospholipases, endonucleases, and proteases (e.g., calpains), which damage cellular structures such as the cytoskeleton, membranes, and components of DNA.

[0059] Neuronal hyperexcitability may be associated with spinal cord injury, stroke, traumatic brain injury, hearing loss (due to noise overexposure or ototoxicity), epilepsy, painful neuropathies, attention deficit hyperactivity disorder, autism, central pain syndromes, neurodegenerative diseases, multiple sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, frontotemporal dementia, schizophrenia, Rasmussen's encephalitis, Huntington's disease, alcoholism or alcohol withdrawal, especially benzodiazepines Other common conditions that can lead to elevated glutamate concentrations around neurons include rapid withdrawal from antidepressants and Huntington's disease. Hypoglycemia is the primary means of removing glutamate from the intersynaptic space at NMDA and AMPA receptor sites.

[0060] As used herein, "non-bicyclic modified sugar moiety" means a modified sugar moiety comprising a modification (eg, a substituent) that does not form a bridge between two atoms of the sugar to form a second ring.

[0061] As used herein, "nucleobase" means an unmodified nucleobase or a modified nucleobase. As used herein, "unmodified nucleobase" is adenine (A), thymine (T), cytosine (C), uracil (U) or guanine (G). As used herein, "modified nucleobase" refers to a group of atoms other than unmodified A, T, C, U or G that can pair with at least one unmodified nucleobase or modified nucleobase. "5-methylcytosine" or "mC" is a modified nucleobase. Universal base is a modified nucleobase that can pair with any of the five unmodified nucleobases. As used herein, "nucleobase sequence" means the order of the continuous nucleobases in a nucleic acid or oligonucleotide that are independent of any sugar or internucleoside linkage modification.

[0062] In some aspects, the oligonucleotide of modification comprises one or more nucleosides, and described nucleosides comprise unmodified core base.In some aspects, the oligonucleotide of modification comprises one or more nucleosides, and described nucleosides comprise modified core base.In some aspects, the oligonucleotide of modification comprises one or more nucleosides that do not comprise core base, referred to as abasic nucleoside.

[0063] In some aspects, the modified nucleobase is selected from the group consisting of: a 5-substituted pyrimidine, a 6-azapyrimidine, an alkyl or alkynyl substituted pyrimidine, an alkyl substituted purine, and N-2, N-6, and O-6 substituted purines. In some aspects, the modified nucleobase is selected from the group consisting of: 2-aminopropyladenine, 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (-C≡C-CH3) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxy, 8-aza and other 8- Substituted purines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, 5-halouracil and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, hybrid bases, size-extended bases and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines such as 1,3-diazaphenoxazine-2-one, 1,3-diazaphenoxazine-2-one, and 9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced by other heterocycles such as 7-deaza-adenine, 7-deazaguanine, 2-aminopyridine, and 2-pyridone. Additional nucleobases include those disclosed in U.S. Pat. No. 3,687,808, disclosed in The Concise Encyclopedia OfPolymerScience AndEngineering , Kroschwitz, JI ed., John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, YS, Chapter 15, Antisense Research andApplications , Crooke, ST and Lebleu, B., eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 15, AntisenseDrug Technology , Crooke ST ed., CRC Press, 2008, 163-166 and 442-443.

[0064] As used herein, "nucleoside" means a compound comprising a core base and a sugar moiety. The core base and sugar moiety are each independently unmodified or modified. As used herein, "modified nucleoside" means a nucleoside comprising a modified core base and / or a modified sugar moiety. Modified nucleosides include abasic nucleosides lacking a core base. "Connected nucleosides" are nucleosides connected in a continuous sequence (i.e., without additional nucleosides between the connected nucleosides).

[0065] As used herein, "oligomeric compound" means an oligonucleotide and optionally one or more additional features, such as conjugate groups or terminal groups. An oligomeric compound may be paired with a second oligomeric compound that is complementary to the first oligomeric compound, or may be unpaired. A "single-stranded oligomeric compound" is an unpaired oligomeric compound. The term "oligoduplex" means a duplex formed by two oligomeric compounds having complementary nucleobase sequences. Each oligomeric compound of an oligoduplex may be referred to as a "duplex oligomeric compound."

[0066] As used herein, "oligonucleotide" means a chain of linked nucleosides connected by internucleoside linkages, wherein each nucleoside and internucleoside linkage can be modified or unmodified. The internucleoside linkage can be any connection described herein. Unless otherwise indicated, an oligonucleotide consists of 8-50 linked nucleosides. As used herein, "modified oligonucleotide" means an oligonucleotide in which at least one nucleoside or internucleoside linkage is modified. As used herein, "unmodified oligonucleotide" means an oligonucleotide that does not comprise any nucleoside modifications or internucleoside modifications.

[0067] "PIKFYVE," also known in the art as "phosphatidylinositol 3-phosphate 5-kinase type III" or "PIPKIII," is a phosphoinositide kinase containing the FYVE finger, encoded by the PIKFYVE gene. PIKFYVE is an evolutionarily highly conserved lipid kinase that also possesses protein kinase activity, regulating endomembrane homeostasis and playing a role in the biogenesis of endosomal carrier vesicles within early endosomes. PIKFYVE-mediated conversion of PI3P to PI(3,5)P2 blocks the recruitment of the protein EEA1. Recruitment is blocked because PIP3 forms a platform with RAB5, enabling EEA1 to anchor to early endosomes. EEA1 then drives fusion with endocytic vesicles and other endosomal vesicles.

[0068] As used herein, "PIKFYVE diseases or conditions" include lysosomal degradation diseases and conditions mediated by PIKFYVE. For example, PIKFYVE diseases or conditions include, but are not limited to, amyloid diseases (such as Alzheimer's disease, Parkinson's disease, Huntington's disease, type 2 diabetes, diabetic amyloidosis, and chronic hemodialysis-associated amyloid), multiple sclerosis, and MPS disorders (such as MPS1, MPS II, MPS IIIA, MPS IIIB, MPS IIIC, MPS HID, MPS IVA, MPS IVB, MPS VI, MPS VII, or MPS IX). In some aspects, these diseases are autoimmune diseases (such as multiple sclerosis, rheumatoid arthritis, juvenile chronic arthritis, ankylosing spondylitis, psoriasis, psoriatic arthritis, adult Still's disease, Behcet's syndrome, familial Mediterranean fever, Crohn's disease, leprosy, osteomyelitis, tuberculosis, chronic bronchiectasis, Castleman's disease) or central nervous system diseases (such as spongiform encephalopathies (Creut. Jakob encephalopathy, kuru, mad cow disease)). The compositions and methods of the present disclosure can be used to treat individuals with lysosomal storage diseases, comprising administering to a subject in need of treatment a therapeutically effective amount of a PIKfyve ASO or pharmaceutical composition described herein. In some aspects, the ASOs and compositions of the present disclosure reduce or inhibit the activity of PIKfyve and alter the biogenesis, function, or dynamics of the endosomal or lysosomal system in a manner that reduces the abundance of substances abnormally stored in lysosomes in lysosomal storage diseases. In some aspects, the ASOs and compositions target, reduce or inhibit the activity of PIKfyve, thereby altering the biogenesis, function or dynamics of the endoplasmic reticulum or Golgi apparatus in a manner that reduces the abundance of substances abnormally stored in lysosomes in lysosomal storage diseases. In some aspects, the disease is a neurological disorder.

[0069] As used herein, "sugar moiety" means an unmodified sugar moiety or a modified sugar moiety. A superscript prime (') is used to describe the numbering of sugars in a nucleoside or nucleotide (nucleobase positions are numbered without a prime). When describing only sugars, a prime is not used. As used herein, "unmodified sugar moiety" means the 2-OH (H) furanosyl moiety found in RNA ("unmodified RNA sugar moiety"), or the 2-H (H) moiety found in DNA ("unmodified DNA sugar moiety"). The unmodified sugar moiety has one hydrogen at positions 1, 3, and 4, one oxygen at position 3, and two hydrogens at position 5. As used herein, "modified sugar moiety" or "modified sugar" means a modified furanosyl sugar moiety or sugar surrogate. As used herein, a modified furanosyl sugar moiety means a furanosyl sugar comprising a non-hydrogen substituent to replace at least one hydrogen of the unmodified sugar moiety. In some aspects, the modified furanosyl sugar moiety is a 2-substituted sugar moiety. Such modified furanosyl sugar moieties include bicyclic and non-bicyclic sugars.

[0070] In some aspects, the modified sugar moiety is a non-bicyclic modified sugar moiety comprising a furanosyl ring with one or more substituents, wherein no substituent bridges the two atoms of the furanosyl ring to form a bicyclic structure. Such non-bridging substituents can be located at any position of the furanosyl group, including but not limited to substituents at positions 2, 4, and / or 5. In some aspects, one or more non-bridging substituents of the non-bicyclic modified sugar moiety are branched. Examples of 2-substituents suitable for non-bicyclic modified sugar moieties include but are not limited to: 2-F, 2-OCH3 ("OMe" or "O-methyl") and 2-O(CH2)2OCH3 ("MOE"). In some aspects, the 2-substituents are selected from: halogen, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, O 1-10 Alkoxy, OC 1-10 Substituted alkoxy, OC 1-10 Alkyl, OC 1-10 Substituted alkyl, S-alkyl, N(R m )-alkyl, O-alkenyl, S-alkenyl, N(R m )-alkenyl, O-alkynyl, S-alkynyl, N(Rm)-alkynyl, O-alkylene-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH2)2SCH3, O(CH2)2ON(R m )(R n ) or OCH2C(=O)-N(R m )(R n ), where each R m and R n are independently H, an amino protecting group, or a substituted or unsubstituted C 1-10 Alkyl, and the 2- substituent may be further substituted by one or more substituents independently selected from the following: hydroxyl, amino, alkoxy, carboxyl, benzyl, phenyl, nitro (NO2), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl. Examples of 4'- substituents suitable for non-bicyclic modified sugar moieties include but are not limited to alkoxy (e.g., methoxy) and alkyl. Examples of 5- substituents suitable for non-bicyclic modified sugar moieties include but are not limited to: 5-methyl (R or S), 5-vinyl and 5-methoxy. In some aspects, the non-bicyclic modified sugar moiety includes more than one non-bridging sugar substituent, such as 2-F-5-methyl sugar moiety.

[0071] In some aspects, the 2'-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2'-substituent selected from the group consisting of F, NH2, N3, OCF3, OCH3, O(CH2)3NH2, CH2CH=CH2, OCH2CH=CH2, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(R m )(R n ), O(CH2)2O(CH2)2N(CH3)2 and N-substituted acetamide (OCH2C(=O)-N(R m )(R n )), where each R m and R n are independently H, an amino protecting group or a substituted or unsubstituted C 1-10 alkyl.

[0072] In some aspects, the 2'-substituted nucleoside is a non-bicyclic modified nucleoside comprising a sugar moiety comprising a non-bridging 2'-substituent selected from the group consisting of F, OCF3, OCH3, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(CH3)2, O(CH2)2O(CH2)2N(CH3)2, and OCH2C(=O)-N(H)CH3 ("NMA").

[0073] In some aspects, the 2'-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2'-substituent group selected from the group consisting of F, OCH3, and OCH2CH2OCH3.

[0074] Certain modified sugar moieties comprise a substituent that bridges two atoms of the furanosyl ring to form a second ring, thereby generating a bicyclic sugar moiety. In some aspects, the bicyclic sugar moiety comprises a bridge between the 4' furanose ring atom and the 2' furanose ring atom. Examples of such 4 to 2 bridged sugar substituents include, but are not limited to, 4-CH2-2, 4-(CH2)2-2, 4-(CH2)3-2, 4-CH2-O-2 ("LNA"), 4-CH2-S-2, 4-(CH2)2-O-2 ("ENA"), 4-CH(CH3)-O-2 (referred to as "constrained ethyl" or "cEt"), 4-CH2-O-CH2-2, 4-CH2-N(R)-2, 4-CH(CH2OCH3)-O-2 ("constrained MOE" or "cMOE") and the like, 4-C(CH3)(CH3)-O-2 and the like, 4-CH2-N(OCH3)-2 and the like, 4-CH2-ON(CH3)-2, 4-CH2-C(H)(CH3)-2, 4-CH2-C(=CH2)-2 and the like, 4-C(Ra R b )-N(R)-O-2、4-C(R a R b )-ON(R)-2, 4-CH2-ON(R)-2 and 4-CH2-N(R)-O-2, where R, R a and R b Each independently is H, a protecting group or C 1-12 alkyl.

[0075] In some aspects, such 4 to 2 bridges independently comprise 1 to 4 linking groups independently selected from: -[C(R a )(R b )] n -、-[C(R a )(R b )] n -O-、-C(R a )=C(R b )-、-C(R a )=N-、-C(=NR a )-, -C(=O)-, -C(=S)-, -O-, -Si(R a )2-、-S(=O) x- and -N(R a )-; wherein: x is 0, 1 or 2; n is 1, 2, 3 or 4; each R a and R b are independently H, a protecting group, a hydroxyl group, a C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkenyl, substituted C 2-12 Alkenyl, C 2-12 Alkynyl, substituted C 2-12 Alkynyl, C 5-20 Aryl, substituted C 5-20 Aryl, heterocyclic, substituted heterocyclic, heteroaryl, substituted heteroaryl, C 5-7 alicyclic group, substituted C5-7 alicyclic group, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl group (C(=O)-H), substituted acyl group, CN, sulfonyl group (S(=O)2-J1) or sulfoxyl group (S(=O)-J1); and J1 and J2 are each independently H, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 2-12 Alkenyl, substituted C 2-12 Alkenyl, C 2-12 Alkynyl, substituted C 2-12 Alkynyl, C 5-20 Aryl, substituted C5-20 Aryl, acyl (C (= O) -H), substituted acyl, heterocyclic, substituted heterocyclic, C 1-12 Aminoalkyl, substituted C 1-12 aminoalkyl, or a protecting group.

[0076] Additional bicyclic sugar moieties are known in the art, see for example: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443, Albaek et al., J. Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 20017, 129, 8362-8379; Wengel et al., U.S. Pat. No. 7,053,2 07; Imanishi et al., U.S. Patent No. 6,268,490; Imanishi et al., U.S. Patent No. 6,770,748; Imanishi et al., USRE 44,779; Wengel et al., U.S. Patent No. 6,794,499; Wengel et al., U.S. Patent No. 6,670,461; Wengel et al., U.S. Patent No. 7,034,133; Wengel et al., U.S. Patent No. 8,080,644; Wengel et al., U.S. Patent No. 8,034,909; Wengel et al., U.S. Patent No. 8,153,365; Wengel et al., U.S. Patent No. 7,572,582; and Ramasamy et al., U.S. Patent No. 6,525,191; Torsten et al., WO 2004 / 106356; Wengel et al., WO 1999 / 014226; Seth et al., WO 2007 / 134181; Seth et al., U.S. Patent No. 7,547,684; Seth et al., U.S. Patent No. 7,666,854; Seth et al., U.S. Patent No. 8,088,746; Seth et al., U.S. Patent No. 7,750,131; Seth et al., U.S. Patent No. 8,030,467; Seth et al., U.S. Patent No. 8,268,980; Seth et al., U.S. Patent No. 8,546,556; Seth et al., U.S. Patent No. 8,530,640; Migawa et al., U.S. Patent No. 9,012,421; Seth et al., U.S. Patent No. 8,501,805; and U.S. Patent Publication Nos. Allerson et al., US2008 / 0039618 and Migawa et al., US2015 / 0191727.

[0077] "Subject" and "patient" used interchangeably herein refer to any vertebrate, including but not limited to mammals (e.g., cows, pigs, camels, llamas, horses, goats, rabbits, sheep, hamsters, guinea pigs, cats, dogs, rats and mice, non-human primates (e.g., monkeys, such as cynomolgus monkeys or rhesus monkeys, chimpanzees, etc.) and humans). In some aspects, the subject can be human or non-human. In some aspects, the subject or patient is human. The subject or patient may be receiving other forms of treatment. In some aspects, the patient suffers from a neurological disease due to a mutation in the C9ORF72 gene (e.g., the patient may be haploinsufficient for the C9ORF72 gene (e.g., resulting in a 50% or greater reduction in C9ORF72 protein activity), or the C9ORF72 gene may contain a GGGGCC repeat expansion (such as (GGGGCC) in C9ORF72). n (SEQ ID NO: 1565) hexanucleotide expansion)). The variable "n" can be at least 30.

[0078] Unless otherwise defined, "therapeutically effective amount" or "effective dose" or "effective amount" used interchangeably herein means a dosage of a drug that is effective to achieve the desired therapeutic result within the necessary time period. The effective dose can be determined by those skilled in the art and can vary according to factors such as the disease state, age, sex and weight of the individual and the ability of the drug to elicit a desired response in the individual. The term as used herein can also refer to an amount that is effective to produce a desired in vivo effect in an animal, mammal or human, such as reducing and / or inhibiting the function of a receptor. A therapeutically effective amount can be given in one or more administrations (e.g., a medicament can be given as a preventive treatment or therapeutically at any stage of disease development, before or after symptoms, etc.), applications or dosages, and is not intended to be limited to a specific formulation, combination or route of administration. The drug can be administered at different times during the course of treatment of the subject, which is within the scope of the present disclosure. The time of administration and the dosage used will depend on several factors, such as the therapeutic goal (e.g., treatment versus prevention), the condition of the subject, etc., and can be easily determined by those skilled in the art.

[0079] As used herein, the terms "treat" or "treating" a subject refer to administering a composition or agent as described herein to a subject such that at least one symptom of a disease or condition is cured, alleviated, mitigated, altered, remedied, reduced, ameliorated, or improved. Treatment includes administering an effective amount to alleviate, mitigate, alter, cure, reduce, ameliorate, and / or improve one or more symptoms associated with a disease or condition. The treatment can inhibit the deterioration or worsening of symptoms associated with a disease or condition.

[0080] The treatment methods described herein may include administering to a subject in need thereof a composition comprising an effective amount of one or more antisense oligonucleotides that treat a neurological disease by inhibiting or suppressing PIKFYVE expression. The one or more antisense oligonucleotides can reduce or suppress neurodegeneration. The one or more antisense oligonucleotides can reduce neuronal hyperexcitability. Reducing PIKFYVE mRNA and PIKFYVE protein levels can suppress neurodegeneration driven by toxic TDP-43 aggregates, DPR aggregates (e.g., in C9ORF72-ALS patients) and promoting the retention of TDP-43 in the nucleus. Delivery of ASOs targeting PIKFYVE mRNA as described herein reduces PIKFYVE protein levels.

[0081] The composition can inhibit kinase activity by inhibiting the expression of kinases. The composition can inhibit PIKFYVE kinase activity or expression. The one or more antisense oligonucleotides can be combined with a small molecule therapeutic agent (such as apilimod and / or YM201636).

[0082] The present disclosure provides oligonucleotides (modified or unmodified) that can be used to modulate PIKFYVE expression. Table 1 provides the universal base sequences (5' to 3') of the PIKFYVE antisense oligonucleotides or inhibitory nucleic acids of the present disclosure. Table 6 provides the modified base sequences (5' to 3') of the PIKFYVE antisense oligonucleotides or inhibitory nucleic acids of the present disclosure. The unmodified sequences of Table 6 are additionally contemplated as universal base sequences of the PIKFYVE antisense oligonucleotides or inhibitory nucleic acids of the present disclosure. It is predicted that the ASO sequences of SEQ ID NOs: 1-49 will effectively skip exon 3 of PIKFYVE and generate a stop codon to result in premature termination of PIKFYVE translation, as Figure 1A and Figure 1B It is expected that the ASO sequences of SEQ ID NO: 50-80 will effectively skip exon 5 of PIKFYVE and cause significant structural changes to the protein, rendering it nonfunctional, such as Figure 2A and Figure 2BAs shown. As determined by the method described in Hagedorn et al., Nucleic Acid Therapeutics, 2022, 32(3): 151-162 (DOI: 10.1089 / nat / 2021 / 0071), ASO sequences SEQ ID NO: 81-328, 330-373, 376-420, 423-479 have lower acute neurotoxicity scores. The unmodified versions of the ASO sequences SEQ ID NO: 307, 308, 330, 424, 480-540, 542-619, 621-685, 688-782 and SEQ ID NO: 1646-2321 have a low number of predicted off-target binding sites, as determined by the GGGenome search tool using 2 mismatches / gaps.

[0083] Table 1

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104] In some aspects, the present disclosure provides a modified oligonucleotide consisting of 12-30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotide bases of any one of the following nucleobase sequences: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782 in Table 1); and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the present disclosure provides a modified oligonucleotide consisting of 12-30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotide bases of any one of the following nucleobase sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781 in Table 1;and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0105] In some aspects, the modified oligonucleotide is at least 80% to 100% (i.e., 80%, 82%, 84%, 86%, 88%, 90%, 92%, 94%, 96%, 98%, or 100%; or any numerical range or value between any of the foregoing values) identical to any sequence comprising or consisting of SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782) and unmodified versions of SEQ ID NOs: 1646-2321. In some aspects, the modified oligonucleotide is at least 80% to 100% (i.e., 80%, 82%, 84%, 86%, 88%, 90%, 92%, 94%, 96%, 98%, or 100%; or any numerical range or value between any of the foregoing values) identical to any sequence comprising or consisting of SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0106] The sequences provided in Table 1 and unmodified versions of the sequences in Table 6 can be used to design antisense molecules for inhibiting PIKFYVE expression. For example, gapmer oligonucleotides can be designed using the sequences in Table 1 and unmodified versions of the sequences in Table 6 and can comprise a 5' flap of about 3-5 nucleotides, a 3' flap of about 3-5 nucleotides, and a gap region comprising 8-12 consecutive deoxyribonucleosides of any of the sequences in Table 1 and the unmodified versions of the sequences in Table 6.

[0107] In some aspects, the oligonucleotides of the present disclosure comprise a gapmer having a gap segment of at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotide bases of any one of the following nucleobase sequences: SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782 in Table 1); and unmodified versions of SEQ ID NOs: 1646-2321; flanked by 5' and 3' wing segments, wherein the gap segment is located between the 5' and 3' wing segments, and wherein each wing segment comprises a modified sugar. In some aspects, the gap segment is 8-10 nucleosides in length and each wing segment is 3-5 modified nucleosides in length. In some aspects, the oligonucleotides of the present disclosure comprise a 5' wing segment comprising a modified sugar and having a nucleobase sequence of the first 3-5 nucleobases of any one of SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782) and an unmodified version of SEQ ID NOs: 1646-2321; followed by a gap of the next 8-12 unmodified nucleotides corresponding to the same sequence of SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782) and an unmodified version of SEQ ID NOs: 1646-2321, followed by a 5' wing segment comprising a modified sugar and having a nucleobase sequence of the first 3-5 nucleobases of any one of SEQ ID NOs: 1-782 (e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782) and an unmodified version of SEQ ID NOs: 1646-2321, NO: 1-782 (eg, 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782) and the 3' wing segment of the nucleobase sequence of the last 3-5 nucleobases of the unmodified versions of the same sequences of SEQ ID NO: 1646-2321. Tables 2 and 6 provide MOE gapmers of the present disclosure.

[0108] In some aspects, the oligonucleotides of the present disclosure comprise a gapmer having a gap segment of at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotide bases of any of the following nucleobase sequences: SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781 in Table 1; NO:1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1 698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837 7. 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097,2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2 In some aspects, the gap segment is 8-10 nucleosides in length and each wing segment is 3-5 modified nucleosides in length. In some aspects, the oligonucleotides of the present disclosure comprise a 5' wing segment comprising a modified sugar and having the nucleobase sequence of the first 3-5 nucleobases of any one of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781, and SEQ ID NOs: NO:1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 17 08-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-177 5. 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892,1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985- 1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104、2106-2109、2111-2116、2118、2120-2121、2123-2124、2126-2128、2130、2132-2143、2146-2147、2149-2164、2166-2177、2179-2183、2185、2187-2205、 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321; followed by a gap of the next 8-12 unmodified nucleotides corresponding to the same sequence as follows: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781 and SEQ ID NO: NO:1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1 692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1 727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811,1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909 , 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031 -2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177 , 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, followed by an unmodified version of a 3' wing segment comprising a modified sugar and having a nucleobase sequence corresponding to the last 3-5 nucleobases of the same sequence: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781 and SEQ ID NO: NO:1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1 692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725,1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955- 1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316 and 2319-2321. Tables 2 and 6 provide the MOE gapmers of the present disclosure.

[0109] The 5' and / or 3' wings can comprise the following chemicals: 2'-OMe, 2'-MOE, LNA, or DNA, used alone or in combination with one another. The backbone linkages of the 5' and / or 3' wings can be phosphorothioates or a mixture of phosphodiester and phosphorothioate. The linkages in the gap region can be phosphorothioates.

[0110] In some aspects, the oligonucleotide is single stranded. In some aspects, the oligonucleotide comprises or is complexed with a moiety that neutralizes the charge on the oligonucleotide to facilitate uptake and transfer across cell membranes.

[0111] In some aspects, the ASO sequences of SEQ ID NOs: 1-80 in Table 1 have the following 5-8-5 motif: 2MOE*2MOE-2MOE-2MOE-2MOE-N*N*N*N*N*N*N*N*2MOE-2MOE-2MOE*2MOE*2MOE, wherein (i) 2MOE is a nucleobase with a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) N is a nucleobase, (iii) an asterisk (*) indicates a phosphorothioate linkage, and (iv) a dash (-) indicates a phosphodiester linkage. Table 2 below shows this motif on SEQ ID NOs: 1-80 (herein, SEQ ID NOs: 783-862). In some aspects, the ASO sequences of SEQ ID NOs: 783-831 effectively skip exon 3 of PIKFYVE. In some aspects, the ASO sequences of SEQ ID NOs: 832-862 effectively skip exon 5 of PIKFYVE.

[0112] In some aspects, the unmodified versions of the ASO sequences of SEQ ID NOs: 1-782 (e.g., 81-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782) and SEQ ID NOs: 1646-2321 in Table 1 have the following 5-10-5 motif: 2MOE*2MOE-2MOE-2MOE-2MOE-N*N*N*N*N*N*N*N*N*N*N*2MOE-2MOE-2MOE*2MOE*2MOE, wherein (i) 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) N is a nucleobase, (iii) asterisks (*) indicate phosphorothioate linkages, and (iv) dashes (-) indicate phosphodiester linkages. Table 2 below shows this motif on SEQ ID NOs: 1-782 (e.g. 81-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782 (here SEQ ID NOs: 783-1564, e.g. 863-1110, 1112-1155, 1158-1202, 1205-1322, 1324-1401, 1403-1467, 1470-1564)).

[0113] In some aspects, SEQ ID NOs: 81-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781 and SEQ ID NOs: NO:1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1 703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762- 1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869 9. 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2 2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183,The unmodified versions of the ASO sequences of 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321 have the following 5-10-5 motif, 2MOE*2MOE-2MOE -2MOE-2MOE-N*N*N*N*N*N*N*N*N*N*2MOE-2MOE-2MOE*2MOE*2MOE, wherein (i) 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) N is a nucleobase, (iii) the asterisk (*) indicates a phosphorothioate linkage, and (iv) the dash (-) indicates a phosphodiester linkage.

[0114] Table 2: Base sequences in PIKFYVE antisense oligonucleotides (ASOs). (Gapmer design: 5' - 5 2'-methoxyethyl ribonucleotides - 10 DNA nucleotides - 5 2'-methoxyethyl ribonucleotides - 3'; uppercase letters are 2'-methoxyethyl ribonucleosides; lowercase letters are DNA nucleosides; asterisks (*) are phosphorothioate linkages; linkages without asterisks are phosphodiester linkages) (Note that the table below provides a 2'MOE wing; however, alternative wings comprising 2'-OMe or LNA (locked nucleic acid) are contemplated).

[0115] Table 2

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136] In some aspects, the ASO sequences of SEQ ID NOs: 1-49 in Table 1 have the following motif: 2MOE*2MOE-2MOE-2MOE-2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE-2MOE*2MOE*2MOE, wherein (i) 2MOE is a nucleobase with a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) an asterisk (*) indicates a phosphorothioate linkage, and (iii) a dash (-) indicates a phosphodiester linkage. Table 3 below shows this motif on SEQ ID NOs: 1-49 (herein, SEQ ID NOs: 1566-1614). In some aspects, the ASO sequences of Table 3 effectively skip exon 3 of PI KFYVE. In some aspects, the ASOs of the sequences shown in SEQ ID NOs: 1-49 effectively skip exon 3 of PIKFYVE.

[0137] Table 3: Base sequences in PIKFYVE antisense oligonucleotides (ASOs). (Design: 5'-eighteen 2'-methoxyethyl ribonucleotides - 3'; capital letters are 2'-methoxyethyl ribonucleosides; asterisks (*) are phosphorothioate linkages; linkages without asterisks are phosphodiester linkages) (Note that the table below provides a 2'MOE wing; however, alternative wings comprising 2'-OMe or LNA (locked nucleic acid) are contemplated).

[0138] Table 3

[0139]

[0140]

[0141] In some aspects, the ASO sequences of SEQ ID NOs: 50-80 in Table 1 have the following motif: 2MOE*2MOE-2MOE-2MOE-2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE*2MOE-2MOE*2MOE*2MOE, wherein (i) 2MOE is a nucleobase with a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) an asterisk (*) indicates a phosphorothioate linkage, and (iii) a dash (-) indicates a phosphodiester linkage. Table 4 below shows this motif on SEQ ID NOs: 50-80 (herein, SEQ ID NOs: 1615-1645). In some aspects, the ASO sequences of Table 4 effectively skip exon 5 of PIKFYVE. In some aspects, the ASOs of the sequences shown in SEQ ID NOs: 50-80 effectively skip exon 5 of PIKFYVE.

[0142] Table 4: Base sequences in PIKFYVE antisense oligonucleotides (ASOs). (Design: 5'-eighteen 2'-methoxyethyl ribonucleotides - 3'; capital letters are 2'-methoxyethyl ribonucleosides; asterisks (*) are phosphorothioate linkages; linkages without asterisks are phosphodiester linkages) (Note that the table below provides a 2'MOE wing; however, alternative wings comprising 2'-OMe or LNA (locked nucleic acid) are contemplated).

[0143] Table 4

[0144]

[0145]

[0146] The PIKFYVE kinase antisense or inhibitory nucleic acids disclosed herein can inhibit expression and thus inhibit activities associated with PIKFYVE. PIKFYVE kinase antisense or inhibitory nucleic acids can include any combination of the oligonucleotides listed in Tables 1, 2, 3, 4, 5, and 6 and sequences 98%-99% identical thereto.

[0147] The PIKFYVE ASOs described herein, e.g., SEQ ID NOs: 783-1564 (e.g., 783-1110, 1112-1155, 1158-1202, 1205-1322, 1324-1401, 1403-1467, 1470-1564, and 1566-2321), can suppress PIKFYVE mRNA expression while minimizing off-target binding.

[0148] In some aspects, the PIKFYVE ASOs described herein, e.g., SEQ ID NOs: 783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, and 1566-2321 (e.g., SEQ ID NOs: NO:863-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563); can suppress PIKFYVE mRNA expression while minimizing off-target binding.

[0149] The therapeutic method may include any number of ways of administering the disclosed compositions. The modes of administration may include aqueous solutions, lipid solutions, oily solutions or other solutions, solutions in simulated cerebrospinal fluid, emulsions such as oil-in-water emulsions, liposomes, aqueous or oily suspensions, and the like. Typically, the ASOs of the present disclosure will be administered directly to the CNS of the subject. Thus, the formulation or composition will be sterile and more preferably suitable for injection. The following formulations and methods are exemplary only and are in no way limiting.

[0150] Preparations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, antibacterial agents, and solutes that make the preparation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents, solubilizers, thickeners, stabilizers, and preservatives. The preparations may be present in unit dose or multi-dose sealed containers, such as ampoules and vials, and may be used as liquids or stored under freeze-dried (lyophilized) conditions, requiring only the addition of sterile liquid excipients, such as water for injection, immediately before use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets. The preparations may be provided in prefilled syringes.

[0151] Additional therapeutic agents can be administered simultaneously or sequentially with one or more antisense or inhibitory nucleic acids and compositions disclosed herein. Sequential administration includes administration before or after the one or more antisense or inhibitory nucleic acids or compositions disclosed herein. In some aspects, additional one or more therapeutic agents can be administered in the same composition as the one or more antisense or inhibitory nucleic acids disclosed herein. In some aspects, there can be a time interval between the administration of additional therapeutic agents and the one or more antisense or inhibitory nucleic acids disclosed herein. In some aspects, administering additional therapeutic agents together with the one or more antisense or inhibitory nucleic acids disclosed herein can allow for lower doses of other therapeutic agents and / or administration at less frequent intervals. When used in combination with one or more other active ingredients, the one or more antisense or inhibitory nucleic acids of the present disclosure and other active ingredients can be used in a lower dose than when each is used alone. Therefore, in addition to the one or more antisense or inhibitory nucleic acids of the present disclosure, pharmaceutical compositions of the present disclosure also include those containing one or more other active ingredients. The above-mentioned combinations include not only combinations of one or more antisense or inhibitory nucleic acids of the present disclosure with one other active compound, but also combinations with two or more other active compounds. For example, the compounds of the present disclosure can be combined with various drugs to treat neurological diseases. The antisense oligonucleotide can be covalently linked to another oligonucleotide, such as an oligonucleotide with a target other than PIKFYVE. The antisense oligonucleotide can be covalently linked to an antibody.

[0152] The disclosed one or more antisense or inhibitory nucleic acids can be used in combination with, but are not limited to, anticholinergics, anticonvulsants, antidepressants, benzodiazepines, Other types of treatment and management include, but are not limited to, digital communication devices, feeding tubes, mechanical ventilation, nutritional support, deep brain stimulation, occupational therapy, physical therapy, and / or speech therapy.

[0153] The disclosed compositions can be incorporated into pharmaceutical compositions suitable for administration to a subject (such as a patient, which can be human or non-human). The pharmaceutical composition can include a carrier (e.g., a pharmaceutically acceptable carrier). Any suitable carrier can be used in the context of the present disclosure, and such carriers are well known in the art. The choice of carrier will be determined in part by the specific use of the composition (e.g., administration to an animal) and the specific method for administering the composition. Thus, the compositions of the present disclosure have a variety of suitable formulations.

[0154] The pharmaceutical composition can include a therapeutically effective amount or a prophylactic effective amount of the antisense oligonucleotide. The therapeutically effective amount of the composition can be determined by those skilled in the art and can vary according to factors such as individual disease state, age, sex and body weight, and the ability of the composition to elicit a desired response in an individual. A therapeutically effective amount is also an amount in which any toxic or deleterious effects of one or more antisense or inhibitory nucleic acids of the present disclosure are exceeded by a therapeutically beneficial effect. A "prophylactic effective amount" refers to an effective amount that achieves the desired preventive effect within the necessary dosage and time period. Typically, because a prophylactic dose is used in a subject before a disease or in the early stages of a disease, the prophylactic effective amount will be less than the therapeutically effective amount.

[0155] The pharmaceutical composition may include one or more pharmaceutically acceptable carriers. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials that may serve as pharmaceutically acceptable carriers, at the discretion of the formulator, are sugars, including but not limited to lactose, glucose, and sucrose; starches, such as but not limited to corn starch and potato starch; cellulose and its derivatives, such as but not limited to sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as but not limited to cocoa butter and suppository waxes; oils, such as but not limited to peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; esters, such as but not limited to ethyl oleate and ethyl laurate; agar; buffers, such as but not limited to magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol and phosphate buffer solutions, and other non-toxic compatible lubricants, such as but not limited to sodium lauryl sulfate and magnesium stearate, and release agents, coating agents, preservatives, and antioxidants may also be present in the composition.

[0156] The route of administration of the disclosed antisense or inhibitory nucleic acid(s) and the form of the composition will dictate the type of vector used.

[0157] The pharmaceutical compositions of the present disclosure can be administered in a variety of ways, depending on whether local or systemic treatment is needed and the area to be treated. Administration can be parenteral, including intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial administration, such as intrathecal, intraventricular or intraventricular administration. In some aspects, the antisense or inhibitory nucleic acid is administered intravenously, intraperitoneally or as a bolus injection, or is directly administered into the target organ. In some aspects, the antisense or inhibitory nucleic acid is administered as a bolus injection intrathecally or intraventricularly.

[0158] Carriers for systemic administration typically include at least one of solvents, diluents, lubricants, binders, disintegrants, colorants, flavorings, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, combinations thereof, etc. All carriers are optional in the composition.

[0159] Suitable diluents include sugars such as glucose, lactose, dextrose and sucrose; glycols such as propylene glycol; calcium carbonate; sodium carbonate; sugar alcohols such as glycerol; mannitol; and sorbitol.

[0160] Suitable lubricants include silicon dioxide, talc, stearic acid and its magnesium and calcium salts, calcium sulfate; and liquid lubricants such as polyethylene glycol, and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter. The amount of lubricant in the systemic or local composition is generally from about 5% to about 10%.

[0161] Suitable binders include polyvinyl pyrrolidone; magnesium aluminum silicate; starches such as corn starch and potato starch; gelatin; gum tragacanth; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, methylcellulose, microcrystalline cellulose, sodium carboxymethylcellulose, etc. The amount of binder in the systemic composition is generally from about 5% to about 50%.

[0162] Suitable disintegrants include agar, alginic acid and its sodium salt, effervescent mixtures, cross-linked carboxymethylcellulose, crospovidone, sodium carboxymethyl starch, sodium starch glycolate, clays and ion exchange resins. The amount of disintegrant in systemic compositions is generally from about 0.1% to about 10%.

[0163] Suitable coloring agents include colorants such as FD&C dyes.When used, coloring agents are generally present in systemic or topical compositions in amounts of from about 0.005 to about 0.1%.

[0164] Suitable flavoring agents include menthol, mint, and fruit flavors.When used, flavoring agents are generally present in systemic or topical compositions at levels of from about 0.1% to about 1.0%.

[0165] Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of antioxidant in the systemic or topical compositions is generally from about 0.1% to about 5%.

[0166] Suitable preservatives include benzalkonium chloride, methylparaben, and sodium benzoate.The amount of preservative in systemic or topical compositions is generally from about 0.01 to about 5%.

[0167] Suitable glidants include silicon dioxide.The amount of glidant in the systemic or topical composition is generally from about 1% to about 5%.

[0168] Suitable solvents include water, isotonic saline, ethyl oleate, glycerol, hydroxylated castor oil, alcohols such as ethanol, and phosphate buffered saline.The amount of solvent in systemic or topical compositions is generally from about 0 to about 100%.

[0169] Suitable suspending agents include AVICEL RC-591 (from FMC Corporation of Philadelphia, PA) and sodium alginate.The amount of suspending agent in systemic or topical compositions is generally from about 1% to about 8%.

[0170] Suitable surfactants include lecithin, polysorbate 80, and sodium lauryl sulfate, as well as TWEENS from Atlas Powder Company of Wilmington, Delaware. Suitable surfactants include those disclosed in the CTFA Cosmetic Ingredient Handbook, 1992, pages 587-592; Remington's Pharmaceutical Sciences, 15th edition, 1975, pages 335-337; and McCutcheon's, Volume 1, Emulsifiers & Detergents, 1994, North American edition, pages 236-239. The amount of surfactant in the systemic or topical composition is generally from about 0.1% to about 5%.

[0171] Compositions and preparations for parenteral, intrathecal, intraventricular or intraventricular administration may include sterile aqueous solutions that may also contain buffers, diluents and other suitable additives, such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients. For example, an intrathecal cerebrospinal fluid (CSF) catheter may be used to deliver the antisense formulations of the present disclosure. The catheter may be inserted into the L3 or L4 vertebra. The distal end of the catheter extends to approximately the L1 vertebra in the intrathecal space. The antisense oligonucleotides are dissolved in saline, sterilized by filtration, and administered at a speed of 0.33 ml / min in a volume of 1.0 ml, followed by rinsing with 0.5 ml of sterile water. The total infusion time is 4.5 minutes.

[0172] Compositions for parenteral administration generally contain from 0.1% to 10% of active substance and from 90% to 99.9% of carrier (including diluent and solvent).

[0173] The amount of the carrier used in conjunction with the disclosed compound is sufficient to provide the actual amount of the composition for each unit dose of the drug to be administered. Techniques and compositions for preparing dosage forms that can be used in the disclosed methods are described in the following references: Modern Pharmaceutics, Chapters 9 and 10, edited by Banker & Rhodes (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, Second Edition (1976).

[0174] In vivo testing of candidate antisense or inhibitory nucleic acids can be performed by methods known to those of ordinary skill in the art. For example, one or more candidate antisense or inhibitory nucleic acids can be administered to a mammal, such as a mouse or rabbit. A dose of the candidate antisense or inhibitory nucleic acid can be administered to the mammal by any route deemed appropriate. Conventional methods and standards can then be used to monitor the animal's motor neuron activity and / or the expression or activity of the PIKFYVE gene or protein for signs of reduction or improvement. If desired, the results obtained in the presence of the candidate antisense or inhibitory nucleic acid can be compared with the results in control animals not treated with the candidate antisense or inhibitory nucleic acid. Dosage studies can be performed in conjunction with the methods described herein for identifying one or more antisense or inhibitory nucleic acids capable of treating neurological diseases and / or in any subsequent testing of candidate antisense or inhibitory nucleic acids in vivo. A person skilled in the art of medicine can determine the appropriate dosage of one or more antisense or inhibitory nucleic acids. The dosage can be determined by monitoring the subject for signs of disease suppression or improvement. The dosage can be increased or decreased to obtain the desired frequency of treatment. In some embodiments, the toxicity of one or more antisense or inhibitory nucleic acids and their efficacy can be determined by the standard drug procedures in cell culture or experimental animals, for example, determining the lethal dose (LD50) to 50% of the colony and the therapeutically effective dose (ED50) to 50% of the colony. The dosage ratio of LD50 / ED50 is a therapeutic index, which represents the ratio between toxicity and the therapeutic effect. Delivery systems can be designed to help prevent toxic side effects by delivering one or more antisense or inhibitory nucleic acids to specific targets (for example, specifically delivered to motor neurons or central nervous system neurons). For example, the optimal dose of one or more antisense or inhibitory nucleic acids can be determined based on the result of clinical electrophysiology or electromyography to analyze the excitability of peripheral nerves.

[0175] The dosage for humans can be determined by evaluating the data obtained from animal studies and cell culture assays. The preferred dosage will have very little toxicity or no toxicity and include ED50. The dosage may vary depending on the dosage form and route of administration. For any antisense or inhibitory nucleic acid used in the methods described herein, the dosage can be initially estimated in cell culture. The dosage can be formulated in an animal model that includes the concentration of the test compound that achieves half the maximum inhibition (LD50) of the symptoms measured in cell culture. Such information obtained from cell culture and animal models can be used to more accurately determine the useful dosage for humans.

[0176] The present disclosure has several aspects, illustrated by the following non-limiting examples.

[0177] Example

[0178] Small-molecule inhibitors of PIKFYVE kinase and antisense oligonucleotides (ASOs) that suppress PIKFYVE expression can prevent neuronal degeneration in humans and mice carrying mutations in the C9ORF72 gene that cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD).

[0179] ASOs are an attractive therapeutic option for neurodegenerative diseases because they are easily delivered to the central nervous system and have relatively low exposure to the periphery. These properties maximize target engagement in the central nervous system and minimize undesired target engagement or off-target effects in the periphery.

[0180] The present disclosure provides novel antisense oligonucleotide (ASO) sequences targeting the PIKFYVE gene that can suppress PIKFYVE expression in human cells. PIKFYVE ASOs can also rescue the survival of motor neurons from sporadic ALS patients. Furthermore, PIKFYVE ASOs can reduce the levels of neurotoxic dipeptide repeat protein aggregates derived from C9ORF72 repeat expansion in mice.

[0181] Example 1

[0182] To identify ASO sequences that suppress PIKFYVE expression in human cells, ASOs were designed (see Tables 2, 3, 4, and 6) and synthesized as MOE gapmers containing sugar and linkage modifications that increased nuclease resistance and melting temperature while maintaining the ability to serve as RNase H substrates. Each ASO was tested for its ability to suppress PIKFYVE RNA levels by transfecting them into human embryonic kidney 293T cells using Lipofectamine 2000 at a concentration of 100 nM and measuring PIKFYVE expression 7 days after transfection. A control (NCASO) was used. The relative PIKFYVE expression shown is the average of three technical replicates and values ​​were calculated by normalization to a GAPDH control.

[0183] Example 2

[0184] In this study, newborn transgenic hPIKFYVE BAC mice received 25 μg of negative control ASO or test compound via intracerebroventricular (ICV) injection at P1 (postnatal day 1), and tissue samples were collected 14 days after treatment.

[0185] Example 3

[0186] ASOs of SEQ ID NOs. 863-1261 were subjected to an in silico study to determine the ASO acute neurotoxicity score for each ASO. The acute neurotoxicity score was generated using the method described by Hagedorn et al., 2022 (DOI: 10.1089 / nat / 2021 / 0071). ASOs of SEQ ID NOs. 863-1261 were determined to have low acute neurotoxicity scores. The results of this study are shown in Table 5 below.

[0187] Table 5

[0188]

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198] In some aspects, the ASOs of the sequences shown in SEQ ID NOs: 81-306, 310-328, 332-373, 376-420, 423, and 425-479 comprise a low acute neurotoxicity score.

[0199] Example 4

[0200] For various ASOs as described herein, computer simulations have predicted the containment of off-target genes (such as CNTN5). Various ASOs have been studied in silico to determine the number of off-target binding sites for various ASOs. The number of off-target binding sites is generated using the GGGenome search tool by 2 mispairings / gaps. The results of this study are shown in Table 6 below.

[0201] The ASOs in Table 6 have the following gapmer design: 5'-five 2'-methoxyethyl ribonucleotides-ten DNA nucleotides-five 2'-methoxyethyl ribonucleotides-3'; wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, eighteenth, and nineteenth linkages are phosphorothioate linkages, and the second, third, fourth, fifth, sixteenth, and seventeenth linkages are phosphodiester linkages (same gapmer design as Table 2). Please note that the table below provides a 2'MOE wing; however, alternative wings comprising 2'-OMe or LNA (locked nucleic acid) are also contemplated. Also note that the table below provides gapmer ASOs, but contemplates ASOs having a complete phosphodiester backbone (unmodified version) of 20 DNA nucleotides (without a 2'MOE wing) and the SEQ IDs of Table 6.

[0202] Table 6

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226] In some aspects, the ASOs of the sequences shown below contain a small number of off-target binding sites: SEQ ID NO: 480-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0227] The present disclosure provides ASOs that suppress PIKFYVE expression in human cells. The accompanying data are consistent with the ability of these ASOs to prevent, inhibit, or suppress neurodegeneration in ALS and FTD patients.

[0228] The above description and accompanying drawings should be regarded as merely illustrative of the principles of the present disclosure. The present disclosure is not intended to be limited in these respects and can be implemented in a variety of ways that are apparent to those skilled in the art. Those skilled in the art will readily appreciate the various applications of the present disclosure. Therefore, it is not intended that the present disclosure be limited to the specific examples disclosed or the precise construction and operation shown and described. On the contrary, all suitable modifications and equivalents falling within the scope of the present disclosure may be adopted. All references cited herein are incorporated herein by reference.

Claims

1. A single-stranded antisense oligonucleotide that inhibits PIKFYVE expression, wherein the antisense oligonucleotide comprises a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any one of the nucleobase sequences shown in SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

2. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide comprises a nucleobase sequence as set forth in any one of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782 -1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 209 7. Unmodified forms of 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

3. The antisense oligonucleotide of claim 1 or 2, wherein the antisense oligonucleotide has 18 to 20 linked nucleosides.

4. The antisense oligonucleotide of any preceding claim, wherein at least one internucleoside linkage is a modified internucleoside linkage.

5. The antisense oligonucleotide of claim 4, wherein at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.

6. The antisense oligonucleotide of claim 4, wherein each modified internucleoside linkage is a phosphorothioate internucleoside linkage.

7. The antisense oligonucleotide of any one of the preceding claims, wherein at least one internucleoside linkage is a phosphodiester internucleoside linkage.

8. The antisense oligonucleotide of claim 7, wherein at least one internucleoside linkage is a phosphorothioate linkage and at least one internucleoside linkage is a phosphodiester linkage.

9. The antisense oligonucleotide of any one of the preceding claims, wherein at least one nucleoside comprises a modified nucleobase.

10. The antisense oligonucleotide of claim 9, wherein the modified nucleobase is 5-methylcytosine.

11. The antisense oligonucleotide of any one of the preceding claims, wherein at least one nucleoside of the antisense oligonucleotide comprises a modified sugar moiety.

12. The antisense oligonucleotide of claim 11, wherein the modified sugar moiety comprises a 2'-O-methoxyethyl group.

13. The antisense oligonucleotide of any one of the preceding claims, wherein the antisense oligonucleotide is a gapmer.

14. The antisense oligonucleotide of claim 13, wherein the antisense oligonucleotide comprises: a gap segment consisting of 8 to 12 linked deoxynucleosides; a 5' wing segment consisting of 3 to 5 linked nucleosides; and a 3' wing segment consisting of 3 to 5 linked nucleosides, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein the nucleosides of each wing segment comprise a modified sugar moiety.

15. The antisense oligonucleotide of claim 14, wherein each nucleoside of each wing segment comprises a modified sugar moiety.

16. The antisense oligonucleotide of claim 14, wherein the nucleosides comprising each wing segment comprise at least two different modified sugar moieties.

17. The antisense oligonucleotide of claim 14, wherein the nucleosides comprising each wing segment comprise the same modified sugar moiety.

18. The antisense oligonucleotide of claim 15, wherein the modified sugar moiety comprises a 2'-O-methoxyethyl group.

19. The antisense oligonucleotide of any one of the preceding claims, wherein the antisense oligonucleotide comprises 15 to 50 nucleosides.

20. The antisense oligonucleotide of any one of the preceding claims, wherein the antisense oligonucleotide comprises a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any one of the nucleobase sequences shown in: SEQ ID NO: NO:783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1 481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677 、1679、1684-1687、1689、1692、1694、1696-1698、1700、1702-1703、1708-1711、1713、1716-1717、1719、1722、1725、1727-1728、1731、1740、1743-1745、1747-1751、1754、1756、1760、1762-1764、1770-1775、1777-1779、1782-1784、1787-1788、1791-1793、1795、1797-1802、1 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 193 0, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109,2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316 and 2319-2321.

21. The antisense oligonucleotide according to any one of the preceding claims, wherein the antisense oligonucleotide comprises a sequence as shown in any one of the following: SEQ ID NO:783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563 ,1650,1652-1654,1660,1666-1667,1671-1672,1674,1677,1679,168 4-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811 , 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943 , 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124,2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316 and 2319-2321.

22. A pharmaceutical composition comprising the antisense oligonucleotide according to any one of the preceding claims and a pharmaceutically acceptable carrier, diluent and / or excipient.

23. The pharmaceutical composition of claim 22, wherein the pharmaceutical composition is formulated for parenteral delivery.

24. The pharmaceutical composition of claim 22, wherein the pharmaceutical composition is formulated for intracerebroventricular injection.

25. A method of treating a subject in need thereof having a neurological disease or a neurodegenerative disease, comprising administering a therapeutically effective amount of the antisense oligonucleotide of any one of claims 1-21 or the pharmaceutical composition of any one of claims 22-24.

26. The method of claim 25, wherein the neurological disorder is associated with neuronal hyperexcitability.

27. The method of claim 25, wherein the neurological disease is associated with abnormal endosomal trafficking.

28. The method of claim 25, wherein the neurological disease is associated with abnormal lysosomal trafficking.

29. The method of claim 25, wherein the neurological disease is selected from the group consisting of familial and sporadic amyotrophic lateral sclerosis (ALS), familial and sporadic frontotemporal dementia (FTD), progressive supranuclear palsy, Alzheimer's disease, chronic traumatic encephalopathy, Parkinson's disease, Charcot-Marie-Tooth disease 2A and 4B, Huntington's disease, dementia, transmissible spongiform encephalopathy, spinobulbar muscular atrophy, dentatorubral pallidum atrophy with Lewy bodies, spinocerebellar ataxia, and Creutzfeldt-Jakob disease.

30. The method of claim 25, wherein the neurological disease is familial amyotrophic lateral sclerosis.

31. The method of claim 25, wherein the neurological disease is sporadic amyotrophic lateral sclerosis.

32. The method of claim 25, wherein the neurological disease is familial frontotemporal dementia.

33. The method of claim 25, wherein the neurological disease is sporadic frontotemporal dementia.

34. The method of claim 25, wherein the neurological disease is frontotemporal dementia with TDP-43 pathology.

35. The method of claim 25, wherein the neurological disease is frontotemporal dementia with tau pathology.

36. The method of any one of claims 25-35, wherein the subject is haploinsufficient for the C9ORF72 gene.

37. The method of any one of claims 25-35, wherein the subject has a GGGGCC repeat expansion in C9ORF72.

38. The method of any one of claims 25-35, wherein the subject has (GGGGCC) in C9ORF72 n (SEQ ID NO: 1565) Hexanucleotide amplification, wherein n is at least 30.

39. The method of any one of claims 25-35, wherein the subject has C9orf72-related frontotemporal dementia.

40. The method of any one of claims 25-35, wherein the subject suffers from microtubule-associated protein tau (MAPT)-related frontotemporal dementia.

41. The method of claim 40, wherein the patient has a V337MMAPT mutation.

42. A method of inhibiting or suppressing PIKFYVE expression in a patient suffering from a neurological disease or a neurodegenerative disease, comprising administering an effective amount of the antisense oligonucleotide of any one of claims 1-21 or the pharmaceutical composition of any one of claims 22-24.

43. An oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the nucleobase sequences shown in SEQ ID NO: NO:1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 7 49-750, 752, 759, 762, 764, 770, 772, 781, 783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 138 2-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1710 11, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953,1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 21 18, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316 and 2319-2321, and SEQ. IDNo:1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689 ,1692,1694,1696-1698,1700,1702-1703,1708-1711,1713,1716-1717,1719,1722,172 5. 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1810 11. 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997,1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2125 6-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316 and 2319-2321. The antisense oligonucleotide of claim 1 , comprising the sequence shown in SEQ ID NO: 1-80.

45. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide suppresses the expression of PIKFYVE by at least 80%.

46. ​​A single-stranded antisense oligonucleotide for suppressing PIKFYVE expression, wherein the antisense oligonucleotide has a nucleobase sequence comprising any one of the following sequences: SEQ ID NO: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NO:1650、1652-1654、1660、1666-1667、1671-1672、1674、1677、1679、168 4-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713 ,1716-1717,1719,1722,1725,1727-1728,1731,1740,1743-1745,1747- 1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-184 3. 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1 917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-201 5. 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2 2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, wherein the first 3-5 nucleosides at the 5' end (the "5' wing segment") comprise a modified sugar,The last 3-5 nucleosides at the 3' end (the "3' wing segment") contain a modified sugar, and the remaining nucleosides contain a gap segment.

47. The antisense oligonucleotide of claim 46, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

48. The antisense oligonucleotide of claim 47, wherein at least one of the modified sugars comprises 2'-OMe.

49. The antisense oligonucleotide of claim 47, wherein at least one of the modified sugars comprises 2'-MOE.

50. The antisense oligonucleotide of claim 47, wherein at least one of the modified sugars comprises LNA.

51. The antisense oligonucleotide of claim 46, wherein the backbone linkages of the 5' wing segment, the 3' wing segment, and the gap segment comprise a mixture of phosphorothioate and phosphodiester linkages.

52. A single-stranded antisense oligonucleotide for suppressing PIKFYVE expression, wherein the antisense oligonucleotide comprises a sequence as shown in any one of the following: SEQ ID NO:863-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563 ,1650,1652-1654,1660,1666-1667,1671-1672,1674,1677,1679,168 4-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811 , 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943 , 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124,2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, wherein the backbone linkages of the antisense oligonucleotide comprise a mixture of phosphorothioate linkages and phosphodiester linkages.

53. The antisense oligonucleotide of claim 52, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, eighteenth, and nineteenth linkages are phosphorothioate linkages, and the second, third, fourth, fifth, sixteenth, and seventeenth linkages are phosphodiester linkages.

54. The antisense oligonucleotide of claim 46, wherein at least one nucleoside comprises a modified nucleobase.

55. The antisense oligonucleotide of claim 46, wherein the modified nucleobase is 5-methylcytosine.

56. The antisense oligonucleotide of claim 46, wherein the antisense oligonucleotide comprises a moiety that neutralizes the charge on the antisense oligonucleotide.

57. The antisense oligonucleotide of claim 46, wherein the antisense oligonucleotide suppresses the expression of PIKFYVE by at least 80%.

58. A pharmaceutical composition comprising the antisense oligonucleotide according to claim 46 and a pharmaceutically acceptable carrier, diluent and / or excipient.

59. A pharmaceutical composition comprising the antisense oligonucleotide according to claim 52 and a pharmaceutically acceptable carrier, diluent and / or excipient.

60. A pharmaceutical composition comprising the antisense oligonucleotide according to claim 53 and a pharmaceutically acceptable carrier, diluent and / or excipient.

61. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition is formulated for parenteral delivery or intracerebroventricular injection.

62. The pharmaceutical composition of claim 61, wherein the pharmaceutical composition is formulated for parenteral delivery or intracerebroventricular injection.

63. The antisense oligonucleotide of claim 14, wherein the antisense oligonucleotide comprises: A gap segment consisting of 8 linked deoxynucleosides; a 5' wing segment consisting of 5 linked nucleosides; and A 3' wing segment consisting of 5 linked nucleosides, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein the nucleosides of each wing segment comprise a modified sugar moiety.

64. The antisense oligonucleotide of claim 14, wherein the antisense oligonucleotide comprises: a gap segment consisting of 10 linked deoxynucleosides; a 5' wing segment consisting of 5 linked nucleosides; and A 3' wing segment consisting of 5 linked nucleosides, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein the nucleosides of each wing segment comprise a modified sugar moiety.

65. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide comprises the nucleobase sequence shown in any one of SEQ ID NOs: 1-49, wherein the antisense oligonucleotide skips exon 3 of the PIKFYVE protein.

66. The antisense oligonucleotide of claim 65, wherein the oligonucleotide comprises one or more modified sugars.

67. The antisense oligonucleotide of claim 66, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

68. The antisense oligonucleotide of claim 67, wherein at least one of the modified sugars comprises 2'-OMe.

69. The antisense oligonucleotide of claim 67, wherein each nucleoside of the antisense oligonucleotide comprises a 2'-MOE modified sugar.

70. The antisense oligonucleotide of claim 67, wherein at least one of the modified sugars comprises 2'-MOE.

71. The antisense oligonucleotide of claim 67, wherein at least one of the modified sugars comprises LNA.

72. The antisense oligonucleotide of claim 65, wherein the backbone linkage oligonucleotide comprises a mixture of phosphorothioate linkages and phosphodiester linkages.

73. The antisense oligonucleotide of claim 72, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, sixteenth, and seventeenth linkages are phosphorothioate linkages, and the second, third, fourth, fifth, fourteenth, and fifteenth linkages are phosphodiester linkages.

74. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide comprises the nucleobase sequence shown in any one of SEQ ID NOs: 50-80, wherein the antisense oligonucleotide skips exon 5 of the PIKFYVE protein.

75. The antisense oligonucleotide of claim 74, wherein the oligonucleotide comprises one or more modified sugars.

76. The antisense oligonucleotide of claim 75, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

77. The antisense oligonucleotide of claim 76, wherein at least one of the modified sugars comprises 2'-OMe.

78. The antisense oligonucleotide of claim 76, wherein each nucleoside of the antisense oligonucleotide comprises a 2'-MOE modified sugar.

79. The antisense oligonucleotide of claim 76, wherein at least one of the modified sugars comprises 2'-MOE.

80. The antisense oligonucleotide of claim 76, wherein at least one of the modified sugars comprises LNA.

81. The antisense oligonucleotide of claim 74, wherein the backbone linkage oligonucleotide comprises a mixture of phosphorothioate linkages and phosphodiester linkages.

82. The antisense oligonucleotide of claim 81, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, sixteenth, and seventeenth linkages are phosphorothioate linkages, and the second, third, fourth, fifth, fourteenth, and fifteenth linkages are phosphodiester linkages.

83. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide comprises the nucleobase sequence set forth in any one of SEQ ID NOs: 81-306, 310-328, 332-373, 376-420, 423, and 425-479, wherein the antisense oligonucleotide comprises a low acute neurotoxicity score.

84. The antisense oligonucleotide of claim 83, wherein the oligonucleotide comprises one or more modified sugars.

85. The antisense oligonucleotide of claim 84, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

86. The antisense oligonucleotide of claim 85, wherein at least one of the modified sugars comprises 2'-OMe.

87. The antisense oligonucleotide of claim 85, wherein at least one of the modified sugars comprises 2'-MOE.

88. The antisense oligonucleotide of claim 85, wherein at least one of the modified sugars comprises LNA.

89. The antisense oligonucleotide of claim 83, wherein the backbone linkage oligonucleotide comprises a mixture of phosphorothioate linkages and phosphodiester linkages.

90. The antisense oligonucleotide of claim 89, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, eighteenth, and nineteenth linkages are phosphorothioate linkages, and the second, third, fourth, fifth, sixteenth, and seventeenth linkages are phosphodiester linkages.

91. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide comprises a nucleobase sequence as set forth in any one of SEQ ID NOs: 480-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781,and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 191 5. 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2 109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and unmodified versions of 2319-2321, wherein the antisense oligonucleotide comprises a low number of off-target binding sites.

92. The antisense oligonucleotide of claim 91, wherein the oligonucleotide comprises one or more modified sugars.

93. The antisense oligonucleotide of claim 92, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

94. The antisense oligonucleotide of claim 93, wherein at least one of the modified sugars comprises 2'-OMe.

95. The antisense oligonucleotide of claim 93, wherein at least one of the modified sugars comprises 2'-MOE.

96. The antisense oligonucleotide of claim 93, wherein at least one of the modified sugars comprises LNA.

97. The antisense oligonucleotide of claim 91, wherein the backbone linkage oligonucleotide comprises a mixture of phosphorothioate linkages and phosphodiester linkages.

98. The antisense oligonucleotide of claim 97, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, eighteenth, and nineteenth linkages are phosphorothioate linkages, and the second, third, fourth, fifth, sixteenth, and seventeenth linkages are phosphodiester linkages.

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