Crystalline compounds, pharmaceutical compositions, and methods for treating disease

Abstract

The present disclosure provides crystalline oxopyrimidinyl-((isobutyryloxy) methyl)tetrahydrofuranyl compounds, pharmaceutical compositions containing crystalline compounds, and methods of treatment using the crystalline compounds.

Description

CRYSTALLINE COMPOUNDS, PHARMACEUTICAL COMPOSITIONS, AND METHODS FOR TREATING DISEASE CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to United States Provisional Patent Application serial number 63 / 582,729, filed September 14, 2023, the contents of which are hereby incorporated by reference. FIELD OF THE INVENTION

[0002] The present disclosure provides crystalline oxopyrimidinyl-((isobutyryloxy) methyl)tetrahydrofuranyl compounds, pharmaceutical compositions containing crystalline compounds, and methods of treatment using the crystalline compounds. BACKGROUND

[0003] Transposable elements (or transposons) are genomic DNA sequences that have the ability to move within the genome which leads to altering its organization, increase its size and creates 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).

[0004] Long Interspersed Nuclear Element 1 (LINE-1) are class I transposable elements in the 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 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, 1 32017260 400807-030WO (212251)Development, 143:4101-4114 (2016)). LINE-1 activity is normally tightly regulated in the germline by DNA methylation, histone modifications, and piRNA.

[0005] Retrotransposons are transposable elements which are associated with the pathogenesis 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).

[0006] Pathogenic interferon production is a characteristic feature of type I interferonopathies. 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 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 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. 2 32017260 400807-030WO (212251)

[0007] Hypomethylated and highly expressed LINE-1 has been found in many patients with autoimmune 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 QUZMXNMXVU i IUL QUZMXNMXVU k) #@[XQ_ITI MZ IS' J. Am. Acad, Dermatol., 84(4):1103-1105 (2020)).

[0008] LINE-1 has also been implicated in neurological disorders such as ataxia telangiectasia (AT), Rett syndrome, 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).

[0009] Cancer continues to be a significant health problem despite the substantial research efforts 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.

[0010] High LINE-1 activity has been found in many tumor tissues. LINE-1 RT uses a procedure 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, 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. 3 32017260 400807-030WO (212251)

[0011] Human endogenous retroviruses (HERVs) comprise nearly 8% of the human genome and are believed to be derived from ancient integrations of retroviruses into the germline. The biology of HERVs is poorly defined, but there is accumulating evidence supporting pathological roles in diverse diseases such as cancer, autoimmune, neurodegenerative diseases, and aging. Functional proteins are produced by HERV-encoded genes including reverse transcriptases (RTs), which could be a contributor to the pathology attributed to aberrant HERV-K expression.

[0012] HERVs play a role in early development by rewiring the gene regulatory network of the preimplantation embryo (Fu et al, Biomolecules, 11(6):829 (2021)). HERV expression appears to be a hallmark of the undifferentiated state, the acquisition of phenotypic plasticity and stem cell character (Balestrieri et al, Frontiers in Microbiology, 9:1448 (2018)); traits associated with aggressive cancer and poor patient outcomes. HERV expression is normally tightly controlled in normal adult tissues but is reported to be aberrantly expressed in cancer (Downey et al, Int. J. Cancer, 137(6):1249-1257 (2015)), inflammatory diseases (Greenig, PeerJ 7:e6711 (2019)), neurological diseases (Kury et al, Trends Mol. Med., 24(4):379-394 (2018)), aging (Gorbunova et al, Nature, 596(7870):43-53 (2021)), and viral disease (Romer, Frontiers in Neuroscience, 15:648629-648629 (2021)). There are numerous reports of upregulation of HERV-K [HML-2 (human endogenous MMTV-like) subtype derived mRNA and protein in a variety of solid and liquid tumor types (Dervan et al, Front. Onc., 11:658489 (2021); Hohn et al, Front. Onc., 3:246 (2013)). The disease association with endogenous retroviruses and the expression of HERV encoded proteins during disease states suggests that anti-retroviral therapy could be explored in the management of these conditions.

[0013] Compounds for treating medical disorders, such as cancer, autoimmune disease, neurological disorders, aging, and diseases associated with aging are needed. Particularly beneficial are compounds in a particular physical form that provide advantages for use in medicine, which may include superior performance properties that are beneficial for the manufacturing process used to make the active ingredient. As appreciated, different crystalline forms of a compound can have different physical and chemical (i.e., physicochemical) properties such as crystal shape, density, hardness, color, chemical stability, melting point, hygroscopicity, suspendability and dissolution rate, and biological availability. 4 32017260 400807-030WO (212251)

[0014] The present disclosure addresses the foregoing needs and provides other related advantages. SUMMARY

[0015] The present disclosure provides crystalline oxopyrimidinyl-((isobutyryloxy) methyl)tetrahydrofuranyl compounds, pharmaceutical compositions containing crystalline compounds, and methods of treatment using the crystalline compounds.

[0016] One aspect of the disclosure provides solid forms of Compound I:

[0017] Another aspect of theform of Compound II:

[0018] Further description ofprovided herein. The compounds may be presented in a pharmaceutical composition containing a pharmaceutically acceptable carrier.

[0019] The disclosed compounds and compositions may be used for treating medical disorders, such as by inhibiting LINE1 reverse transcriptase and / or HERV-K reverse transcriptase. For example, one aspect of the disclosure provides a method of treating a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder, comprising 5 32017260 400807-030WO (212251)administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein to treat the disorder.

[0020] Another aspect of the disclosure provides a method of inhibiting LINE1 reverse transcriptase 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. In certain embodiments, the method further comprises inhibiting HERV-K reverse transcriptase activity in the subject.

[0021] Another aspect of the disclosure provides a method of inhibiting HERV-K reverse transcriptase activity in a subject, comprising contacting a HERV-K reverse transcriptase with an effective amount of a compound disclosed herein to inhibit the activity of said HERV-K reverse transcriptase. BRIEF DESCRIPTION OF FIGURES

[0022] FIG.1 depicts an XRPD diffractogram of Compound II.

[0023] FIG.2 depicts an XRPD pattern overlay of crystalline forms A, B, C, and D of Compound I.

[0024] FIG.3 depicts an XRPD diffractogram of crystalline Form A of Compound I.

[0025] FIG.4 depicts TGA and DSC thermograms of crystalline Form A of Compound I.

[0026] FIG.5 depicts a DVS analysis of crystalline Form A of Compound I.

[0027] FIG.6 depicts an XRPD diffractogram used in indexing crystalline Form A of Compound I.

[0028] FIG.7 depicts an XRPD diffractogram of crystalline Form C of Compound I.

[0029] FIG.8 depicts TGA and DSC thermograms of crystalline Form C of Compound I.

[0030] FIG.9 depicts an XRPD diffractogram used in indexing of crystalline Form B of Compound I.

[0031] FIG.10 depicts TGA and DSC thermograms of crystalline Form B of Compound I.

[0032] FIG.11 depicts a DVS analysis of crystalline Form B of Compound I.

[0033] FIG.12 depicts an XRPD diffractogram of crystalline Form D of Compound I. 6 32017260 400807-030WO (212251)

[0034] FIG.13 depicts TGA and DSC thermograms of crystalline Form D of Compound I.

[0035] FIG.14 is a graph depicting normalized interferon levels in THP1-DualTMKO- TREX1 xenografts from mice treated once daily, for four days with decitabine (DAC) and varying doses of Compound A. The graph depicts data obtained on day 5, with tumor harvested 24 hours after the final decitabine dosing on day 4. DETAILED DESCRIPTION

[0036] The present disclosure provides crystalline oxopyrimidinyl-((isobutyryloxy) methyl)tetrahydrofuranyl compounds, pharmaceutical compositions containing crystalline compounds, and methods of treatment using the crystalline compounds.

[0037] Differences between solid forms of an active pharmaceutical compound can have profound effects on the properties of the compound. For example, differences can arise in the crystallinity, solubility, intrinsic dissolution rate, bioavailability, stability to mechanical grinding, storage stability, and stability in aqueous and other media of a polymorphic form of a compound as compared to the amorphous and other polymorphic forms of the same compound.

[0038] Polymorphism refers to the ability of a solid substance to exist in one or more distinct crystal structures (i.e., with one or more distinct arrangements of molecules relative to each other in the crystal lattice). Different polymorphs of a substance may have different physical properties such as bioavailability, solubility, intrinsic dissolution rate and calorimetric behavior (e.g., melting point). Different polymorphs may also exhibit differences in stability (e.g., differences in stability with respect to conversion to other crystalline or amorphous forms or differences in stability to grinding). The physical properties of an active pharmaceutical ingredient may affect the drug product safety performance and efficacy. It is therefore advantageous to identify polymorphic forms of a drug substance which have pharmaceutically acceptable properties.

[0039] Various aspects of the invention are set forth below in sections; however, aspects of the invention 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. 7 32017260 400807-030WO (212251)Definitions

[0040] The terms “a” and “an” as used herein mean “one or more” and include the plural, unless the context is inappropriate.

[0041] As used herein, the terms “subject” and “patient” are used interchangeably and refer to organisms to be treated by the methods of the present invention. 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.

[0042] The term “inhibit”, “inhibition”, or “inhibiting” includes a decrease in the baseline activity of a biological activity or process.

[0043] As used herein, the term “effective amount” refers to the amount of a compound that will elicit a biological or medical response in a subject, for example, the reduction or inhibition of enzyme or protein activity relative to the baseline levels before treatment. Typically, an “effective amount” of a compound is one which is sufficient to effect some beneficial change 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, reducing, modulating, ameliorating, or eliminating, that results in the improvement of the condition, disease, disorder, and the like, or ameliorating a symptom thereof. For example, treatment of cancer may mean prolonging the period of time where tumor burden does not increase (progression-free survival), reduction of the tumor burden, extension of the overall survival time of a patient, amelioration of symptoms associated with the cancer, prevention of metastasis, slowing of metastasis, and the like. Treatment of an autoimmune disease includes reduction of the symptoms of the disease, extension of time between disease flare-ups, remission of disease, prevention of worsening of the disease, and the like. Treatment of neurological disease may include improvement of cognitive function, reduction of the rate of cognitive loss, reduction of symptoms, and the like.

[0044] In some embodiments, treatment is administered after one or more symptoms have developed. In some embodiments, treatment is administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and / or in light of genetic or other susceptibility 8 32017260 400807-030WO (212251)factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.

[0045] As used herein, the term “pharmaceutical composition” refers to the combination of an active agent with a carrier, adjuvant, or vehicle, inert or active, making the composition especially suitable for diagnostic or therapeutic use in vivo or ex vivo.

[0046] As used herein, the term “pharmaceutically acceptable carrier, adjuvant, and / or vehicle” refers to any non-toxic carrier, adjuvant, and / or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants, and / or vehicles that are used in the compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as potassium sorbate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, 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 / or preservatives. For examples of carriers, vehicles and adjuvants, see e.g., Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA

[1975] .

[0047] The term “combination” refers to simultaneous, separate, or sequential administration. In one aspect of the disclosure, “combination” refers to simultaneous administration. In another aspect of the disclosure, “combination” refers to separate administration. In another aspect of the disclosure, “combination” refers to sequential administration. Where the administration is separate or sequential, the delay in administering the one or more additional therapeutic agents is done at an interval designed such as not to lose the beneficial effect of the combination.

[0048] Throughout the description, where compositions are described as having, including, or comprising 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 invention that consist essentially of, or consist of, the recited 9 32017260 400807-030WO (212251)components, and that there are processes and methods according to the present invention that consist essentially of, or consist of, the recited processing steps.

[0049] As used herein, the term “comprising” or “comprises” is used in reference to compounds, uses, compositions, methods, and respective component(s) thereof that are essential to the method or composition, yet open to the inclusion of unspecified elements, whether essential or not. The term “consisting of” refers to compounds, uses, compositions, methods, and respective component(s) thereof as described herein which are exclusive of any element not recited in that description of the embodiment. The term “consisting essentially of” refers to those elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic and novel functional characteristic(s) of that embodiment.

[0050] As a general matter, compositions specifying a percentage are by weight unless otherwise specified.

[0051] “About,” as used herein, means approximately, in the region of, roughly, or around. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term “about” is used herein to modify a numerical value above and below the stated value by a variance of 10%. Therefore, about 50% means in the range of 45%-55%. Numerical ranges recited herein by endpoints include all numbers and fractions subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.90, 4, and 5). It is also to be understood that all numbers and fractions thereof are presumed to be modified by the term “about.”

[0052] “Anhydrate” or “anhydrous polymorph” or “anhydrous crystalline form” refers to a crystalline form that does not have water bound in the crystal lattice. However, the crystals may contain trace amount of water or other solvents not bound in the crystal lattice.

[0053] “Amorphous” refers to a solid state wherein the molecules are present in a disordered arrangement and do not form a distinguishable crystal lattice or unit cell.

[0054] “Chemical purity” refers to the overall level of a desired product. If a compound is present in enantiomeric forms, “chemical purity” as used herein would include both enantiomeric forms in the calculation of the overall level of the desired product. If a compound is present in solvate forms, “chemical purity” as used herein would include the solvate in the calculation of 10 32017260 400807-030WO (212251)the overall level of the desired product. Impurities may be in the form of, e.g., the presence of unwanted process reagents, process intermediates, degradation products or oxidation products. In particular embodiments, the chemical purity is high, that is greater than 90% chemical purity, especially above 92.5%, 95%, 96%, 97%, 98%, 99%, 99.9, and includes 100%. Purity may be measured by a variety of techniques, including HPLC analysis.

[0055] “Enantiomer” and “enantiomeric” refers to a molecule that cannot be superimposed on its mirror image and hence is optically active, wherein the enantiomer rotates the plane of polarized light in one direction and its minor image compound rotates the plane of polarized light to the same degree, in the opposite direction.

[0056] “Enantiomeric excess” or “e.e.” refers to a difference between the amount of one enantiomer and the amount of the other enantiomer that is present in the product mixture. The enantiomeric excess value in each example given below gives an indication of the relative amount of each enantiomer. The value is defined as the difference between the relative percentages for the two enantiomers. Thus, for example, when the percentage of the (S)- enantiomer of the compound of the invention is 97.5% and the percentage for the (R)-enantiomer is 2.5%, the enantiomeric excess for the (S)-enantiomer is 95%.

[0057] “Enantiomerically pure” or “enantiomeric purity” is a measure of how much more of one enantiomer there is than the other enantiomer in a mixture of enantiomers. For example, a mixture of 99% (S)-enantiomer and 1% (R)-enantiomer has 99% enantiomeric purity of the (S)- enantiomer. Enantiomerically pure is preferably at least 95% or at least 98% enantiomeric purity, more preferably at least about 99%. In another embodiment enantiomerically is about 99.90% to about 100% enantiomeric purity.

[0058] “Isolated”, referring to solid state forms of compounds of the present disclosure, corresponds to a solid state form of the compound that is physically separated from the solution in which it is formed.

[0059] “Volume of solvent” refers to the volume of solvent, expressed in liters at ambient temperature, used in a process to dissolve 1 kg of solid material. For example, 5 volumes of solvent used in a process starting with 1 kg of starting material would equal 5 liters of solvent. 11 32017260 400807-030WO (212251)

[0060] “Non-solvate polymorph” or “non-solvate crystalline form” refer to a crystalline form that does not have a solvent bound in the crystal lattice, for example an anhydrous polymorph. However, the crystals may contain trace amount of solvent not bound in the crystal lattice.

[0061] “Polymorph” refers to the different crystal structures (of solvated or non-solvated forms) in which a compound can crystallize.

[0062] “Racemic” or “racemate”, and other like terms refer to generally equimolar proportions of a (S)-compound and a (R)- compound.

[0063] “Solvate”, “solvate polymorph” or “solvate crystalline form” refer to a crystalline form that has solvate bound in the crystal lattice.

[0064] “Substantially pure crystal form”, unless otherwise specified is to be understood as a substance free of other crystal forms, or amorphous form, at amounts detectable with typical analytical methods such as X-ray powder diffraction and / or solid state infrared absorption, i.e., containing less than 10% of other crystal forms. Preferably, there is less than 5%, more preferably less than 2%, and even more preferably less than l% of any other crystal form, or amorphous form, of the compound present.

[0065] When used in reference to a diffractogram, a spectrum or data presented in a graph, the term “substantially similar” means that the subject diffractogram, spectrum or data presented in a graph encompasses all diffractograms, spectra or data presented in graphs that vary within acceptable boundaries of experimentation that are known to a person of skill in the art. Such boundaries of experimentation will vary depending on the type of the subject diffractogram, spectrum or data presented in a graph, but will nevertheless be known to a person of skill in the art. I. Crystalline Compounds

[0066] One aspect of the disclosure provides crystalline compounds. The compounds may be used in the pharmaceutical compositions and therapeutic methods described herein. For example, one aspect of the disclosure provides a crystalline Compound I: 12 32017260 400807-030WO (212251)

[0067] Multiple crystalline forms of Compound I have been identified, and are described herein.

[0068] Another aspect of the disclosure provides a crystalline Compound II:.

[0069] Crystalline Compounds I and II are described in more detail below. Part A: Crystalline Form A of Compound I

[0070] One aspect of the disclosure provides crystalline Form A of Compound I. The crystalline form may be characterized according to, for example, XRPD, differential scanning calorimetry, and thermogravimetric analysis. Exemplary features are described in more detail below.

[0071] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.6 ± 0.2, 7.6 ± 0.2, 9.3 ± 0.2, 14.0 ± 0.2, 15.3 ± 0.2, 19.2 ± 0.2, and 23.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 5.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 6.7 ± 0.2. In certain embodiments, the X-ray powder diffraction 13 32017260 400807-030WO (212251)pattern further comprises a peak at the following diffraction angle (2u): 7.4 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.2 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 17.8 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.3 ± 0.2.

[0072] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.61 ± 0.20, 7.64 ± 0.20, 9.26 ± 0.20, 13.98 ± 0.20, 15.33 ± 0.20, 19.20 ± 0.20, and 23.10 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 5.06 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 6.73 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.35 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.19 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 17.82 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.30 ± 0.20.

[0073] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 7.64 ± 0.20, 9.26 ± 0.20, 13.98 ± 0.20, 15.33 ± 0.20, 19.20 ± 0.20, and 23.10 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 4.61 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 5.06 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 6.73 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.35 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.19 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the 14 32017260 400807-030WO (212251)following diffraction angle (2u): 17.82 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.30 ± 0.20.

[0074] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 7.6 ± 0.2, 9.3 ± 0.2, 14.0 ± 0.2, 15.3 ± 0.2, 19.2 ± 0.2, and 23.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 4.6 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 5.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 6.7 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.4 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.2 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 17.8 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.3 ± 0.2.

[0075] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising at least six peaks selected from the following diffraction angles (2u): 4.6 ± 0.2, 5.1 ± 0.2, 6.7 ± 0.2, 7.4 ± 0.2, 7.6 ± 0.2, 9.2 ± 0.2, 9.3 ± 0.2, 14.0 ± 0.2, 15.3 ± 0.2, 17.8 ± 0.2, 19.2 ± 0.2, 19.3 ± 0.2, and 23.1 ± 0.2. In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising at least seven peaks selected from said diffraction angles (2u).

[0076] In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 20%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 15%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 10%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 5%.

[0077] In certain embodiments, the crystalline compound is characterized by the following X- ray powder diffraction pattern expressed in terms of diffraction angle 2u, inter-planar distances d, and relative intensity (expressed as a percentage with respect to the most intense peak): 15 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%] 461 ± 020 19153 ± 0830 10016 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%] 1930 ± 020 4595 ± 0047 11.

[0078] In certain embodiments, the compound is characterized by having an x-ray powder diffraction (XRPD) pattern comprising three, four, five, six or seven or more peaks having an 17 32017260 400807-030WO (212251)approximate 2u ± 0.20 value selected from the group consisting of: 4.61, 5.06, 6.73, 7.35, 7.64, 9.19, 9.26, 13.98, 15.33, 17.82, 19.20, 19.30, and 23.10.

[0079] In certain embodiments, the X-ray powder diffraction pattern is substantially as shown in FIG.3.

[0080] In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 124 degrees Celsius to about 128 degrees Celsius. In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry at about 126 degrees Celsius.

[0081] In certain embodiments, the compound has a peak melting temperature of about 130 degrees Celsius measured by differential scanning calorimetry (DSC). In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a peak at a temperature of about 130.2°C, and an onset at about 126.4°C, measured with a heating rate of 10°C / min. In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a heat of fusion of about 49.2 J / g.

[0082] In certain embodiments, the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.4. Part B: Crystalline Form B of Compound I

[0083] One aspect of the disclosure provides crystalline Form B of Compound I. The crystalline form may be characterized according to, for example, XRPD, differential scanning calorimetry, and thermogravimetric analysis. Exemplary features are described in more detail below.

[0084] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.4 ± 0.2, 5.8 ± 0.2, 7.5 ± 0.2, 9.7 ± 0.2, 15.1 ± 0.2, 18.1 ± 0.2, and 22.7 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.3 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.7 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.8 ± 0.2. In certain embodiments ,the X-ray powder diffraction pattern further comprises a peak at the 18 32017260 400807-030WO (212251)following diffraction angle (2u): 8.9 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.6 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 12.43 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 13.9 ± 0.2.

[0085] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.44 ± 0.20, 5.78 ± 0.20, 7.51 ± 0.20, 9.65 ± 0.20, 15.08 ± 0.20, 18.05 ± 0.20, and 22.71 ± 0.20. In certain embodiments, the X- ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.31 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.71 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.79 ± 0.20. In certain embodiments ,the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.89 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.57 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 12.43 ± 0.20. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 13.89 ± 0.20.

[0086] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising at least six peaks selected from the following diffraction angles (2u): 4.4 ± 0.2, 5.8 ± 0.2, 7.3 ± 0.2, 7.5 ± 0.2, 8.7 ± 0.2, 8.8 ± 0.2, 8.9 ± 0.2, 9.6 ± 0.2, 9.7 ± 0.2, 12.4 ± 0.2, 13.9 ± 0.2, 15.1 ± 0.2, 18.1 ± 0.2, and 22.7 ± 0.2. In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising at least seven peaks selected from said diffraction angles (2u).

[0087] In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 20%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 15%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 10%. 19 32017260 400807-030WO (212251)

[0088] In certain embodiments, the crystalline compound is characterized by the following X- ray powder diffraction pattern expressed in terms of diffraction angle 2u, inter-planar distances d, and relative intensity (expressed as a percentage with respect to the most intense peak): Angle [2u]d-spacing [Å] Relative Intensity [%]20 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%] .

[0089] In certain embodiments, the compound is characterized as having an x-ray powder diffraction (XRPD) pattern comprising three, four, five, six or seven or more peaks having an approximate 2u ± 0.20 value selected from the group consisting of: 4.44, 5.78, 7.31, 7.51, 8.71, 8.79, 8.89, 9.57, 9.65, 12.43, 13.89, 15.08, 18.05, and 22.71. In certain embodiments, the X-ray powder diffraction pattern is substantially as shown in FIG.9.

[0090] In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 125 degrees Celsius to about 129 degrees Celsius. In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry at about 127 degrees Celsius. In certain embodiments, the compound has a peak melting temperature of about 132 degrees Celsius measured by differential scanning calorimetry (DSC).

[0091] In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a peak at a temperature of about 132.0°C, and an onset at about 127.1°C, measured with a heating rate of 10°C / min. In certain embodiments, 21 32017260 400807-030WO (212251)the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a heat of fusion of about 50.1 J / g.

[0092] In certain embodiments, the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.10. Part C: Crystalline Form C of Compound I

[0093] One aspect of the disclosure provides crystalline Form C of Compound I. The crystalline form may be characterized according to, for example, XRPD, differential scanning calorimetry, and thermogravimetric analysis. Exemplary features are described in more detail below.

[0094] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.4 ± 0.3, 5.0 ± 0.3, 6.5 ± 0.3, 7.2 ± 0.3, 9.1 ± 0.3, 13.9 ± 0.3, and 17.8 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 18.2 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.2 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 22.1 ± 0.3.

[0095] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.4 ± 0.2, 5.0 ± 0.2, 6.5 ± 0.2, 7.2 ± 0.2, 9.1 ± 0.2, 13.9 ± 0.2, and 17.8 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 18.2 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.2 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 22.1 ± 0.2.

[0096] In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 20%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 15%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 10%. 22 32017260 400807-030WO (212251)

[0097] In certain embodiments, the X-ray powder diffraction pattern is substantially as shown in FIG.7.

[0098] In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 127 degrees Celsius to about 131 degrees Celsius. In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry at about 129 degrees Celsius.

[0099] In certain embodiments, the compound has a peak melting temperature of about 132 degrees Celsius measured by differential scanning calorimetry (DSC). In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a peak at a temperature of about 131.9°C, and an onset at about 128.5°C, measured with a heating rate of 10°C / min. In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a heat of fusion of about 49.9 J / g.

[0100] In certain embodiments, the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.8. Part D: Crystalline Form D of Compound I

[0101] One aspect of the disclosure provides crystalline Form D of Compound I. The crystalline form may be characterized according to, for example, XRPD, differential scanning calorimetry, and thermogravimetric analysis. Exemplary features are described in more detail below.

[0102] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.2 ± 0.3, 5.7 ± 0.3, 7.3 ± 0.3, 9.5 ± 0.3, 10.4 ± 0.3, 13.9 ± 0.3, and 22.3 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.8 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 12.2 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 15.9 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.1 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 29.9 ± 0.3. 23 32017260 400807-030WO (212251)

[0103] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.2 ± 0.2, 5.7 ± 0.2, 7.3 ± 0.2, 9.5 ± 0.2, 10.4 ± 0.2, 13.9 ± 0.2, and 22.3 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.8 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 12.2 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 15.9 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 29.9 ± 0.2.

[0104] In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 20%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 15%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 10%. .

[0105] In certain embodiments, the X-ray powder diffraction pattern is substantially as shown in FIG.12.

[0106] In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 126 degrees Celsius to about 130 degrees Celsius. In certain embodiments, the compound has a melting point onset as determined by differential scanning calorimetry at about 128 degrees Celsius. In certain embodiments, the compound has a peak melting temperature of about 132 degrees Celsius measured by differential scanning calorimetry (DSC).

[0107] In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a peak at a temperature of about 132.2°C, and an onset at about 127.7°C, measured with a heating rate of 10°C / min. In certain embodiments, the compound is characterized as having a differential scanning calorimetry (DSC) thermogram having a heat of fusion of about 49.9 J / g.

[0108] In certain embodiments, the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.13. 24 32017260 400807-030WO (212251)Part E: Crystalline Compound II

[0109] One aspect of the disclosure provides a crystalline Compound II. The crystalline form may be characterized according to, for example, XRPD, differential scanning calorimetry, and thermogravimetric analysis. Exemplary features are described in more detail below.

[0110] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 8.9 ± 0.3, 13.3 ± 0.2, 19.6 ± 0.3, 20.7 ± 0.3, 21.6 ± 0.3, 26.8 ± 0.3, and 35.7 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 16.7 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 18.1 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 27.7 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 29.1 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 33.9 ± 0.3. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 35.0 ± 0.3.

[0111] In certain embodiments, the crystalline compound exhibits an X-ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 8.9 ± 0.2, 13.3 ± 0.2, 19.6 ± 0.2, 20.7 ± 0.2, 21.6 ± 0.2, 26.8 ± 0.2, and 35.7 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 16.7 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 18.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 27.7 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 29.1 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 33.9 ± 0.2. In certain embodiments, the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 35.0 ± 0.2. 25 32017260 400807-030WO (212251)

[0112] In certain embodiments, the compound is characterized as having an x-ray powder diffraction (XRPD) pattern comprising three, four, five, six or seven or more peaks having an approximate 2u value selected from the group consisting of: 8.9, 13.3, 16.7, 18.1, 19.6, 20.7, 21.6, 26.8, 27.7, 29.1, 33.9, 35.0, and 35.7. In certain embodiments, the compound is characterized as having an x-ray powder diffraction (XRPD) pattern comprising three, four, five, six or seven or more peaks having an 2u value selected from the group consisting of: 8.9 ± 0.3, 13.3 ± 0.2, 16.7 ± 0.3, 18.1 ± 0.3, 19.6 ± 0.3, 20.7 ± 0.3, 21.6 ± 0.3, 26.8 ± 0.3, 27.7 ± 0.3, 29.1 ± 0.3, 33.9 ± 0.3, 35.0 ± 0.3, and 35.7 ± 0.3.

[0113] In certain embodiments, the compound is characterized as having an x-ray powder diffraction (XRPD) pattern comprising three, four, five, six or seven or more peaks having an 2u value selected from the group consisting of: 8.9 ± 0.2, 13.3 ± 0.2, 16.7 ± 0.2, 18.1 ± 0.2, 19.6 ± 0.2, 20.7 ± 0.2, 21.6 ± 0.2, 26.8 ± 0.2, 27.7 ± 0.2, 29.1 ± 0.2, 33.9 ± 0.2, 35.0 ± 0.2, and 35.7 ± 0.2.

[0114] In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 20%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 15%. In certain embodiments, the relative intensity of the peak at said diffraction angles (2u) is at least 10%.

[0115] In certain embodiments, the X-ray powder diffraction pattern is substantially as shown in FIG.1. Part F: General Additional Features

[0116] Crystalline forms in Parts A through E above may be further characterized. For example, in certain embodiments, the crystalline form is characterized according to chemical purity. For example, in certain embodiments, the crystalline form has a chemical purity of at least about 95%. In certain embodiments, the crystalline form has a chemical purity of at least about 98%. In certain embodiments, the crystalline form has a chemical purity of at least about 99%. In certain embodiments, the crystalline form has a chemical purity in the range of about 98.00% to about 99.00%. In certain embodiments, the crystalline form has a chemical purity in the range of about 99.00% to about 99.95%. In certain embodiments, the crystalline form is a substantially pure crystal form. 26 32017260 400807-030WO (212251)

[0117] In certain embodiments, the crystalline form is isolated. II. Therapeutic Applications

[0118] It is contemplated that crystalline forms in Section 1 above provide therapeutic benefits to subjects suffering from a medical disorder, such as cancer, autoimmune disorders, and / or neurological disorders.

[0119] One aspect of the disclosure provides a method of treating a disorder selected from the group 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.

[0120] Another aspect of the disclosure provides a method of inhibiting LINE1 reverse transcriptase 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. In certain embodiments, the method further comprises inhibiting HERV-K reverse transcriptase activity in the subject.

[0121] Another aspect of the disclosure provides a method of inhibiting HERV-K reverse transcriptase activity in a subject, comprising contacting a HERV-K reverse transcriptase with an effective amount of a compound disclosed herein to inhibit the activity of said HERV-K reverse transcriptase.

[0122] In certain embodiments, the subject has a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder. In certain embodiments, the disorder is cancer.

[0123] 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 crystalline Compound I) to treat the viral infection.

[0124] In certain embodiments of each of the foregoing methods, the compound is crystalline Compound I. Cancer 27 32017260 400807-030WO (212251)

[0125] In certain embodiments, the disorder is cancer. In certain embodiments, the cancer is a 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.

[0126] 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.

[0127] 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, or bladder cancer.

[0128] In certain embodiments, the cancer has (i) expression of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; (ii) activity of LINE1 reverse transcriptase; (iii) expression of HERV-K RNA, and / or (iv) activity of HERV-K reverse transcriptase.

[0129] In certain embodiments, the cancer has (i) expression of LINE1 RNA, LINE1 ORF1 polypeptide, 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.

[0130] In certain embodiments, the cancer has (i) expression of HERV-K RNA, and / or (ii) activity of HERV-K reverse transcriptase. In certain embodiments, the cancer has expression of HERV-K RNA. In certain embodiments, the cancer has activity of HERV-K reverse transcriptase. 28 32017260 400807-030WO (212251)

[0131] In certain embodiments, the cancer has elevated (i) levels of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; (ii) activity of LINE1 reverse transcriptase; (iii) levels of HERV-K RNA, and / or (iv) activity of HERV-K reverse transcriptase.

[0132] In certain embodiments, the cancer has elevated (i) levels of LINE1 RNA, LINE1 ORF1 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.

[0133] In certain embodiments, the cancer has elevated (i) levels of HERV-K RNA, and / or (ii) activity of HERV-K reverse transcriptase. In certain embodiments, the cancer has elevated levels of HERV-K RNA. In certain embodiments, the cancer has elevated activity of HERV-K reverse transcriptase.

[0134] 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 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. 29 32017260 400807-030WO (212251)

[0135] In certain embodiments, the cancer is a preneoplastic or early cancer lesion. In certain embodiments, 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.

[0136] In certain embodiments, the cancer has elevated levels of pericentrometric human satellite 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.

[0137] In aspects of any of the embodiments, the cancer is associated with long interspersed nuclear 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.

[0138] In certain embodiments, the cancer is selected from B cell lymphomas (e.g., B cell chronic 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-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 30 32017260 400807-030WO (212251)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.

[0139] In some embodiments, the cancer is a virus-associated cancer. As used herein, the term “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.

[0140] In some embodiments, the cancer is a cancer other than a virus-associated cancer. In certain 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.

[0141] 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 31 32017260 400807-030WO (212251)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.

[0142] In some embodiments, the cancer is hepatocellular carcinoma (HCC). In some embodiments, 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 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.

[0143] In certain embodiments, the cancer is a leukemia (e.g., acute leukemia, acute lymphocytic 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 32 32017260 400807-030WO (212251)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).

[0144] 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.

[0145] 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, 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 33 32017260 400807-030WO (212251)

[0146] In certain embodiments, the disorder is an autoimmune disorder. As used herein, the terms “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.

[0147] In certain embodiments, the invention provides for treatment of an autoimmune disorder. 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 and / or HERV-K RNA protein expression.

[0148] One embodiment of the invention is a method of treating type I interferonopathies. In one 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 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. 34 32017260 400807-030WO (212251)

[0149] In one embodiment, the invention provides a method of treating an autoimmune disease 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. In another aspect of this embodiment, the autoimmune disease is associated with elevated HERV-K RNA activity and / or expression.

[0150] In certain embodiments, the autoimmune disorder is selected from the group consisting 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 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 35 32017260 400807-030WO (212251)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.

[0151] In certain embodiments, the autoimmune disorder is selected from Aicardi-Goutières 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.

[0152] 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. 36 32017260 400807-030WO (212251)

[0153] In certain embodiments, the autoimmune disorder is a type 1 interferonopathy. In certain 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).

[0154] In certain embodiments, the autoimmune disorder is type 1 diabetes. In certain embodiments, 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.

[0155] 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.

[0156] In certain embodiments, the autoimmune disorder is osteoarthritis, nonalcoholic steatohepatitis (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.

[0157] In certain embodiments, the autoimmune disorder is Reiter's syndrome, exfoliative psoriatic 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, 37 32017260 400807-030WO (212251)malabsorption syndrome, pernicious anemia, hepatitis, hypopituitarism, diabetes insipidus, or sicca syndrome. Neurological Disorders

[0158] In certain embodiments, the disorder is a neurological disorder. In certain embodiments, 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. 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.

[0159] 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.

[0160] In certain embodiments, the neurological disorder is Alzheimer’s disease. In other embodiments, the neurological disorder is amyotrophic lateral sclerosis (ALS). In another embodiment, the neurological disorder is multiple sclerosis. In a further embodiment, the 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. Viral Infection

[0161] In certain embodiments, the viral infection is an infection by human immunodeficiency viruses 1 or 2 (HIV-1 or HIV-2), human T-cell leukemia viruses 1 or 2 (HTLV-1 or HTLV-2), 38 32017260 400807-030WO (212251)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 flavivirus selected from Yellow Fever virus, Dengue virus, Japanese Encephalitis, and West Nile virus.

[0162] In certain embodiments, the viral infection is an infection by human immunodeficiency 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.

[0163] In certain embodiments, the viral infection is an infection by an adenovirus. In certain embodiments, 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 39 32017260 400807-030WO (212251)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.

[0164] In certain embodiments, the viral infection is an infection by a coronavirus. In some embodiments, 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).

[0165] In certain embodiments, the viral infection is an infection by an influenza virus. In certain 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.

[0166] In certain embodiments, the viral infection is an infection by a poliovirus. In certain embodiments, 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

[0167] In certain embodiments, the subject has (i) expression of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; (ii) activity of LINE1 reverse transcriptase; (iii) expression of HERV-K RNA, and / or (iv) activity of HERV-K reverse transcriptase.

[0168] In certain embodiments, the subject has (i) expression of LINE1 RNA, LINE1 ORF1 polypeptide, 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. 40 32017260 400807-030WO (212251)

[0169] In certain embodiments, the subject has (i) expression of HERV-K RNA, and / or (ii) activity of HERV-K reverse transcriptase. In certain embodiments, the subject has expression of HERV-K RNA. In certain embodiments, the subject has activity of HERV-K reverse transcriptase.

[0170] In certain embodiments, the subject has elevated (i) levels of LINE1 RNA, LINE1 ORF1 polypeptide, and / or LINE1 ORF2 polypeptide; (ii) activity of LINE1 reverse transcriptase; (iii) levels of HERV-K RNA, and / or (iv) activity of HERV-K reverse transcriptase.

[0171] In certain embodiments, the subject has elevated (i) levels of LINE1 RNA, LINE1 ORF1 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.

[0172] In certain embodiments, the subject has elevated (i) levels of HERV-K RNA, and / or (ii) activity of HERV-K reverse transcriptase. In certain embodiments, the subject has elevated levels of HERV-K RNA. In certain embodiments, the subject has elevated activity of HERV-K reverse transcriptase.

[0173] In certain embodiments, the subject is a human. In certain embodiments, the subject is an 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

[0174] Another aspect of the disclosure provides for the use of a compound described herein (such as a crystalline Compound I, or other compounds in Section I) for treating a medical disorder, such as a medical disorder described herein.

[0175] Another aspect of the disclosure provides for the use of a compound described herein (such as a crystalline Compound I), in the manufacture of a medicament. In certain 41 32017260 400807-030WO (212251)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. Pharmaceutical Compositions and Dosing Considerations

[0176] As indicated above, the invention provides pharmaceutical compositions, which comprise 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.

[0177] In certain embodiments, the invention provides a pharmaceutical composition comprising a compound described herein (e.g., a crystalline Compound I) and a pharmaceutically acceptable carrier.

[0178] The phrase “therapeutically effective amount” as used herein means that amount of a compound, material, or composition comprising a compound of the present invention 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.

[0179] The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, 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. 42 32017260 400807-030WO (212251)

[0180] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium 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.

[0181] Examples of pharmaceutically acceptable antioxidants include: (1) water soluble antioxidants, 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.

[0182] Formulations of the present invention 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.

[0183] Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0184] Formulations of the invention suitable for oral administration may be in the form of capsules, 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 43 32017260 400807-030WO (212251)(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 invention as an active ingredient. A compound of the present invention may also be administered as a bolus, electuary or paste.

[0185] In solid dosage forms of the invention 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.

[0186] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or 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.

[0187] The tablets, and other solid dosage forms of the pharmaceutical compositions of the present invention, such as dragees, capsules, pills and granules, may optionally be scored or 44 32017260 400807-030WO (212251)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.

[0188] Liquid dosage forms for oral administration of the compounds of the invention include 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.

[0189] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0190] 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.

[0191] Formulations of the pharmaceutical compositions of the invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more 45 32017260 400807-030WO (212251)compounds of the invention 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 room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.

[0192] Formulations of the present invention which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0193] Dosage forms for the topical or transdermal administration of a compound of this invention 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.

[0194] The ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, 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.

[0195] Powders and sprays can contain, in addition to a compound of this invention, 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.

[0196] Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving 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.

[0197] Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of this invention. 46 32017260 400807-030WO (212251)

[0198] Pharmaceutical compositions of this invention suitable for parenteral administration comprise one or more compounds of the invention 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.

[0199] Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the invention 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.

[0200] 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.

[0201] When the compounds of the present invention are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99% (more preferably, 10 to 30%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0202] The preparations of the present invention 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. 47 32017260 400807-030WO (212251)

[0203] The phrases “parenteral administration” and “administered parenterally” as used herein 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.

[0204] The phrases “systemic administration,” “administered systemically,” “peripheral administration” 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.

[0205] These compounds may be administered to humans and other animals for therapy by any 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.

[0206] Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention 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.

[0207] The selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the 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.

[0208] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the 48 32017260 400807-030WO (212251)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.

[0209] In general, a suitable daily dose of a compound of the invention will be that amount of 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.

[0210] If desired, the effective daily dose of the active compound may be administered as two, 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.

[0211] The invention further provides a unit dosage form (such as a tablet or capsule) comprising censavudine or a related compound described herein in a therapeutically effective amount for the treatment of a medical disorder described herein. EXAMPLES

[0212] The invention now being generally described, will be more readily understood by reference 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. Starting materials described herein can be obtained from commercial sources or may be readily prepared from commercially available materials using transformations known to those of skill in the art.

[0213] Techniques referred to in the Examples were carried out under the following conditions:

[0214] Computational Techniques: XRPD Indexing: Within the figure referenced for a given indexed XRPD pattern, agreement between the allowed peak positions, marked with red bars, and the observed peaks indicates a consistent unit cell determination. Successful indexing of a pattern indicates that the sample is composed primarily of a single crystalline phase unless 49 32017260 400807-030WO (212251)otherwise stated. Space groups consistent with the assigned extinction symbol, unit cell parameters, and derived quantities are tabulated below the figure. To confirm the tentative indexing solution, the molecular packing motifs within the crystallographic unit cells must be determined. No attempts at molecular packing were performed. Indexing of XRPD patterns was conducted using TRIADS®(United States Patent No.8,576,985), X'Pert High-Score Plus 2.2a (2.2.1), and TOPAS (Coelho, A.A., J. Appl. Cryst., 2018, 51, 210-218).

[0215] Differential Scanning Calorimetry (DSC): DSC was performed using a Mettler-Toledo DSC3+ differential scanning calorimeter. A tau lag adjustment is performed with indium, tin, and zinc. The temperature and enthalpy are adjusted with octane, phenyl salicylate, indium, tin and zinc. The adjustment is then verified with octane, phenyl salicylate, indium, tin, and zinc. The sample was placed into a hermetically sealed aluminum DSC pan, the weight was accurately recorded, the lid was pierced, and the sample was inserted into the DSC cell. A weighed aluminum pan configured as the sample pan was placed on the reference side of the cell. The WIU SQL ]IY WQMXKML WXQVX ZV YITWSM IUIS_YQY) GPM YITWSMY ]MXM IUIS_`ML NXVT h-* a: ZV - / * a: at 10 °C / min.

[0216] Dynamic Vapor Sorption (DVS): Automated vapor sorption (VS) data were collected on a Surface Measurement System DVS Intrinsic instrument. Samples were not dried prior to analysis. Sorption and desorption data were collected over a range from 5% to 95% RH at 10% RH increments under a nitrogen purge. The equilibrium criterion used for analysis was less than 0.0100% weight change in 5 minutes with a maximum equilibration time of 3 hours. Data were not corrected for the initial moisture content of the samples.

[0217] Thermogravimetric Analysis (TGA): TG analyses were performed using a Mettler- Toledo TGA / DSC3+ analyzer. Temperature and enthalpy adjustments were performed using indium, phenyl salicylate, aluminum, gold, tin, and zinc, and then verified with indium. The balance was verified with a certified standard weight. The samples were placed in an open aluminum pan. The pan was hermetically sealed, the lid pierced, then inserted into the TG furnace. A weighed aluminum pan configured as the sample pan was placed on the reference platform. The furnace was heated under nitrogen. The samples were analyzed from 25 °C to 350 °C at 10 °C / min. 50 32017260 400807-030WO (212251)

[0218] Proton Nuclear Magnetic Resonance Spectroscopy (1H NMR): The solution NMR spectra were acquired with an Avance 600 MHz NMR The samples were prepared by dissolving a given amount of sample in DMSO-d6TMS. The data acquisition parameters are displayed in the first plot of each spectrum in the Data Section of this report.

[0219] X-ray Powder Diffraction (XRPD): XRPD pattern was collected with a PANalytical X'Pert PRO MPD or PANalytical Empyrean diffractometer using an incident beam of Cu radiation produced using a long, fine-focus source. An elliptically graded multilayer mirror was [YML ZV NVK[Y :[ @i H(XI_Y ZPXV[OP ZPM YWMKQTMU IUL VUZV ZPM LMZMKZVX) DXQVX ZV ZPM IUIS_YQY' I silicon specimen (NIST SRM 640f) was analyzed to verify the observed position of the Si 111 peak is consistent with the NIST-certified position. A specimen of the sample was sandwiched JMZ]MMU -(mT(ZPQKR NQSTY IUL IUIS_`ML QU ZXIUYTQYYQVU OMVTMZX_) 8 JMIT(YZVW' YPVXZ IUZQYKIZZMX extension, and antiscatter knife edge were used to minimize the background generated by air. Soller slits for the incident and diffracted beams were used to minimize broadening and asymmetry from axial divergence. Diffraction patterns were collected using a scanning position- sensitive detector (X'Celerator) located 240 mm from the specimen and Data Collector software v.5.5. EXAMPLE 1 – Preparation of 4-amino-1-((2R,3S,4R,5R)-5-(chloromethyl)-3-fluoro-4- hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-5-fluoropyrimidin-2(1H)-one (Compound II)2N O II

[0220] Step 1: To a stirred solution of (2R,3S,4R,5R)-5-((benzoyloxy)methyl)-3- fluorotetrahydrofuran-2,4-diyl dibenzoate (1700 g) in DCM (1700 mL, 1 V) was added HBr in AcOH (40%) (1700 mL, 1 V) at RT. (endothermic, clear solution) Stirred at RT for 3 h. HPLC showed all starting material was fully consumed. (SM< 1%) The reaction was quenched by water (1.7 L, 1 V). Organic phase was collected and the water phase was extracted by DCM (500 ml) twice. Organic phase was combined and washed by water (2 L) for three times and sat. NaHCO3 (2 L) for three times. HPLC showed no benzoic acid residue in the organic phase and PH for the latest water phase was basic. The organic phase was dried by Na2SO4and concentrated in vacuo 52 32017260 400807-030WO (212251)to give crude bromide 1 (1510 g, 97.5%) as a yellow oil, which would be used in next step without further purification.

[0221] Step 2: Into a stirred solution of fluorouracil (725.9 g, 2.0 eq) in Dioxane (9.44 L, 8 V) was added BSA (2270.6 g, 4.0 eq) in portions. (Heterogeneous) Stirred at 80oC for 3 h. (Homogeneous, clear solution) The reaction was cooled down to room temperature. A Dioxane (2.36 L, 2 V) solution of bromide 1 (1181 g, 1.0 eq) was added. Stirred at 80oC for another 16 h. HPLC showed the SM was less than 3%. The reaction was cooled down and added into a stirred heptane solution (28 L, around two times of the reaction liquid) for precipitation. The resulting suspension was stirred for additional 4 h before the filtration. Filtration cake was collected and slurry with water (11.8 L, 10 V) to remove the residual fluorouracil. Filtration again and the cake was dried under an IR lamp overnight to give Compound 2 (1162 g, 88.2%, Ratio>50: 1, KF<1%).

[0222] Step 3: To a stirred solution of Compound 2 (1157 g, 1.0 eq) in MeOH (11.6 L, 10 V) was added NaOH (97.96 g, 1.0 eq) in portions at 0oC. The mixture was moved to room temperature and stirred for additional 3 h. HPLC showed the completion of the reaction. The reaction mixture was filtrated and the filtrate was collected. (Removal of the insoluble inorganic salt) The resulting solution was directly concentrated in vacuo to afford a viscous white solid. (High MeOH residue) The solid was added to a stirred solution of MTBE (6 L) for slurry and stirred for 16 h. The slurry mixture was filtrated and the filtration cake was collected (quickly, otherwise become viscous due to the hydroscopic nature.) The solid was further dried in vacuo. HNMR showed the MeOH residue was less than 10% mmol. Crude diol 3 (888 g, Crude) was obtained, which would be used in next step without further purification.

[0223] Step 4: To a stirred solution of diol 3 (785 g, 1.0 eq) and imidazole (606.87 g, 3.0 eq) in DMF (7.9 L, 10 V) was added TBSCl (671.77 g, 1.5 eq) in portions at 0oC under N2. The reaction mixture was then moved to room temperature and stirred for 16 h. HPLC showed the completion of the reaction. (Notably, 100 g and 785 g batch were worked up together) The reaction was quenched by H2O (15 L, 20 V) at 0oC. (exothermic) The resulting solution was then extracted with EA (10 L) for 3 times. Organic phase was combined and washed with H2O (10 L) twice, sat. NaHCO3 (aq.) (10 L) and brine (10 L). (Around 2 g, 0.2% lost) Dried by 53 32017260 400807-030WO (212251)Na2SO4and concentrated in vacuo to give a pale-brown solid Slurry by n-hexane (5.4 L, 6 V) to afford pure 4 (584 g, 63% for 2 steps) as a white solid. (Around 0.87 g, 0.1% Lost in slurry).

[0224] Step 5: To a stirred solution of alcohol 4 (553 g, 1.0 eq) in DCM (5.5 L, 10 V) was added AgNO3(496.44 g, 2.0 eq) and collidine (579.39 mL, 3.0 eq) in one portion at room temperature. The reaction system was cooled to 0oC. MMTrCl (676.84 g, 1.5 eq.) was added in one portion. (No obvious heat release) Moved to room temperature and stirred for additional 3 h. HPLC showed the completion of the reaction. (LCAP= 93.8%) (Notably, 84 g and 553 g batch were worked up together) The reaction was filtrated and the filtrate was collected. MTBE (12.8 L, 20 V) was added and the resulting solution was washed with H2O (10 L), 1M critic acid (10 L) for three times. (No lost) HPLC showed no collidine residue in the organic phase. Organic phase was further washed by sat. NaHCO3(10 L) and brine (10 L). (No lost) Dried by Na2SO4and concentrated in vacuo. Purified by silica column (PE: EA= 10:1 to 1:1) to give compound 5 (971 g, HPLC= 97.2%, IY= 89%) as a pale-yellow foam and some mixture including around 60 g compound 5 (5%).

[0225] Step 6: To a stirred solution of compound 5 (971 g, 1.0 eq) in MeOH (9.7 L, 10 V) was added NH4F (552.60 g, 10.0 eq) in one portion at room temperature. (Most of the NH4F dissolved) Then moved to 60oC and stirred for 16 h. HPLC showed the completion of the reaction. (5 fully consumed) The reaction was cooled down to room temperature and concentrated directly. The residue was dissolved in EA (10 L, 10 V) and H2O (10 L, 10 V). Organic phase was collected and washed with H2O (10 L, 10 V) for additional four times. (0.6 g lost, 0.1%) Organic phase was dried in Na2SO4and concentrated to afford alcohol 6 (750 g, 94%, HPLC= 96.3%) as a white foam.

[0226] Step 7: To a stirred solution of 6 (460 g, 1 equiv) in MeCN (9.20 L, 20 V) was added IBX (480.15 g, 2.00 equiv) in one portion at room temperature. (No obvious heat release) The reaction system was then move to 65 oC and stirred for 4h. HPLC showed completion of the reaction. (SM less than 3%) The reaction was cooled down to room temperature. The mixture was filtrated and the filtration cake was washed with MeCN (1 L). The filtrate was collected and concentrated directly in vacuo to afford crude 7 (463 g, 101.03%) as a yellow foam, which would be directly used in the next step without further purification. 54 32017260 400807-030WO (212251)

[0227] Step 8: Into a stirred solution of 7 (463 g, 1 eq.) and HCHO (926 mL, 37%, 2 V) in dioxane (4.6 L, 10 V) was added NaOH (926 mL, 2 M in H2O, 2 V) at 0oC within 10 min. (no obvious heat release) The reaction mixture was then moved to room temperature and stirred for 16 h. HPLC / LCMS showed the completion of reaction, stage I. The reaction was cooled to 0oC. NaBH4 (65.54 g, 1732.408 mmol, 2 eq.) was added in portions. (Obvious heat release observed) The reaction mixture was then moved to room temperature and stirred for 2 h. HPLC / LCMS showed the completion of reaction, stage II. (HPLC= 75.3%) The reaction was cooled to 0oC and quenched by NH4Cl (4.6 L, 10 V) (PH changed from 10 to 8). The organic phase was collected and the water phase was extracted with EA (10 V once and 5 V twice). The organic phase was combined and further washed by water (10 L) and brine (10 L). (2 g lost, 0.4%) The resulting solution was dried by Na2SO4and concentrated directly to afford crude 8 (454 g, IY= 92.5% for two steps, HPLC= 78.8%) as a pale-yellow foam, which would be used in next step without further purification.

[0228] Step 9: Into a stirred solution of 8 (452 g, 1 eq.) in Pyridine (2260 mL, 5 V) and DCM (6.78 L, 15 V) was added Tf2O (562.71 g, 2.5 eq.) at -30oC dropwise in 30 min. (Obvious heat release observed). The cold bath was then removed, making the reaction temperature return to room temperature naturally. HPLC then showed the completion of the reaction after 1 h. (SM<1%, HPLC= 60.8%, -10oC at this time) The reaction was quenched by water (10 L) at 0oC. MTBE (13 L, double of DCM) was added. The organic phase was washed by 1 M critic acid (10 L twice). HPLC showed no pyridine residue in the organic phase. The organic phase was further washed by 10% NaCl (10 L). (0.8 g lost, 0.1%) The organic phase was dried by Na2SO4and concentration directly to afford crude 9 (616 g, IY= 93.0%, HPLC= 77.4%) as a pale-yellow foam, which need to be directly used in the next step ASAP to maintain stability.

[0229] Step 10: Into a stirred solution of 9 (616 g, 1 equiv) in THF (6.2 L, 10 V) was added TEA (150.08 g, 2 equiv) in one portion at room temperature. (No heat release) The reaction was moved to 60oC and stirred for 2 h. HPLC showed the completion of stage I. (SM< 1%) LiCl (314.35 g, 10 equiv) was added directly at 60oC in one portion and reaction was further stirred at 60oC for 12 h. HPLC then showed the completion of stage II. (10A< 1%) NaOH (1 M in H2O, 1.48 L, 2 equiv) was added directly at 60oC. (No heat release) The reaction was stirred at 50oC for 2h. HPLC and LCMS indicated the completion of stage III. (10B< 1%, MS of 10B disappeared, HPLC= 83%) The reaction was then cooled down to 0oC, and neutralized with 1M 55 32017260 400807-030WO (212251)critic acid to the PH of 7-8. H2O (6.1 L, 10 V) was added and organic phase was collected. The water phase was extracted with EA (6 L,10 V, twice). (1.2 g lost, 0.3%) The organic phase was combined, washed with 10% brine (6 L), dried by Na2SO4 and concentrated in vacuo. (No lost) The product was purified by silica gel column (PE:THF=10: 1 to 1:1) to afford alcohol 10 (242 g, IY= 56%, HPLC= 95.8%) as a pale-yellow foam.

[0230] Step 11: Into a stirred solution of Compound 10 (160 g, 1 eq.) and Imidazole (37.24 g, 2 eq.) in DCM (3.20 L, 20 V) was added TBSCl (49.47 g, 1.2 eq.) in one portion under 0 oC. (No obvious heat release) The reaction was moved to room temperature and stirred for additional 16 h. HPLC showed the completion of the reaction. H2O (3.2 L, 20 V) was added to the reaction and organic phase was collected. The organic phase was washed by extra H2O (3.2 L twice) and 10% brine (3.2 L). (0.5 g lost, 0.3%) The organic phase was dried by Na2SO4and concentrated in vacuo to afford crude 11 (166 g, IY= 86.8%, HPLC= 98.1%) as a pale-yellow foam.

[0231] Step 12: Into a stirred solution of crude 11 (166 g, 1 eq.), DMAP (87.01 g, 3 eq.) and TEA (96.09 g, 4 eq.) in ACN (3.3 L, 20 V) was added TsCl (135.77 g, 3 eq.) in one portion at room temperature. (No obvious heat release) The reaction was stirred for 2 h. HPLC then indicated the completion of the reaction, stage I. The reaction was cooled down to 0oC and NH4OH (498 mL, 3 V) was added dropwise within 10 min. (heat release observed) The reaction was moved to room temperature and stirred for additional 2 h. HPLC then showed the completion of the reaction, stage II. The reaction was concentrated directly, and the resulting residue was dissolved in MTBE (3.3 L, 20 V) and H2O (3.3 L, 20 V). Organic phase was collected and the water phase was further extracted with MTBE (800 ml, 5 V). The organic phase was combined and washed with H2O (1.6 L, 10 V) for three times, NH4Cl (1.6 L, 10 V) once and brine (1.6 L, 10 V) once. (No lost) After drying by Na2SO4, the organic phase was directly concentrated in vacuo to afford crude 12 (168 g, 101.35%) as an orange foam, which would be used in the next step without further purification.

[0232] Step 13: Into a stirred solution of crude 12 (80 g, 1 eq.) in MeOH (800 mL, 10 V) was added HCl (2.0 M in H2O, 229.13 mL, 4 eq.) in one portion under 0oC. (No obvious heat release) The reaction was then moved to 60oC and stirred for 3 h. HPLC then showed the completion of the reaction. The reaction was concentrated in vacuo directly, then H2O (160 ml, 56 32017260 400807-030WO (212251)2 V) and EA (240 ml, 3 V) was added. EA phase was separated and the water phase was further extracted with EA (160 ml, 2 V). Then NH4OH was added to the water phase to adjust the PH from 1 to 9. The water phase was then concentrated in vacuo to get a mixture of desired product and NH4Cl as a pale-yellow solid. (60 g) The solid was dissolved in acetone (6 L, 100 V) and stirred for 5 h. The resulting solution was then filtrated and the filtration cake was washed with acetone (500 ml). The filtrate was collected and concentrated in vacuo to give a crude product (22.9 g) as a white solid. The crude product was further purified by slurry in ACN (130 ml, 5 V) to afford Compound II (22.4 g, IY= 64% for 2 steps, 99.0% HPLC, KF= 0.02%, Crystal) as a white solid. (0.5 g, 2.2%wt lost in ML, HPLC= 75.1%).

[0233] Physical characterization of the white solid product was performed. The XRPD spectrum obtained is provided in FIG.1. EXAMPLE 2 – Preparation of (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)- yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl)tetrahydrofuran-3-yl isobutyrate (Compound I)a - - hydroxy-5-(hydroxymethyl) oxolan-2-yl]-5-fluoropyrimidin-2-one (5.5 g, 17.68 mmol) and DMAP (4 g, 35.36 mmol) in pyridine (50 mL) was added isobutyric anhydride (17 g, 106.11 mmol) dropwise at 0oC under nitrogen atmosphere. The mixture was stirred overnight at 80oC 57 32017260 400807-030WO (212251)and cooled to room temperature. The mixture was concentrated under reduced pressure. The residue was diluted with EtOAc, washed with aqueous citric acid and 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 (PE: EtOAc = 1:1) to afford [(2R,3R,4S,5R)-2-(chloromethyl)-4-fluoro-5-[5-fluoro-4-(2-methylpropanamido)-2- oxopyrimidin-1-yl]-3-[(2-methylpropanoyl) oxy] oxolan-2-yl] methyl 2-methylpropanoate (6.6 g, 12.65 mmol, 71.55%) as a white solid. LC-MS (ES, m / z): 522 / 524[M+H]+.

[0235] Step 2: A solution of [(2R,3R,4S,5R)-2-(chloromethyl)-4-fluoro-5-[5-fluoro-4-(2- methylpropanamido)-2-oxopyrimidin-1-yl]-3-[(2-methylpropanoyl) oxy] oxolan-2-yl] methyl 2- methylpropanoate (6.6 g, 12.65 mmol) in EtOH (660 mL) was stirred for 20 h at 100oC under nitrogen atmosphere. The resulting mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH=10 / 1. The fraction was collected and concentrated under vacuum to afford [(2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1-yl)-2-(chloromethyl)-4-fluoro-3- [(2-methylpropanoyl) oxy] oxolan-2-yl] methyl 2-methylpropanoate (5.2 g, 11.53 mmol, 89.46%) as an off-white solid. LC-MS(ES, m / z): 452 / 454[M+H]+, 98.3% purity. Conditions for the LCMS: (Column: Shim‐WIKR FKMWZMX :+2' --%-)* TT5 -)* mA WIXZQKSMY5 AVJQSM DPIYM 84 water / 5mM NH4HCO3; Mobile Phase B: Acetonitrile; Flow rate: 1.500 mL / min; Gradient: 20% B to 60% B in 1.70 min, 60% B to 95% B in 0.60 min, 95% B to 95% B in 0.50 min, 95% B to 10% B in 0.03 min; Wave Length: 254 nm; RT(min): 1.546).1H NMR (400 MHz, DMSO-d6$ j 7.99 (s, 1H), 7.80 – 7.72 (m, 2H), 6.34 (dd, J = 16.7, 4.2 Hz, 1H), 5.66 (dd, J = 19.4, 2.7 Hz, 1H), 5.38 (ddd, J = 51.6, 4.3, 2.7 Hz, 1H), 4.48 – 4.24 (m, 2H), 3.93 (dd, J = 63.0, 11.9 Hz, 2H), 2.64 (dp, J = 30.2, 7.0 Hz, 2H), 1.19 – 1.03 (m, 12H). EXAMPLE 3 - Preparation of Crystalline (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxo- pyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran- 3-yl isobutyrate Form A (Compound I, Form A) Part I - Fast Evaporation (FE) Procedure

[0236] A solution of (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate from <^ITWSM , QU LX_ AMC> ]MXM NQSZMXML ]QZP *), mT U_SVU VX DG=< Y_XQUOM NQSZMXY QUZV KSMIU \QISY) 58 32017260 400807-030WO (212251)The vials were left open at ambient temperature for evaporation for 1 day to give white dendritic needle solids of crystalline Form A of Compound I. Part II – Alternative Procedure for Preparation of Crystalline (2R,3R,4S,5R)-5-(4-amino-5- fluoro-2-oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy) methyl) tetrahydrofuran-3-yl isobutyrate Form A (Compound I, Form A)was to reactor kg, 1.00eq) and additional dichloromethane (13.25 kg, 2V). The temperature was adjusted to 25 ± 5°C. Pyridine (4.40 kg, 5.00eq) was added dropwise. The temperature was then adjusted to -20 ± 10°C. Charged dropwise trifluoromethanesulfonic anhydride to reactor A. The mixture was stirred at least 1 hour at -20 ± 10°C. The reaction mixture was then quenched with citric acid solution (15% in H2O) (26.75 kg, 5V) twice and the temperature was adjusted to 0~25°C. The organic layer was then washed with sodium chloride (10% in H2O) (54.00 kg, 10V). The organic layers were then concentrated under reduce pressure. 59 32017260 400807-030WO (212251)

[0238] Step 2: Tetrahydrofuran (17.80 kg, 4V) was added to reactor A. The temperature was adjusted to 20 ± 5°C. Potassium phosphate (4.70 kg, 2.00 eq., Batch) and softened water (0.50eq.) were added to reactor. The temperature was adjusted to 40 ± 5°C. A solution of THF and 2 ]IY ZPMU ILLML LXVW]QYM ZV XMIKZVX 8 IZ .* c / o V\MX + PV[X) GPM YVS[ZQVU ]IY YZQXXML IZ least 2 hours at 40 ± 5°C, and then cooled to room temperature. The reaction mixture was filtered through diatomaceous earth and the filter cake was washed with tetrahydrofuran (8.90 kg, 2V). The filtrate was added to reactor A and charged with lithium chloride (1.41 kg, 3.00 eq.). The reaction was warmed to 50 ± 5°C and stirred at least 5 hours. The reaction was then cooled to 5 ± 5°C and charged with sodium hydroxide solution (4% in H2O) (11.10 kg, 1.00 eq.) dropwise over 60 minutes. The reaction was then stirred at least 1 hour at 15 ± 5°C. The reaction was then cooled to 0 ± 5°C and quenched by dropwise addition of a citric acid solution (15% in H2O) (53.50 kg, 10V) over 60 minutes. Methyl tert-butyl ether (37.00 kg, 10V) was then added dropwise and stirred at least 0.5 hours at 0 ± 5°C. The organic layer was washed with a 10% sodium chloride solution. The organic layers were used directly in the next step.

[0239] Step 3: The crude material from step 2 was added to a rotary evaporator at 40 ± 5°C and concentrated until the volume was 2.5V. Methanol (23.09 kg, 7.0 vol.) was added to the rotary evaporator and the mixture was concentrated until the volume was 2.5V. Methanol (23.09 kg, 7.0 vol.) was added to the rotary evaporator again and allowed to cool to room temperature. Under nitrogen protection, the solution was transferred to reactor A. Potassium fluoride (2.58 kg, 5.00 eq.) was added to the reaction. It was then warmed to 60 ± 10°C and stirred at least 8 hours and then cooled. The reaction mixture was rotary evaporated until the volume was 2.5V. Reaction was acidified with citric acid and separated. The aqueous layer was washed with 2- methyltetrahydrofuran and sodium chloride solution (10% in H2O). The organic layers were combined and concentrated until the volume was 2.5V. Isopropyl acetate (18.20 kg, 5.0 vol) was charged to rotary evaporator and concentrated until the volume was 2.5V. Isopropyl acetate (18.20 kg, 5.0 vol) was charged to the rotary evaporator again and concentrated until the volume was 2.5V. N-heptane (42.06 kg, 15.0 vol.) was added to reactor A followed by dropwise addition of the reaction solution over 4 hours. The reaction was stirred at least 5 hours at room temperature. The slurry was discharged from the reactor to a centrifuge, filtered, and the cake was washed with n- heptane (5.61 kg, 2.0 V). The filter cake was dried at least 12 hours and used crude in the next reaction. 60 32017260 400807-030WO (212251)

[0240] Step 4: Methyl tert-butyl ether (38.52 kg, 13V) was added to reactor A and warmed to 25 ± 5°C.5 (5.71kg, 1.00eq) was then added into reactor A followed by 4-dimethyl- aminopyridine (0.156 kg, 0.10eq). Triethylamine (3.87 kg, 3.00eq) was then added dropwise. The reaction was cooled to 10 ± 5°C. Isobutyric anhydride (5.06 kg, 2.50eq) was then added dropwise over 30 minutes. The reaction was then warmed to 25 ± 5°C, stirred at least 0.5 hours and then cooled to 10 ± 10°C. The reaction was then quenched with citric acid solution (15% in H2O) (42.80 kg, 10V) and the organic phase was washed with a sodium bicarbonate solution (5% in H2O) (21.60 kg, 5V, Batch: N / A). The organic phased was combined with silica gel (4.00 kg) stirred at least 2 hours, and filtered through diatomite (0.80 kg) wetted with methyl tert-butyl ether (2.96 kg, 1V). The filter caked was rinsed with methyl tert-butyl ether (5.92 kg, 2V) and the filter liquor was charged to a rotary evaporator and concentrated until the volume was 3.0-4.0V. The organic phase was loaded onto a silica gel column and eluted with methyl tert-butyl ether (14.8 kg, 5V). Organics were then concentrated until the volume was 3.0-4.0V. N-heptane (54.72 kg, 20.0 vol.) was added to reactor A followed by a seed crystal. The reaction solution was then added dropwise into reactor A over 2 hours and stirred at least 2 hours. The slurry was discharged from the reactor to a centrifuge, filtered, and the cake was washed with n-heptane (5.47 kg, 2.0 V). Finally the filter cake was dried at least 12 hours and used crude in the next reaction.

[0241] Step 5: Acetonitrile (21.33 kg, 6V) was added to reactor A and warmed to 25 ± 5°C. Triazole (4.46 kg, 6.50eq) and ethylbis(propan-2-yl)amine (11.57 kg, 9.00eq) were then added to reactor A and the mixture was cooled to 0 ± 5oC. POCl3(2.56 kg, 1.70eq.) was added slowly over 0.5 hours. The reaction was then stirred at least 1 hour at 0 ± 5°C.6 in acetonitrile was then added slowly over 0.5 hour. The reaction was then warmed to 25 ± 5°C and stirred at least 1 hour. The reaction was then cooled to 5 ± 5°C followed by addition of NH4OH (3.49 kg, 2.5eq) into reactor over 1h. the resulting mixture was stirred at least 3 hours at 10 ± 5°C. The reaction was then quenched with a citric acid solution (15% in H2O) (72.23 kg, 15V) added dropwise followed by isopropyl acetate and separated. The organics were then co-distilled two times with isopropyl acetate (39.15 kg, 10V). Charged 2-methyltetrahydrofuran (27.09 kg, 7V) to reactor A. Washed organics with water x2. Transferred the organic phase into reactor A. Vacuum distillation until the volume was 5.0V. Charged isopropyl acetate (39.15 kg, 10V). Vacuum distillation until the volume was 5.0V. Adjusted to 80 ± 5°C. Stirred until the solid dissolved. Cooled the solution to 60 ± 5oC. Charged n-heptane (15.30 kg, 5V) dropwise to reactor A at least 1 hour. Stirred at 61 32017260 400807-030WO (212251)least 2 hours at 60 ± 5°C. Adjusted to 20 ± 5°C for over 4 hours. Stirred at least 3 hours. Filtered, washed the cake with heptane. Charged isopropyl acetate (15.66 kg, 4V) to reactor A at 20 ± 5 °C. Started agitating. Charged wet cake to reactor A. Adjusted to 80 ± 5°C. Stirred until the solid dissolved. Cooled the solution to 60 ± 5oC. Charged n-heptane (15.30 kg, 5V) dropwise to reactor A at least 1 hour. Stirred at least 2 hours at 60 ± 5°C. Adjusted to 20 ± 5°C for over 4 hours. Stirred at least 3 hours. Filtered, washed the cake with heptane: IPAC=3:2 (3 V). Dried the cake under vacuum at 45°C for over 10 hours. Charged ethanol (23.70 kg) to reactor A. Charged soft water (60.00 kg) to reactor A. Agitated at least 30 min. Charged dried cake to reactor A at 20 ± 5 °C. Adjusted to 50 ± 5°C. Stirred at least 6 hours. Cooled to 20 ± 5oC. Stirred at least 6 hours. Filtered, washed the cake with ethanol: water =1:2 (18.00 L, 4V). Dried the cake under vacuum at 45°C for over 10 hours to give crystalline Form A of Compound I (2.82 kg, 62.8%). EXAMPLE 4 – Physical Characterization of Crystalline (2R,3R,4S,5R)-5-(4-amino-5- fluoro-2-oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate Form A (Compound I, Form A)

[0242] A sample of crystalline (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)- 2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate Form A that contained a minor amount of crystalline (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3- yl isobutyrate Form B was subjected to physical characterization by XRPD, TGA, DSC and DVS analysis. The XRPD spectrum is provided in FIG.3. The TGA and DSC thermograms are provided in FIG.4.

[0243] DVS testing of this material provided a 0.6% weight gain from 5% to 95% RH, equivalent to 0.2 mol / mol water with a subsequent 0.6% weight loss from the 95% to 5% (FIG. 5.). This material overall exhibited limited hygroscopicity. Post-DVS XRPD of the material showed that it remained Form A + minor Form B when it was collected after DVS sorption / desorption cycle at 5% RH (FIG.5).

[0244] Peaks observed in the XRPD from Crystalline Form A of Compound I are listed in Table 1 below. Exemplary prominent peaks for crystalline Form A of Compound I are provided in Table 2 below. 62 32017260 400807-030WO (212251)Table 1. Observed Peaks for Form A of Compound I Angle [2u]d-spacing [Å] Relative Intensity [%]63 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%]Table 2. Exemplary Prominent Peaks for Form A of Compound I Angle [2u]d-spacing [Å] Relative Intensity [%]

[0245] An XRPD diffractogram used in indexing for crystalline Form A of Compound I is provided in FIG.6. 64 32017260 400807-030WO (212251)EXAMPLE 4 - Preparation of Crystalline (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate Form B (Compound I, Form B)

[0246] The title compound was prepared according to the following procedures. Part I - Vapor Stress (VS) Procedure

[0247] (2R,3R,4S,5R)-5-(4-Amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4- fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate from Example 2 was weighed out and placed inside 1-dram vial. The 1-dram vial was placed, uncapped, in 20-mL vials containing dry MeOH. After 7 days, white solids were isolated and dried under N2 gas to give crystalline Form B of Compound I. The white solids product was analyzed by XRPD, TGA, DSC and DVS as described herein.

[0248] The XRPD spectrum used in indexing is provided FIG.9. TGA and DSC thermograms are provided in FIG.10. A DVS isotherm plot is provided in FIG.11.

[0249] Peaks observed in the XRPD from Crystalline Form B of Compound I are listed in Table 3 below. Exemplary prominent peaks for crystalline Form B of Compound I are provided in Table 4 below. Table 3. Observed Peaks for Crystalline Form B of Compound I Angle [2u]d-spacing [Å] Relative Intensity [%]65 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%] 14.68 ± 0.20 6.029 ± 0.082 466 32017260 400807-030WO (212251)Table 4. Exemplary Prominent Peaks for Crystalline Form B of Compound I Angle [2u]d-spacing [Å] Relative Intensity [%]Part II – Alternative Procedure for Preparation of Crystalline Form B of Compound I

[0250] Crystalline Form B of Compound I was also made according to the following procedure.

[0251] To a solution of 1H-1,2,4-triazole(247 g, 3.59 mol, 6.50 eq) in ACN (1.50 L) was added DIEA (642 g, 4.97 mol, 9.00 eq) at 0oC. POCl3(143 g, 938 mmol, 1.70 eq) was added dropwise into reaction at 0oC over 30 mins. Hold contents of reaction at 0oC over 1 h. A solution of 1 (250 g, 552 mmol, 1.00 eq) in ACN (250 mL) was added dropwise into reaction at 0oC over 0.5 h. The mixture was warmed to 25oC and stirred at 25oC for 1 h. NH3.H2O (193 g, 1.38 mol, 25% purity, 2.50 eq) was added dropwise into the mixture for over 1 h at 5oC. The mixture was stirred at 10oC for 3 h. LCMS (0.488 min) showed that the reaction was completed. 2 reactions were combined to work up. Charge citric acid (15%, 500 mL) and 67 32017260 400807-030WO (212251)MTBE (1.80 L) into reaction. Separated and collected the organic phase. The organic phase was washed with brine (1.50 L), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to give a residue. The crude product was triturated with MTBE (200 mL) at 25oC for 30 mins to give Compound I (302 g, 668 mmol, 60.5% yield) as a white solid.

[0252] A mixture of Compound I (302 g, 670 mmol, 1.00 eq) in ACN (480 mL) and H2O (720 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 55 °C for 12 h under N2atmosphere. The reaction was stop after 12 h, the reaction was concentrated under reduced pressure to 2 V. Charge EtOH (300 mL) to the vessel. Hold contents of vessel at 20 °C for 1 h. Filtered and the filter cake was washed with EtOH / H2O (v / v = 1 / 2, 600 mL), the filter cake was dried under vacuum to give crystalline Form B of Compound I (265 g, 586 mmol, 87.4% yield) as a white solid.1H NMR: 400 MHz DMSO-d6j 1)32 #Y' +>$' 1)1 / (1)11 #T' ,>$' 6.31-6.36 (m, 1H), 5.63-5.68 (m, 1H), 5.30-5.45 (m, 1H), 4.36-4.46 (m, 2H), 4.00 (d, J = 11.6 Hz, 1H), 3.84 (d, J = 12.0 Hz, 1H), 2.58-2.69 (m, 2H), 1.10-1.16 (m, 12H). EXAMPLE 5 - Preparation of Crystalline (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate Form C (Compound I, Form C)

[0253] The title compound was prepared according to the following procedures. Slow Cool (SC) Procedure

[0254] A solution of (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate in dry QDXC8K ]IY NQSZMXML IZ / / (0* a: ]QZP ]IXTML *), mT U_SVU Y_XQUOM NQSZMXY QUZV WXM(PMIZML' KSMIU vials. The heat was turned off and samples slowly cooled to ambient temperature for 1 day. Samples were placed in refrigerator at 2-8°C for 3 days. The vials were centrifuged to decant, and solids were dried under N2gas to produce a white solid, Compound I, Form C. The white solid product was analyzed by XRPD, TGA and DSC as described herein. The XRPD spectrum for Form C is provided in FIG.7. TGA and DSC thermograms for Form C are provided in FIG. 8. 68 32017260 400807-030WO (212251)EXAMPLE 6 - Preparation of Crystalline (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2- oxopyrimidin-1(2H)-yl)-2-(chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate Form D (Compound I, Form D)

[0255] A solution of (2R,3R,4S,5R)-5-(4-amino-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2- (chloromethyl)-4-fluoro-2-((isobutyryloxy)methyl) tetrahydrofuran-3-yl isobutyrate in 35:65 dry THF / H2O (aw= 0.99) was stirred at 5 °C for 14 days, then centrifuged to decant, and dried under N2gas, resulting in crystalline Form D of Compound I as a white solid. The white solid product was analyzed by XRPD, TGA and DSC as described herein. The XRPD spectrum for Form D is provided in FIG.12. TGA and DSC thermograms for Form D are provided in FIG.13. EXAMPLE 7 - Stable Cellular Assay for Inhibiting LINE1 Reverse Transcriptase

[0256] Compound A and Compound B were tested for ability to inhibit LINE1 reverse transcriptase using a stable artificial-intron Cis LINE1 reporter assay. Assay procedures and results are described below. Compound Id ntifi r Chemical StructurePart I – Procedure for Stable Artificial-Intron Cis LINE1 Reporter Assay

[0849] A stable HeLa Tet-On 3G (Takara, cat no 631183) cell line expressing a bi-directional inducible 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 69 32017260 400807-030WO (212251)clones were screened for high Luciferase expression and the highest expression Firefly expressing clone was chosen for compound testing.

[0850] 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-Aldrich cat #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 #E2940) following the manufacturer’s instructions. The Firefly Luciferase activity was used to report LINE1 activity. Part II – Results

[0851] Compound A was determined to have an IC50\IS[M d *)* / * fA) :VTWV[UL 9 PIL IU IC50 \IS[M QU ZPM XIUOM VN NXVT 7 *), / * fA ZV d +fA) EXAMPLE 8 – THP1 TREX1 KO Inhibition Assay

[0257] Compound A and Compound B were tested for their ability to alter the type 1 interferon XMYWVUYM QU G>D+ ;[IS GE<H+ @C KMSSY ZXMIZML ]QZP / (I`I(,n(LMV^_K_ZQLQUM) 8YYI_ WXVKML[XMY and results are described below. Compound I ifi Chemical Structure70 32017260 400807-030WO (212251)Part I - Procedure

[0258] THP1 Dual TREX1 KO cells were purchased from Invivogen. The THP1-Dual™ KO- TREX1 cells were cultured in RPMI 1640, 10% heat-inactivated fetal bovine serum, 25 mM HEPES, 10µg / mL blasticidin, and 100µg / mL zeocin. THP1-Dual™ KO-TREX1 cells were ZXMIZML ]QZP I LVYM ZQZXIZQVU VN I ZMYZ KVTWV[UL ISVUM VX QU ZPM WXMYMUKM VN +fA / (I`I(,n( deoxycytidine (Sigma-Aldrich, cat# 189825). Type 1 Interferon and cell viability were assessed after six days of treatment.

[0259] Stock solution of the test compound was prepared in DMSO followed by a three-fold dilution in DMSO. Additional 50x dilution was prepared in cell culture media for each dilution. 10µL of diluted test compound was then added to a 96-well plate.

[0260] G>D+(;[ISb @C(GE<H+ KMSSY ]MXM ZXMIZML ]QZP MQZPMX +^D9F VX +fA / (I`I(,n( deoxycytidine.190µL of the THP1-Dual™ KO-TREX1 cells from either treatment were added to each well of the 96-well plate containing the test compound titration at 50,000 cells / well. Cells were incubated at 37 ºC, 5% CO2in a humidified incubator for six days. Cells treated with 1µM / (I`I(,n(LMV^_K_ZQLQUM ]MXM ILLML IZ ZPM YITM LMUYQZ_ IY IJV\M IUL QUK[JIZML IZ -1 g:' / " :C2 in a humidified incubator for six days. On day six, 25µL of cell supernatant was transferred to a 96-well white-walled plate, followed by addition to each well of 50µL of QUANTI-LUC solution containing stabilizer. Luminescence was detected on a plate reader according to the manufacturer’s instructions.

[0261] The remaining cells were assessed for cell viability by adding 25µL of CellTiter-Glo (Promega, G9683) solution to each well, and being placed on a shaker for 10 minutes at room temperature. Luminescence was detected on a plate reader, according to the manufacturer’s instructions. Part II - Results

[0262] Compound A was determined to have an IC50 value in the range of from > 0.250 µM to d + fA) :VTWV[UL 9 PIL IU ?:50 value >1 µM. 71 32017260 400807-030WO (212251)EXAMPLE 9 – Analysis of UVB-Induced pTBK1 in HaCaT Cells

[0263] Compound A was evaluated in an UVB-Induced pTBK1 assay in HaCaT Cells. Experimental procedures and results are provided below. Compound Identifier Chemical StructurePart I – Procedure

[0264] pTBK1 is an important signaling molecule in regulating inflammatory responses. HaCaT cells were plated in 6-well plates at a density of ~100 k / well in HaCaT media (DMEM, optimized 1x (Addex Bio) + 1 % pen strep (Gibco) + 5 % heat inactivated fetal bovine serum (Gibco)). The cells were then cultured at 37 ºC overnight. The next day, the cells were treated with the test compounds. The test compound was diluted and added to media aliquots to provide desired concentrations. To add the test compound+media mixture, an equivalent amount of media from each well was aspirated and then replenished with the media dosed with the test compound. The cells were then cultured for an additional 72 hours with compound treatment prior to UVB exposure.

[0265] The media was then aspirated from the wells, with the remaining cells at least 80% confluent in each well. One mL of PBS was then added to each well, and the plate was then placed under a UVB lamp. A UVB sensor was positioned near the plate to register the plate’s exposure. The cells were exposed to the UVB light until they reached 0.1 mJ / cm2. Then the plate was covered and transferred to a sterile hood for processing.

[0266] The PBS was aspirated out of the wells, and the wells were replenished with 3 mL fresh culture media. The cells were then cultured for an additional 24 hours, and samples were processed 24 hours post-UVB exposure. To process the samples, the media was aspirated, the plate placed on ice, and the cells washed with cold PBS, which was then aspirated off. Another 1 mL of cold PBS was added to each well. The cells were then scraped in the cold PBS solution 72 32017260 400807-030WO (212251)IUL ZXIUYNMXXML ZV KVUQKIS Z[JMY VU QKM) GPM KMSSY ]MXM ZPMU YW[U IZ e+*** E:= IZ . g: NVX / minutes. The cells were then resuspended in 1 mL of cold PBS and transferred to a TQKXVKMUZXQN[OM Z[JM) GPM KMSSY ]MXM YW[U IZ e+*** E:= IZ .g: NVX IUVZPMX / TQU[ZMY' IUL ZPM PBS was aspirated off. The cell pellet was prepared for lysis.

[0267] A RIPA lysis buffer (#BP-115, Boston BioProducts) was added to a Halt protease and phosphate inhibitor cocktail (#78440, ThermoFisher), and the mixture was cooled on ice. About 30µL of the lysis buffer mix was added to the cells. The samples were briefly vortexed and then QUK[JIZML VU QKM NVX IZ SMIYZ + / TQU[ZMY) GPM KMSSY ]MXM ZPMU YW[U e+*** E:= IZ .g: NVX / minutes and the supernatant was transferred to a clean tube. The protein concentration of the cell lysate was measured using Pierce™ Rapid Gold BCA Protein Assay Kit #AF3225 (ThermoFisher). ELISA analysis was run on select samples using one of the following kits: a. FastScan™ Phospho-TBK1 / NAK (Ser172) ELISA Kit #46948 (Cell Signaling Technologies) b. FastScan™ Total TBK1 / NAK ELISA Kit #15816 (Cell Signaling Technologies) c. FastScan™ Phospho-STING (Ser366) ELISA Kit #82083 (Cell Signaling Technologies) d. FastScan™ Phospho-IRF-3 (Ser396) ELISA Kit #50386 (Cell Signaling Technologies) e. FastScan™ Total IRF-3 ELISA Kit #29771 (Cell Signaling Technologies) f. Phospho-NAK / TBK1 (S172) ELISA Kit (ab279952) (Abcam) g. Phospho-IRF3 (S386) ELISA Kit (ab279833) (Abcam) Part II – Results

[0268] An IC50value was determined based on a response curve. Compound A has an IC50value in the range > 2.0 µA ZV d / )* µM. EXAMPLE 10 – Assay for Altering IFN Production in THP1-DualTMKO-TREX1 Mouse Xenografts

[0269] Compound A was tested for its ability to alter IFN levels in THP1-DualTMKO-TREX1 xenografts in mice. Assay procedures are described below. 73 32017260 400807-030WO (212251)Compound Identifier Chemical StructurePart I – Procedure for Altering IFN Production in THP1-DualTMKO-TREX1 Xenografts

[0270] CB-17 SCID female mice were inoculated subcutaneously with 10 million THP1- DualTM@C(GE<H+ KMSSY QU ,** mS D9F ]QZP AIZXQOMS #+4+$ IUL OXV[WML IZ B6 / WMX ZXMIZTMUZ per sampling timepoint when tumor volume reached an average of 340 mm3. Six groups were co- administered: (1) decitabine (DAC) at 5mg / kg SC, once daily, for 5 days, and (2) test Compound A at one of six doses (0.3mg / kg PO, 1mg / kg PO, 3mg / kg PO, 10mg / kg PO, 30mg / kg PO, 100mg / kg PO) once daily, for 5 days. Two additional groups were administered (1) decitabine and test compound vehicle control SC+PO, once daily, for 5 days, and (2) decitabine at 5mg / kg SC, once daily, for 5 days. Decitabine was formulated in sterile PBS, pH 7.4. Test compound was formulated in 20% HPBCD in 50 mM citrate buffer (pH 4.5).

[0271] Tumors were harvested 4h post-last dose administered on day 5, grinded in PBS at 50 Hz for 5 min, kept on ice for 30 min while being vortexed every 5 min. Tumors were then centrifuged, and PierceTMBCA Protein Assay Kit was used to measure protein concentration. Equal amounts of proteins were added to 96-well black plates, and luciferase signal was measured using the QUANTI-LucTMdetection medium according to manufacturer’s instructions. Luminescence was measured using the EnVision® 2105 Multimode Plate Reader. Part II – Results

[0272] Experimental results are depicted in Fig.14. Data was normalized relative to interferon signal from vehicle group. In Fig.14, IFN signal was measured in tumors harvested from mice 4 hours after the final decitabine and test compound dosing on day 5.

[0273] Compound A showed a dose-response inhibition of DAC-induced IFN signal in THP1 tumors. 74 32017260 400807-030WO (212251)INCORPORATION BY REFERENCE

[0274] The entire disclosure of each of the patent documents and scientific articles referred to herein is incorporated by reference for all purposes. EQUIVALENTS

[0275] The invention may be embodied in other specific forms without departing from the spirit 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. 75 32017260 400807-030WO (212251)

Claims

Claims:

1. A crystalline compound I:

2. The crystalline compound of claim 1, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.6 ± 0.2, 7.6 ± 0.2, 9.3 ± 0.2, 14.0 ± 0.2, 15.3 ± 0.2, 19.2 ± 0.2, and 23.1 ± 0.

2.

3. The crystalline compound of claim 2, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 5.1 ± 0.

2.

4. The crystalline compound of claim 2 or 3, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 6.7 ± 0.

2.

5. The crystalline compound of any one of claims 2-4, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.4 ± 0.

2.

6. The crystalline compound of any one of claims 2-5, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.2 ± 0.

2.

7. The crystalline compound of any one of claims 2-6, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 17.8 ± 0.

2.

8. The crystalline compound of any one of claims 2-7, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.3 ± 0.

2.

9. The crystalline compound of claim 1, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising at least six peaks selected from the following 76 32017260 400807-030WO (212251)diffraction angles (2u): 4.6 ± 0.2, 5.1 ± 0.2, 6.7 ± 0.2, 7.4 ± 0.2, 7.6 ± 0.2, 9.2 ± 0.2, 9.3 ± 0.2, 14.0 ± 0.2, 15.3 ± 0.2, 17.8 ± 0.2, 19.2 ± 0.2, 19.3 ± 0.2, and 23.1 ± 0.

2.

10. The crystalline compound of claim 9, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising at least seven peaks selected from said diffraction angles (2u).

11. The crystalline compound of any one of claims 2-10, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 10%.

12. The crystalline compound of any one of claims 2-10, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 5%.

13. The crystalline compound of claim 1 characterized by the following X-ray powder diffraction pattern expressed in terms of diffraction angle 2u, inter-planar distances d, and relative intensity (expressed as a percentage with respect to the most intense peak): Angle [2u]d-spacing [Å] Relative Intensity [%]77 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%] 1431 ± 020 6184 ± 0086 478 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%] 2605 ± 020 3418 ± 0026 2 .

14. The crystalline compound of claim 1, wherein the X-ray powder diffraction pattern is substantially as shown in FIG.

3.

15. The crystalline compound of any one of claims 1-14, wherein the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 124 degrees Celsius to about 128 degrees Celsius.

16. The crystalline compound of any one of claims 1-14, wherein the compound has a melting point onset as determined by differential scanning calorimetry at about 126 degrees Celsius.

17. The crystalline compound of any one of claims 1-16, wherein the compound has a peak melting temperature of about 130 degrees Celsius measured by differential scanning calorimetry (DSC).

18. The crystalline compound of any one of claims 1-14, wherein the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.

4.

19. The crystalline compound of claim 1, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.4 ± 0.2, 5.8 ± 0.2, 7.5 ± 0.2, 9.7 ± 0.2, 15.1 ± 0.2, 18.1 ± 0.2, and 22.7 ± 0.

2. 79 32017260 400807-030WO (212251)20. The crystalline compound of claim 19, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 7.3 ± 0.

2.

21. The crystalline compound of claim 19 or 20, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.7 ± 0.

2.

22. The crystalline compound of any one of claims 19-21, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.8 ± 0.

2.

23. The crystalline compound of any one of claims 19-22, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.9 ± 0.

2.

24. The crystalline compound of any one of claims 19-23, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 9.6 ± 0.

2.

25. The crystalline compound of any one of claims 19-24, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 12.4 ± 0.

2.

26. The crystalline compound of any one of claims 19-25, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 13.9 ± 0.

2.

27. The crystalline compound of claim 1, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising at least six peaks selected from the following diffraction angles (2u): 4.4 ± 0.2, 5.8 ± 0.2, 7.3 ± 0.2, 7.5 ± 0.2, 8.7 ± 0.2, 8.8 ± 0.2, 8.9 ± 0.2, 9.6 ± 0.2, 9.7 ± 0.2, 12.4 ± 0.2, 13.9 ± 0.2, 15.1 ± 0.2, 18.1 ± 0.2, and 22.7 ± 0.

2.

28. The crystalline compound of claim 27, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising at least seven peaks selected from said diffraction angles (2u).

29. The crystalline compound of any one of claims 19-28, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 15%.

30. The crystalline compound of any one of claims 19-28, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 10%. 80 32017260 400807-030WO (212251)31. The crystalline compound of claim 1 characterized by the following X-ray powder diffraction pattern expressed in terms of diffraction angle 2u, inter-planar distances d, and relative intensity (expressed as a percentage with respect to the most intense peak): Angle [2u]d-spacing [Å] Relative Intensity [%]81 32017260 400807-030WO (212251)Angle [2u] d-spacing [Å] Relative Intensity [%]32. The crystalline compound of claim 1, wherein the X-ray powder diffraction pattern is substantially as shown in FIG.

9.

33. The crystalline compound of any one of claims 19-32, wherein the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 125 degrees Celsius to about 129 degrees Celsius.

34. The crystalline compound of any one of 19-32, wherein the compound has a melting point onset as determined by differential scanning calorimetry at about 127 degrees Celsius.

35. The crystalline compound of any one of claims 19-34, wherein the compound has a peak melting temperature of about 132 degrees Celsius measured by differential scanning calorimetry (DSC).

36. The crystalline compound of any one of claims 19-32, wherein the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.

10. 82 32017260 400807-030WO (212251)37. The crystalline compound of claim 1, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.4 ± 0.3, 5.0 ± 0.3, 6.5 ± 0.3, 7.2 ± 0.3, 9.1 ± 0.3, 13.9 ± 0.3, and 17.8 ± 0.

3.

38. The crystalline compound of claim 37, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 18.2 ± 0.

3.

39. The crystalline compound of claim 37 or 38, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.2 ± 0.

3.

40. The crystalline compound of any one of claims 37-39, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 22.1 ± 0.

3.

41. The crystalline compound of any one of claims 37-40, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 15%.

42. The crystalline compound of any one of claims 37-40, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 10%.

43. The crystalline compound of claim 1, wherein the X-ray powder diffraction pattern is substantially as shown in FIG.

7.

44. The crystalline compound of any one of claims 37-43, wherein the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 127 degrees Celsius to about 131 degrees Celsius.

45. The crystalline compound of any one of claims 37-43, wherein the compound has a melting point onset as determined by differential scanning calorimetry at about 129 degrees Celsius.

46. The crystalline compound of any one of claims 37-45, wherein the compound has a peak melting temperature of about 132 degrees Celsius measured by differential scanning calorimetry (DSC).

47. The crystalline compound of any one of claims 37-43, wherein the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.

8. 83 32017260 400807-030WO (212251)48. The crystalline compound of claim 1, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 4.2 ± 0.3, 5.7 ± 0.3, 7.3 ± 0.3, 9.5 ± 0.3, 10.4 ± 0.3, 13.9 ± 0.3, and 22.3 ± 0.

3.

49. The crystalline compound of claim 48, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 8.8 ± 0.

3.

50. The crystalline compound of claim 48 or 49, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 12.2 ± 0.

3.

51. The crystalline compound of any one of claims 48-50, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 15.9 ± 0.

3.

52. The crystalline compound of any one of claims 48-51, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 19.1 ± 0.

3.

53. The crystalline compound of any one of claims 48-52, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 29.9 ± 0.

3.

54. The crystalline compound of any one of claims 48-53, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 15%.

55. The crystalline compound of any one of claims 48-53, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 10%. .

56. The crystalline compound of claim 1, wherein the X-ray powder diffraction pattern is substantially as shown in FIG.

12.

57. The crystalline compound of any one of claims 48-56, wherein the compound has a melting point onset as determined by differential scanning calorimetry in the range of from about 126 degrees Celsius to about 130 degrees Celsius.

58. The crystalline compound of any one of claims 48-56, wherein the compound has a melting point onset as determined by differential scanning calorimetry at about 128 degrees Celsius. 84 32017260 400807-030WO (212251)59. The crystalline compound of any one of claims 48-58, wherein the compound has a peak melting temperature of about 132 degrees Celsius measured by differential scanning calorimetry (DSC).

60. The crystalline compound of any one of claims 48-56, wherein the compound has a differential scanning calorimetry curve substantially the same as shown in FIG.

13.

61. A crystalline compound II:

62. The crystalline compound of claim 61, wherein the crystalline compound exhibits an X- ray powder diffraction pattern comprising peaks at the following diffraction angles (2u): 8.9 ± 0.3, 13.3 ± 0.2, 19.6 ± 0.3, 20.7 ± 0.3, 21.6 ± 0.3, 26.8 ± 0.3, and 35.7 ± 0.

3.

63. The crystalline compound of claim 61, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 16.7 ± 0.

3.

64. The crystalline compound of claim 62 or 63, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 18.1 ± 0.

3.

65. The crystalline compound of any one of claims 62-64, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 27.7 ± 0.

3.

66. The crystalline compound of any one of claims 62-65, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 29.1 ± 0.

3.

67. The crystalline compound of any one of claims 62-66, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 33.9 ± 0.

3.

68. The crystalline compound of any one of claims 62-67, wherein the X-ray powder diffraction pattern further comprises a peak at the following diffraction angle (2u): 35.0 ± 0.

3. 85 32017260 400807-030WO (212251)69. The crystalline compound of any one of claims 62-68, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 15%.

70. The crystalline compound of any one of claims 62-68, wherein the relative intensity of the peak at said diffraction angles (2u) is at least 10%.

71. The crystalline compound of claim 62, wherein the X-ray powder diffraction pattern is substantially as shown in FIG.

1.

72. A pharmaceutical composition comprising a compound of any one of claims 1-71 and a pharmaceutically acceptable carrier.

73. A method of treating a disorder selected from the group 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 according to any one of claims 1-71 to treat the disorder.

74. A method of inhibiting LINE1 reverse transcriptase activity in a subject, comprising contacting a LINE1 reverse transcriptase with an effective amount of a compound according to any one of claims 1-71 to inhibit the activity of said LINE1 reverse transcriptase.

75. The method of claim 74, wherein the method further comprises inhibiting HERV-K reverse transcriptase activity in the subject.

76. A method of inhibiting HERV-K reverse transcriptase activity in a subject, comprising contacting a HERV-K reverse transcriptase with an effective amount of a compound according to any one of claims 1-71 to inhibit the activity of said HERV-K reverse transcriptase.

77. The method of any one of claims 74-76, wherein the subject has a disorder selected from the group consisting of cancer, an autoimmune disorder, and a neurological disorder.

78. The method of claim 73 or 77, wherein the disorder is cancer.

79. The method of claim 78, wherein 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. 86 32017260 400807-030WO (212251)80. The method of claim 73 or 77, wherein the disorder is an autoimmune disorder.

81. The method of claim 80, wherein the autoimmune disorder is selected from Aicardi- Goutières 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.

82. The method of claim 80, wherein the autoimmune disorder is systemic lupus erythematosus (SLE).

83. The method of claim 73 or 77, wherein the disorder is a neurological disorder.

84. The method of claim 83, 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.

85. The method of any one of claims 73-84, 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.

86. The method of any one of claims 73-84, wherein the subject has (i) expression of HERV- K RNA and / or (ii) activity of HERV-K reverse transcriptase.

87. The method of any one of claims 73-86, wherein the subject is a human. 87 32017260 400807-030WO (212251)

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