Compounds and their use in treating medical conditions

Abstract

Compounds, pharmaceutical compositions, their use for inhibiting LINE1 reverse transcriptase activity, and their use in the treatment of medical disorders, such as cancer, are disclosed herein.

Description

COMPOUNDS AND THEIR USE IN TREATING MEDICAL CONDITIONS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to United States Provisional PatentApplication serial number 63 / 709,162, filed October 18, 2024, and United States Provisional Patent Application serial number 63 / 655,675, filed June 4, 2024, the contents of each of which are hereby incorporated by reference in their entirety. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] This application contains a Sequence Listing which has been submitted electronicallyvia Patent Center in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on June 3, 2025, is named 218484_seqlist.xml and is 2,671 bytes in size. FIELD OF THE DISCLOSURE

[0003] The present disclosure provides compounds, compositions, and methods for treatingmedical disorders, such as cancer, autoimmune disorders, and / or neurological disorders, and modulating LINE1 reverse transcriptase using a compound according to Formula I or a pharmaceutically acceptable salt thereof, or a related compound provided herein. BACKGROUND

[0004] Transposable elements (or transposons) are genomic DNA sequences that have theability to move within the genome which leads to altering its organization, increasing its size and creating duplications and redundancy. (Ukadike and Mustelin, J. Clin. Med., 10:856 (2021)). These genomic sequences are believed to have been introduced into the human genome by either an infection by exogenous retroviruses that infected human ancestors millions of years ago or ancient descendants of retroviruses which retained the ability to embed and replicate in human germline genome. (Ukadike and Mustelin, 2021).

[0005] Long Interspersed Nuclear Element 1 (LINE-1) are class I transposable elements inthe DNA of some organisms and comprise about 17% of the human genome. LINE-1 harbors two open reading frames, ORF1 and ORF2, which in turn respectively encode ORF1p, which has nucleic acid chaperone activity, and ORF2p, with reverse transcriptase (RT) and endonuclease 1 33049332.1 400807-032WO (218484)activities. (Reviewed in Babushok and Kazazian, Hum. Mut.28:527-539, (2007)). LINE-1 retrotransposition activity is mediated by ORF2p. The majority of LINE-1 elements in the human genome contain inactivating mutations but a small percentage of LINE-1 elements are intact and have retained the ability to retrotranspose. This ability varies both among individuals and among cell types within an individual. Active LINE-1 elements are thought to disrupt the genome through insertions, deletions, rearrangements and recombinations. (Garcia-Perez et al, Development, 143:4101-4114 (2016)). LINE-1 activity is normally tightly regulated in the germline by DNA methylation, histone modifications, and piRNA.

[0006] Retrotransposons are transposable elements which are associated with thepathogenesis of many diseases such as cancer, autoimmune disease, neurological disorders and aging, among others. (Zhang, et al, Frontiers in Cell and Dev. Bio., 8:657 (Aug.2020); Kuriyama et al, Nature: Scientific Reports, 11:23146 (2021)). LINE-1 RNA and protein overexpression can promote apoptosis, DNA damage and repair, and cellular plasticity, which can promote tumor progression. Furthermore, genomic hypomethylation can induce expression of repetitive sequences which can drive a pro-inflammatory response characterized by overproduction of type 1 interferon. (Zhang, 2020).

[0007] Pathogenic interferon production is a characteristic feature of type Iinterferonopathies. These include rare genetic diseases with occurrence rates from 1:10,000 to 1:1,000,000. Pathological induction of type I interferon causes immune system hyperactivation that leads to systemic inflammation which can affect the nervous system, lung and blood vessels, among other organ systems. (Nesterova et al. "Congenital and Acquired Interferonopathies: Differentiated Approaches to Interferon Therapy". Innate Immunity in Health and Disease, Ed. Saxena and Prakash, IntechOpen, 2020). Aicardi-Goutières Syndrome (AGS) is a monogenic inflammatory encephalomyopathy driven by mutations in genes that are critical in maintaining homeostatic cytosolic nucleic acid oligomers. As a result, increased level of cytoplasmic nucleic acid accumulation leads to heightened interferon response. The double stranded DNA products of LINE-1 reverse transcription are potential triggers of DNA sensing receptors such as cGAS, which is a DNA sensor that activates the STING pathway leading to type I interferon production. (Zhao, J. Autoimmunity, 90:105-115 (2018)). LINE-1 reverse transcriptase products have been implicated as a primary source of pro-inflammatory nucleic acids in AGS patients. Administering a combination of three nucleoside reverse transcriptase inhibitors to AGS patients 2 33049332.1 400807-032WO (218484)for 12 months effectively reduced their systemic interferon response. The pathogenic interferon response responsible for AGS has also been implicated in the pathogenesis of SLE ,with several case studies identifying monogenic forms of SLE driven by hypomorphic alleles of nucleic acid metabolizing enzymes such as TREX1.

[0008] Hypomethylated and highly expressed LINE-1 has been found in many patients withautoimmune diseases such as systemic lupus erythematosus (SLE), cutaneous lupus, Sjögren’s syndrome (SS) and psoriasis. (Zhang et al). LINE-1 has also been found to be significantly upregulated in patients with dermatomyositis (DM), with significantly elevated levels of interferon α and interferon β. (Kuriyama et al, J. Am. Acad, Dermatol., 84(4):1103-1105 (2020)).

[0009] LINE-1 has also been implicated in neurological disorders such as ataxiatelangiectasia (AT), Rett syndrome, Parkinson's disease, Friederichs's ataxia, perisupranuclear palsy, amyotrophic lateral sclerosis, frontotemporal dementia and schizophrenia. Increased retrotransposition as well as elevated levels of type 1 interferon have been identified in each of these diseases. LINE-1 is also implicated in the aging process and frontotemporal lobe degeneration. (Zhang, 2020).

[0010] Cancer continues to be a significant health problem despite the substantial researchefforts and scientific advances reported in the literature for treating this disease. Solid tumors, including prostate cancer, breast cancer, and lung cancer remain highly prevalent among the world population. Leukemias and lymphomas also account for a significant proportion of new cancer diagnoses. Current treatment options for these cancers are not effective for all patients and / or can have substantial adverse side effects. New therapies are needed to address this unmet need in cancer therapy.

[0011] High LINE-1 activity has been found in many tumor tissues. LINE-1 RT uses aprocedure termed target-site-primed reverse transcription (TPRT) which involves nicking of the genomic DNA followed by reverse transcription and insertion of LINE-1 into the genome. LINE-1 mediated gene rearrangement can trigger oncogene amplification. Additionally, LINE-1 can mediate the deletion of tumor suppressor genes (Zhang, 2020). Inhibition of LINE-1 RT in cancer cells, either via RNA interference-dependent silencing of active LINE-1 elements or using RT inhibitory compounds can reduce cancer cell proliferation, promote cancer cell differentiation and can retard tumor progression in certain animal models. (Sciamann et al, 3 33049332.1 400807-032WO (218484)Frontiers in Chemistry, 4:6 (Feb.2016)). LINE1 has also been shown to promote tumor metastasis. Furthermore, chronic production of type 1 interferon in the tumor microenvironment has been linked to resistance to immunosurveillance with therapeutic blockade of interferon signaling increased anti-cancer immune responses.

[0012] Compounds for treating medical disorders, such as cancer, autoimmune disease,neurological disorders, aging, and diseases associated with aging are needed. SUMMARY

[0013] Provided herein are compounds and compositions which are useful for, among otheruses, the treatment of cancer, an autoimmune disorder, and / or a neurological disorder. The compounds inhibit LINE1 reverse transcriptase activity. The compounds may be formulated in a pharmaceutical composition. Therapeutic methods and methods of inhibiting LINE1 reverse transcriptase activity are provided.

[0014] One aspect of the disclosure provides compounds having a superior combination ofproperties including potent inhibitory activity towards LINE1 reverse transcriptase, selectivity for inhibiting LINE1 reverse transcriptase, and superior physical properties that render the compounds well-suited for use as an orally administered medicine. Selectivity for inhibiting LINE1 reverse transcriptase can be characterized according to ability of the compounds to inhibit LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNApolymerases (e.g., ^, ^ and ^). In part because inhibition of DNA polymerases, such as DNApolymerase gamma, can result in toxicity in the form of mitotoxicity, compounds demonstrating potent inhibitory activity towards LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNA polymerases are an important discovery and significant scientific advance. The potent inhibitory activity towards LINE1 reverse transcriptase in combination with low inhibitory activity towards DNA polymerases contributes to a high therapeutic index for subject compounds, thereby providing a superior performance profile for the compound in medical therapy.

[0015] Another aspect of the disclosure provides compounds having a superior combinationof properties including potent inhibitory activity towards LINE1 reverse transcriptase, selectivity for inhibiting LINE1 reverse transcriptase, potent inhibition of pathogenic interferon response in inflammatory tissues, and superior physical properties that render the compounds well-suited for 4 33049332.1 400807-032WO (218484)use as an orally administered medicine. As described above, selectivity for inhibiting LINE1 reverse transcriptase can be characterized according to ability of the compounds to inhibit LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNApolymerases (e.g., ^, ^ and ^). In part because inhibition of DNA polymerases, such as DNApolymerase gamma, can result in toxicity in the form of mitotoxicity, compounds demonstrating potent inhibitory activity towards LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNA polymerases are an important discovery and significant scientific advance. Compounds having potent inhibition of pathogenic interferon response in inflammatory tissues are useful for treating cancer, autoimmune disease (e.g., SLE and CLE), neurological disorders, aging, and diseases associated with aging. The potent inhibitory activity towards LINE1 reverse transcriptase in combination with low inhibitory activity towards DNApolymerases (e.g., ^, ^ and ^) contributes to a high therapeutic index for subject compounds,thereby providing a superior performance profile for the compound in medical therapy.

[0016] Accordingly, one aspect of the disclosure provides compounds represented byFormula I: or a pharmaceutically acceptable, ables are as defined in the detailed description.

[0017] Another aspect of the disclosure provides compounds represented by Formula II:33049332.1400807-032WO (218484)or a pharmaceutically acceptable salt thereof, where the variables are as defined in the detailed description.

[0018] Also provided herein are pharmaceutical compositions comprising a compounddescribed herein, such as a compound of Formula I. The disclosed compounds and compositions are designed for treating medical disorders by inhibiting LINE1 reverse transcriptase using the disclosed compounds. In particular, one aspect of the disclosure provides the compounds according to Formula I, or a pharmaceutically acceptable salt thereof, in a method of treating a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. Another aspect of the disclosure provides the compounds according to Formula II, or a pharmaceutically acceptable salt thereof, in a method of treating a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. Additional features of the methods are described in the detailed description.

[0019] Another aspect of the disclosure provides the compounds according to Formula I, or apharmaceutically acceptable salt thereof, in a method of inhibiting LINE1 reverse transcriptase activity in a subject, such as a subject suffering from a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. The method comprises contacting a LINE1 reverse transcriptase with an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof, in order to inhibit the activity of said LINE1 reverse transcriptase. Another aspect of the disclosure provides the compounds according to Formula II, or a pharmaceutically acceptable salt thereof, in a method of inhibiting LINE1 reverse transcriptase activity in a subject, such as a subject suffering from a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. The method comprises contacting a LINE1 reverse transcriptase with an effective amount of a compound of Formula II, or a pharmaceutically acceptable salt thereof, in order to inhibit the activity of said LINE1 reverse transcriptase. Additional features of the methods are described in the detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 depicts the effects of Compounds I-1, I-2, and I-39 on expression of theinterferon-stimulated genes TNFα and CXCL10 in the hearts of TREX1 KO mice, as further described in Example 25. 6 33049332.1 400807-032WO (218484)

[0021] Figure 2 depicts the effects of Compounds I-1, I-2, and I-39 on expression of theinterferon-stimulated genes TNFα and CXCL10 in the kidneys of TREX1 KO mice, as further described in Example 25.

[0022] Figure 3 depicts the effects of Compounds I-1, I-2, and I-39 on the cardiac histologyscore of TREX1 KO mice, as further described in Example 25. DETAILED DESCRIPTION

[0023] Compounds of Formula I and related compounds, pharmaceutical compositions, theiruse for inhibiting LINE1 reverse transcriptase, and their use in the treatment of medical disorders are disclosed herein. The practice of the present disclosure employs, unless otherwise indicated, conventional techniques of organic chemistry, pharmacology, molecular biology (including recombinant techniques), cell biology, biochemistry, and immunology. Such techniques are explained in the literature, such as in “Comprehensive Organic Synthesis” (B.M. Trost & I. Fleming, eds., 1991-1992); “Comprehensive Organic Synthesis” (P. Knochel & G.A. Molander, eds., 2014); “Handbook of Experimental Immunology” (D.M. Weir & C.C. Blackwell, eds.); “Current Protocols in Molecular Biology” (F.M. Ausubel et al., eds., 1987, and periodic updates); and “Current Protocols in Immunology” (J.E. Coligan et al., eds., 1991), each of which is herein incorporated by reference in its entirety.

[0024] Various aspects of the disclosure are set forth below in sections; however, aspects ofthe disclosure described in one particular section are not to be limited to any particular section. Further, when a variable is not accompanied by a definition, the previous definition of the variable controls. Definitions

[0025] Compounds of the present disclosure include those described generally herein, and arefurther illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. These definitions apply regardless of whether a term is used by itself or in combination with other terms, unless otherwise indicated. Hence, the definition of “alkyl” applies to “alkyl” as well as the “alkyl” portions of “- O-alkyl” etc. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th7 33049332.1 400807-032WO (218484)Ed. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5thEd., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0026] The term “aliphatic” or “aliphatic group”, as used herein, means a straight-chain (i.e.,unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “cycloaliphatic”), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, “cycloaliphatic” refers to a monocyclic C3-C6 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0027] As used herein, the term “bicyclic ring” or “bicyclic ring system” refers to anybicyclic ring system, i.e. carbocyclic or heterocyclic, saturated or having one or more units of unsaturation, having one or more atoms in common between the two rings of the ring system. Thus, the term includes any permissible ring fusion, such as ortho-fused or spirocyclic. As used herein, the term “heterobicyclic” is a subset of “bicyclic” that requires that one or more heteroatoms are present in one or both rings of the bicycle. Such heteroatoms may be present at ring junctions and are optionally substituted, and may be selected from nitrogen (including N- oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as phosphates), boron, etc. In some embodiments, a bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. As used herein, the term “bridged bicyclic” refers to any bicyclic ring system, i.e. carbocyclic or heterocyclic, saturated or partially unsaturated, having at least one bridge. As 8 33049332.1 400807-032WO (218484)defined by IUPAC, a “bridge” is an unbranched chain of atoms or an atom or a valence bond connecting two bridgeheads, where a “bridgehead” is any skeletal atom of the ring system which is bonded to three or more skeletal atoms (excluding hydrogen). In some embodiments, a bridged bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include those groups set forth below where each group is attached to the rest of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, a bridged bicyclic group is optionally substituted with one or more substituents as set forth for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of a bridged bicyclic group is optionally substituted. Exemplary bicyclic rings include:

[0028] ExemplaryNH N H9 33049332.1 400807-032WO (218484)O HN O O ONHNH NH NH .

[0029] up. Exemplarylower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0030] The term “lower haloalkyl” refers to a C1-4 straight or branched alkyl group that issubstituted with one or more halogen atoms.

[0031] The term “heteroatom” means one or more of oxygen, sulfur, nitrogen, phosphorus, orsilicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR+(as in N-substituted pyrrolidinyl)).

[0032] The term “unsaturated,” as used herein, means that a moiety has one or more units ofunsaturation.

[0033] As used herein, the term “bivalent C1-8 (or C1-6) saturated or unsaturated, straight orbranched, hydrocarbon chain”, refers to bivalent alkylene, alkenylene, and alkynylene chains that are straight or branched as defined herein.

[0034] The term “alkylene” refers to a bivalent alkyl group. An “alkylene chain” is apolymethylene group, i.e., –(CH2)n–, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.

[0035] The term “-(C0 alkylene)-“ refers to a bond. Accordingly, the term “-(C0-3 alkylene)-”encompasses a bond (i.e., C0) and a -(C1-3alkylene)- group. 10 33049332.1 400807-032WO (218484)

[0036] The term “alkenylene” refers to a bivalent alkenyl group. A substituted alkenylenechain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.

[0037] The term “halogen” or “halo” means F, Cl, Br, or I.

[0038] The term “aryl” used alone or as part of a larger moiety as in “aralkyl,” “aralkoxy,” or“aryloxyalkyl,” refers to monocyclic or bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term “aryl” may be used interchangeably with the term “aryl ring.” In certain embodiments of the present disclosure, “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term “aryl,” as it is used herein, is a group in which an aromatic ring is fused to one or more non–aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like. The term “phenylene” refers to a multivalent phenyl group having the appropriate number of open valences to account for groups attached to it. For example, “phenylene” is a bivalent h l h i h h i “ h l ” is a e term

[0039] The terms “heteroaryl” and “heteroar–,” used alone or as part of a larger moiety, e.g.,“heteroaralkyl,” or “heteroaralkoxy,” refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 ^ electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. The term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and “heteroar–”, as used herein, also 11 33049332.1 400807-032WO (218484)include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where unless otherwise specified, the radical or point of attachment is on the heteroaromatic ring or on one of the rings to which the heteroaromatic ring is fused. Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H–quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, and tetrahydroisoquinolinyl. A heteroaryl group may be mono– or bicyclic. The term “heteroaryl” may be used interchangeably with the terms “heteroaryl ring,” “heteroaryl group,” or “heteroaromatic,” any of which terms include rings that are optionally substituted. The term “heteroaralkyl” refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.

[0040] The term “heteroarylene” refers to a multivalent heteroaryl group having theappropriate number of open valences to account for groups attached to it. For example, “heteroarylene” is a bivalent heteroaryl group when it has two groups attached to it; “heteroarylene” is a trivalent heteroaryl group when it has three groups attached to it. The term “pyridinylene” refers to a multivalent pyridine radical having the appropriate number of open valences to account for groups attached to it. For example, “pyridinylene” is a bivalent pyridine radical when it has two groups attached to it (e.g ); “pyridinylene” is a trivalentpyridine radical when it has three groups attached to it (e.g ).

[0041] As used herein, the terms “heterocycle,” “heteroy y , rocyclic radical,” and“heterocyclic ring” are used interchangeably and refer to a stable 5– to 7–membered monocyclic or 7–10–membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0–3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4– dihydro–2H–pyrrolyl), NH (as in pyrrolidinyl), or+NR (as in N–substituted pyrrolidinyl). 12 33049332.1 400807-032WO (218484)

[0042] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbonatom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, 2-oxa-6- azaspiro[3.3]heptane, and quinuclidinyl. The terms “heterocycle,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclic group,” “heterocyclic moiety,” and “heterocyclic radical,” are used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H–indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. A heterocyclyl group may be mono– or bicyclic. The term “heterocyclylalkyl” refers to an alkyl group substituted by a heterocyclyl, wherein the alkyland heterocyclyl portions independently are optionally substituted. The term “oxo-heterocyclyl”refers to a heterocyclyl substituted by an oxo group. The term “heterocyclylene” refers to a multivalent heterocyclyl group having the appropriate number of open valences to account for groups attached to it. For example, “heterocyclylene” is a bivalent heterocyclyl group when it has two groups attached to it; “heterocyclylene” is a trivalent heterocyclyl group when it has three groups attached to it.

[0043] As used herein, the term “partially unsaturated” refers to a ring moiety that includes atleast one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as herein defined.

[0044] As described herein, compounds of the disclosure may contain “optionallysubstituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are preferably those that result in theformation of stable or chemically feasible compounds. The term “stable,” as used herein, refers13 33049332.1 400807-032WO (218484)to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0045] Each optional substituent on a substitutable carbon is a monovalent substituentindependently selected from halogen; –(CH2)0–4R°; –(CH2)0–4OR°; -O(CH2)0-4Ro, –O–(CH2)0– 4C(O)OR°; –(CH2)0–4CH(OR°)2; –(CH2)0–4SR°; –(CH2)0–4Ph, which may be substituted with R°; –(CH2)0–4O(CH2)0–1Ph which may be substituted with R°; –CH=CHPh, which may be substituted with R°; –(CH2)0–4O(CH2)0–1-pyridyl which may be substituted with R°; –NO2; –CN; – N3; -(CH2)0–4N(R°)2; –(CH2)0–4N(R°)C(O)R°; –N(R°)C(S)R°; –(CH2)0–4N(R°)C(O)NR°2; -N(R°)C(S)NR°2; –(CH2)0–4N(R°)C(O)OR°; –N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; –(CH2)0–4C(O)R°; –C(S)R°; –(CH2)0–4C(O)OR°; –(CH2)0–4C(O)SR°; -(CH2)0–4C(O)OSiR°3; –(CH2)0–4OC(O)R°; –OC(O)(CH2)0–4SR–, SC(S)SR°; –(CH2)0–4SC(O)R°; –(CH2)0–4C(O)NR°2; –C(S)NR°2; –C(S)SR°; –SC(S)SR°, -(CH2)0–4OC(O)NR°2;-C(O)N(OR°)R°; –C(O)C(O)R°; –C(O)CH2C(O)R°; –C(NOR°)R°; -(CH2)0–4SSR°; –(CH2)0–4S(O)2R°; –(CH2)0–4S(O)2OR°; –(CH2)0–4OS(O)2R°; –S(O)2NR°2; –S(O)(NR°)R°; – S(O)2N=C(NR°2)2; -(CH2)0–4S(O)R°; -N(R°)S(O)2NR°2; –N(R°)S(O)2R°; –N(OR°)R°; – C(NH)NR°2; –P(O)2R°; -P(O)R°2; -OP(O)R°2; –OP(O)(OR°)2; SiR°3; –(C1–4straight or branched alkylene)O–N(R°)2; or –(C1–4 straight or branched alkylene)C(O)O–N(R°)2.

[0046] Each R° is independently hydrogen, C1–6 aliphatic, –CH2Ph, –O(CH2)0–1Ph, -CH2-(5-6membered heteroaryl ring), or a 5–6–membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3–12–membered saturated, partially unsaturated, or aryl mono– or bicyclic ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted by a divalent substituent on a saturated carbon atom of R° selected from =O and =S; or each R° is optionally substituted with a monovalent substituent independently selected from halogen, –(CH2)0–2Rl, –(haloRl), –(CH2)0–2OH, –(CH2)0–2ORl, – (CH2)0–2CH(ORl)2; -O(haloRl), –CN, –N3, –(CH2)0–2C(O)Rl, –(CH2)0–2C(O)OH, –(CH2)0– 2, –8484)

[0047] Each Rl is independently selected from C1–4 aliphatic, –CH2Ph, –O(CH2)0–1Ph, or a 5–6–membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each Rlis unsubstituted or where preceded by halo is substituted only with one or more halogens; or wherein an optional substituent on a saturated carbon is a divalent substituent independently selected from =O, =S, =NNR*2, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)2R*, =NR*, =NOR*, –O(C(R*2))2–3O–, or – S(C(R*2))2–3S–, or a divalent substituent bound to vicinal substitutable carbons of an “optionallysubstituted” group is –O(CR*2)2–3O–, wherein each independent occurrence of R* is selectedfrom hydrogen, C1–6 aliphatic or an unsubstituted 5–6–membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0048] When R* is C1–6 aliphatic, R* is optionally substituted with halogen, –Rl, -(haloRl), -OH, –ORl, –O(haloRl), –CN, –C(O)OH, –C(O)ORl, –NH2, –NHRl, –NRl2, or – NO2, wherein each Rlis independently selected from C1–4 aliphatic, –CH2Ph, –O(CH2)0–1Ph, or a 5–6–membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each Rlis unsubstituted or where preceded by halo is substituted only with one or more halogens.

[0049] An optional substituent on a substitutable nitrogen is independently –R†, –NR†2, –C(O)R†, –C(O)OR†, –C(O)C(O)R†, –C(O)CH2C(O)R†, -S(O)2R†, -S(O)2NR†2, –C(S)NR†2, – C(NH)NR†2, or –N(R†)S(O)2R†; wherein each R†is independently hydrogen, C1–6 aliphatic, unsubstituted –OPh, or an unsubstituted 5–6–membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, two independent occurrences of R†, taken together with their intervening atom(s) form an unsubstituted 3–12–membered saturated, partially unsaturated, or aryl mono– or bicyclic ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; wherein when R†is C1–6 aliphatic, R†is optionally substituted with halogen, –Rl, -(haloRl), -OH, –ORl, – O(haloRl), –CN, –C(O)OH, –C(O)ORl, –NH2, –NHRl, –NRl2, or –NO2, wherein each Rlis independently selected from C1–4aliphatic, –CH2Ph, –O(CH2)0–1Ph, or a 5–6–membered saturated, partially unsaturated, or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each Rlis unsubstituted or where preceded by halo is substituted only with one or more halogens. 15 33049332.1 400807-032WO (218484)

[0050] As used herein, the term "pharmaceutically acceptable salt" refers to those salts whichare, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1–19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2–hydroxy–ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2–naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3–phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p–toluenesulfonate, undecanoate, valerate salts, and the like.

[0051] Further, acids which are generally considered suitable for the formation ofpharmaceutically useful salts from basic pharmaceutical compounds are discussed, for example, by P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food & Drug Administration, Washington, D.C. on their website). These disclosures are incorporated herein by reference.

[0052] Salts derived from appropriate bases include alkali metal, alkaline earth metal,ammonium and N+(C1–4alkyl)4salts. Representative alkali or alkaline earth metal salts include 16 33049332.1 400807-032WO (218484)sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

[0053] Unless otherwise stated, structures depicted herein are also meant to include allisomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the disclosure. Unless otherwise stated, all tautomeric forms of the compounds of the disclosure are within the scope of the disclosure. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a13C- or14C-enriched carbon are within the scope of this disclosure. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present disclosure.

[0054] Diastereomeric mixtures can be separated into their individual diastereomers on thebasis of their physical chemical differences by methods known to those skilled in the art, such as, for example, by chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Alternatively, a particular enantiomer of a compound of the present disclosure may be prepared by asymmetric synthesis. Still further, where the molecule contains a basic functional group (such as amino) or an acidic functional group (such as carboxylic acid) diastereomeric salts are formed with an appropriate optically- active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means known in the art, and subsequent recovery of the pure enantiomers. 17 33049332.1 400807-032WO (218484)

[0055] Individual stereoisomers of the compounds of the disclosure may, for example, besubstantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. Chiral center(s) in a compound of the present disclosure can have the S or R configuration as defined by the IUPAC 1974 Recommendations. Further, to the extent a compound described herein may exist as a atropisomer (e.g., substituted biaryls), all forms of such atropisomer are considered part of this disclosure.

[0056] Chemical names, common names, and chemical structures may be usedinterchangeably to describe the same structure. If a chemical compound is referred to using both a chemical structure and a chemical name, and an ambiguity exists between the structure and the name, the structure predominates. It should also be noted that any carbon as well as heteroatom with unsatisfied valences in the text, schemes, examples and tables herein is assumed to have the sufficient number of hydrogen atom(s) to satisfy the valences.

[0057] The terms “a” and “an” as used herein mean “one or more” and include the pluralunless the context is inappropriate.

[0058] The term “alkyl” refers to a saturated straight or branched hydrocarbon, such as astraight or branched group of 1-12, 1-10, or 1-6 carbon atoms, referred to herein as C1-C12 alkyl, C1-C10alkyl, and C1-C6alkyl, respectively. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3- methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1- butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, etc.

[0059] The term “cycloalkyl” refers to a monovalent saturated cyclic, bicyclic, or bridgedcyclic (e.g., adamantyl) hydrocarbon group of 3-12, 3-8, 4-8, or 4-6 carbons, referred to herein, e.g., as “C3-C6 cycloalkyl,” derived from a cycloalkane. Exemplary cycloalkyl groups include cyclohexyl, cyclopentyl, cyclobutyl, and cyclopropyl. The term “cycloalkylene” refers to a bivalent cycloalkyl group.

[0060] The term “haloalkyl” refers to an alkyl group that is substituted with at least onehalogen. Exemplary haloalkyl groups include -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, and the 18 33049332.1 400807-032WO (218484)like. The term “haloalkylene” refers to a bivalent haloalkyl group. The term “halomethyl” refers to a haloalkyl group containing a single carbon atom.

[0061] The term “hydroxyalkyl” refers to an alkyl group that is substituted with at least onehydroxyl. Exemplary hydroxyalkyl groups include -CH2CH2OH, -C(H)(OH)CH3, -CH2C(H)(OH)CH2CH2OH, and the like.

[0062] The terms “alkenyl” and “alkynyl” are art-recognized and refer to unsaturatedaliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.

[0063] The term “carbocyclylene” refers to a multivalent carbocyclyl group having theappropriate number of open valences to account for groups attached to it. For example, “carbocyclylene” is a bivalent carbocyclyl group when it has two groups attached to it; “carbocyclylene” is a trivalent carbocyclyl group when it has three groups attached to it.

[0064] The terms “alkoxyl” or “alkoxy” are art-recognized and refer to an alkyl group, asdefined above, having an oxygen radical attached thereto. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, tert-butoxy and the like. The term “haloalkoxyl” refers to an alkoxyl group that is substituted with at least one halogen. Exemplary haloalkoxyl groups include -OCH2F, -OCHF2, -OCF3, -OCH2CF3, -OCF2CF3, and the like.

[0065] The term “oxo” is art-recognized and refers to a “=O” substituent. For example, acyclopentane substituted with an oxo group is cyclopentanone.

[0066] The symbol “ ” indicates a point of attachment.

[0067] When a chemical structure containing a ring is depicted with a substituent having abond that crosses a ring bond, the substituent may be attached at any available position on the ring. For example, the chemical structur encompass ,and . In the context of a polycyclic fused ring, when a chemical structure containing a polycyclic fused ring is depicted with one or more substituent(s) having a bond that crosses multiple rings, the one or more substituent(s) may be independently attached to any of the rings 19 33049332.1 400807-032WO (218484)crossed by the bond. To illustrate, the chemical structur encompasses, forexample, .

[0068] ime in any constituent or thecompound of the disclosure, its definition on each occurrence is independent of its definition at every other occurrence, unless otherwise indicated.

[0069] One or more compounds of the disclosure may exist in unsolvated as well as solvatedforms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the disclosure embrace both solvated and unsolvated forms. “Solvate” means a physical association of a compound of this disclosure with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. “Solvate” encompasses both solution-phase and isolatable solvates. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. “Hydrate” is a solvate wherein the solvent molecule is H2O.

[0070] As used herein, the terms “subject” and “patient” are used interchangeable and refer toorganisms to be treated by the methods of the present disclosure. Such organisms preferably include, but are not limited to, mammals (e.g., murines, simians, equines, bovines, porcines, canines, felines, and the like), and most preferably includes humans.

[0071] The term “IC50” is art-recognized and refers to the concentration of a compound that isrequired to achieve 50% inhibition of the target.

[0072] As used herein, the term “effective amount” refers to the amount of a compoundsufficient to effect beneficial or desired results (e.g., a therapeutic, ameliorative, inhibitory or preventative result). An effective amount can be administered in one or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route. As used herein, the term “treating” includes any effect, e.g., lessening, 20 33049332.1 400807-032WO (218484)reducing, modulating, ameliorating or eliminating, that results in the improvement of the condition, disease, disorder, and the like, or ameliorating a symptom thereof.

[0073] As used herein, the term “pharmaceutical composition” refers to the combination of anactive agent with a carrier, inert or active, making the composition especially suitable for diagnostic or therapeutic use in vivo or ex vivo.

[0074] As used herein, the term “pharmaceutically acceptable carrier” refers to any of thestandard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions (e.g., such as an oil / water or water / oil emulsions), and various types of wetting agents. The compositions also can include stabilizers and preservatives. For examples of carriers, stabilizers and adjuvants, see e.g., Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA

[1975] .

[0075] For therapeutic use, salts of the compounds of the present disclosure are contemplatedas being pharmaceutically acceptable. However, salts of acids and bases that are non- pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

[0076] In addition, when a compound of the disclosure contains both a basic moiety (such as,but not limited to, a pyridine or imidazole) and an acidic moiety (such as, but not limited to, a carboxylic acid) zwitterions (“inner salts”) may be formed. Such acidic and basic salts used within the scope of the disclosure are pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts. Such salts of the compounds of the disclosure may be formed, for example, by reacting a compound of the disclosure with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.

[0077] Throughout the description, where compositions are described as having, including, orcomprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions of the present disclosure that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present disclosure that consist essentially of, or consist of, the recited processing steps. 21 33049332.1 400807-032WO (218484)

[0078] As a general matter, compositions specifying a percentage are by weight unlessotherwise specified. I. Cytosine Nucleoside Analogs and Related Compounds

[0079] Cytosine nucleoside analogs and related compounds are disclosed herein. Thecompounds may be used in the pharmaceutical compositions and therapeutic methods described herein. Exemplary compounds are described in the following sections, along with exemplary procedures for making the compounds.

[0080] One aspect of the disclosure provides a compound represented by Formula I:or a pharmaceutically ac: R1is -(C0-4 alkylene)-(phenyl substituted by m occurrences of R6), C10-C20 alkenyl, C1-2 alkyl, C3-4 alkyl, C5-6 alkyl, C7-10 alkyl, or -(C0-4 alkylene)-C3-6 cycloalkyl; R2is halomethyl, C1-3alkyl, C2-4alkenyl, or C2-4alkynyl; R3is hydrogen, -C(O)-(C1-2alkyl), -C(O)-(C3-6alkyl), -C(O)-(C7-10alkyl), -C(O)-(C0-4alkylene)-(phenyl substituted by n occurrences of R7), -C(O)-(C0-4 alkylene)-C3-6 cycloalkyl, or - C(O)-(C0-4 alkylene)-(3-7 membered saturated heterocyclyl containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur); R4is fluoro, Cl, bromo, iodo, hydrogen, -OH, C2-4 alkynyl, or C1-3 alkyl; R5is fluoro, Cl, bromo, iodo, or hydrogen; R6represents independently for each occurrence halo or C1-3alkyl; R7represents independently for each occurrence halo or C1-3 alkyl; m is 0, 1, or 2; and 22 33049332.1 400807-032WO (218484)n is 0, 1, or 2.

[0081] The definitions of variables in Formula I above encompass multiple chemical groups.The application contemplates embodiments where, for example, i) the definition of a variable is a single chemical group selected from those chemical groups set forth above, ii) the definition of a variable is a collection of two or more of the chemical groups selected from those set forth above, and iii) the compound is defined by a combination of variables in which the variables are defined by (i) or (ii).

[0082] In certain embodiments, the compound is a compound of Formula I.

[0083] As defined generally above, R1 is -(C0-4 alkylene)-(phenyl substituted by moccurrences of R6), C10-C20alkenyl, C1-2alkyl, C3-4alkyl, C5-6alkyl, C7-10alkyl, or -(C0-4alkylene)-C3-6cycloalkyl. In certain embodiments, R1is -(C0-4alkylene)-phenyl substituted by m occurrences of R6. In certain embodiments, R1is C10-C20 alkenyl. In certain embodiments, R1is C1-2 alkyl. In certain embodiments, R1is C3-4 alkyl. In certain embodiments, R1is C5-6 alkyl. In certain embodiments, R1is C7-10alkyl. In certain embodiments, R1is -(C0-4alkylene)-C3-6cycloalkyl. In certain embodiments, R1i . In certain embodiments, R1is isopropyl. In certain embodiments, R1is selected fepicted in Table 1, below.

[0084] As defined generally above, R2 is halomethyl, C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl.In certain embodiments, R2is halomethyl. In certain embodiments, R2is C1-3 alkyl. In certain embodiments, R2is C2-4alkenyl. In certain embodiments, R2is C2-4alkynyl. In certain embodiments, R2is -CH2Cl. In certain embodiments, R2is ethyl. In certain embodiments, R2is ethenyl. In certain embodiments, R2is ethynyl. In certain embodiments, R2is selected from those depicted in Table 1, below.

[0085] As defined generally above, R3 is hydrogen, -C(O)-(C1-2 alkyl), -C(O)-(C3-6 alkyl), -C(O)-(C7-10 alkyl), -C(O)-(C0-4 alkylene)-(phenyl substituted by n occurrences of R7), -C(O)-(C0-4 alkylene)-C3-6 cycloalkyl, or -C(O)-(C0-4 alkylene)-(3-7 membered saturated heterocyclyl containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur). In certain embodiments, R3is hydrogen. In certain embodiments, R3is -C(O)-(C1-2 alkyl). In certain embodiments, R3is -C(O)-(C3-6 alkyl). In certain embodiments, R3is -C(O)-(C7-10 alkyl). In 23 33049332.1 400807-032WO (218484)certain embodiments, R3is -C(O)-(C0-4alkylene)-phenyl substituted by n occurrences of R7. In certain embodiments, R3is -C(O)-(phenyl). In certain embodiments, R3is -C(O)-(C0-4 alkylene)- C3-6 cycloalkyl. In certain embodiments, R3is -C(O)-(C0-4 alkylene)-(3-7 membered saturated heterocyclyl containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur). In certain embodiments, R3is . In certain embodiments, R3is -C(O)- O (isopropyl). In certain embodiments In certain embodiments, R3is selected from those depicted in Table 1, belo

[0086] As defined generally above, R4 is fluoro, Cl, bromo, iodo, hydrogen, -OH, C2-4alkynyl, or C1-3 alkyl. In certain embodiments, R4is fluoro. In certain embodiments, R4is Cl. In certain embodiments, R4is bromo. In certain embodiments, R4is iodo. In certain embodiments, R4is hydrogen. In certain embodiments, R4is -OH. In certain embodiments, R4is C2-4alkynyl . In certain embodiments, R4is C1-3 alkyl. In certain embodiments, R4is hydrogen or -OH. In certain embodiments, R4is fluoro, Cl, bromo, iodo, C2-4 alkynyl, or C1-3 alkyl. In certain embodiments, R4is fluoro, Cl, bromo, or iodo. In certain embodiments, R4is selected from those depicted in Table 1, below.

[0087] As defined generally above, R5 is fluoro, Cl, bromo, iodo, or hydrogen. In certainembodiments, R5is fluoro. In certain embodiments, R5is Cl. In certain embodiments, R5is bromo. In certain embodiments, R5is iodo. In certain embodiments, R5is hydrogen. In certain embodiments, R5is selected from those depicted in Table 1, below.

[0088] As defined generally above, R6 represents independently for each occurrence halo orC1-3alkyl. In certain embodiments, R6represents independently for each occurrence halo. In certain embodiments, R6represents independently for each occurrence C1-3 alkyl. In certain embodiments, R6is halo. In certain embodiments, R6is C1-3 alkyl. In certain embodiments, R6is selected from those depicted in Table 1, below.

[0089] As defined generally above, R7 represents independently for each occurrence halo orC1-3 alkyl. In certain embodiments, R7represents independently for each occurrence halo. In 24 33049332.1 400807-032WO (218484)certain embodiments, R7represents independently for each occurrence C1-3alkyl. In certain embodiments, R7is halo. In certain embodiments, R7is C1-3 alkyl. In certain embodiments, R7is selected from those depicted in Table 1, below.

[0090] As defined generally above, m is 0, 1, or 2. In certain embodiments, m is 1 or 2. Incertain embodiments, m is 0. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is selected from those depicted in Table 1, below.

[0091] As defined generally above, n is 0, 1, or 2. In certain embodiments, n is 1 or 2. Incertain embodiments, n is 0. In certain embodiments, m is 1. In certain embodiments, n is 2. In certain embodiments, n is selected from those depicted in Table 1, below.

[0092] In certain embodiments, the compound of Formula I is represented by Formula Ia orIb, or a pharmaceutically acceptable salt thereof: wherein all vaa es e e a e as e e o o u a a ove.

[0093] In certain embodiments, the compound of Formula I is represented by Formula Ic or apharmaceutically acceptable salt thereof:wherein all variables therein are as defined for Formula I, above. 25 33049332.1 400807-032WO (218484)

[0094] In certain embodiments, the compound of Formula I is represented by Formula Id or apharmaceutically acceptable salt thereof: wherein all variables therein areve.

[0095] Another aspect of the disclosure provides a compound represented by Formula II:or a pharmaceutically accep, ein: R1is -C(O)-(C1-2alkyl), -C(O)-(C3-6alkyl), or -C(O)-(C7-10alkyl); and R2is C1-3 alkyl or C2-4 alkenyl.

[0096] The definitions of variables in Formula II above encompass multiple chemical groups.The application contemplates embodiments where, for example, i) the definition of a variable is a single chemical group selected from those chemical groups set forth above, ii) the definition of a variable is a collection of two or more of the chemical groups selected from those set forth above, and iii) the compound is defined by a combination of variables in which the variables are defined by (i) or (ii).

[0097] In certain embodiments, the compound is a compound of Formula II.

[0098] As defined generally above, R2 is C1-3 alkyl or C2-4 alkenyl. In certain embodiments,R2is halomethyl. In certain embodiments, R2is C1-3alkyl. In certain embodiments, R2is C2-4alkenyl. In certain embodiments, R2is ethyl. In certain embodiments, R2is ethenyl. In certain 26 33049332.1 400807-032WO (218484)embodiments, R2is selected from those depicted in Table 2, below.

[0099] As defined generally above, R3 is -C(O)-(C1-2 alkyl), -C(O)-(C3-6 alkyl), or -C(O)-(C7-10 alkyl). In certain embodiments, R3is -C(O)-(C1-2 alkyl). In certain embodiments, R3is -C(O)- (C3-6alkyl). In certain embodiments, R3is -C(O)-(C7-10alkyl). In certain embodiments, R3is - C(O)-(isopropyl). In certain embodiments, R3is selected from those depicted in Table 2, below.

[0100] Another aspect of the disclosure provides a compound in Table 1, below, or apharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound in Table 1, below.

[0101] Another aspect of the disclosure provides a compound selected from I-1 to I-38 inTable 1, below, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound selected from I-1 to I-38 in Table 1, below.

[0102] Another aspect of the disclosure provides a compound selected from I-1 to I-39 inTable 1, below, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound selected from I-1 to I-39 in Table 1, below. TABLE 1. Compound No. Chemical Structure27 33049332.1 400807-032WO (218484)Compound No. Chemical Structure233049332.17-032WO (218484)Compound No. Chemical StructureO 2 33049332.17-032WO (218484)Compound No. Chemical Structure230 33049332.1 400807-032WO (218484)Compound No. Chemical Structure31 33049332.1 400807-032WO (218484)Compound No. Chemical Structure33049332.17-032WO (218484)Compound No. Chemical Structure33 33049332.1 400807-032WO (218484)Compound No. Chemical Structure33049332.17-032WO (218484)Compound No. Chemical Structure

[0103] Another aspect of the disclosure provides a compound in Table 2, below, or apharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound in Table 2, below. TABLE 2. Compound No. Chemical Structure35 33049332.1 400807-032WO (218484)Compound No. Chemical Structure

[0104] Methods for preparing compounds described herein are illustrated in the Examplesbelow.

[0105] It is understood by one skilled in the art of organic synthesis that the functionalitypresent on various portions of the molecule should be compatible with the reagents and reactions proposed. Substituents not compatible with the reaction conditions will be apparent to one skilled in the art, and alternate methods are therefore indicated (for example, use of protecting groups or alternative reactions). Protecting group chemistry and strategy is well known in the art, for example, as described in detail in “Protecting Groups in Organic Synthesis”, T. W. Greene and P. G. M. Wuts, 3rdedition, John Wiley & Sons, 1999, the entire contents of which are hereby incorporated by reference. For example, the exocyclic nitrogen atom of cytosine, and 5- substituted cytosine, nucleobases may be protected using, for example, a substituted trityl protecting group.

[0106] The modular synthetic routes described herein and in the foregoing references can alsobe readily modified by one of skill in the art of organic synthesis to provide additional compounds using strategies and reactions well known in the art, as described in, for example, “Comprehensive Organic Synthesis” (B.M. Trost & I. Fleming, eds., 1991-1992) or “Comprehensive Organic Synthesis” (P. Knochel & G.A. Molander, eds., 2014). 36 33049332.1 400807-032WO (218484)Scheme 1. Diester Compound General Synthesis[01a g o a uc eos e suc as sc e e e es e co pou can be madeby the following general method outlined in scheme 1. First the hydroxyl groups in compound A are protected as silyl ethers using TBS-Cl in DMF with imidazole as base. Next the nucleobase nitrogen is protected as the monomethoxytrityl amine using MMTrCl in DCM with collidine and silver nitrate. The hydroxyl groups are then deprotected using a solution of TBAF in THF. The hydroxyl groups are then bis-esterified using an excess of acid chloride in acetonitrile with DMAP as base. Compound 4 is deprotected using an aqueous solution of 80% formic acid to provide the desired compound 5. 37 33049332.1 400807-032WO (218484)Scheme 2. Monoester Compound General Synthesisg ,made by the following general method outlined in scheme 2. With the nucleobase nitrogen protected as the monomethoxytrityl amine, the primary hydroxyl group is selectively esterified using one equivalent of the acid chloride in acetonitrile with a base. Compound 6 is subjected to deprotection conditions using a solution of 80% aqueous formic acid to provide the desired compound 7. II. Methods of Treating Medical Disorders

[0109] One aspect of the disclosure provides a method of treating a disorder selected from thegroup consisting of cancer, an autoimmune disorder, and a neurological disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein to treat the disorder.

[0110] Another aspect of the disclosure provides a method of inhibiting LINE1 reversetranscriptase activity in a subject, comprising contacting a LINE1 reverse transcriptase with an effective amount of a compound disclosed herein to inhibit the activity of said LINE1 reverse transcriptase. 38 33049332.1 400807-032WO (218484)

[0111] In certain embodiments, the subject has a disorder selected from the group consistingof cancer, an autoimmune disorder, and a neurological disorder. In certain embodiments, the disorder is cancer.

[0112] Yet another aspect of the disclosure provides a method of treating a viral infection.The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound described herein (such as a compound of Formula I) to treat the viral infection.

[0113] In certain embodiments of each of the foregoing methods, the compound is acompound of Formula I. In certain embodiments of each of the foregoing methods, the compound is a compound of Formula II.

[0114] In certain embodiments, the compound is administered in a pharmaceuticalcomposition comprising the compound and a carrier, excipient, and / or vehicle, as further described in Section IV, below.

[0115] In certain embodiments, the method further comprises administering an effectiveamount of one or more additional therapeutic agents, as further described in Section IV, below. Cancer

[0116] In certain embodiments, the disorder is cancer. In certain embodiments, the cancer isa solid tumor or leukemia. In certain embodiments, the cancer is a solid tumor. In certain embodiments, the cancer is a carcinoma or melanoma. In certain embodiments, the cancer is a carcinoma. In certain embodiments, the cancer is a sarcoma. In certain embodiments, the cancer is a melanoma. In certain embodiments, the cancer is a lymphoma. In certain embodiments, the cancer is a leukemia.

[0117] In certain embodiments, the cancer is breast cancer, ovarian cancer, uterine cancer,cervical cancer, prostate cancer, testicular cancer, lung cancer, leukemia, head and neck cancer, oral cancer, esophageal cancer, stomach cancer, bile duct cancer, gallbladder cancer, bladder cancer, urinary tract cancer, colon cancer, rectal cancer, thyroid cancer, pancreatic cancer, kidney cancer, liver cancer, brain cancer, skin cancer, or eye cancer.

[0118] In certain embodiments, the cancer is breast cancer, ovarian cancer, uterine cancer,cervical cancer, prostate cancer, testicular cancer, lung cancer, leukemia, head and neck cancer, 39 33049332.1 400807-032WO (218484)oral cancer, esophageal cancer, stomach cancer, bile duct cancer, gallbladder cancer, or bladder cancer.

[0119] In certain embodiments, the cancer has (i) expression of LINE1 RNA, LINE1 ORF1polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase.

[0120] In certain embodiments, the cancer has (i) expression of LINE1 RNA, LINE1 ORF1polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase. In certain embodiments, the cancer has expression of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide. In certain embodiments, the cancer has expression of LINE1 RNA. In certain embodiments, the cancer has expression of LINE1 ORF1 polypeptide. In certain embodiments, the cancer has expression of LINE1 ORF2 polypeptide. In certain embodiments, the cancer has activity of LINE1 reverse transcriptase.

[0121] In certain embodiments, the cancer has elevated (i) levels of LINE1 RNA, LINE1ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase.

[0122] In certain embodiments, the cancer has elevated (i) levels of LINE1 RNA, LINE1ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase. In certain embodiments, the cancer has elevated levels of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide. In certain embodiments, the cancer has elevated levels of LINE1 RNA. In certain embodiments, the cancer has elevated levels of LINE1 ORF1 polypeptide. In certain embodiments, the cancer has elevated levels of LINE1 ORF2 polypeptide. In certain embodiments, the cancer has elevated activity of LINE1 reverse transcriptase.

[0123] In certain embodiments, the cancer is an epithelial cancer. In certain embodiments,the epithelial cancer is pancreatic cancer, colorectal cancer, breast cancer, prostate cancer, esophageal cancer, head and neck cancer, renal cancer, ovarian cancer, or lung cancer. In certain embodiments, the cancer is pancreatic cancer, colorectal cancer, breast cancer, prostate cancer, renal cancer, ovarian cancer, or lung cancer. In certain embodiments, the cancer is pancreatic cancer. In certain embodiments, the cancer is pancreatic adenocarcinoma. In certain embodiments, the cancer is colorectal cancer. In certain embodiments, the cancer comprises 40 33049332.1 400807-032WO (218484)microsatellite instable (MSI) colorectal cancer or microsatellite stable (MSS) colorectal cancer. In certain embodiments, the cancer is breast cancer. In certain embodiments, the cancer is prostate cancer. In certain embodiments, the cancer is esophageal cancer. In certain embodiments, the cancer is head and neck cancer. In certain embodiments, the cancer is renal cancer. In certain embodiments, the cancer is ovarian cancer. In certain embodiments, the cancer is lung cancer. In certain embodiments, the lung cancer is non-small cell lung carcinoma or small cell lung carcinoma. In certain embodiments, the cancer is non-small cell lung carcinoma. In certain embodiments, the cancer is small cell lung carcinoma.

[0124] In certain embodiments, the cancer is a preneoplastic or early cancer lesion. In certainembodiments, the cancer is intraductal papillary mucinous neoplasm (IPMN), pancreatic intraepithelial neoplasia (PanIN), ductal carcinoma in situ (DCIS), or Barrett’s Esophagus. In certain embodiments, the cancer intraductal papillary mucinous neoplasm (IPMN). In certain embodiments, the cancer is pancreatic intraepithelial neoplasia (PanIN). In certain embodiments, the cancer is ductal carcinoma in situ (DCIS). In certain embodiments, the cancer is Barrett’s Esophagus.

[0125] In certain embodiments, the cancer has elevated levels of pericentrometric humansatellite II (HSATII) RNA. In some embodiments, the cancer is a microsatellite instable (MSI) cancer. In some embodiments, the cancer is a microsatellite stable (MSS) cancer.

[0126] In aspects of any of the embodiments, the cancer is associated with long interspersednuclear element-1 (LINE-1) reverse transcriptase (RT). In further aspects of these embodiments, the cancer is associated with high levels of LINE-1 RT activity.

[0127] In certain embodiments, the cancer is selected from B cell lymphomas (e.g., B cellchronic lymphocytic leukemia, B cell non-Hodgkin lymphoma, cutaneous B cell lymphoma, diffuse large B cell lymphoma), basal cell carcinoma, bladder cancer, blastoma, brain metastasis, breast cancer, Burkitt lymphoma, carcinoma (e.g., adenocarcinoma (e.g., of the gastroesophageal junction)), cervical cancer, colon cancer, colorectal cancer (colon cancer and rectal cancer), endometrial carcinoma, esophageal cancer, Ewing sarcoma, follicular lymphoma, gastric cancer, gastroesophageal junction carcinoma, gastrointestinal cancer, glioblastoma (e.g., glioblastoma multiforme, e.g., newly diagnosed or recurrent), glioma, head and neck cancer (e.g., head and neck squamous cell carcinoma), hepatic metastasis, Hodgkin' s and non- 41 33049332.1 400807-032WO (218484)Hodgkin' s lymphoma, kidney cancer (e.g., renal cell carcinoma and Wilms' tumors), laryngeal cancer, leukemia (e.g., chronic myelocytic leukemia, hairy cell leukemia), liver cancer (e.g., hepatic carcinoma and hepatoma), lung cancer (e.g., non-small cell lung cancer and small-cell lung cancer), lymphoblastic lymphoma, lymphoma, mantle cell lymphoma, metastatic brain tumor, metastatic cancer, myeloma (e.g., multiple myeloma), neuroblastoma, ocular melanoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer (e.g., pancreatic ductal adenocarcinoma), prostate cancer (e.g., hormone refractory (e.g., castration resistant), metastatic, metastatic hormone refractory (e.g., castration resistant, androgen independent)), renal cell carcinoma (e.g., metastatic), salivary gland carcinoma, sarcoma (e.g., rhabdomyosarcoma), skin cancer (e.g., melanoma (e.g., metastatic melanoma)), soft tissue sarcoma, solid tumor, squamous cell carcinoma, synovia sarcoma, testicular cancer, thyroid cancer, transitional cell cancer (urothelial cell cancer), uveal melanoma (e.g., metastatic), verrucous carcinoma, vulval cancer, and Waldenstrom macroglobulinemia.

[0128] In some embodiments, the cancer is a virus-associated cancer. As used herein, theterm “virus-associated cancer” means any cancer in which a virus is known to play a role. For example, Epstein-Barr virus (EBV) has been reported to be associated with the endemic variant of Burkitt lymphoma and certain other lymphomas. Infection by human papilloma virus (HPV) is believed to be responsible for certain types of cervical and / or genital cancer. Human T-cell leukemia virus 1 (HTLV-1) has been reported to be linked adult T-cell leukemia / lymphoma (ATLL). Human T-cell leukemia virus 2 (HTLV-2) has been reported to be linked to cutaneous T-cell lymphoma. Human herpes virus 8 (HHV-8) is believed to cause Kaposi’s sarcoma in patients with AIDS. In certain embodiments, the cancer is a cancer associated with EBV, HPV, HTLV-1, HTLV-2, or HHV-8. In certain embodiments, the cancer is Burkitt lymphoma, cervical cancer, genital cancer, adult T-cell leukemia / lymphoma, cutaneous T-cell lymphoma, or Kaposi’s sarcoma.

[0129] In some embodiments, the cancer is a cancer other than a virus-associated cancer. Incertain embodiments, the cancer is a cancer other than a cancer associated with EBV, HPV, HTLV-1, HTLV-2, or HHV-8. In certain embodiments, the cancer is a cancer other than Burkitt lymphoma, cervical cancer, genital cancer, adult T-cell leukemia / lymphoma, cutaneous T-cell lymphoma, or Kaposi’s sarcoma. In one embodiment, the cancer is a tumor associated with Li_Fraumeni syndrome. 42 33049332.1 400807-032WO (218484)

[0130] In some embodiments, the cancer is renal cell carcinoma, or kidney cancer,mesothelioma, hepatobiliary (hepatic and biliary duct), bone cancer, rhabdomyosarcoma, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (gastric, colorectal, and duodenal), uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin’s Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, adrenocortical carcinoma, sarcoma of soft tissue, soft tissue and bone synovial sarcoma, cancer of the urethra, cancer of the penis, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, acute myeloid leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, non-Hodgkin’s lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocortical cancer, gall bladder cancer, multiple myeloma, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma, squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST); and medulloblastoma, or a combination of one or more of the foregoing cancers.

[0131] In some embodiments, the cancer is hepatocellular carcinoma (HCC). In someembodiments, the cancer is hepatoblastoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is rectal cancer. In some embodiments, the cancer is ovarian cancer, or ovarian carcinoma. In some embodiments, the cancer is ovarian epithelial cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is papillary serous cystadenocarcinoma. In some embodiments, the cancer is uterine papillary serous carcinoma (UPSC). In some embodiments, the cancer is hepatocholangiocarcinoma. In some embodiments, the cancer is soft tissue and bone synovial sarcoma. In some embodiments, the cancer is rhabdomyosarcoma. In some embodiments, the cancer is osteosarcoma. In some embodiments, the cancer is anaplastic thyroid cancer. In some embodiments, the cancer is adrenocortical carcinoma. In some embodiments, the cancer is pancreatic cancer, or pancreatic ductal carcinoma. In some embodiments, the cancer is pancreatic adenocarcinoma. In some embodiments, the cancer is glioma. In some embodiments, the cancer is malignant peripheral 43 33049332.1 400807-032WO (218484)nerve sheath tumors (MPNST). In some embodiments, the cancer is neurofibromatosis-1 associated MPNST. In some embodiments, the cancer is Waldenstrom macroglobulinemia. In some embodiments, the cancer is medulloblastoma.

[0132] In certain embodiments, the cancer is a leukemia (e.g., acute leukemia, acutelymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin disease or non-Hodgkin disease), Waldenstrom macroglobulinemia, multiple myeloma, or heavy chain disease. In one embodiment, the cancer is a solid tumor such as a sarcoma or carcinoma (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing’s tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, and retinoblastoma).

[0133] In some embodiments, the cancer is glioma, astrocytoma, glioblastoma multiforme(GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, or retinoblastoma.

[0134] In some embodiments, the cancer is acoustic neuroma, astrocytoma (e.g. Grade I –Pilocytic Astrocytoma, Grade II – Low-grade Astrocytoma, Grade III – Anaplastic Astrocytoma, or Grade IV – Glioblastoma (GBM)), chordoma, CNS lymphoma, 44 33049332.1 400807-032WO (218484)craniopharyngioma, brain stem glioma, ependymoma, mixed glioma, optic nerve glioma, subependymoma, medulloblastoma, meningioma, metastatic brain tumor, oligodendroglioma, pituitary tumors, primitive neuroectodermal (PNET) tumor, or schwannoma. In some embodiments, the cancer is a type found more commonly in children than adults, such as brain stem glioma, craniopharyngioma, ependymoma, juvenile pilocytic astrocytoma (JPA), medulloblastoma, optic nerve glioma, pineal tumor, primitive neuroectodermal tumors (PNET), or rhabdoid tumor. Autoimmune Diseases and Disorders

[0135] In certain embodiments, the disorder is an autoimmune disorder. As used herein, theterms “autoimmune disorders” and “autoimmune diseases” are used interchangeably, and include those diseases and disorders which are traditionally classified as autoimmune disorders, as well as inflammatory disorders and immune disorders (excluding viral infections). The intention with the terms “autoimmune disease” and “autoimmune disorder” is to include all diseases and disorders which are driven by innate immune responses and adaptive immune responses which initiate some sort of innate immune inflammatory response. It is the applicant’s intent to have the terms “autoimmune disease” and “autoimmune disorder” include the full scope of diseases and disorders which are driven by innate inflammation, with the exception of viral infections.

[0136] In certain embodiments, the disclosure provides for treatment of an autoimmunedisorder. Traditional autoimmune disorders commonly occur when the immune system attacks normal cells and / or tissues in the body. Inflammatory disorders often present with chronic inflammation (among other symptoms) in the absence of infection. Autoimmune disorders also include symptoms which arise when the cellular immune system reacts against the body’s autoantigens. There may also be autoimmune and / or inflammation manifestation associated with a range of primary immunodeficiency diseases. In further aspects of these embodiments, the autoimmune disease or disorder is associated with high levels of LINE-1 protein expression.

[0137] One embodiment of the disclosure is a method of treating type I interferonopathies. Inone aspect of this embodiment, the type I interferonopathy is a congenital disorder associated with type I interferon overexpression. In one aspect of this embodiment, the congenital type I interferonopathy is selected from Aicardi-Goutières syndrome (AGS), Singleton-Merten 45 33049332.1 400807-032WO (218484)syndrome, proteasome-associated autoinflammatory syndromes, chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature (CANDLE), STING-associated vasculopathy with onset in infancy (SAVI), Japanese autoinflammatory syndrome with lipodystrophy (JASL), spondyloenchondrodysplasia (SPENCD), ISG15 deficiency, Ubiquitin- Specific Peptidase 18 deficiency (pseudo-TORCH syndrome), chronic atypical neurophilic dermatitis with lipodystrophy, DNA II deficiency, trichoheptoenteric syndrome 2, retinal vasculopathy with cerebral leukodystrophy, familial chilblain lupus, and X-linked reticulate pigmentary disorder (XLPDR). In another embodiment, the type I interferonopathy is an acquired disorder in the IFN system.

[0138] In one embodiment, the disclosure provides a method of treating an autoimmunedisease which results in an overproduction of interferon. In one aspect of this embodiment, the interferon expressed includes type I interferon. In another aspect of this embodiment, the autoimmune disease is associated with elevated LINE-1 activity and / or expression.

[0139] In certain embodiments, the autoimmune disorder is selected from the groupconsisting of achalasia, Addison’s disease, adult Still’s disease, agammaglobulinemia, alopecia areata, amyloidosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, antiphospholipid syndrome, autoimmune angioedema, autoimmune dysautonomia, autoimmune encephalitis, autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune myocarditis, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal & neuronal neuropathy (AMAN), Balo disease, Behcet’s disease, benign mucosal pemphigoid, bullous pemphigoid, Castleman disease (CD), celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic recurrent multifactorial osteomyelitis (CRMO), Churg-Strauss syndrome or eosinophilic granulomatosis, cicatricial pemphigoid, Cogan’s syndrome, cold agglutinin disease, complex regional pain syndrome (previously called reflex sympathetic dystrophy), congenital heart block, coxsackie myocarditis, CREST syndrome, Crohn’s disease, cutaneous lupus erythematosus (CLE), dermatitis herpetiformis, dermatomyositis, Devic’s disease (neuromyelitis optica), discoid lupus, Dressler’s syndrome, endometriosis, eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum, essential mixed cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, Goodpasture’s syndrome, granulomatosis with 46 33049332.1 400807-032WO (218484)polyangiitis, graft versus host disease, Graves’ disease, Guillain-Barre syndrome, Hashimoto’s thyroiditis, hemolytic anemia, Henoch-Schonlein purpura (HSP), herpes gestationis or pemphigold gestationis (PG), hidradenitis suppurativa (acne inversa), inflammatory bowel disease, IgA nephropathy, IgG4-related sclerosing disease, immune thrombocytopenic pupura (ITP), inclusion body myositis (IBM), interstitial cystitis (IC), juvenile arthritis, juvenile diabetes (type I diabetes), juvenile myositis (JM), Kawasaki disease, Lambert-Eaton syndrome, lichen planus, lichen sclerosus, ligneous conjunctivitis, linear IgA disease (LAD), lupus nephritis, lyme disease (chronic), Meniere’s disease, microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), multifocal motor neuropathy, multiple sclerosis, myasthenia gravis, myelin oligodendrocyte glycoprotein antibody disorder, myositis, narcolepsy, neonatal lupus, neutropenia, ocular cicatricial pemphigoid, optic neuritis, palindromic rheumatism, pediatric autoimmune neuropsychiatric disorders associated with streptococcus infections (PANDAS), paraneoplastic cerebellar degeneration (PCD), paroxysmal nocturnal hemoglobinuria (PNH), pars planitis (peripheral uveitis), Parsonage-Turner syndrome, pemphigus, peripheral neuropathy, perivenous encephalomyelitis, pernicious anemia (PA), POEMS syndrome, polyarteritis rheumatica, polymyositis, postmyocardial infarction syndrome, postpericardiotomy syndrome, primary biliary cholangitis, primary sclerosing cholangitis, progesterone dermatitis, progressive hemifacial atrophy (Parry Romberg syndrome), psoriasis, psoriatic arthritis, pure red cell aplasia, pyoderma gangrenosum, Raynoud’s phenomena, reactive arthritis, relapsing polychondritis, restless leg syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis (RA), sarcoidosis, Schmidt syndrome (autoimmune polyendocrine syndrome type II), scleritis, scleroderma, Sjögren’s disease, stiff person syndrome, Susac’s syndrome, sympathetic ophthalmia, systemic lupus erythematosus (SLE), Takayasu’s arteritis, thrombotic thrombocytopenic pupura, thyroid eye disease, Tolosa-Hunt syndrome, transverse myelitis, ulcerative colitis, undifferentiated connective tissue disease, uveitis, vasculitis, vitiligo, Vogt-Koyanagi-Harada disease, and warm autoimmune hemolytic anemia.

[0140] In certain embodiments, the autoimmune disorder is selected from Aicardi-Goutièressyndrome, rheumatoid arthritis, psoriasis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), graft versus host disease, scleroderma, type I diabetes, dermatomyositis, 47 33049332.1 400807-032WO (218484)inflammatory bowel disease, ulcerative colitis, Crohn’s disease, vasculitis, and Sjögren’s syndrome.

[0141] In certain embodiments, the autoimmune disorder is Aicardi-Goutières syndrome(AGS). In another embodiment, the autoimmune disorder is systemic lupus erythematosus (SLE). In another embodiment, the autoimmune disease is lupus nephritis. In a further embodiment, the autoimmune disease is cutaneous lupus erythematosus (CLE). In another embodiment, the autoimmune disease is dermatomyositis.

[0142] In certain embodiments, the autoimmune disorder is a type 1 interferonopathy. Incertain embodiments, the autoimmune disorder is type 1 diabetes, Aicardi-Goutières syndrome (AGS), systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), familial chilblain lupus, systemic sclerosis, STING-associated vasculopathy with onset in infancy (SAVI), Sjögren’s syndrome, or dermatomyositis. In certain embodiments, the immune disorder is a type 1 interferonopathy, type 1 diabetes, Aicardi-Goutières syndrome (AGS), systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), dermatomyositis, or Sjogren’s syndrome. In certain embodiments, the autoimmune disorder is systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), or familial chilblain lupus. In certain embodiments, the immune disorder is systemic lupus erythematosus (SLE).

[0143] In certain embodiments, the autoimmune disorder is type 1 diabetes. In certainembodiments, the autoimmune disorder is familial chilblain lupus. In certain embodiments, the autoimmune disorder is systemic sclerosis. In certain embodiments, the autoimmune disorder is STING-associated vasculopathy with onset in infancy (SAVI). In certain embodiments, the autoimmune disorder is Sjögren’s syndrome.

[0144] In certain embodiments, the autoimmune disorder is inflammatory bowel disease,Crohn’s disease, or ulcerative colitis. In certain embodiments, the autoimmune disorder is inflammatory bowel disease. In certain embodiments, the autoimmune disorder is Crohn’s disease. In certain embodiments, the autoimmune disorder is ulcerative colitis. In one embodiment, the autoimmune disorder is drug-induced colitis, such as colitis associated with the administration of checkpoint inhibitors to cancer patients. 48 33049332.1 400807-032WO (218484)

[0145] In certain embodiments, the autoimmune disorder is osteoarthritis, nonalcoholicsteatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cholestatic liver disease, sclerosing cholangitis, asthma, bronchitis, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary hypertension, pericarditis, gout, or myositis.

[0146] In certain embodiments, the autoimmune disorder is Reiter's syndrome, exfoliativepsoriatic dermatitis, pemphigus vulgaris, autoimmune uveitis, pulmonary hemosiderosis, amyloidosis, aphthous stomatitis, thyroiditis, gastritis, adrenalitis (Addison's disease), ovaritis, primary biliary cirrhosis, myasthenia gravis, gonadal failure, hypoparathyroidism, alopecia, malabsorption syndrome, pernicious anemia, hepatitis, hypopituitarism, diabetes insipidus, or sicca syndrome. Neurological Disorders

[0147] In certain embodiments, the disorder is a neurological disorder. In certainembodiments, the neurological disorder is Alzheimer’s disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis, Parkinson’s disease, Huntington’s disease, peripheral neuropathy, age- related macular degeneration, Creutzfeldt-Jacob disease, stroke, prion disease, frontotemporal dementia, Pick’s disease, progressive supranuclear palsy, spinocerebellar ataxias, Lewy body disease, dementia, multiple system atrophy, autism, other tauopathies, Fragile X-associated tremor / ataxia syndrome, epilepsy, bipolar disorder, schizophrenia, an anxiety disorder, or major depression. In certain embodiments, the neurological disorder is Alzheimer’s disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis, Parkinson’s disease, Huntington’s disease, or dementia. In another embodiment, the neurological disorder is ALS or progressive supranuclear palsy.

[0148] In certain embodiments, the neurological disorder is peripheral neuropathy, age-related macular degeneration, Creutzfeldt-Jacob disease, stroke, prion disease, frontotemporal dementia, Pick’s disease, progressive supranuclear palsy, spinocerebellar ataxias, Lewy body disease, dementia, multiple system atrophy, epilepsy, bipolar disorder, schizophrenia, an anxiety disorder, or major depression.

[0149] In certain embodiments, the neurological disorder is Alzheimer’s disease. In otherembodiments, the neurological disorder is amyotrophic lateral sclerosis (ALS). In another embodiment, the neurological disorder is multiple sclerosis. In a further embodiment, the 49 33049332.1 400807-032WO (218484)neurological disorder is Parkinson’s disease. In another embodiment, the neurological disorder is Huntington’s disease. In another embodiment, the neurological disorder is dementia. In certain embodiments, the neurological disorder is age-related macular degeneration. In a further embodiment, the neurological disorder is progressive supranuclear palsy. In certain embodiments, the neurological disorder is stroke.

[0150] There is compelling evidence that increased LINE-1 expression and activity contributeto neuroinflammation and neurodegeneration in chronic neurodegenerative diseases4,5,6.

[0151] First, LINE-1 expression is normally repressed in healthy somatic cells but can bereactivated in disease settings due to exogenous stress conditions and / or epigenetic changes, and with aging10, a major risk factor for PD. Genetic studies have demonstrated a correlation between increased LINE-1 insertion polymorphisms, a measure of LINE-1 activity, and PD progression, as observed in the Parkinson's Progression Marker Initiative (PPMI) dataset11,12. LINE-1 protein expression also increases with mitochondrial stress, a hallmark of PD, in neurons of both human and murine models13,14.

[0152] Second, as LINE-1 increases in tissues with age, including the brain, it may correlatewith increased inflammatory and senescent gene signatures15that have been suggested as pathophysiologic mechanisms of PD. The neuroinflammatory disease Aicardi-Goutières Syndrome (AGS) is driven by innate immune signaling through the cGAS-STING pathway, that can be induced by loss of the exonuclease TREX1. In a neuronal model of TREX1 loss, accumulation of cytoplasmic LINE-1 derived nucleic acids correlates with increased activation of the cGAS-STING pathway. This leads to neurotoxicity, that is rescued by treatment with LINE-1 NRTIs16.

[0153] Third, published work has shown the protective role of LINE-1 RT inhibition in bothin vitro and in vivo models of acute toxin-induced PD. Using oxidative stressors (such as hydrogen peroxide and 6-OHDA) in murine neuronal cultures or following intracranial injection, dopaminergic cell loss is prevented both by genetic knockdown of LINE-1 and by enzymatic inhibition with an NRTI14.

[0154] Fourth, NRTIs were originally developed as therapeutic treatments for HIV-1. Inepidemiological studies of HIV patients treated with NRTIs, lower incidences of multiple diseases, including neurodegenerative diseases such as PD, have been documented17,18,19,20,21. 50 33049332.1 400807-032WO (218484)While these HIV therapies were developed for potency against HIV-1 RT, we now know that many of these drugs are also weak to moderate LINE-1 RT inhibitors.

[0155] In ALS it has been shown that increased expression of repetitive elements, includingLINE-1, occurs in human cases. Experimental support includes association of de-condensation of LINE1 genomic loci and increased RNA expression with loss of nuclear TDP43 in post-mortem nuclei7. Elevation of retrotransposon expression including LINE-1 can be detected in post- mortem frontal and motor cortex samples of c9orf72-positive patients8and defines a molecular subclass of sporadic ALS associated with TDP43 pathology9. Putative ALS mouse models have also demonstrated the association of LINE-1 with neurodegenerative and inflammatory disease. In the SOD1 G93A transgenic model, Line-1 RNA and Orf1p protein expression is increased and maintained during disease progression in spinal cord neurons10. References 4. Frost B, Dubnau J. The Role of Retrotransposons and Endogenous Retroviruses in Age- Dependent Neurodegenerative Disorders. Annu Rev Neurosci.2024 5. Copley KE, Shorter J. Repetitive elements in aging and neurodegeneration. Trends in Genetics.2023 6. Ravel-Godreuil C, Znaidi R, Bonnifet T, Joshi RL, Fuchs J. Transposable elements as new players in neurodegenerative diseases. FEBS Lett.2021 10. Bonnifet T, Sinnassamy S, Massiani-Beaudoin O, Mailly P, Monnet H, Loew D, Lombard B, Servant N, Joshi RL, Fuchs J. Steady-state neuron-predominant LINE-1 encoded ORF1p protein and LINE-1 RNA increase with aging in the mouse and human brain. eLife.2024 11. Fröhlich A, Pfaff AL, Bubb VJ, Quinn JP, Koks S. Reference LINE-1 insertion polymorphisms correlate with Parkinson’s disease progression and differential transcript expression in the PPMI cohort. Sci Rep.2023 12. Kõks S, Pfaff AL, Singleton LM, Bubb VJ, Quinn JP. Non-reference genome transposable elements (TEs) have a significant impact on the progression of the Parkinson's disease. Exp Biol Med (Maywood).2022 13. Baeken MW, Moosmann B, Hajieva P. Retrotransposon activation by distressed mitochondria in neurons. BBRC.2020 51 33049332.1 400807-032WO (218484)14. Blaudin de Thé FX, Rekaik H, Peze-Heidsieck E, Massiani-Beaudoin O, Joshi RL, Fuchs J, Prochiantz A. Engrailed homeoprotein blocks degeneration in adult dopaminergic neurons through LINE-1 repression. EMBO J.2018 15. De Cecco M, Ito T, Petrashen AP, Elias AE, Skvir NJ, Criscione SW, Caligiana A, Brocculi G, Adney EM, Boeke JD, Le O, Beauséjour C, Ambati J, Ambati K, Simon M, Seluanov A, Gorbunova V, Slagboom PE, Helfand SL, Neretti N, Sedivy JM. L1 drives IFN in senescent cells and promotes age-associated inflammation. Nature 2019 16. Thomas CA, Tejwani L, Trujillo CA, Negraes PD, Herai RH, Mesci P, Macia A, Crow YJ, Muotri AR. Modeling of TREX1-Dependent Autoimmune Disease using Human Stem Cells Highlights L1 Accumulation as a Source of Neuroinflammation. Cell Stem Cell.2017 17. Coghill AE, Engels EA, Schymura MJ, Mahale P, Shiels MS. Risk of Breast, Prostate, and Colorectal Cancer Diagnoses Among HIV-Infected Individuals in the United States. J Natl Cancer Inst.2018 18. Tan J, Pina A, Borges-Costa J. Skin Diseases in the Era of Highly Active Antiretroviral Therapy: A Retrospective Study of 534 Patients. J Int Assoc Provid AIDS Care.2018 19. Gold J, Goldacre R, Maruszak H, Giovannoni G, Yeates D, Goldacre M. HIV and lower risk of multiple sclerosis: beginning to unravel a mystery using a record-linked database study. J Neurol Neurosurg Psychiatry.2015 20. Yang JJ, Tsai MS, Sun HY, Hsieh SM, Chen MY, Sheng WH, Chang SC. Autoimmune diseases-related arthritis in HIV-infected patients in the era of highly active antiretroviral therapy. J Microbiol Immunol Infect.2015 21. Siangphoe U, Archer KJ, Nguyen C, Lee KR. Associations of antiretroviral therapy and comorbidities with neurocognitive outcomes in HIV-1-infected patients. AIDS.2020 Viral Infection

[0156] In certain embodiments, the viral infection is an infection by humanimmunodeficiency viruses 1 or 2 (HIV-1 or HIV-2), human T-cell leukemia viruses 1 or 2 (HTLV-1 or HTLV-2), respiratory syncytial virus (RSV), human papilloma virus (HPV), adenovirus, hepatitis B virus (HBV), hepatitis C virus (HCV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), cytomegalovirus (CMV), herpes simplex viruses 1 or 2 (HSV-1 or HSV-2), human herpes virus 8 (HHV-8, also known as Kaposi's sarcoma-associated virus), or a 52 33049332.1 400807-032WO (218484)flavivirus selected from Yellow Fever virus, Dengue virus, Japanese Encephalitis, and West Nile virus.

[0157] In certain embodiments, the viral infection is an infection by humanimmunodeficiency viruses 1 or 2 (HIV-1 or HIV-2). In certain embodiments, the viral infection is an infection by human immunodeficiency virus 1 (HIV-1). In certain embodiments, the viral infection is an infection by human immunodeficiency virus 2 (HIV-2). In certain embodiments, the viral infection is an infection by human T-cell leukemia viruses 1 or 2 (HTLV-1 or HTLV- 2). In certain embodiments, the viral infection is an infection by respiratory syncytial virus (RSV). In certain embodiments, the viral infection is an infection by human papilloma virus (HPV). In certain embodiments, the viral infection is an infection by adenovirus. In certain embodiments, the viral infection is an infection by hepatitis B virus (HBV). In certain embodiments, the viral infection is an infection by hepatitis C virus (HCV). In certain embodiments, the viral infection is an infection by Epstein-Barr virus (EBV). In certain embodiments, the viral infection is an infection by varicella zoster virus (VZV). In certain embodiments, the viral infection is an infection by cytomegalovirus (CMV). In certain embodiments, the viral infection is an infection by herpes simplex viruses 1 or 2 (HSV-1 or HSV-2). In certain embodiments, the viral infection is an infection by human herpes virus 8 (HHV-8, also known as Kaposi's sarcoma-associated virus). In certain embodiments, the viral infection is an infection by a flavivirus selected from Yellow Fever virus, Dengue virus, Japanese Encephalitis, and West Nile virus.

[0158] In certain embodiments, the viral infection is an infection by an adenovirus. In certainembodiments, the viral infection is an infection by a herpesvirus. In certain embodiments, the viral infection is an infection by a poxvirus. In certain embodiments, the viral infection is an infection by a parvovirus. In certain embodiments, the viral infection is an infection by a reovirus. In certain embodiments, the viral infection is an infection by a picornavirus. In certain embodiments, the viral infection is an infection by a rhinovirus or enterovirus. In certain embodiments, the viral infection is an infection by a togavirus. In certain embodiments, the viral infection is an infection by an orthomyxovirus. In certain embodiments, the viral infection is an infection by a rhabdovirus. In certain embodiments, the viral infection is an infection by a retrovirus. In certain embodiments, the viral infection is an infection by a hepadnavirus. 53 33049332.1 400807-032WO (218484)

[0159] In certain embodiments, the viral infection is an infection by a coronavirus. In someembodiments, the coronavirus is an alpha, beta, gamma, or delta coronavirus. In certain embodiments, the viral infection is an infection by a coronavirus selected from 229E (alpha coronavirus), NL63 (alpha coronavirus), OC43 (beta coronavirus), HKU1 (beta coronavirus), MERS-CoV (beta coronavirus), SARS-CoV (beta coronavirus), and SARS-CoV-2 (coronavirus disease 2019, or COVID-19).

[0160] In certain embodiments, the viral infection is an infection by an influenza virus. Incertain embodiments, the viral infection is an infection by a type A or type B influenza virus. In certain embodiments, the viral infection is an infection by an influenza virus selected from H5N1, H1N1, and H3N2.

[0161] In certain embodiments, the viral infection is an infection by a poliovirus. In certainembodiments, the viral infection is an infection by a type 1 poliovirus. In certain embodiments, the viral infection is an infection by a type 2 poliovirus. In certain embodiments, the viral infection is an infection by a type 3 poliovirus. Subjects

[0162] In certain embodiments, the subject has (i) expression of LINE1 RNA, LINE1 ORF1polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase.

[0163] In certain embodiments, the subject has (i) expression of LINE1 RNA, LINE1 ORF1polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase. In certain embodiments, the subject has (i) elevated expression of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) elevated activity of LINE1 reverse transcriptase. In certain embodiments, the subject has expression of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide. In certain embodiments, the subject has expression of LINE1 RNA. In certain embodiments, the subject has expression of LINE1 ORF1 polypeptide. In certain embodiments, the subject has expression of LINE1 ORF2 polypeptide. In certain embodiments, the subject has activity of LINE1 reverse transcriptase. 54 33049332.1 400807-032WO (218484)

[0164] In certain embodiments, the subject has elevated (i) levels of LINE1 RNA, LINE1ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase.

[0165] In certain embodiments, the subject has elevated (i) levels of LINE1 RNA, LINE1ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) activity of LINE1 reverse transcriptase. In certain embodiments, the subject has elevated levels of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide. In certain embodiments, the subject has elevated levels of LINE1 RNA. In certain embodiments, the subject has elevated levels of LINE1 ORF1 polypeptide. In certain embodiments, the subject has elevated levels of LINE1 ORF2 polypeptide. In certain embodiments, the subject has elevated activity of LINE1 reverse transcriptase.

[0166] In certain embodiments, the subject is a human. In certain embodiments, the subject isan adult human. In certain embodiments, the subject is a pediatric human. In certain embodiments, the subject is a companion animal. In certain embodiments, the subject is a canine, feline, or equine. Uses of Compounds

[0167] Another aspect of the disclosure provides for the use of a compound described herein(such as a compound of Formula I, or other compounds in Section I) for treating a medical disorder, such as a medical disorder described herein.

[0168] Another aspect of the disclosure provides for the use of a compound described herein(such as a compound of Formula I), in the manufacture of a medicament. In certain embodiments, the medicament is for treating a disorder described herein, such as cancer, an autoimmune disorder, and / or a neurological disorder. In certain embodiments the disorder is aging, or a disease associated with aging. III. Methods of Inhibiting LINE1 Reverse Transcriptase Activity

[0169] Another embodiment of the disclosure provides a method of inhibiting LINE1 reversetranscriptase activity in a subject suffering from a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. The method comprises contacting a LINE1 reverse transcriptase with an effective amount of a compound of Formula I, in order to 55 33049332.1 400807-032WO (218484)inhibit the activity of said LINE1 reverse transcriptase. In certain embodiments, the method comprises contacting a LINE1 reverse transcriptase with an effective amount of a compound of Formula II, in order to inhibit the activity of said LINE1 reverse transcriptase. Additional compounds useful in the method are further described in Section I. In certain embodiments, the method includes administration of any compound in Table 1, or a pharmaceutically acceptable salt thereof, and / or a pharmaceutical composition comprising the same. In certain embodiments, the method includes administration of any compound in Table 2, or a pharmaceutically acceptable salt thereof, and / or a pharmaceutical composition comprising the same.

[0170] Another embodiment of the disclosure provides a method of inhibiting LINE1 reversetranscriptase activity in a subject, the method comprising contacting a LINE1 reverse transcriptase with an effective amount of a compound according to Formula I, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound according to Formula I, in order to inhibit the activity of said LINE1 reverse transcriptase.

[0171] Another embodiment of the disclosure provides a method of inhibiting LINE1 reversetranscriptase activity in a subject, the method comprising contacting a LINE1 reverse transcriptase with an effective amount of a compound according to Formula II, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound according to Formula II, in order to inhibit the activity of said LINE1 reverse transcriptase.

[0172] In certain embodiments, the disorder is a disorder defined by one of the embodimentsdescribed above, such as cancer, an autoimmune disorder, and / or a neurological disorder. In certain embodiments the disorder is aging, or a disease associated with aging.

[0173] Another embodiment of the disclosure provides a method of inhibiting LINE1 reversetranscriptase activity in a subject suffering from a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. The method comprises contacting a LINE1 reverse transcriptase with an effective amount of a compound described herein, such as a compound of Formula I or a compound in Table 1 in order to inhibit the activity of said LINE1 reverse transcriptase.

[0174] In certain embodiments, the disorder is a disorder defined by one of the embodimentsdescribed above, such as cancer, an autoimmune disorder, and / or a neurological disorder. 56 33049332.1 400807-032WO (218484)

[0175] In certain embodiments, the particular compound is a compound described for any ofthe embodiments herein, in any and all combinations of the various embodiments, and aspects of embodiments, described. For example, in certain embodiments, the compound is a compound of Formula I, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound in Table 1 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is administered in a pharmaceutical composition comprising the compound and a carrier, excipient, and / or vehicle, as further described in Section IV, below.

[0176] Another embodiment of the disclosure provides a method of inhibiting LINE1 reversetranscriptase activity. The method comprises contacting a LINE1 reverse transcriptase with a therapeutically effective amount of a compound described herein, such as a compound in Table 1 to inhibit the activity of said LINE1 reverse transcriptase. In certain embodiments, the particular compound is a compound described for any of the embodiments herein, in any and all combinations of the various embodiments, and aspects of embodiments, described. For example, in certain embodiments, the compound is a compound of Formula I, or a pharmaceutically acceptable salt thereof.

[0177] In certain embodiments, the compound is administered in a pharmaceuticalcomposition comprising the compound and a carrier, excipient, and / or vehicle, as further described in Section IV, below.

[0178] In certain embodiments, the disorder is a disorder defined by one of the embodimentsdescribed in Section I, above, such as cancer, an autoimmune disorder, and / or a neurological disorder. In certain embodiments the disorder is aging, or a disease associated with aging.

[0179] In certain embodiments, the method further comprises inhibiting LINE1 reversetranscriptase activity in the subject.

[0180] In certain embodiments, the disorder is a disorder defined by one of the embodimentsdescribed above, such as cancer, an autoimmune disorder, and / or a neurological disorder.

[0181] In certain embodiments, the particular compound is a compound described for any ofthe embodiments herein, in any and all combinations of the various embodiments, and aspects of embodiments, described. In certain embodiments, the compound is administered in a 57 33049332.1 400807-032WO (218484)pharmaceutical composition comprising the compound and a carrier, excipient, and / or vehicle, as further described in Section IV, below.

[0182] One aspect of the disclosure provides compounds having a superior combination ofproperties including potent inhibitory activity towards LINE1 reverse transcriptase, selectivity for inhibiting LINE1 reverse transcriptase, and superior physical properties that render the compounds well-suited for use as an orally administered medicine. Selectivity for inhibiting LINE1 reverse transcriptase can be characterized according to ability of the compounds to inhibit LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNApolymerases (e.g., ^, ^ and ^). In part because inhibition of DNA polymerases, such as DNApolymerase gamma, can result in toxicity in the form of mitotoxicity, compounds demonstrating potent inhibitory activity towards LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNA polymerases are an important discovery and significant scientific advance. The potent inhibitory activity towards LINE1 reverse transcriptase in combination with low inhibitory activity towards DNA polymerases contributes to a high therapeutic index for subject compounds, thereby providing a superior performance profile for the compound in medical therapy.

[0183] Another aspect of the disclosure provides compounds having a superior combinationof properties including potent inhibitory activity towards LINE1 reverse transcriptase, selectivity for inhibiting LINE1 reverse transcriptase, potent inhibition of pathogenic interferon response in inflammatory tissues, and superior physical properties that render the compounds well-suited for use as an orally administered medicine. As described above, selectivity for inhibiting LINE1 reverse transcriptase can be characterized according to ability of the compounds to inhibit LINE1 reverse transcriptase while having substantially less inhibitory activity towards DNApolymerases (e.g., ^, ^ and ^). In part because inhibition of DNA polymerase, such as DNApolymerase gamma, can result in toxicity in the form of mitotoxicity, compounds demonstrating potent inhibitory activity towards LINE1 reverse transcriptase while having substantially lessinhibitory activity towards DNA polymerases (e.g., ^, ^ and ^) are an important discovery andsignificant scientific advance. Compounds having potent inhibition of pathogenic interferon response in inflammatory tissues are useful for treating cancer, autoimmune disease (e.g., SLE and CLE), neurological disorders, aging, and diseases associated with aging. The potent 58 33049332.1 400807-032WO (218484)inhibitory activity towards LINE1 reverse transcriptase in combination with low inhibitoryactivity towards DNA polymerases (e.g., ^, ^ and ^) contributes to a high therapeutic index forsubject compounds, thereby providing a superior performance profile for the compound in medical therapy.

[0184] In certain embodiments, the compound is administered in a pharmaceuticalcomposition comprising the compound and a carrier, excipient, and / or vehicle, as further described in Section IV, below.

[0185] Compounds may be tested for ability to inhibit LINE1 reverse transcriptase activity,for example, as described in the Examples. IV. Pharmaceutical Compositions and Dosing Considerations

[0186] As indicated above, the disclosure provides pharmaceutical compositions, whichcomprise a therapeutically effective amount of one or more of the compounds described above, formulated together with one or more pharmaceutically acceptable carriers, adjuvant, and / or vehicle. The pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; (3) topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin; (4) intravaginally or intrarectally, for example, as a pessary, cream or foam; (5) sublingually; (6) ocularly; (7) transdermally; or (8) nasally.

[0187] In certain embodiments, the disclosure provides a pharmaceutical compositioncomprising a compound described herein (e.g., a compound of Formula I) and a pharmaceutically acceptable carrier.

[0188] The phrase “therapeutically effective amount” as used herein means that amount of acompound, material, or composition comprising a compound of the present disclosure which is effective for producing some desired therapeutic effect in at least a sub-population of cells in an animal at a reasonable benefit / risk ratio applicable to any medical treatment. 59 33049332.1 400807-032WO (218484)

[0189] The phrase “pharmaceutically acceptable” is employed herein to refer to thosecompounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0190] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0191] Examples of pharmaceutically acceptable antioxidants include: (1) water solubleantioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0192] Formulations of the present disclosure include those suitable for oral, nasal, topical(including buccal and sublingual), rectal, vaginal and / or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated and / or the particular mode of administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0193] Methods of preparing these formulations or compositions include the step of bringinginto association a compound of the present disclosure with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and 60 33049332.1 400807-032WO (218484)intimately bringing into association a compound of the present disclosure with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0194] Formulations of the disclosure suitable for oral administration may be in the form ofcapsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound of the present disclosure as an active ingredient. A compound of the present disclosure may also be administered as a bolus, electuary or paste.

[0195] In solid dosage forms of the disclosure for oral administration (capsules, tablets, pills,dragees, powders, granules, troches and the like), the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds and surfactants, such as poloxamer and sodium lauryl sulfate; (7) wetting agents, such as, for example, cetyl alcohol, glycerol monostearate, and non-ionic surfactants; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, zinc stearate, sodium stearate, stearic acid, and mixtures thereof; (10) coloring agents; and (11) controlled release agents such as crospovidone or ethyl cellulose. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-shelled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0196] A tablet may be made by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or 61 33049332.1 400807-032WO (218484)hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0197] The tablets, and other solid dosage forms of the pharmaceutical compositions of thepresent disclosure, such as dragees, capsules, pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be formulated for rapid release, e.g., freeze-dried. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0198] Liquid dosage forms for oral administration of the compounds of the disclosureinclude pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0199] Besides inert diluents, the oral compositions can also include adjuvants such aswetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents. 62 33049332.1 400807-032WO (218484)

[0200] Suspensions, in addition to the active compounds, may contain suspending agents as,for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0201] Formulations of the pharmaceutical compositions of the disclosure for rectal orvaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the disclosure with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at RT, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.

[0202] Formulations of the present disclosure which are suitable for vaginal administrationalso include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0203] Dosage forms for the topical or transdermal administration of a compound of thisdisclosure include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically-acceptable carrier, and with any preservatives, buffers, or propellants which may be required.

[0204] The ointments, pastes, creams and gels may contain, in addition to an activecompound of this disclosure, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0205] Powders and sprays can contain, in addition to a compound of this disclosure,excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0206] Transdermal patches have the added advantage of providing controlled delivery of acompound of the present disclosure to the body. Such dosage forms can be made by dissolving 63 33049332.1 400807-032WO (218484)or dispersing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

[0207] Ophthalmic formulations, eye ointments, powders, solutions and the like, are alsocontemplated as being within the scope of this disclosure.

[0208] Pharmaceutical compositions of this disclosure suitable for parenteral administrationcomprise one or more compounds of the disclosure in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0209] Examples of suitable aqueous and nonaqueous carriers which may be employed in thepharmaceutical compositions of the disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0210] These compositions may also contain adjuvants such as preservatives, wetting agents,emulsifying agents and dispersing agents. Prevention of the action of microorganisms upon the subject compounds may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

[0211] When the compounds of the present disclosure are administered as pharmaceuticals, tohumans and animals, they can be given per se or as a pharmaceutical composition containing, for 64 33049332.1 400807-032WO (218484)example, 0.1 to 99% (more preferably, 10 to 30%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0212] The preparations of the present disclosure may be given orally, parenterally, topically,or rectally. They are of course given in forms suitable for each administration route. For example, they are administered in tablets or capsule form, by injection, inhalation, eye lotion, ointment, suppository, etc. administration by injection, infusion or inhalation; topical by lotion or ointment; and rectal by suppositories. Oral administrations are preferred.

[0213] The phrases “parenteral administration” and “administered parenterally” as usedherein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

[0214] The phrases “systemic administration,” “administered systemically,” “peripheraladministration” and “administered peripherally” as used herein mean the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient’s system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

[0215] These compounds may be administered to humans and other animals for therapy byany suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginally, parenterally, intracisternally and topically, as by powders, ointments or drops, including buccally and sublingually.

[0216] Actual dosage levels of the active ingredients in the pharmaceutical compositions ofthis disclosure may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0217] The selected dosage level will depend upon a variety of factors including the activityof the particular compound of the present disclosure employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the 65 33049332.1 400807-032WO (218484)particular compound being employed, the rate and extent of absorption, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0218] A physician or veterinarian having ordinary skill in the art can readily determine andprescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the disclosure employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0219] In general, a suitable daily dose of a compound of the disclosure will be that amountof the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above. Preferably, the compounds are administered at about 0.01 mg / kg to about 200 mg / kg, more preferably at about 0.1 mg / kg to about 100 mg / kg, even more preferably at about 0.5 mg / kg to about 50 mg / kg. When the compounds described herein are co-administered with another agent (e.g., as sensitizing agents), the effective amount may be less than when the agent is used alone.

[0220] If desired, the effective daily dose of the active compound may be administered astwo, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. Preferred dosing is one administration per day. EXAMPLES

[0221] The invention now being generally described, will be more readily understood byreference to the following examples, which are included merely for purposes of illustration of certain aspects and embodiments of the present invention, and are not intended to limit the invention. EXAMPLE 1 - Synthesis of (2R,3S,5R)-5-(4-amino-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate (I-1) 66 33049332.1 400807-032WO (218484), , -5-(hydroxymethyl) tetrahydrofuran-2-yl) pyrimidin-2(1H)-one (9) (See, WO 2023 / 178133) (6 g, 21.8 mmol) in pyridine (50 mL) were added DMAP (3.79 g, 43.5 mmol) and isobutyric anhydride (20.7 g, 130 mmol) at 0 °C under an inert atmosphere. The resulting mixture was stirred for 16 h at 50 °C. The resulting mixture was cooled to room temperature and concentrated under reduced pressure. The resulting mixture was diluted with water at 0 °C and extracted with EA (3 x 250 mL). The combined organic layers were washed with saturated NaHCO3solution and citric acid solution, dried over anhydrous Na2SO4 and concentrated under reduced pressure. Purification by silica gel column chromatography, eluted with PE / EA (3:1) afforded (2R,3S,5R)- 2-(chloromethyl)-5-(4-isobutyramido-2-oxopyrimidin-1(2H)-yl)-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate (9 g, 77%) as a white solid. LC / MS ES, m / z): 486 / 488 [M+H]+.

[0223] Step 2: A solution of (2R,3S,5R)-2-(chloromethyl)-5-(4-isobutyramido-2-oxopyrimidin-1(2H)-yl)-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate (10) (9 g, 18.5 mmol) in EtOH (300 mL) and MeOH (300 mL) was stirred for 5 days at 100 ℃ under a nitrogen atmosphere. The resulting mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatographywith DCM / MeOH (10:1) to afford the crude product. The crude product was re- crystallized from MTBE / Hexane (10:1) to afford (2R,3S,5R)-5-(4-amino-2-oxopyrimidin-1(2H)- 67 33049332.1 400807-032WO (218484)yl)-2-(chloromethyl)-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate (6.61 g, 86%) as a white solid.

[0224] LC / MS- (ES, m / z): 416 / 418 [M+H]+; 99.9% purity. Conditions for the LC / MS:(Column: Shim‐pack Scepter C18, 33*3.0 mm, 3.0 μm; Mobile Phase A: water / 5 mM NH4HCO3, Mobile Phase B: ACN; Flow rate: 1.50 mL / min; Gradient: 10% B to 95% B in 1.20 min, 95% B to 95% B in 0.60 min, 95% B to 10% B in 0.02 min; Wavelength: 254 / 220 nm; RT1(min): 0.884).

[0225] 1H NMR (400 MHz, DMSO-d6) δ 7.61 (d, J = 7.4 Hz, 1H), 7.24 (d, J = 14.1 Hz, 2H),6.25 (t, J = 6.9 Hz, 1H), 5.76 (d, J = 7.4 Hz, 1H), 5.50 (dd, J = 6.8, 3.6 Hz, 1H), 4.35 – 4.23 (m, 2H), 3.95 – 3.72 (m, 2H), 2.70 – 2.52 (m, 3H), 2.44 – 2.30 (m, 1H), 1.20 – 1.05 (m, 12H). EXAMPLE 2 – Synthesis of [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethyl- 3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (I-2)

[0226] Step 1: To a solution of 4-amino-1-[(2R,4S,5R)-5-ethyl-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-5-fluoropyrimidin-2-one (11) (See, WO 2023 / 178133) (100 mg,0.36 mmol) and DMAP (89 mg, 0.73 mmol) in ACN (4 mL) was added 2-methylpropanoyl 2- methylpropanoate (116 mg, 0.73 mmol) dropwise at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at 0 °C was after which point, it was diluted with EtOAc. The solution was washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 28% B to 58% B in 10 min; wavelength: 254 nm / 220 nm; RT 1 (min): 9.22)) to afford [(2R,3S,5R)-5-(4-amino-5- 68 33049332.1 400807-032WO (218484)fluoro-2-oxopyrimidin-1-yl)-2-ethyl-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2- methylpropanoate (45.9 mg, 30 %) as a white solid.

[0227] LC / MS (ES, m / z): 414 (M+H+). Conditions for the HPLC: (Column: Shim‐packScepter C18, 33*3.0 mm, 3.0 μm; Mobile phase A:5 mM NH4HCO3 / 10%ACN, 90%water, Mobile PhaseB: CAN; Flow rate: 1.5 mL / min; Gradient: 0% B to 0% B in 0.01 min, 0% B to 60% B in 0.69 min, 60% B to 90% B in 0.60 min,90% B to 90% B in 0.50min; 90% B to 0% B in 0.03min; Wavelength: 254 nm; RT1 (min): 1.528).

[0228] 1H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 7.0 Hz, 1H), 7.59 (br s, 1H), 6.09 (t, J =6.4 Hz, 1H), 5.37 – 5.35 (m, 1H), 4.20 – 4.12 (m, 2H), 2.68 – 2.55 (m, 2H), 2.47 – 2.45 (m, 1H), 2.35 – 2.30 (m, 1H), 1.73 – 1.68 (m, 2H), 1.59 – 1.53 (m, 1H), 1.14 – 1.09 (m, 12H), 0.88 (t, J = 7.6 Hz, 3H). EXAMPLE 3 – Synthesis of ((2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-3-hydroxytetrahydrofuran-2-yl) methyl 2-propylpentanoate (I-3)84)

[0229] Step 1: To a stirred solution of 4-amino-1-((2R,4S,5R)-5-(chloromethyl)-4-hydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl)-5-fluoropyrimidin-2(1H)-one (12) (See, WO 2023 / 178133 A1) (500 mg, 1.71 mmol) and imidazole (579 mg, 8.51 mmol) in DMF (5 mL) was added TBS-Cl (769 mg, 5.11 mmol) at rt under an inert atmosphere. The resulting mixture was stirred for 16 h at rt while being monitored by LC / MS. The reaction was then quenched by adding water (20 mL) and extracted with EtOAc (20 mL). The combined organic extracts were washed with water (2x20 mL), brine (2x20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by Prep-TLC (EA) to afford 4-amino-1-((2R,4S,5R)-4-((tert-butyldimethylsilyl) oxy)-5-(((tert- butyldimethylsilyl) oxy) methyl)-5-(chloromethyl) tetrahydrofuran-2-yl)-5-fluoropyrimidin- 2(1H)-one (550 mg, 62%) as a white solid. LC-MS- (ES, m / z): 522 / 524 [M+H]+.

[0230] Step 2: To a stirred solution of 4-amino-1-((2R,4S,5R)-4-((tert-butyldimethylsilyl)oxy)-5-(((tert-butyldimethylsilyl) oxy) methyl)-5-(chloromethyl) tetrahydrofuran-2-yl)-5- fluoropyrimidin-2(1H)-one (13) (550 mg, 1.05 mmol) in DCM (10 mL) was treated with MMTrCl (390 mg, 1.26 mmol) and collidine (382 mg, 3.15 mmol) at rt under an inert atmosphere followed by the addition of AgNO3 (357 mg, 2.11 mmol) portionwise at rt. The mixture was heated to 60 °C for 16 h at under an inert atmosphere. The resulting mixture was cooled to rt and concentrated in vacuo. The residue was purified by Prep-TLC (PE / EA=4:1) to afford 1-((2R,4S,5R)-4-((tert-butyldimethylsilyl) oxy)-5-(((tert-butyldimethylsilyl) oxy) methyl)- 5-(chloromethyl) tetrahydrofuran-2-yl)-5-fluoro-4-(((4-methoxyphenyl) diphenylmethyl) amino) pyrimidin-2(1H)-one (810 mg, 97%) as a yellow solid. LC / MS (ES, m / z): 792 / 794 [M-H]-.

[0231] Step 3: To a stirred solution of 1-((2R,4S,5R)-4-((tert-butyldimethylsilyl) oxy)-5-(((tert-butyldimethylsilyl) oxy) methyl)-5-(chloromethyl) tetrahydrofuran-2-yl)-5-fluoro-4-(((4- methoxyphenyl) diphenylmethyl) amino) pyrimidin-2(1H)-one (14) (810 mg, 1.02 mmol) in THF (10 mL) was added TBAF (3.4 mL, 3.39 mmol, 1M in THF) at rt. The resulting mixturewas stirred overnight at rt under an inert atmosphere. The resulting mixture was concentratedunder reduced pressure. The residue was purified by Prep-TLC (EA) to afford 1-((2R,4S,5R)-5- (chloromethyl)-4-hydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl)-5-fluoro-4-(((4- methoxyphenyl) diphenyl methyl) amino) pyrimidin-2(1H)-one (400 mg, 60%) as an off-white solid. LC / MS- (ES, m / z): 564 / 566 [M-H]-. 70 33049332.1 400807-032WO (218484)

[0232] Step 4: A solution of 1-((2R,4S,5R)-5-(chloromethyl)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-5-fluoro-4-(((4-methoxyphenyl) diphenyl methyl) amino) pyrimidin- 2(1H)-one (15) (150 mg, 260 µmol) in acetonitrile (3 mL) was treated with DMAP (97 mg, 790 µmol) at rt under an inert atmosphere followed by the addition of 2-propylpentanoyl chloride (43 mg, 0.26 mmol) portionwise at rt. The mixture was stirred for 16 h after which point, it was concentrated under reduced pressure. The residue was purified by Prep-TLC (DCM / MeOH=20:1) to afford ((2R,3S,5R)-2-(chloromethyl)-5-(5-fluoro-4-(((4-methoxyphenyl) diphenylmethyl) amino)-2-oxopyrimidin-1(2H)-yl)-3-hydroxytetrahydrofuran-2-yl) methyl 2- propylpentanoate (60 mg, 33%) as a light yellow solid. LC / MS (ES, m / z): 690 / 692 [M-H]-.

[0233] Step 5: A solution of ((2R,3S,5R)-2-(chloromethyl)-5-(5-fluoro-4-(((4-methoxy-phenyl)diphenylmethyl) amino)-2-oxopyrimidin-1(2H)-yl)-3-hydroxytetrahydrofuran-2-yl) methyl 2-propylpentanoate (16) (20 mg, 20 µmol) in CH3COOH (5 mL, 80% in H2O) was stirred at rt for 24 h under an inert atmosphere. The resulting mixture was concentrated in vacuo. The residue was purified by following conditions (Column: XBridge Prep OBD RP18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 40% B to 60% B in 10 min; Wavelength: 254 nm / 220 nm; RT 1 (min): 7.72). The desired fractions were collected and concentrated in vacuo then re-dissolved in ACN and H2O, prior to being lyophilized to afford ((2R,3S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-3-hydroxytetrahydrofuran-2-yl) methyl 2- propylpentanoate (7.4 mg, 20%) as a white solid.

[0234] LC / MS (ES, m / z): 420 / 422 [M+H]+. Conditions for the LC / MS: (Column: HALOC182.0 um 3.0*30mm; Mobile Phase A: water / 0.05%TFA; B: ACN / 0.05%TFA; Flow rate: 1.50 mL / min; Gradient: 30% B to 60% B in 1.70 min, 60% B to 100% B in 0.80 min, 100% B to 100% B in 0.50 min, 100% B to 5% B in 0.03 min; Wavelength: 254 nm; RT 1 (min): 0.750).

[0235] 1H NMR (400 MHz, DMSO-d6) δ7.85 (s, 1H), 7.78 (d, J = 7.0 Hz, 1H), 7.60 (s, 1H),6.27 (m, 1H), 5.68 (d, J = 4.9 Hz, 1H), 4.35 – 4.25 (m, 2H), 4.20 (d, J = 11.6 Hz, 1H), 3.85 –3.72 (m, 2H), 2.47 – 2.30 (m, 2H), 2.34 – 2.23 (m, 1H), 1.59 – 1.35 (m, 4H), 1.25 (d, J = 7.4 Hz, 4H), 0.86 – 0.82 (m, 6H). 71 33049332.1 400807-032WO (218484)EXAMPLE 4 - Synthesis of (2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-2-(((2-propylpentanoyl)oxy)methyl)tetrahydrofuran-3-yl 2- propylpentanoate (I-4)

[0236] Step 1: To a solution of 1-((2R,4S,5R)-5-(chloromethyl)-4-hydroxy-5-(hydroxyethyl)tetrahydrofuran-2-yl)-5-fluoro-4-(((4-methoxyphenyl) diphenylmethyl) amino) pyrimidin-2(1H)-one (15) (See, Example 3) (80 mg, 140 µmol) in acetonitrile (2 mL) was added DMAP at 0°Cfollowed by the addition of 2-propylpentanoyl chloride (51 mg, 420 µmol) portionwise under an inert atmosphere. The mixture was stirred for 15 min prior to being concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EA=3:1) to afford (2R,3S,5R)-2- (chloromethyl)-5-(5-fluoro-4-(((4-methoxyphenyl) diphenylmethyl) amino)-2-oxopyrimidin- 1(2H)-yl)-2-(((2-propylpentanoyl) oxy) methyl) tetrahydrofuran-3-yl 2-propylpentanoate (60 mg, 52%) as a light yellow oil. LC / MS (ES, m / z): 816 / 818 [M-H]-.

[0237] Step 2: A solution of (2R,3S,5R)-2-(chloromethyl)-5-(5-fluoro-4-(((4-methoxyphenyl)diphenylmethyl) amino)-2-oxopyrimidin-1(2H)-yl)-2-(((2-propylpentanoyl) oxy) methyl) 72 33049332.1 400807-032WO (218484)tetrahydrofuran-3-yl 2-propylpentanoate (17) (60 mg, 70 µmol) in CH3COOH (5 mL, 80% in H2O) was stirred at rt for 24 h under an inert atmosphere. The resulting mixture was concentrated in vacuo. The residue was purified by following conditions (Column: XBridge Prep OBD RP18 Column, 30*150 mm, 5 μm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 35% B to 55% B in 10 min; Wavelength: 254 nm / 220 nm; RT 1 (min): 8.45). The fraction was collected and concentrated in vacuo, the residue was re- dissolved in ACN and H2O, and then was lyophilized to afford (2R,3S,5R)-5-(4-amino-5-fluoro- 2-oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-2-(((2-propylpentanoyl) oxy) methyl) tetrahydrofuran-3-yl 2-propylpentanoate (4.1 mg, 10%) as a grey solid. LC / MS (ES, m / z): 546 / 548 [M+H]+. EXAMPLE 5 - Synthesis of ((2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-3-hydroxytetrahydrofuran-2-yl) methyl (E)-octadec-9-enoate (I-5)

[0238] Step 1: To a stirred mixture of elaidic acid (26 mg, 86 μmol), 1-1-((2R,4S,5R)-5-(chloromethyl)-4-hydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl)-5-fluoro-4-(((4- 73 33049332.1 400807-032WO (218484)methoxyphenyl) diphenylmethyl) amino) pyrimidin-2(1H)-one (15) (See Example 3) (60 mg, 95 µmol) and 1-methylimidazole (107 mg, 1.30 mmol) in ACN (2 mL) was added BOP-Cl (66 mg, 0.26 mmol) at 0oC under an inert atmosphere. The resulting mixture was stirred for 1.5 h at RT. The resulting mixture was filtered; the filter cake was washed with EtOAc (3 x 5 mL). The filtrate was washed with brine (1 x 5 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EA = 3:1) to afford ((2R,3S,5R)-2-(chloromethyl)-5-(5-fluoro-4-(((4-methoxyphenyl) diphenylmethyl) amino)-2-oxopyrimidin-1(2H)-yl)-3-hydroxytetrahydrofuran-2-yl) methyl (E)- octadec-9-enoate (39 mg, 54 %) as a colorless oil. LC / MS (ES, m / z): 828 / 830 [M-H]-.

[0239] Step 2: A solution of (2R,3S,5R)-2-(chloromethyl)-5-(5-fluoro-4-(((4-methoxyphenyl)diphenylmethyl) amino)-2-oxopyrimidin-1(2H)-yl)-3-hydroxytetrahydrofuran-2-yl) methyl (E)- octadec-9-enoate (24) (34 mg, 41 μmol) in 80% AcOH was stirred for 12 h at rt. The resulting mixture was concentrated in vacuo. The crude product was purified by Prep-HPLC with the following conditions: Column: XSelect CSH F-Phenyl OBD column 19*250 mm, 5 μm; Mobile Phase A: Water (0.05% FA), Mobile Phase B: THF; Flow rate: 20 mL / min; Gradient: 65% B to 95% B in 8 min; Wavelength: 254nm / 220nm; RT1 (min): 7.5. The fraction was collected and concentrated in vacuo, the residue was re-dissolved in ACN and H2O, and then was lyophilized to afford ((2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-3-hydroxytetrahydrofuran-2-yl) methyl (E)-octadec-9-enoate (2.4 mg, 11%) as a white solid.

[0240] LC / MS (ES, m / z): 558 / 560 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.05 (d, J =7.0 Hz, 1H), 7.84 (s, 1H), 7.61 (s, 1H), 6.24 (t, J = 7.2 Hz, 1H), 5.57 – 5.26 (m, 4H), 3.83 – 3.61 (m, 4H), 2.45 – 2.20 (m, 8H), 1.36 – 1.21 (m, 22H), 0.87 (t, J = 6.6 Hz, 3H). 74 33049332.1 400807-032WO (218484)EXAMPLE 6 – Synthesis of [(2R,3R,4S,5R)-5-(4-amino-2-oxopyrimidin-1-yl)-2- (chloromethyl)-4-fluoro-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2- methylpropanoate (I-6)

[0241] Step 1: To a solution of 4-amino-1-[(2R,3S,4R,5R)-5-(chloromethyl)-3-fluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one (25) (WO 2023 / 178133) (200 mg, 0.68mmol) and DMAP (250 mg, 2.05 mmol) in ACN (20 mL) were added 2-methylpropanoyl 2- methylpropanoate (215 mg, 1.4 mmol) dropwise at 0°C. The resulting mixture was stirred at 0°C for 30 min and then diluted with EtOAc. The organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (10:1). The product-containing fractions were combined and evaporated partially to remove the solvents, and lyophilized overnight to afford [(2R,3R,4S,5R)-5-(4-amino-2- oxopyrimidin-1-yl)-2-(chloromethyl)-4-fluoro-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (122.7 mg41%) as a white solid. LC / MS (ES, m / z): 434 (M+H+) 99.7% purity.

[0242] Conditions for the HPLC: (Column: Shim‐pack Scepter C18, 33*3.0 mm, 3.0 μm;Mobile phase A: 5mM NH4HCO3in H2O / Acetonitrile (95:5, v / v), Mobile phase B: Acetonitrile; Flow rate: 1.5 mL / min; Gradient: 0% B to 5% B in 0.01 min, 5% B to 90% B in 1.19 min, 90% B to 90% B in 0.60 min,90% B to 5% B in 0.02min; 5% B to 0% B in 0.18 min; Wave Length: 254 nm; RT 1 (min): 0.955).1H NMR (400 MHz, DMSO-d6) δ 7.55 (d, J = 7.2 Hz, 1H), 7.35 – 7.-7.30 (m, 2H), 6.40 – 6.35 (m, 1H), 5.77 (d, J = 7.6 Hz, 1H), 5.66 – 5.61 (m, 1H), 5.42 – 5.28 (m, 1H), 4.41 – 4.34 (m, 2H), 4.02 (d, J = 12.0 Hz, 1H), 3.84 (d, J = 12.0 Hz, 1H), 2.71 – 2.65 (m, 1H), 2.63 – 2.59 (m, 1H), 1.17 – 1.10 (m, 12H). 75 33049332.1 400807-032WO (218484)EXAMPLE 7 – Synthesis of [(2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2- ethenyl-4-fluoro-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (I-7) [0243y-5-(hydroxymethyl)oxolan-2-yl]-5-fluoropyrimidin-2-one (27) (WO 2023 / 178133) (100 mg, 0.35 mmol) and DMAP (127 mg, 1.04 mmol) in ACN (3 mL) was added 2-methylpropanoyl 2- methylpropanoate (300 mg, 1.90 mmol) dropwise at 0°C under an inert atmosphere. The resulting mixture was stirred at rt for 12 h. The resulting mixture was diluted with water. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (10:1). The product-containing fractions were combined and evaporated partially to remove the solvents, then lyophilized overnight to afford [(2R,3R,4S,5R)-5-(4-amino-5-fluoro- 2-oxopyrimidin-1-yl)-2-ethenyl-4-fluoro-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2- methylpropanoate (51 mg, 34%) as a white solid.

[0244] LC / MS (ES, m / z): 430 (M+H+). Conditions for the HPLC: (Column: Shim‐pack Scepter C18, 33*3.0 mm, 3.0 µm; Mobile Phase A: 5mM NH4HCO3in H2O / Acetonitrile(95:5, v / v), Mobile Phase B: Acetonitrile; Flow rate: 1.5 mL / min; Gradient: 5% B to 90% B in 1.19 min, 90% B to 90% B in 0.60 min, 90% B to 5% B in 0.02 min. Wave Length: 254 nm; RT1 (min): 0.967).

[0245] 1H NMR (400 MHz, DMSO-d6) δ 7.99 (br s, 1H), 7.80 – 7.75 (m, 2H), 6.20 – 6.15 (m,1H), 5.82 – 5.75 (m, 1H), 5.55 – 5.46 (m, 2H), 5.38 – 5.22 (m, 2H), 4.38 – 4.35 (m, 1H), 4.27 – 4.24 (m, 1H), 2.68 – 2.55 (m, 2H), 1.15 – 1.09 (m, 12H). 76 33049332.1 400807-032WO (218484)EXAMPLE 8 – Synthesis of [(2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-eth yl-4-fluoro-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (I-8)[024(hydroxymethyl)oxolan-2-yl]-5-fluoropyrimidin-2-one (28) (WO 2023 / 178133) (200 mg, 69 mmol) and DMAP (252 mg, 2.0 mmol) in ACN (20 mL) were added 2-methylpropanoyl 2- methylpropanoate (217 mg, 1.4 mmol) dropwise at 0°C under an inert atmosphere. The resulting mixture was stirred at 0°C for 30 min and then quenched by the addition of ice-water. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the resulting filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions: Column: X-Select Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 30% B to 70% B in 9 min; Wavelength: 254 nm / 220 nm; RT1 (min): 6.56. The product-containing fractions were combined and evaporated partially to remove the solvents, then lyophilized overnight to afford [(2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)- 2-ethyl-4-fluoro-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (61 mg, 21%) as a white solid. LC / MS (ES, m / z): 432 (M+H+)

[0247] Conditions for the HPLC: (Column: Shim‐pack Scepter C18, 33*3.0 mm, 3.0 μm;Mobile phase A: 5 mM NH4HCO3in H2O / Acetonitrile (95:5,v / v), Mobile phase B: Acetonitrile; Flow rate: 1.5 mL / min; Gradient: 0% B to 5% B in 0.01 min, 5% B to 90% B in 1.19 min, 90% B to 90% B in 0.60 min, 90% B to 5% B in 0.02 min; 5% B to 0% B in 0.2 min; Wave Length: 254 nm; RT 1 (min): 0.994). 77 33049332.1 400807-032WO (218484)

[0248] 1H NMR (400 MHz, DMSO-d6) δ 7.95 (br s,1H), 7.75 – 7.70 (m, 2H), 6.16 – 6.10 (m,1H), 5.52 – 5.47 (m, 1H), 5.42 – 5.27 (m, 1H), 4.24 – 4.21 (m, 2H), 2.70 – 2.58 (m, 2H), 1.78 – 1.73 (m, 1H), 1.60 – 1.55 (m, 1H), 1.15 – 1.11 (m, 12H), 0.89 (t, J = 7.2 Hz, 3H). EXAMPLE 9 – Synthesis of [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethyn yl-3-[(2-methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (I-39)

[0249] To a stirred solution of 4-amino-1-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (120 mg, 0.45 mmol) and DMAP (109 mg, 0.89 mmol) in MeCN (2.5 mL) was added 2-methylpropanoyl 2-methylpropanoate (148 mg, 0.94 mmol) dropwise at 0 °C under -inert atmosphere. The resulting mixture was stirred at 0°C for 2 h and then quenched by the addition of ice water at 0°C. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (10:1). The resulting solution was concentrated under reduced pressure and the reissue was lyophilized overnight to afford [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethynyl-3-[(2- methylpropanoyl)oxy]oxolan-2-yl]methyl 2-methylpropanoate (67.4 mg, 36.9%, 99.8%purity) as a white solid. LC-MS-(ES, m / z): 410(M+H+) 99.8% purity.

[0250] 1H NMR (400 MHz, DMSO-d6) δ 7.92 (brs, 1H), 7.88 (d, J = 7.2 Hz, 1H), 7.67 (brs,1H), 6.22 (t, J = 6.0 Hz, 1H), 5.44 – 5.41 (m, 1H), 4.38 – 4.35 (m, 1H), 4.29 – 4.26 (m, 1H), 3.77 (s, 1H), 2.68 – 2.61 (m, 3H), 2.40 – 2.35 (m, 1H), 1.19 – 1.12 (m, 12H). 78 33049332.1 400807-032WO (218484)EXAMPLE 10 - Synthesis of (2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethyl- 2-(hydroxymethyl) oxolan-3-yl 2-methylpropanoate (II-1)(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (100 mg, 0.37 mmol) in Py (2 mL) was added 1-[chloro(4-methoxyphenyl) benzyl]-4-methoxybenzene (136 mg, 0.40 mmol) portion wise at 0°C under an inert atmosphere. The resulting mixture was stirred at room temperature for 12h, then the mixture was neutralized to pH 7 with 0.1M aq. HCl. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with CH2Cl2 / MeOH (10:1) to afford 4-amino-1-[(2R,4S,5R)-5-{[bis(4-methoxyphenyl) (phenyl)methoxy] methyl}-5-ethyl-4-hydroxyoxolan-2-yl]-5-fluoropyrimidin-2-one (130 mg, 0.23 mmol, 61.71%) as a light yellow solid. LC-MS(ES, m / z): 576(M+H+).

[0252] Step 2 -To a stirred solution of 4-amino-1-[(2R,4S,5R)-5-{[bis(4-methoxyphenyl)(phenyl)methoxy] methyl}-5-ethyl-4-hydroxyoxolan-2-yl]-5-fluoropyrimidin-2-one (130 mg, 0.23 mmol) and DMAP (41 mg, 0.34 mmol) in ACN (1.5 mL) was added 2-methylpropanoyl 2- methylpropanoate (38 mg, 0.24 mmol) dropwise at 0°C under an inert atmosphere. The resulting mixture was stirred at room temperature for 0.5 h and then quenched by the addition of ice-water at 0°C. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. This resulted in (2R,3S,5R)-5-(4-amino-5- fluoro-2-oxopyrimidin-1-yl)-2-{[bis(4-methoxyphenyl) (phenyl)methoxy] methyl}-2- 79 33049332.1 400807-032WO (218484)ethyloxolan-3-yl 2-methylpropanoate (140 mg, crude) as a white solid. The crude product was used in the next step directly without further purification. LC-MS(ES, m / z): 646 (M+H+).

[0253] Step 3 - A solution of (2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-{[bis(4-methoxyphenyl) (phenyl)methoxy] methyl}-2-ethyloxolan-3-yl 2-methylpropanoate (130 mg, crude) in AcOH (1.5 mL, 80%) was stirred at room temperature for 12 h. The mixture was neutralized to pH 7 with saturated NaHCO3 (aq.). The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure giving rise to a residue which was purified via Prep-TLC (CH2Cl2 / MeOH 10:1). The product was dissolved in MeOH and lyophilized to afford (2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1- yl)-2-ethyl-2-(hydroxymethyl) oxolan-3-yl 2-methylpropanoate (40.8 mg, 0.12 mmol, 59.02%yield in two steps) as a white solid.

[0254] LC-MS(ES, m / z): 344 (M+H+). 98.7% purity.

[0255] 1H NMR (400 MHz, DMSO) δ 8.11 (d, J = 7.2 Hz, 1H), 7.77 (brs, 1H), 7.54 (brs, 1H),6.14-6.06 (m, 1H), 5.37-5.29 (m, 2H), 3.58-3.44 (m, 2H), 2.68-2.53 (m, 1H), 2.42-2.30 (m, 1H), 2.26-2.16 (m, 1H), 1.70-1.56 (m, 1H), 1.56-1.43 (m, 1H), 1.16-1.09 (m, 6H), 0.86 (t, J = 7.5 Hz, 3H). EXAMPLE 11 – Synthesis of [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2- ethyl-3-hydroxyoxolan-2-yl] methyl 2-methylpropanoate (I-41)484)

[0256] Step 1 - To a stirred solution of 4-amino-1-[(2R,4S,5R)-5-ethyl-4-hydroxy-5-(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (100 mg, 0.37 mmol) in Py (2 mL) was added 1-[chloro(4-methoxyphenyl) benzyl]-4-methoxybenzene (136 mg, 0.40 mmol) portion wise at 0°C under an inert atmosphere. The resulting mixture was stirred at room temperature for 20 h and then neutralized to pH 7 with 0.1M. HCl. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with CH2Cl2 / MeOH (10:1) to afford 4-amino-1-[(2R,4S,5R)-5-{[bis(4-methoxyphenyl) (phenyl)methoxy] methyl}-5-ethyl- 4-hydroxyoxolan-2-yl]-5-fluoropyrimidin-2-one (130 mg, 0.23 mmol, 61.71%) as a light yellow solid. LC-MS(ES, m / z): 576(M+H+).

[0257] Step 2 - To a stirred solution of 4-amino-1-[(2R,4S,5R)-5-{[bis(4-methoxyphenyl)(phenyl)methoxy] methyl}-5-ethyl-4-hydroxyoxolan-2-yl]-5-fluoropyrimidin-2-one (130 mg, 0.23 mmol) and Imidazole (77 mg, 1.13 mmol) in DMF (2 mL) was added TBS-Cl (136 mg, 0.90 mmol) portion wise at 0°C under an inert atmosphere. The resulting mixture was stirred at room temperature for 2 days and then quenched by the addition of ice-water at 0°C. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 10:1) to afford 4-amino-1-[(2R,4S,5R)-5-{[bis(4-methoxyphenyl) (phenyl)methoxy]methyl}-4- [(tert-butyldimethylsilyl) oxy]-5-ethyloxolan-2-yl]-5-fluoropyrimidin-2-one (155 mg, 0.23 mmol, 99.48%) as a white solid. LC-MS(ES, m / z): 690(M+H+).

[0258] Step 3 - A solution of 4-amino-1-[(2R,4S,5R)-5-{[bis(4-methoxyphenyl)(phenyl)methoxy] methyl}-4-[(tert-butyldimethylsilyl) oxy]-5-ethyloxolan-2-yl]-5- fluoropyrimidin-2-one (155 mg, 0.23 mmol) in AcOH (2 mL, 80%) was stirred at room temperature for 12h. The mixture was neutralized to pH 7 with saturated NaHCO3 (aq.). The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 10:1) to afford 4-amino-1-[(2R,4S,5R)-4-[(tert-butyldimethylsilyl) oxy]-5-ethyl-5- 81 33049332.1 400807-032WO (218484)(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (85 mg, 0.22 mmol, 97.63%) as a white solid. LC-MS(ES, m / z): 388(M+H+).

[0259] Step 4 - To a stirred solution of 4-amino-1-[(2R,4S,5R)-4-[(tert-butyldimethylsilyl)oxy]-5-ethyl-5-(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (85 mg, 0.22 mmol) and DMAP (40 mg, 0.33 mmol) in ACN (1 mL) was added 2-methylpropanoyl 2-methylpropanoate (36 mg, 0.23 mmol) dropwise at 0°C under an inert atmosphere. The resulting mixture was stirred at room temperature for 0.5 h and then quenched by the addition of ice-water at 0°C. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure to afford [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin- 1-yl)-3-[(tert-butyldimethylsilyl) oxy]-2-ethyloxolan-2-yl] methyl 2-methylpropanoate (95 mg, crude) as a white solid. The crude product was used in the next step directly without further purification. LC-MS(ES, m / z): 458(M+H+).

[0260] Step 5 - To a stirred solution of [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-3-[(tert-butyldimethylsilyl) oxy]-2-ethyloxolan-2-yl] methyl 2-methylpropanoate (95 mg, crude) in THF (1 mL) was added TBAF (27 mg, 0.10 mmol). The resulting mixture was stirred at room temperature for 1h. The reaction was quenched by the addition of sat. NH4Cl (aq.) at 0°C. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions (Column: XSelect CSH Prep C18 OBD Column, 19*250 mm, 5μm; Mobile Phase A: Water(0.1%FA), Mobile Phase B: ACN; Flow rate: 20 mL / min; Gradient (B%): 10% B to 40% B in 8 min; Wavelength: 254nm / 220nm; RT1(min): 7.8). The product- containing fractions were combined and evaporated partially to remove the solvents, then lyophilized to afford [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethyl-3- hydroxyoxolan-2-yl] methyl 2-methylpropanoate (46.4 mg, 0.10 mmol, 65.10% yield in two steps) as a white solid.

[0261] LC-MS(ES, m / z): 344(M+H+). 99.9% purity.82 33049332.1 400807-032WO (218484)

[0262] 1H NMR (400 MHz, DMSO) δ 7.77 (d, J = 7.0 Hz, 2H), 7.52 (brs, 1H), 6.09-6.01 (m,1H), 5.31 (d, J = 4.5 Hz, 1H), 4.26-4.18 (m, 1H), 4.16-4.05 (m, 2H), 2.66-2.51 (m, 1H), 2.30- 2.13 (m, 2H), 1.75-1.65 (m, 1H), 1.65-1.51 (m, 1H), 1.13-1.08 (m, 6H), 0.92-0.85 (m, 3H).EXAMPLE 12 - Synthesis of [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethenyl-3-[(2-methylpropanoyl) oxy] oxolan-2-yl] methyl 2-methylpropanoate (Compound I-42)

[0263] , ,(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (200 mg, 0.74 mmol) and DMAP (135 mg, 1.11 mmol) in ACN (3 mL) was added 2-methylpropanoyl 2-methylpropanoate (198 mg, 1.25 mmol) dropwise at 0°C under an inert atmosphere. The resulting mixture was stirred at room temperature for 1h, then quenched by the addition of a solution of 0.1M. HCl (aq.) at 0°C. The resulting mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and filtered to remove solid. The resulting filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 10:1). The product-containing fractions were combined and evaporated partially to remove the solvents, then lyophilized to afford [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2- ethenyl-3-[(2-methylpropanoyl) oxy] oxolan-2-yl] methyl 2-methylpropanoate (109.2 mg, 35.9% yield) as a white solid.

[0264] LC-MS(ES, m / z): 412(M+H+). 99.5% purity

[0265] 1H NMR (400 MHz, DMSO) δ 7.92-7.86 (m, 2H), 7.62 (s, 1H), 6.15 (t, J = 1.8 Hz,1H), 5.83-5.72 (m, 1H), 5.49-5.37 (m, 2H), 5.35-5.28 (m, 1H), 4.27-4.15 (m, 2H), 2.65-2.50 (m, 2H), 2.50-2.41(m, 1H), 2.34-2.23 (m, 1H), 1.14-1.08 (m, 12H). 83 33049332.1 400807-032WO (218484)EXAMPLE 13 and 14 - Synthesis of [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethenyl-3-hydroxyoxolan-2-yl] methyl 2-methylpropanoate (I-43) and (2R,3S,5R)-5-(4-am ino-5-fluoro-2-oxopyrimidin-1-yl)-2-ethenyl-2-(hydroxymethyl) oxolan-3-yl 2-methylpropa noate (II-2)

[0266] yrimidin-1-yl)-2-ethenyl-3-[(2-methylpropanoyl) oxy] oxolan-2-yl] methyl 2-methylpropanoate (310 mg, 0.75 mmol) in MeOH (6 mL) was added TEA (305 mg, 3.01 mmol) dropwise. The resulting mixture was stirred for 2 days, and then concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions (Column: X-Select Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(0.1%FA), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient (B%): 17% B to 45% B in 10 min; Wavelength: 254 nm; RT1(min): 7.80; RT2(min): 8.22). The product-containing fractions were combined and evaporated partially to remove the solvents, then lyophilized to afford [(2R,3S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1-yl)-2-ethenyl-3-hydroxyoxolan-2-yl] methyl 2-methylpropanoate (I-43, 47.7 mg, 0.14 mmol, RT1(min): 7.80, 18.5% yield) as a white solid and (2R,3S,5R)-5-(4-amino-5-fluoro- 2-oxopyrimidin-1-yl)-2-ethenyl-2-(hydroxymethyl) oxolan-3-yl 2-methylpropanoate (II-2, 42.5 mg, 16.5% yield) as a white solid.

[0267] LC-MS I-43 (ES, m / z): 342(M+H+). 99.9% purity

[0268] 1H NMR I-43 (400 MHz, DMSO) δ 7.86 (d, J = 7.0 Hz, 1H), 7.78 (brs, 1H), 7.53 (brs,1H), 6.05 – 6.02 (m, 1H), 5.95 – 5.88 (m, 1H), 5.47 – 5.46 (m, 1H), 5.42 – 5.37 (m, 1H), 5.31 – 84 33049332.1 400807-032WO (218484)5.28 (m, 1H), 4.36 – 4.35 (m, 1H), 4.22 – 4.12 (m, 2H), 2.62 – 2.55 (m, 1H), 2.18 – 2.09 (m, 2H), 1.10 – 1.08 (m, 6H).

[0269] LC-MS II-2 (ES, m / z): 342(M+H+). 99.0% purity

[0270] 1H NMR II-2 (400 MHz, DMSO) δ 8.21 (d, J = 7.2 Hz, 1H), 7.81 (brs, 1H), 7.57 (brs,1H), 6.19 – 6.16 (m, 1H), 5.76 – 5.69 (m, 1H), 5.55 (brs, 1H), 5.42 – 5.36 (m, 2H), 5.22 – 5.19 (m, 1H), 3.62 – 3.59 (m, 1H), 3.44 – 3.41 (m, 1H), 2.59 – 2.53 (m, 1H), 2.38 – 2.31 (m, 1H), 2.22 – 2.16 (m, 1H), 1.10 – 1.07 (m, 6H). EXAMPLE 15 – Synthesis of (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)- 2-ethynyl-4-hydroxy-2-((isobutyryloxy)methyl)tetrahydrofuran-3-yl isobutyrate (I-45)

[0271] Step 1 - To a stirred mixture of 4-amino-1-((2R,3S,4S,5R)-4-(benzyloxy)-5-((benzyloxy)methyl)-5-ethynyl-3-hydroxytetrahydrofuran-2-yl)-5-fluoropyrimidin-2(1H)-one (300 mg, 0.644 mmol) and tert-butyl (chloro) dimethylsilane (291mg, 1.93 mmol) in DMF (20 mL) were added imidazole (263 mg, 3.86 mmol) at room temperature under nitrogen 85 33049332.1 400807-032WO (218484)atmosphere. The resulting mixture was stirred at 80°C for overnight under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 90% gradient in 30 min; detector, UV 254 nm. The combined organic layers was concentrated under reduced pressure to afford 4-amino-1-((2R,3S,4S,5R)-4-(benzyloxy)-5-((benzyloxy)methyl)-3- ((tert-butyldimethylsilyl)oxy)-5-ethynyltetrahydrofuran-2-yl)-5-fluoropyrimidin-2(1H)-one (230 mg, 58.4% yield) as a light yellow solid. LC-MS (ES, m / z): 580 [M+H]+.

[0272] Step 2 - To a stirred mixture of 4-amino-1-((2R,3S,4S,5R)-4-(benzyloxy)-5-((benzyloxy)methyl)-3-((tert-butyldimethylsilyl)oxy)-5-ethynyltetrahydrofuran-2-yl)-5- fluoropyrimidin-2(1H)-one (220 mg, 0.379 mmol) in DCM (10 mL) was added boron trichloride (2.4 mL, 1.0 M in methylene chloride) at -78°C under nitrogen atmosphere. The resulting mixture was stirred at -30°C for 3h under nitrogen atmosphere. The reaction was quenched by the addition of MeOH(5mL) at -30°C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 90% gradient in 30 min; detector, UV 254 nm. The combined organic layers was concentrated under reduced pressure to afford 4-amino-1-((2R,3S,4S,5R)-3-((tert-butyldimethylsilyl)oxy)-5- ethynyl-4-hydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl)-5-fluoropyrimidin-2(1H)-one (140 mg, 83.1% yield) as a light yellow solid. LC-MS (ES, m / z): 400 [M+H]+.

[0273] Step 3 - To a stirred mixture of 4-amino-1-((2R,3S,4S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-5- fluoropyrimidin-2(1H)-one (130 mg, 0.325 mmol) and DMAP (67 mg, 0.552 mmol) in MeCN (10 mL) was added isobutyric anhydride (77 mg, 0.488 mmol) at 0°C under nitrogen atmosphere. The resulting mixture was stirred at 0°C for 30 min under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1). The combined organic layers was concentrated under reduced pressure to afford (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1(2H)-yl)-4-((tert-butyldimethylsilyl) oxy)-2-ethynyl-2- ((isobutyryloxy)methyl)tetrahydrofuran-3-yl isobutyrate (150 mg, 76.8% yield) as a white solid. LC-MS (ES, m / z): 540 [M+H]+86 33049332.1 400807-032WO (218484)

[0274] Step 4 - To a stirred mixture of (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-4-((tert-butyldimethylsilyl)oxy)-2-ethynyl-2-((isobutyryloxy)methyl)tetrahydrofuran- 3-yl isobutyrate (70 mg, 0.130 mmol) in THF (10 mL) was added Et3N.3HF (209 mg, 1.30 mmol) at room temperature under air atmosphere. The resulting mixture was stirred at 60°C for 2h under air atmosphere. The resulting mixture was concentrated under reduced pressure. The crude product (60 mg) was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10mmol / L NH4HCO3+0.05% NH3H2O), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient (B%): 27% B to 47% B in 8min; Wave Length: 254nm / 220nm; RT1(min): 7.34). The fraction was collected and concentrated under vacuum, the residue was re-dissolved in CH3CN and H2O, and then was lyophilized to afford (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- ethynyl-4-hydroxy-2-((isobutyryloxy) methyl)tetrahydrofuran-3-yl isobutyrate (29.2 mg, 52.7 % yield) as a white solid.

[0275] LC-MS (ES, m / z): 426 [M+H]+.

[0276] 1H NMR (400 MHz, DMSO) δ 7.85 (s, 1H), 7.63 (d, J = 7.2 Hz, 2H), 6.19 – 6.07 (m,2H), 5.15 (d, J = 2.9 Hz, 1H), 4.50 – 4.30 (m, 2H), 4.28 – 4.20 (m, 1H), 3.80 (s, 1H), 2.70 – 2.54 (m, 2H), 1.22 – 1.03 (m, 12H). EXAMPLE 16 – Synthesis of ([(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2- (difluoromethyl)-3-[(2-methylpropanoyl) oxy]oxolan-2-yl]methyl 2-methylpropanoate (I- 46)

[0277] Step 1 - To a stirred mixture of 4-amino-1-[(2R,4S,5R)-5-(difluoromethyl)-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-5-fluoropyrimidin-2-one (100 mg, 0.3 mmol) and 2- methylpropanoyl 2-methylpropanoate (91 mg, 0.5 mmol) in MeCN (5 mL) was added DMAP 87 33049332.1 400807-032WO (218484)(62 mg, 0.5 mmol) in portions at 0 °C under nitrogen atmosphere and stirred at 0°C for 1 h. The resulting mixture was concentrated under reduced pressure. The crude product was purified by Prep-HPLC. The fractions with the desired product were collected and concentrated under vacuum, the residue was re-dissolved in CH3CN and H2O, and then was lyophilized to afford [(2R,3S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-(difluoromethyl)-3-[(2- methylpropanoyl) oxy] oxolan-2-yl]methyl 2-methylpropanoate (59.4 mg, 40.3% yield) as a white solid.

[0278] LC-MS (ES, m / z): 436 [M+H]+.

[0279] 1H NMR (400 MHz, DMSO) δ 7.91 (d, J = 7.0 Hz, 2H), 7.67 (s, 1H), 6.44 – 6.14 (m,2H), 5.75 – 5.67 (m, 1H), 4.37 – 4.26 (m, 2H), 2.68 – 2.52 (m, 3H), 2.42 – 2.31 (m, 1H), 1.16 – 1.06 (m, 12H). EXAMPLE 17- Synthesis of (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)- 4-hydroxy-2-((isobutyryloxy) methyl)-2-vinyltetrahydrofuran-3-yl isobutyrate (I-47)

[0280] Step 1 - To a stirred solution of (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-ethynyl-4-hydroxy-2-((isobutyryloxy)methyl)tetrahydrofuran-3-yl isobutyrate (35 mg, 0.082 mmol) in EtOH (10 mL) was added carbonic acid calcium palladium plumbane (6 mg, 0.016 mmol) at room temperature under air atmosphere. The resulting mixture was stirred at room temperature for 30 min under hydrogen atmosphere (1 atm). The resulting mixture was filtered and the filter cake was washed with EtOH (2x10 mL). The filtrate was concentrated under reduced pressure. The crude product (32 mg) was purified by Prep-HPLC. The fractions containing the product were collected and concentrated under vacuum, the residue was re- dissolved in CH3CN and H2O, and then was lyophilized to afford (2R,3S,4S,5R)-5-(4-amino-5- fluoro-2-oxopyrimidin-1(2H)-yl)-4-hydroxy-2-((isobutyryloxy)methyl)-2-vinyltetrahydrofuran- 3-yl isobutyrate (11.9 mg, 33.6% yield) as a white solid. 88 33049332.1 400807-032WO (218484)

[0281] LC-MS (ES, m / z): 428 [M+H]+.

[0282] 1H NMR (400 MHz, DMSO) δ 7.90 – 7.47 (m, 3H), 6.15 – 6.03 (m, 2H), 5.78 – 5.65(m, 1H), 5.45 – 5.36 (m, 1H), 5.31 – 5.24 (m, 1H), 5.17 (d, J = 3.1 Hz, 1H), 4.45 (d, J = 11.5 Hz, 1H), 4.18 (d, J = 11.4 Hz, 2H), 2.63 – 2.53 (m, 2H), 1.19 – 1.03 (m, 12H). EXAMPLE 18 – Synthesis of (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)- 2-ethyl-4-hydroxy-2-((isobutyryloxy) methyl)tetrahydrofuran-3-yl isobutyrate (I-48)[02midin-1(2H)-yl)-2-ethynyl-4-hydroxy-2-((isobutyryloxy)methyl)tetrahydrofuran-3-yl isobutyrate (24 mg, 0.056 mmol) in MeOH (10 mL) was added Pd / C (2 mg, 0.023 mmol) at room temperature under air atmosphere. The resulting mixture was stirred at room temperature for 30 min under hydrogen atmosphere (5 atm). The resulting mixture was filtered, the filter cake was washed with MeOH (2x10 mL). The filtrate was concentrated under reduced pressure. The crude product (22 mg) was purified by Prep-HPLC. The fractions containing the product were collected and concentrated under vacuum, the residue was re-dissolved in CH3CN and H2O, and then was lyophilized to afford (2R,3S,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-ethyl-4- hydroxy-2-((isobutyryloxy)methyl)tetrahydrofuran-3-yl isobutyrate (12.1 mg, 49.7 % yield) as a white solid.

[0284] LC-MS (ES, m / z): 430 [M+H]+.

[0285] 1H NMR (400 MHz, DMSO) δ 7.84 – 7.47 (m, 3H), 6.13 – 5.97 (m, 2H), 5.14 (d, J =2.4 Hz, 1H), 4.40 (d, J = 11.6 Hz, 1H), 4.23 – 4.13 (m, 2H), 2.70 – 2.53 (m, 2H), 1.77 – 1.49 (m, 2H), 1.20 – 1.02 (m, 12H), 0.85 (t, J = 7.4 Hz, 3H). 89 33049332.1 400807-032WO (218484)

[0286] Another aspect of the disclosure provides a compound selected from I-1 to I-38 inTable 1, below, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound selected from I-1 to I-38 in Table 1, below.

[0287] Another aspect of the disclosure provides a compound selected from I-1 to I-38 inTable 1, below, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound selected from I-1 to I-38 in Table 1, below.EXAMPLE 19 – Synthesis of Additional Compounds

[0288] Compounds I-9, I-10, I-11, I-12, I-13, I-14, I-15, I-16, I-17, I-18, I-19, I-20, I-21, I-22, I-24, I-25, I-26, I-27, I-28, I-29, I-30, I-31, I-32, I-33, I-34, I-35, I-36, I-37, and I-38 were synthesized in procedures analogous to those presented in Examples 1-9 and in Examples 10-18.EXAMPLE 20 – Stable Cellular Assay for Inhibiting LINE1 Reverse Transcriptase

[0289] Exemplary compounds were tested for ability to inhibit LINE1 reverse transcriptaseusing a stable artificial-intron Cis LINE1 reporter assay. Assay procedures and results are described below. Part I – Procedure for Stable Artificial-Intron Cis LINE1 Reporter Assay

[0290] A stable HeLa Tet-On 3G (Takara, cat no 631183) cell line expressing a bi-directionalinducible LINE1 construct was generated as described in Xie, Y. et al. “Cell division promotes efficient retrotransposition in a stable L1 reporter cell line,” Mobile DNA (2013) 4:10. Single cell clones were screened for high Luciferase expression and the highest expression Firefly expressing clone was chosen for compound testing.

[0291] Test compounds were serially diluted in DMSO and spotted in 96-well plates.Subsequently the HeLa L1 artificial-intron reporter cells were plated into the compound- containing wells (8,000 cells / well), and the cells were induced for reporter expression with doxycycline (Sigma cat no D9891) at a final concentration of 500 ng / mL. Luminescence was measured 72 h after plating using the Dual-Glo Luciferase Assay System (Promega cat no E2940) following the manufacturer’s instructions. The Firefly Luciferase activity (normalized against its activity in a control well without test compound) was used to report LINE1 activity. 90 33049332.1 400807-032WO (218484)Part II – Results

[0292] Experimental results are provided in Table 2, below. The symbol “***” indicates anIC50 less than or equal to 0.05 µM. The symbol “**” indicates an IC50 in the range of greater than 0.05 µM to less than or equal to 0.5 µM. The symbol “*” indicates an IC50greater than 0.5 µM. TABLE 2. Compound IC50 (µM) **Part III – Results for Additional Compounds

[0293] Experimental results obtained with additional compounds are provided in Table 2-A,below. The symbol “***” indicates an IC50less than or equal to 0.05 µM. The symbol “**” indicates an IC50in the range of greater than 0.05 µM to less than or equal to 0.5 µM. The symbol “*” indicates an IC50 greater than 0.5 µM. 91 33049332.1 400807-032WO (218484)TABLE 2-A. Compound IC50 (µM)EXAMPLE 21 – Biochemical Assay for Inhibiting LINE1 Reverse Transcriptase

[0294] Exemplary compounds may be tested for ability to inhibit LINE1 reverse transcriptaseusing a homogeneous time-resolved fluorescence (HTRF) assay. Assay procedures are described below. Part I – Procedure for Homogeneous Time-Resolved Fluorescence LINE1 RT Assay

[0295] The LINE1 reverse transcriptase homogeneous time-resolved fluorescence (HTRF)assay is performed with recombinant MBP-tagged LINE1 protein (238-1061) (generated and purified according to procedures in Dai L. et al. BMC Biochemistry 2011; 12:18) in a 384-well format. Test compound is serially diluted in DMSO and further diluted in the assay buffer (50 mM Tris-HCl, 50 mM KCl, 10 mM MgCl2, 10 mM DTT, pH 8.1) to achieve a final DMSO concentration of 1%. The serially diluted compound is mixed with 64 ng / well of LINE1 enzyme, 5 nM of pre-annealed template / biotin-primer pair (synthesized at Geneway Biotechnology), 10 nM of Fluorescein-12-dCTP fluorescent probe (PerkinElmer), and 1 mM dGTP, dATP, and DTTP (ThermoFisher). in the assay buffer. The template / biotin-primer sequences are as follows: (SEQ ID NO:1) (SEQ ID NO:2) . 92 33049332.1 400807-032WO (218484)

[0296] After incubating at 25°C for 60 minutes, the detection reagent (20 mM EDTA withstreptavidin-terbium cryptate, Cisbio Bioassay) in the PPI buffer (Cisbio Bioassay) is added, and the mixture is incubated at 25°C for 30 minutes. At the end of the incubation, fluorescence is read at ex / em=337 / 485 nm and ex / em=337 / 520 nm on an Envision 2104 plate reader (Perkin Elmer). The fluorescence ratio at 520 / 485 nm is used for the calculation. Percent inhibition is calculated with the DMSO sample as 0% inhibition and no enzyme as 100% inhibition. The IC50 is calculated by fitting the compound dose inhibition curve with a 4-parameter non-linear regression equation. Part II- Results

[0297] The following triphosphate analogs were made and tested in this assay. Thetriphosphates were synthesized from the parent nucleoside at NuBlocks LLC (Oceanside, CA) or Pharmaron (Beijing, China). Compound Triphosphat IC50 Number e Structure ( M)93 33049332.1 400807-032WO (218484)EXAMPLE 22– Cellular Assay for Altering IFN Production in THP1 TREX1 KO Cells

[0298] Exemplary compounds were tested for their ability to alter the type 1 interferonresponse in THP1 Dual TREX1 KO cells treated with 5-aza-2ʹ-deoxycytidine. Assay procedures and results are described below. Part I - Procedure

[0299] THP1-Dual™ KO-TREX1 cells were purchased from Invivogen (cat# thpd-kotrex).The THP1-Dual™ KO-TREX1 cells were cultured in RPMI 1640, 10% heat-inactivated fetal bovine serum, 25 mM HEPES, 10 µg / mL Blasticidin, and 100 µg / mL Zeocin. THP1-Dual™ KO-TREX1 cells were treated with a dose titration of test compound in the presence of 1 µM 5- aza-2ʹ-deoxycytidine (Sigma, cat# 189825). Type 1 Interferon and cell viability were assessed after five days of treatment.

[0300] Stock solution of test compound was prepared in DMSO followed by a three-folddilution in DMSO. Additional 50x dilution was prepared in cell culture media for each dilution. 10 µL of diluted test compound was then added to a 384-well plate.

[0301] THP1-Dual™ KO-TREX1 cells were treated with 1 µM 5-aza-2ʹ-deoxycytidine.THP1-Dual™ KO-TREX1 cells (50 µL) were added to each well of the 384-well plate 94 33049332.1 400807-032WO (218484)containing test compound titration at 10,000 cells / well. Cells were incubated at 37ºC, 5% CO2in a humidified incubator for five days. On day five, 20 µL of cell supernatant was transferred to a 384-well, white-walled plate, followed by addition to each well of 50 µL of QUANTI-LUC solution containing stabilizer. Luminescence was detected on a plate reader according to manufacturer’s instructions.

[0302] For certain compounds, the assay was run in a 96-well format, with the followingmodifications: ^190 mL, instead of 50 mL, of cells were added to each well, to provide 50,000 cells / well,instead of 10,000 cells / well, ^Cells were incubated for 6 days, instead of 5 days,^ 25 mL, instead of 20 mL, of cell supernatant was added to the white-walled plate forQUANTI-LUC treatment; and ^25 mL, instead of 30 mL, of CellTiter-Glo was added to assess cell viability.

[0303] Percent inhibition of interferon was calculated using the following analysis: (AverageDMSO-Sample) / (Average DMSO-Average 30 µM control reagent)*100. The control reagent for inhibition of interferon was a specific nucleoside reverse-transcriptase inhibitor with molecular weight < 600 a.m.u. Percent induction of interferon was calculated using the following analysis: (Sample-Average DMSO) / (10 µM control reagent-Average DMSO)*100. The control reagent for induction of interferon was a specific nucleoside reverse-transcriptase inhibitor with molecular weight < 600 a.m.u.

[0304] The remaining cells were assessed for cell viability by adding 30 µl of CellTiter-Glo(Promega, G9683) solution to each well, and placed on a shaker for 10 minutes at RT. Luminescence was detected on a plate reader, according to manufacturer’s instructions. Percent inhibition of cell viability using CellTiter-Glo was calculated using the following analysis: (Average DMSO-Sample) / (Average DMSO-Average 20 µM control reagent)*100. The control reagent was Z-Leu-Leu-leucinal (see, for example, https: / / pubchem.ncbi.nlm.nih.gov / compound / 462382). 95 33049332.1 400807-032WO (218484)Part II - Results

[0305] Results are shown in Table 3, below. In the table, “A” represents values ≤ 0.050 µM;“B” represents values from > 0.050 µM to ≤ 0.100 µM; “C” represents values from > 0.100 µM to ≤ 0.250 µM; “D” represents values from > 0.250 µM to ≤ 1 µM; “E” represents values >1 µM; and “NT” = not tested: TABLE 3. Compound IC50 (µM) I 1 DEXAMPLE 23 – In vivo Decitabine Challenge Model

[0306] Twenty 9-11 week old C57BL / 6 mice are acclimated to the lab for at least 5 days.Test compound is prepared for p.o. administration. Decitabine (Sigma) is dissolved in sterile PBS (pH 7.4) and dosed within 30 minutes of preparation of the solution. Doses of both test compound and decitabine are administered once a day, every day from Day 0 to Day 4.

[0307] On Day 0, mice are split into four groups of five mice and given their first dose ofdecitabine (i.p., 5 mg / kg) and test compound. Dosing groups were: Group Test Compound Dose (mg / kg)96 33049332.1 400807-032WO (218484)

[0308] Decitabine and test compound are administered daily from Day 0 to Day 4. All miceare euthanized 1 hour after the last dose administration on Day 4. Spleens, liver, and terminal colon are collected, along with plasma from each animal. The fold changes in interferon- stimulated gene (ISG) expression are calculated by first normalizing to GAPDH gene using the Delta CT method. The CT (gene of interest) – CT (reference gene) is calculated to generate a delta CT for all samples. The fold change is then calculated by taking the Log2(Delta CT(control) – Delta CT (experimental). The control in this example is the PBS control animal group. The Taqman duplex assay (Thermo Fisher 4331182 and 4448489) is used according to the manufacturer’s instructions to determine levels of GAPDH v. IFIT2.EXAMPLE 24 – Assay for Altering IFN Production in THP1 TREX1 KO Xenografts

[0309] Exemplary compounds may be tested for their ability to alter IFN levels in THP1-DualTMKO-TREX1 xenografts in mice (produced according to the procedure described in the preceding Example). Assay procedures are described below.

[0310] CB-17 SCID female mice are inoculated subcutaneously with 10 million THP1-DualTMKO-TREX1 cells in 200 μl PBS with Matrigel (1:1) and grouped when tumor volume reaches 350-400 mm3. Mice bearing THP1-DualTMKO-TREX1 xenograft tumors are then separated into 5 groups. Three groups are administered: (1) decitabine (DAC) at 5mg / kg IP, once daily, for 4 days, and (2) test compound at one of three doses, once daily, for 4 days. One group is administered decitabine (DAC) at 5 mg / kg IP, once daily, for 4 days, and the test compound vehicle control. The final group is administered the vehicle control from both the test compound and the vehicle control from decitabine. Decitabine is formulated in sterile PBS, pH 7.4.

[0311] Tumors are harvested daily for 5 days starting on day 2, lysed with RIPA lysis buffercontaining protease and phosphatase inhibitors, and grinded at 50 Hz for 5 min. Tumors are then centrifuged, and PierceTMBCA Protein Assay Kit is used to measure protein concentration. Equal amounts of proteins are added to 96-well black plates, and luciferase signal is measured using the QUANTI-LucTMdetection medium according to manufacturer’s instructions. Luminescence is measured using the EnVision® 2105 Multimode Plate Reader. 97 33049332.1 400807-032WO (218484)EXAMPLE 25 – Assay for Reducing Expression of Interferon-Stimulated Genes in TREX1KO mice

[0312] Exemplary compounds were tested for their ability to reduce the expression ofinterferon-stimulated genes, as well as to reduce markers of inflammation and fibrosis in heart tissue, in TREX1 KO mice. Assay procedures and results are described below. Part I - Procedure

[0313] TREX1 is a 3’–5’ DNA exonuclease, loss of which results in Aicardi-Goutieressyndrome, a multi-system interferonopathy (Crow, et al). Deletion of TREX1 in mice results in a similar inflammatory phenotype (Morita, et al). TREX1 is involved in degradation of cytosolic DNAs; in the absence of TREX1 these DNAs accumulate and activate the cGAS:STING DNA sensing pathway, thereby triggering production of Type 1 interferons (Stetson, et al). Endogenous retroelements have been shown to be substrates of TREX1 and to accumulate in the cytosol in cells lacking TREX1 (Stetson, et al., and Thomas, et al.). Therefore, reducing the levels of retroelement-derived cytosolic DNA in TREX1 KO mice via inhibition of LINE-1 RT activity would be expected to reduce expression of interferon and reduce inflammation in these mice.

[0314] Each of Compounds I-1, I-2, I-39, and a vehicle control were administered to 6-weekold female TREX1 KO mice for 28 days. Compound I-1 was administered at 200 mg / kg, BID. Compound I-2 was administered at 100 mg / kg, BID. Compound I-39 was administered at 100 mg / kg, QD. For the wild-type group, N = 6. For each of the TREX1 KO mouse groups, N = 14- 15. At study termination, heart and kidney tissues were collected for histological assessment of inflammation and qPCR for interferon-stimulated genes, including TNFα and CXCL10. Part II - Results

[0315] Treatment with each of Compounds I-1, I-2, and I-39 reduced expression ofinterferon-stimulated genes as well as reduced inflammation and fibrosis in the heart. H&E staining was used to detect inflammatory scores, myocardial changes, and fibrosis in the mouse heart. A composite score was determined by adding the individual scores for inflammation, myocardial changes, and fibrosis. Scoring criteria for the heart are provided in the following table: 98 33049332.1 400807-032WO (218484)

[0316] compounds on expression of the interferon-stimulated genes TNFα and CXCL10 in the hearts of TREX1 KO mice. Figure 2 depicts the effects of the compounds on expression of the interferon- stimulated genes TNFα and CXCL10 in the kidneys of TREX1 KO mice. Figure 3 depicts the effects of the compounds on the cardiac histology score of TREX1 KO mice. In each of Figures 1-3, significance was determined compared to the vehicle group using a one-way ANOVA with Dunnet’s post-test.EXAMPLE 26 – Plasma and Brain Pharmacokinetics

[0317] Plasma and brain pharmacokinetics were determined for exemplary compounds, andtheir corresponding hydrolyzed parent compounds (depicted in Table 4, below). Assay procedures and results are described below. Part I - Procedure

[0318] C57BL6 male mice, approximately 6-8 weeks old, were dosed with either:^ Exemplary compounds from Table 1 at 30 mg / kg by oral gavage (PO), or^ The corresponding hydrolyzed parent compound at 10 mg / kg intravenously (IV).

[0319] For plasma PK analysis, approximately 0.03 mL blood was collected at each indicatedtime point, using EDTA-K2 as the anti-coagulant. Samples were centrifuged at 4000 g for 5 minutes in a 4℃ centrifuge and stored in a freezer at -75±15°C prior to analysis.

[0320] For brain PK analysis, the mice were fully exsanguinated prior to brain collectionusing the following procedure: open chest cavity, cut ventricle and perform a gentle iv saline 99 33049332.1 400807-032WO (218484)flush (saline flush volume ~ 10 ml) with the animal placed head down at a 45-degree angle to facilitate blood removal. All brain samples were collected at the indicated time points, quick frozen in ice box and kept at -75±15°C. All brain samples were weighed and homogenized with water by tissue weight (g) to water volume (mL) ratio 1:4 before analysis. The actual concentration was calculated as the detected value multiplied by the dilution factor.

[0321] The concentrations of exemplary compounds from Table 1 and their correspondinghydrolyzed parent compounds in both plasma and brain samples were analyzed using LC- MS / MS. The binding measurements in plasma and brain were conducted by using Rapid Equilibrium Dialysis Device. Cfree plasma and Cfree brain were represented with total concentration corrected with the unbound fraction in the study (PPB and BTB data). The unbound partition coefficient between plasma and brain (Kp,uu) was calculated according to standard techniques. Part II - Results

[0322] Results are presented in Table 4, below.

[0323] For exemplary compounds from Table 1 dosed orally, the result reported in Table 4 isthe ratio of the brain concentration at 24 hours vs. the IC50 of the compound in the Stable Cellular Assay for Inhibiting LINE1 Reverse Transcriptase described in Example 20. Both the brain concentration, and the IC50 value for determining the ratio, are the values for the corresponding hydrolyzed parent compound (i.e., neither value is for the for the compound from Table 1, itself, that was dosed orally). In the table, the symbol “**” represents a brain concentration of the parent compound that is greater than the IC50of the parent compound, while the symbol “*” represents a brain concentration of the parent compound that is less than the IC50 of the parent compound.

[0324] For the compounds dosed by IV (i.e., the hydrolyzed parent compounds correspondingto the exemplary compound from Table 1), the result reported in Table 4 is the Kp,uu at 4.5 hours after dosing. In the table, the symbol “+++” represents a Kp,uu > 1, the symbol “++” represents a Kp,uu between 0.5-1.0, and the symbol “+” represents a Kp,uu < 0.5. 100 33049332.1 400807-032WO (218484)TABLE 4. Chemical Structure Brain Conc. Parent Compound Kp,uu & Compound No. vs. IC50 Chemical Structure101 33049332.1 400807-032WO (218484)Chemical Structure Brain Conc. Parent Compound ound No. vs. I Kp,uu & Comp C50 Chemical Structure102 33049332.1 400807-032WO (218484)EXAMPLE 27 – Mitochondria Toxicity Assay with PC3 Cells

[0325] Exemplary compounds were tested for inhibition of expression of mitochondria DNAencoded proteins COX-1 (cyclooxygenase-1). SDH-A (succinate dehydrogenase-A) is a control mitochondria protein encoded by nuclear DNA. Reduction in COX-1 expression indicates off target inhibition of mitochondria DNA synthesis which leads to mitochondrial toxicity. Assay procedures and results are described below. Part I – Procedure

[0326] PC3 cells were purchased from ATCC and cultured in F-12 Kaighn's modifiedmedium (Hyclone), 10% heat-inactivated fetal bovine serum (Invitrogen), 1% Penicillin- Streptomycin (Invitrogen). PC3 cells were treated with a dose titration of a test compound on 96- well plates for 5 days at 37 °C, 5% CO2 in a humidified incubator. The treatment plates were duplicated, one for in-cell ELISA for mitochondria proteins and one for cytotoxicity.

[0327] In-cell ELISA was performed to measure mitochondrial proteins usingMitoBiogenesis In-Cell ELISA kit (Abcam) per manufacturer's instruction. The medium was aspirated from the compound treated cells, 100 μl per well of 4% paraformaldehyde (Solarbio) was added and cells were incubated at room temperature for 20 minutes. The cells were washed three times with PBS.100 μl of 0.5% acetic acid was added per cell, and the cells were incubated at room temperature for 5 minutes. Cells were washed once with PBS.100 μl per well of Permeabilization Buffer was added, and the cells incubate at room temperature for 30 minutes. The permeabilization buffer was removed and 200 μl per well 2× Blocking Buffer was added, and the cells were incubated at room temperature for 2 hours. After removing the Blocking Buffer 100 μl per well of primary antibody to SDH-A or COX-1 was added, and the cells were incubated at 4 °C overnight. The primary antibody solution was removed, and cells were washed 3 times with the Wash Buffer.100 μl per well of AP- and HRP-labelled secondary antibodies was added, and the cells were incubated at room temperature for 1 hour. The cells were then washed 4 times with the Wash Buffer. For SDH-A detection, 100 μl per well AP Development Solution was added, and the cells were incubated at room temperature for 10 minutes. Cells were then read on CLARTO starplusmicroplate reader (BMG Labtech) for kinetic readings at 405 nm for 5 minutes. For COX-1 detection, AP development Solution was removed, and 100 μl per well 103 33049332.1 400807-032WO (218484)HRP Development Solution was added, and the cells were incubated at room temperature for 2 minutes. The cells were then read on CLARTO starplusmicroplate reader (BMG Labtech) for kinetic readings at 600 nm for 15 minutes.

[0328] For the cytotoxicity assay, the medium from the cell viability plate was removed, andthen 50 μL per well of Cell Titer Glo (Promega, G9683) was added. Then the cells were incubated for 30 minutes at room temperature. Luminescence was read by Envision (PerkinElmer 2105).

[0329] Results for compounds tested using the above procedure appear in Table 5 below. Inthe table “A” represents values <1.0 μM; “B” represents values from >1.0 μM to ≤10.0 μM; “C” represents values from >10.0 μM to ≤50.0 μM; “D” represents values from >50.0 μM to ≤100.0 μM; and “E” represents values>100.0 μM. TABLE 5. IC50 IC50 IC50 C m nd COX 1 SDH A CTG104 33049332.1 400807-032WO (218484)IC50 IC50 IC50 Compound COX-1 SDH-A CTGEXAMPLE28 – 6-Hydroxydopamine (6-OHDA) Mouse Model for Parkinson’s Disease

[0330] Exemplary compounds may be tested for their effect in the 6-hydroxydopamine (6-OHDA) mouse model for Parkinson’s disease. Assay procedures are described below.

[0331] 6-OHDA injury was performed on 8-week-old male Crl:OF1 mice after 6 days oforally administering an exemplary compound. Mice are anesthetized and placed in a stereotactic frame. The anesthesia is maintained by isoflurane (4 % for induction, 2 % for maintenance) with a face mask coupled to the isoflurane vaporizer and oxygen concentrator machine. The skull is exposed, and a hole is drilled at the following coordinates: AP: -3.3 mm anterior; ML: 1 mm; DV: 4 mm from the surface of the skull, as described in Blaudin de Thé et al., 2018. A volume of 2 µL of 6-OHDA (0.5 mg / mL, 0.5 µL / min) is injected unilaterally (right hemisphere). In positive control groups, a volume of 2 µL of stavudine (10 µM, 0.5 µL / min, in NaCl 0.9 %) is injected unilaterally (right hemisphere), and 30 minutes later, 2 µL of 6-OHDA / stavudine solution (6-OHDA at 0.5 mg / mL and stavudine at 10 µM, 0.5 µL / min, in NaCl 0.9 %, ascorbic acid, 0.02 %) is injected unilaterally (right hemisphere). Depth of anesthesia and rectal temperature are verified every 5 minutes. After surgery, mice are allowed to recover before being placed back in the cage.

[0332] At the end of the experiment (on day 7, 24 hr after 6-OHDA injury), mice (n = 10 pergroup) are deeply anesthetized with a mix of Ketamine (100 mg / kg) : Xylasine (10 mg / kg), 1 hour after the last administration of exemplary compound. The whole blood is collected by heart puncture on lithium heparin tube and centrifuged at 1500 g, 10 minutes, 4 °C. Plasma is collected (2 samples of 120 µL) and placed in low-binding propylene tubes, snap frozen and stored at -80 °C until future use. 105 33049332.1 400807-032WO (218484)

[0333] Directly after blood sampling, mice are subjected to transcardial perfusion with coldPBS 1X (5 minutes), and cold paraformaldehyde (PFA) 4 % in PBS (5 minutes). Immunostaining is performed with n = 8-9 per group.

[0334] Brains are dissected and further fixed in PFA 4 % overnight at 4 °C. Afterwards,brains are placed in 30 % sucrose in PBS 1X solution at 4 °C. Coronal sections, including the substantia nigra pars compacta (SNpc), of 20 µm-thickness are cut.

[0335] For immunostaining, free-floating sections are incubated in PBS 1X with 5.0 %bovine serum albumin, and 0.3 % Triton X-100, for 1 hour at room temperature. This incubation blocks unspecific binding sites and permeabilized cell membranes.

[0336] Four (n = 4) brain sections per animal are processed and incubated overnight at roomtemperature with an antibody to tyrosine hydroxylase (TH) and appropriate secondary antibody. Images are acquired with confocal laser-scanning microscopy and number of TH positive cells are quantitated in the SNpc.EXAMPLE 29 – TDP-43-^ NLS Mouse Model for Amyotrophic Lateral Sclerosis

[0337] Exemplary compounds may be tested for their effect in the TDP-43-^ NLS mousemodel for amyotrophic lateral sclerosis. Assay procedures are described below.

[0338] Transgenic (Tg) rNLS8 mice are generated by breeding [B6;C3-Tg(tetO-TARDBP*)4Vle / J] mice expressing mutant hTDP-43-ΔNLS with [B6;C3-Tg(NEFH- tTA)8Vle / J] mice directing tTA expression using the human neurofilament heavy polypeptide promoter, creating “Tet-Off” offspring. Genotype is confirmed by PCR analysis of ear punch DNA prior to randomizing into experimental groups. To the extent possible, male and female mice are balanced across groups. During breeding and for the first 8-10 weeks-of-life, the animals are maintained on a high-dose doxycycline (DOX) diet (“Full DOX”). Disease induction begins by removal of the Full DOX diet (including a complete cage change) and placing mice on standard laboratory chow for 1 week. After 1 week, the animals are switched to a proprietary low DOX protocol (“Low DOX”) to extend the timing of clinical deterioration.

[0339] All mice receive test article or vehicle daily via oral gavage (p.o.) from 5 days prior todisease initiation until 8 weeks post disease initiation. Disease initiation begins by full removal of doxycycline diet for one week (Week 0), and then the mice are maintained on a low dose 106 33049332.1 400807-032WO (218484)protocol of doxycycline for seven weeks (eight weeks total). The mice are clinically assessed three times per week for the duration of their time off Full DOX diet (8 weeks). Clinical symptoms are scored in a blinded manner; signs of tremor, hindlimb clasping, hindlimb paralysis, grill agility, and overall well-being are scored from 0-3; 0 (not present), 1 (mild), 2 (moderate), or 3 (severe). All mice also undergo grip strength assessments at Baseline, Week 4, and Week 8, and in-life plasma collection at Week 4 and one hour prior to the last dose on Day 56. Following terminal CSF and blood collection, mice are perfused with 20-40 mL of cold PBS with heparin. The brains are immediately extracted and dissected along the midline. The left hemisphere is further dissected into forebrain and hindbrain. The right brain hemisphere is immersion-fixed in 10% neutral-buffered formalin. After 24 hours, the fixed right hemibrains are transferred to 30% sucrose until sunken (one or two days). The tissue is then embedded in the sagittal orientation in OCT and frozen on dry ice with a super-cooled bath of 2-methylbutate, and stored at -80°C prior to sectioning. The gastrocnemius muscles from both hindlimbs are also removed (with the soleus muscle still attached) and placed into separate tubes. Left hemi-brain regions and muscles are weighed, snap frozen in separate tubes, and stored at -80°C for furtherbiochemical analysis. Frozen brain hemispheres are sectioned at 20 μm thickness per section fordownstream immunofluorescence imaging.EXAMPLE 30 – Colony Forming Cellular Assay

[0340] Exemplary compounds may be tested for their effect in a colony forming cellularassay. Assay procedures are described below.

[0341] Primary human bone marrow cells were used to assess the effects of exemplarycompounds on erythroid and myeloid colony formation. To assess the effect of the test compounds on human myeloid and erythroid progenitor proliferation, a methylcellulose-based media formulation containing 25% FBS, 2% BSA, rhIL-3 (10 ng / mL), rhGM-CSF (10 ng / mL), Epo (3U / mL) and SCF (50 ng / mL) is used. Exemplary compounds are added directly to the methyl-cellulose-based matrix and vortexed to ensure an equal distribution of the compound throughout the medium. Cells from a pre-qualified donor are added to the matrix and the tubes vortexed once again to ensure equal distribution of the cells throughout the matrix. A standard tube containing the appropriate media formulation and a solvent control tube containing the solvent of choice are also included. 107 33049332.1 400807-032WO (218484)

[0342] Three replicates are used for each control and for each concentration of the testcompounds. The replicate dishes are placed at 37 °C, 5% CO2 for 14-16 days. Myeloid and erythroid colonies are evaluated microscopically in situ. The number of colonies in each treatment condition is counted. IC50values for the numbers of myeloid and erythroid colonies are determined using GraphPad Prism by non-linear regression with log(inhibitor) vs response – 4 parameters.EXAMPLE 31 – Rat Cellular Cytotoxicity Assay

[0343] Exemplary compounds may be tested for their toxicity in a rat cellular toxicity assay.Assay procedures are described below.

[0344] Rat cell lines GSL-9 (800 cells / well) and PC12 (1,000 cells / well) are plated in 384-well plates in 50 µL growth medium. Following an overnight incubation at 37 degrees and 5% CO2, compounds were added to the wells starting at 10 µM with 3-fold dilutions. Cells were incubated at 37 degrees for 72 hours. Cell viability was assessed using Cell Titer Glo, according to the manufacturer’s instructions. INCORPORATION BY REFERENCE

[0345] The entire disclosure of each of the patent documents and scientific articles referred toherein is incorporated by reference for all purposes. EQUIVALENTS

[0346] The invention may be embodied in other specific forms without departing from thespirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting the invention described herein. Scope of the invention is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein. 108 33049332.1 400807-032WO (218484)

Claims

Claims:

1. A compound represented by Formula I:or a pharmaceutically ac: R1is -(C0-4alkylene)-(phenyl substituted by m occurrences of R6), C10-C20alkenyl, C1-2alkyl, C3-4 alkyl, C5-6 alkyl, C7-10 alkyl, or -(C0-4 alkylene)-C3-6 cycloalkyl; R2is halomethyl, C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl; R3is hydrogen, -C(O)-(C1-2alkyl), -C(O)-(C3-6alkyl), -C(O)-(C7-10alkyl), -C(O)-(C0-4alkylene)-(phenyl substituted by n occurrences of R7), -C(O)-(C0-4alkylene)-C3-6cycloalkyl, or - C(O)-(C0-4 alkylene)-(3-7 membered saturated heterocyclyl containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur); R4is fluoro, chloro, bromo, iodo, hydrogen, -OH, C2-4alkynyl, or C1-3alkyl; R5is fluoro, chloro, bromo, iodo, or hydrogen; R6represents independently for each occurrence halo or C1-3 alkyl; R7represents independently for each occurrence halo or C1-3alkyl; m is 0, 1, or 2; and n is 0, 1, or 2.

2. The compound of claim 1, wherein the compound is a compound of Formula I.

3. The compound of claim 1 or 2, wherein R1 is -(C0-4 alkylene)-phenyl substituted by moccurrences of R6.

4. The compound of claim 1 or 2, wherein R1 is C10-C20 alkenyl.

5. The compound of claim 1 or 2, wherein R1 is C7-10 alkyl.109 33049332.1 400807-032WO (218484)6. The compound of claim 1 or 2, wherein R1 is .

7. The compound of claim 1 or 2, wherein R1 is C3-4 alkyl.

8. The compound of claim 1 or 2, wherein R1 is isopropyl.

9. The compound of any one of claims 1-8, wherein R2 is halomethyl.

10. The compound of any one of claims 1-8, wherein R2 is -CH2Cl, -CH2Br, -CH2I, or -CH2F.

11. The compound of any one of claims 1-8, wherein R2 is -CH2Cl.

12. The compound of any one of claims 1-8, wherein R2 is C1-3 alkyl.

13. The compound of any one of claims 1-8, wherein R2 is ethyl.

14. The compound of any one of claims 1-8, wherein R2 is C2-4 alkenyl.

15. The compound of any one of claims 1-8, wherein R2 is ethenyl.

16. The compound of any one of claims 1-8, wherein R2 is C2-4 alkynyl.

17. The compound of any one of claims 1-8, wherein R2 is ethynyl.

18. The compound of claim 1, wherein the compound is represented by Formula Ia or Ib, ora pharmaceutically acceptable salt thereof:

19. The compound of claim 1, wherein the compound is represented by Formula Ic or apharmaceutically acceptable salt thereof: 110 33049332.1 400807-032WO (218484)20. The comId or apharmaceutically acceptable salt thereof:

21. The compound of any on, is 1 or 2.

22. The compound of any one of claims 1-20, wherein n is 1 or 2.

23. The compound of any one of claims 1-20, wherein R3 is hydrogen.

24. The compound of any one of claims 1-20, wherein R3 is -C(O)-(C3-6 alkyl).

25. The compound of any one of claims 1-20, wherein R3 is -C(O)-(isopropyl).

26. The compound of any one of claims 1-20, wherein R3 is -C(O)-(C4-10 alkyl).O27. The compound of any one of claims 1-20, wherei.

28. The compound of any one of claims 1-20, wherein R is -C(O)-(phenyl).

29. The compound of any one of claims 1-20, wherein R3 is -C(O)-(C0-4 alkylene)-(3-7membered saturated heterocyclyl containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur). 111 33049332.1 400807-032WO (218484)30. The compound of any one of claims 1-20, wherein R3 .

31. The compound of any one of claims 1-30, wherein R4loro, bromo, or iodo.

32. The compound of any one of claims 1-30, wherein R4 is fluoro.

33. The compound of any one of claims 1-30, wherein R4 is hydrogen.

34. The compound of any one of claims 1-33, wherein R5 is fluoro, chloro, bromo, or iodo.

35. The compound of any one of claims 1-33, wherein R5 is fluoro.

36. The compound of any one of claims 1-33, wherein R5 is hydrogen.

37. A compound in Table 1 herein, or a pharmaceutically acceptable salt thereof.

38. A compound represented by Formula II:or a pharmaceutically accep, ein: R1is -C(O)-(C1-2alkyl), -C(O)-(C3-6alkyl), or -C(O)-(C7-10alkyl); and R2is C1-3alkyl or C2-4alkenyl.

39. The compound of claim 38, wherein the compound is a compound of Formula II.

40. The compound of claim 38 or 39, wherein R1 is -C(O)-(C3-6 alkyl).

41. The compound of claim 38 or 39, wherein R1 is -C(O)-(isopropyl).

42. The compound of any one of claims 38-41, wherein R2 is C1-3 alkyl.

43. The compound of any one of claims 38-41, wherein R2 is C2-4 alkenyl.

44. A compound in Table 2 herein, or a pharmaceutically acceptable salt thereof.112 33049332.1 400807-032WO (218484)45. A pharmaceutical composition comprising a compound according to any one of claims 1-44 and a carrier, excipient, and / or vehicle.

46. A method of treating a disorder selected from the group consisting of cancer, anautoimmune disorder, and a neurological disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-44 to treat the disorder.

47. The method of claim 46, wherein the disorder is cancer.

48. The method of claim 47, wherein the cancer is breast cancer, ovarian cancer, uterinecancer, cervical cancer, prostate cancer, testicular cancer, lung cancer, leukemia, head and neck cancer, oral cancer, esophageal cancer, stomach cancer, bile duct cancer, gallbladder cancer, bladder cancer, urinary tract cancer, colon cancer, rectal cancer, thyroid cancer, pancreatic cancer, kidney cancer, liver cancer, brain cancer, skin cancer, or eye cancer.

49. The method of claim 46, wherein the disorder is an autoimmune disorder.

50. The method of claim 49, wherein the autoimmune disorder is selected from Aicardi-Goutieres syndrome, rheumatoid arthritis, psoriasis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), graft versus host disease, scleroderma, type I diabetes, dermatomyositis, inflammatory bowel disease, ulcerative colitis, Crohn’s disease, vasculitis, and Sjögren’s syndrome.

51. The method of claim 49, wherein the autoimmune disorder is systemic lupuserythematosus (SLE).

52. The method of claim 46, wherein the disorder is a neurological disorder.

53. The method of claim 52, wherein the neurological disorder is Alzheimer’s disease,amyotrophic lateral sclerosis (ALS), multiple sclerosis, Parkinson’s disease, Huntington’s disease, peripheral neuropathy, age-related macular degeneration, Creutzfeldt-Jacob disease, stroke, prion disease, frontotemporal dementia, Pick’s disease, progressive supranuclear palsy, spinocerebellar ataxias, Lewy body disease, dementia, multiple system atrophy, epilepsy, bipolar disorder, schizophrenia, an anxiety disorder, or major depression.

54. A method of inhibiting LINE1 reverse transcriptase activity in a subject, the methodcomprising contacting a LINE1 reverse transcriptase with an effective amount of a compound 113 33049332.1 400807-032WO (218484)according to any one of claims 1-44 in order to inhibit the activity of said LINE1 reverse transcriptase.

55. The method of claim 54, wherein the subject has (i) elevated expression of LINE1 RNA,LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; and / or (ii) elevated activity of LINE1 reverse transcriptase. 114 33049332.1 400807-032WO (218484)

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