Modulators of pannexin-1 activity

Abstract

Provided herein are compounds and compositions thereof for modulating pannexin-1. In some embodiments, the compounds and compositions are provided for treatment of diseases or conditions, such as inflammatory conditions, epilepsy, seizure, or pain. In some embodiments, the compounds have the structural formulas (I), (II), and (III), wherein values for the variables (e.g., structure A, structure B, structure C, L, L', X1, X2, X3, X4, R1, R2, R3, R4, R6, R7, R8, m) are as described herein.

Description

Attorney Docket No. 01270-0017-00PCTCOMPOUNDS AND USES THEREOF FOR THE TREATMENT OF DISEASES CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of US Provisional Application No. 63 / 695,058, filed September 16, 2024, and US Provisional Application No. 63 / 777,059, filed March 25, 2025, each of which is incorporated herein by reference in its entirety for any purpose.FIELD

[0002] The present disclosure relates generally to compounds, compositions, and methods for their preparation and use for treating diseases or conditions.BACKGROUND

[0003] Pannexins were first discovered in flatworm by Panchin et al. (et al. Curr. Biol. 2000; 10: R473-R474) as a class of innexin-like membrane channels that form gap junctions in invertebrate animals. The pannexin (Panx) family consists of 3 members: Pannexin-1 (Panxl) is ubiquitously expressed in most tissues (e.g., the eye, thyroid, prostate, kidney, liver, immune cells, and central nervous system (CNS)); Pannexin-2 (Panx2) is primarily restricted to the CNS; and Pannexin-3 (Panx3) is predominantly found in the skin and skeletal tissues.

[0004] They were named as pannexins (‘pan’, meaning ‘all’ in Greek) to reflect their widespread expression in many vertebrate species (Panchin Y, et al. Curr. Biol. 2000; 10: R473-R474). The vertebrate pannexins assemble into hexameric hemichannels that allow ATP release from many types of cells (Chiu et al., J. Gen. Physiol. 2018; 150: 19-39, Valdebenito S, et al., Biochim. Biophys. Acta. 2018; 1860: 154-165).

[0005] Activation of Panxl channels has been associated with the onset and propagation of cell death (Penuela, S. et al., (2009) Mol. Biol. Cell 20 (20), 4313-4323). Panxl forms large and non-selective channels, and its irreversible opening is detrimental. Prolonged opening of Panxl channels leads to the liberation of signaling molecules that are recognized as ‘danger signals’ and trigger inflammatory responses (D'Hondt, C., et al. (2011). Cell. Signal. 23 (2), 305-316). ATP can also be released from the injured cells undergoing necrosis and from infiltrating leukocytes through Panxl channels. ATP release leads to an important loss of energy and stimulates necrotic cell death. (Makarenkova, H. P, and Shestopalov, V. I. (2014). Front. Physiol. 5, 63).

[0006] Under physiological conditions, Panxl hemichannels remain largely closed, as their C- terminal tails may physically plug up the hemichannel pore from the cytoplasmic side. InAttorney Docket No. 01270-0017-00PCT response to inflammatory stimuli, the C-terminal tail of Panxl could be cleaved by several caspases, resulting in a progressive opening of Panxl hemichannel and extracellular release of larger molecules like ATP (Chekeni FB, et al 2010; 467:863-867), which activates the purinergic receptor P2X7R to trigger inflammasome activation (Yang D et al 2015; 43:923-932).

[0007] Upon engagement of Toll-like receptors by external stimuli (Poltorak A, et al. 282:2085- 2088), macrophages and monocytes release a wide array of “early” (e.g., TNF, IL-ip, IFN-y, and CIRP) proinflammatory mediators.

[0008] An overexpression of the Pannexin-1 channel has been proposed to be responsible for the strong inflammatory response, high serum ATP level, systemic cellular energy crisis and histopathological damages in inflammatory conditions like sepsis. Inhibiting Pannexin-1 channel-mediated release of intracellular ATP could decrease the above-mentioned injuries, and Pannexin-1 is expected to be a potential therapeutic target in inflammatory disorders like sepsis and sepsis-induced conditions.

[0009] Pannexins are also involved in the development of neuropathic pain, particularly resulting from inflammatory sensory nerve disorders, as inflammatory pain in peripheral nerves commonly results from the interaction between glia and neurons along the pannexin-l / ATP- signaling axis. Pannexin-1 channel inhibition has therefore been proposed as a method for mitigating neuropathic pain in humans.

[0010] Pannexin-1 channel activity is also involved in the generation and maintenance of epileptiform neuron firing and may be a valuable target for the treatment of epilepsy. Pannexin-1 expression is elevated in both animal seizure models and in resected human epileptic brain tissue. Many of the cellular factors that contribute to seizure generation and maintenance, including potassium, ATP, and glutamate, are transported by open pannexin-1 channels. This has led to the hypothesis that pannexin-1 modulation may represent an effective treatment strategy for epileptic humans.

[0011] There are currently few options for treating inflammatory diseases, epilepsy, or other conditions associated with aberrant Pannexin-1 activity or overexpression. Thus, there remains a need for compounds and methods for treatment of pannexin-1 -associated diseases and conditions.SUMMARY

[0012] Provided herein are compounds for inhibiting pannexin-1, e.g., for treating an inflammatory condition in a subject. Other example uses of the compounds disclosed herein include pain (e.g., neuropathic pain) and epileptic seizures.Attorney Docket No. 01270-0017-00PCT

[0013] Some embodiments provide a compound of the following structural formula:pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein values for the variables (e.g., structure A, L, L', X1, X2, X3, X4, R1, R2, R3R4, m) are as described herein.

[0014] Some embodiments provide a compound of the following structural formula:pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein values for the variables (e.g., structure B, L, L', X1, X2, X3, X4, R2, R3, R4, R6, R7, m) are as described herein.

[0015] Some embodiments provide a compound of the following structural formula:pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein values for the variables (e.g., structure C, L, X1, X2, X3, X4, R2, R3, R4, R8, m) are as described herein.

[0016] Also provided herein is a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of formula(I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof), and a pharmaceutically acceptable carrier.

[0017] Also provided herein is a method of inhibiting pannexin-1 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula(I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof), or a pharmaceutical composition thereof.Attorney Docket No. 01270-0017-00PCT

[0018] Also provided herein is a method of treating an inflammatory condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula(I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof), or a pharmaceutical composition thereof.

[0019] Also provided herein is a method of treating epilepsy and / or seizure or seizures in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula (I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof), or a pharmaceutical composition thereof.

[0020] Also provided herein is a method of treating pain in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula(I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof), or a pharmaceutical composition thereof.

[0021] In some embodiments, a compound disclosed herein (e.g., a compound of Formula (I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof) or a pharmaceutical composition comprising a compound disclosed herein is for use as a medicament, e.g., to inhibit Pannexin-1 and / or treat an inflammatory condition, epilepsy, seizure or seizures, and / or pain in a subject (e.g., a subject in need thereof). Also provided herein is use of a compound disclosed herein (e.g., a compound of Formula (I), (II), or (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof) or a pharmaceutical composition thereof for the manufacture of a medicament for a use described herein, e.g., to inhibit Pannexin-1 and / or treat an inflammatory condition, epilepsy, seizure or seizures, and / or pain in a subject (e.g., a subject in need thereof).DETAILED DESCRIPTIONDefinitions

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs. In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments of the disclosure. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. To the extent any materialAttorney Docket No. 01270-0017-00PCT incorporated herein by reference is inconsistent with the express content of this disclosure, the express content controls. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an”, and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and / or” unless stated otherwise. Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting.

[0023] Unless the context requires otherwise, throughout the present specification and claims, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to”.

[0024] In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. Also, any number range recited herein relating to any physical feature, such as polymer subunits, size, or thickness, are to be understood to include any integer within the recited range, unless otherwise indicated. As used herein, the terms “about” and “approximately” mean ± 20%, ± 10%, ± 5%, or ± 1% of the indicated range, value, or structure, unless otherwise indicated.

[0025] Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0026] “Amino” refers to the -NH2 radical.

[0027] “Carboxy” or “carboxyl” refers to the -CO2H radical.

[0028] “Cyano” refers to the -CN radical.

[0029] “Hydroxy” or “hydroxyl” refers to the -OH radical.

[0030] “Nitro” refers to the -NO2 radical.

[0031] “ Oxo” refers to the =0 substituent.

[0032] “ Thioxo” refers to the =S substituent.

[0033] “ Thiol” refers to the -SH substituent.Attorney Docket No. 01270-0017-00PCT

[0034] “Alkyl” refers to an unbranched or branched saturated hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, having from one to twelve carbon atoms ((Ci- Ci2)alkyl), preferably one to eight carbon atoms ((Ci-C8)alkyl), one to six carbon atoms ((Ci- Ce)alkyl), one to five carbon atoms ((Ci-Cs)alkyl), or one to three carbon atoms ((Ci-C3)alkyl), and which is attached to the rest of the molecule by a single bond. Examples of alkyl include, e.g., methyl, ethyl, zz-propyl, 1 -methylethyl (z.w-propyl), zz-butyl, zz-pentyl, 1,1 -dimethylethyl ( / -butyl), 3 -methylhexyl, 2-methylhexyl and the like.

[0035] “Alkenyl” refers to an unbranched or branched unsaturated hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, which contains one or more carbon-carbon double bonds, having from two to twelve carbon atoms (C2-C12 alkenyl), preferably two to eight carbon atoms (C2-C8 alkenyl) or two to six carbon atoms (C2-C6 alkenyl), and which is attached to the rest of the molecule by a single bond. Examples of alkenyl include, e.g., ethenyl, prop-l-enyl, but-l-enyl, pent-l-enyl, penta- 1,4-dienyl, and the like.

[0036] “Alkynyl” refers to an unbranched or branched unsaturated hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, which contains one or more carbon-carbon triple bonds, having from two to twelve carbon atoms (C2-C12 alkynyl), preferably two to eight carbon atoms (C2-C8 alkynyl) or two to six carbon atoms (C2-C6 alkynyl), and which is attached to the rest of the molecule by a single bond. Examples of alkynyl include, e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

[0037] “Alkoxy” refers to a radical of the formula -ORa where Ra is alkyl as defined herein. Preferred alkoxy groups have one to six carbon atoms (z.e., Ci-Ce alkoxy), one to five carbon atoms (z.e., C1-C5 alkoxy), or one to three carbon atoms (z.e., C1-C3 alkoxy) in the alkyl radical. Examples of alkoxy groups include, e.g., methoxy, ethoxy, propoxy, and butoxy. Unless stated otherwise specifically in the specification, an alkoxy is optionally substituted.

[0038] “Aromatic ring” refers to a cyclic planar portion of a molecule (z.e., a radical) with a ring of resonance bonds that exhibits increased stability relative to other connective arrangements with the same sets of atoms. Generally, aromatic rings contain a set of covalently bound coplanar atoms and comprise a number of 7t-electrons (for example, alternating double and single bonds) that is even but not a multiple of 4 (z.e., 4n + 2 7t-electrons, where n = 0, 1, 2, 3, etc.).Aromatic rings include, but are not limited to, phenyl, naphthenyl, imidazolyl, pyrrolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridonyl, pyridazinyl, pyrimidonyl.

[0039] “Aryl” refers to a carbocyclic ring system radical comprising 6 to 18 carbon atoms and at least one aromatic ring (i.e., Ce-Ci8 aryl), preferably having 6 to 10 carbon atoms (z.e., Ce-Cio aryl). For purposes of embodiments of this disclosure, the aryl radical is a monocyclic, bicyclic,Attorney Docket No. 01270-0017-00PCT tricyclic or tetracyclic ring system, which may include fused, spiro, or bridged ring systems. Aryl includes, but is not limited to, radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, a.s-indacene, 5-indacene, indane, indene, naphthalene, phenalene, phenanthrene, phenyl, pleiadene, pyrene, triphenylene.

[0040] “Arylalkyl” refers to a radical of the formula -Rb-Rc where Rb is an alkylene chain and Rcis one or more aryl as defined above, for example, benzyl, diphenylmethyl and the like. An arylalkyl may contain a C1-C10 alkylene chain connected to a Ce-Cio aryl radical (z.e., Ce-Cio arylalkyl).

[0041] “Cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclic carbocyclic radical consisting solely of carbon and hydrogen atoms, which may include fused, spiro, or bridged ring systems, having from three to fifteen carbon atoms (z.e., C3-C15 cycloalkyl), preferably having from three to ten carbon atoms (z.e., C3-C10 cycloalkyl), six to ten carbon atoms (z.e., Ce-Cio cycloalkyl), three to six carbon atoms (z.e., C3-C6 cycloalkyl), three to five carbon atoms (z.e., C3-C5 cycloalkyl), and which is saturated or unsaturated and attached to the rest of the molecule by a single bond. In some embodiments, cycloalkyl is saturated. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.

[0042] “Halo” or “halogen” refers to bromo, chloro, fluoro, or iodo. In some embodiments, halo is bromo, chloro, or fluoro. In some embodiments, halo is fluoro or chloro. In some embodiments, halo is fluoro.

[0043] “Haloalkyl” refers to alkyl, as defined herein, that is substituted by one or more halo, as defined herein. A preferred haloalkyl group includes an alkyl group having one to six carbon atoms and that is substituted by one or more halo radicals (z.e., Ci-Ce haloalkyl). Examples of Ci-Ce haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, hexafluoroisopropyl and the like. The halo radicals may be all the same or the halo radicals may be different.

[0044] “Heteroaryl” refers to a cyclic ring system radical comprising at least one aromatic ring consisting of carbon ring atoms and one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur. As used herein, heteroaryl includes 1 to 10 ring carbon atoms and 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur within the ring. Preferred heteroaryl groups have a 5- to 10-membered ring system containing one to fourAttorney Docket No. 01270-0017-00PCT heteroatoms selected from nitrogen, oxygen, and sulfur (z.e., a 5- to 10-membered heteroaryl) and a 5- to 6-membered ring system containing one to four heteroatoms selected from nitrogen, oxygen, and sulfur (z.e., a 5- to 6-membered heteroaryl). For purposes of embodiments of this disclosure, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused, spiro, and / or bridged ring systems. Examples of heteroaryl groups include pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, dihydroquinolynyl, dihydronaphthyridinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl and thiophenyl (i.e., thienyl).

[0045] “Heterocyclyl” refers to a saturated or unsaturated cycloalkyl group, wherein one or more ring carbon atoms has been replaced with a heteroatom independently selected from nitrogen, oxygen and sulfur. The term “heterocyclyl” includes heterocycloalkenyl groups (z.e., the heterocyclyl group having at least one double bond), bridged heterocyclyl groups, fused heterocyclyl groups and spiroheterocyclyl groups. A heterocyclyl may be a single ring or multiple rings wherein the multiple rings may be fused, bridged or spiro, and may comprise one or more oxo (C=O) moieties. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of the attachment (z.e., can be bound through a carbon atom or a heteroatom). As used herein, heterocyclyl has 1 to 10 ring carbon atoms, e.g., 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms, and 1 to 5 ring heteroatoms, e.g., 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 to 2 heteroatoms independently selected from nitrogen, sulfur and oxygen. Preferred heterocyclyls have five to 10 members in the ring system including one to four heteroatoms selected from nitrogen and oxygen (z.e., 5- to 10-membered heterocyclyl) or five to eight members in the ring system including one to four heteroatoms selected from nitrogen and oxygen (z.e., 5- to 8-membered heterocyclyl). Examples of heterocyclyl groups include dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl and 1,1-dioxo-thiomorpholinyl.

[0046] As used herein, “N-linked spiroheterocyclyl” refers to a spirocyclyl as defined herein wherein at least one ring atom is a heteroatom selected from nitrogen, oxygen, and sulfur, and which is linked to the rest of the molecule by a nitrogen atom in one of the rings. Examples ofAttorney Docket No. 01270-0017-00PCTN-linked spiroheterocyclyl groups include, but are not limited to:

[0047] As used herein, “N-linked fused heterocyclyl” refers to a fused cyclyl as defined herein wherein at least one ring atom is a heteroatom selected from nitrogen, oxygen, and sulfur, and which is linked to the rest of the molecule by a nitrogen atom in one of the rings. Examples ofN-linked fused heterocyclyl groups include, but are not limited to:

[0048] As used herein, “N-linked bridged heterocyclyl” refers to a bridged cyclyl as defined herein wherein at least one ring atom is a heteroatom selected from nitrogen, oxygen, and sulfur, and which is linked to the rest of the molecule by a nitrogen atom in one of the rings. Examples of N-linked bridged heterocyclyls include, but are not limited to:

[0049] As used herein, “spirocyclyl” refers to a cycloalkyl or heterocyclyl, as those terms are defined herein, comprising at least two rings, wherein at least two rings share a single common atom. In some embodiments, spirocyclyl is spiroheterocyclyl. In some embodiments, spirocyclyl is spirocycloalkyl. Preferred spirocyclyls have 5 to 12 members, e.g., 6 to 10 members, in the ring system.

[0050] As used herein, “fused cyclyl” refers to a cyclic group comprising at least two rings, wherein at least two of the at least two rings share two adjacent atoms. In some embodiments, the cyclic group is a cycloalkyl or heterocyclyl, as those terms are defined herein. Examples ofAttorney Docket No. 01270-0017-00PCT fused cyclyls include, but are not limited to:'N—, and. Preferred fused cyclyls have 5 to 14 members, e.g., 6 to 10 members, in the ring system.

[0051] As used herein, “bridged cyclyl” refers to a cycloalkyl or heterocyclyl, as those terms are defined herein, comprising at least two rings, wherein at least two rings share three or more atoms. Examples of bridged cyclyl include, but are not limited to: LQ^J, FI, L0_B 'O_Band LQ^J.Preferred bridged cyclyls have 5 to 14 members, e.g., 6 to 10 members, in the ring system.

[0052] As used herein, “monocyclyl” refers to a cycloalkyl or heterocyclyl, as those terms are defined herein, consisting of a single ring. Examples of monocyclyl include, but are not limited to: cyclopropanyl, cyclobutanyl, cyclopentenyl, cyclohexyl, cycloheptyl, and piperazinyl. Unless stated otherwise specifically in the specification, a monocyclyl is optionally substituted.Preferred monocyclyls have 3 to 8 members, e.g., 3 to 6 members, in the ring system.

[0053] As used herein, “C-linked spirocyclyl” refers to a spirocylyl, as defined herein that is linked to the rest of the molecule by a carbon ring atom. Examples of C-linked spirocyclyl groups include, but are not limited to:, and

[0054] As used herein, “C-linked fused cyclyl” refers to a fused cyclyl group as defined herein, which is connected to the rest of the molecule by a carbon ring atom.

[0055] As used herein, “C-linked bridged cyclyl” refers to a bridged cyclyl group as defined herein, which is linked to the rest of the molecule by a carbon ring atom.

[0056] As used herein, “C-linked monocyclyl” refers to a monocyclyl group as defined herein, which is linked to the rest of the molecule by a carbon ring atom. Examples of C-linkedAttorney Docket No. 01270-0017-00PCT monocyclyl include, but are not limited to:

[0057] When a group (e.g., Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C12 cycloalkyl, aryl, and heteroaryl) or other substituent is described herein as “substituted” at least one hydrogen atom (e.g., 1, 2, 3, or all hydrogen atoms) of the group is replaced by a non-hydrogen atom such as, but not limited to: a halogen atom such as F, Cl, Br, and I (i.e., “halo”); an oxygen atom in groups such as hydroxyl groups or alkoxy groups (e.g., alkoxy or haloalkoxy); a nitrogen atom in groups such as amines (e.g., -NH2), amides (e.g., -(C=O)NH2), and nitro; alkyl groups including one or more halogen, such as F, Cl, Br, and I (e.g., haloalkyl); and cyano. The term “optionally substituted” includes unsubstituted and substituted. In some embodiments, a group that is optionally substituted is unsubstituted. In some embodiments, a group that is optionally substituted is substituted.

[0058] “Effective amount” or “therapeutically effective amount” of a compound or a composition refers to that amount of the compound or the composition that results in an intended result as desired based on the disclosure herein. Effective amounts can be determined by standard pharmaceutical procedures in cell cultures or experimental animals including, without limitation, by determining the ED50 (the dose therapeutically effective in 50% of the population) and the LD50 (the dose lethal to 50% of the population). In some embodiments, an effective amount of a compound results in reduction or inhibition of symptoms or a prolongation of survival in a subject (i.e., a human patient). The results may require multiple doses of the compound.

[0059] “Treating” or “treatment” of a disease in a subject refers to: 1) inhibiting a disease or arresting its development in a patient; or 2) ameliorating or causing regression of the disease in the patient. As used herein, “treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. For the purposes of this disclosure, beneficial or desired results include, but are not limited to, one or more of the following: decreasing one or more symptoms resulting from the disease or disorder, diminishing the extent of the disease or disorder, stabilizing the disease or disorder (e.g., preventing or delaying the worsening of the disease or disorder), delaying the occurrence or recurrence of the disease or disorder, delay or slowing the progression of the disease or disorder, ameliorating the disease or disorder state, providing a remission (whether partial or total) of the disease or disorder, decreasing the dose ofAttorney Docket No. 01270-0017-00PCT one or more other medications required to treat the disease or disorder, enhancing the effect of another medication used to treat the disease or disorder, delaying the progression of the disease or disorder, increasing the quality of life, and / or prolonging survival of a subject. Also encompassed by “treatment” is a reduction of pathological consequence of the disease or disorder. The methods of this disclosure contemplate any one or more of these aspects of treatment.

[0060] As used herein, the terms “individual(s)”, “subject(s)” and “patient(s)” mean any mammal. Examples include, but are not limited to, mice, rats, hamsters, guinea pigs, pigs, rabbits, cats, dogs, goats, sheep, cows, and humans. In some embodiments, the mammal is a human.

[0061] A “therapeutic effect”, as that term is used herein, encompasses a therapeutic benefit and / or a prophylactic benefit as described herein. A therapeutic effect includes delaying or eliminating the appearance of a disease or condition; delaying or eliminating the onset of symptoms of a disease or condition; slowing, halting, or reversing the progression of a disease or condition; causing partial or complete regression of a disease or condition; or any combination thereof.

[0062] The terms “co-administration”, “administered in combination with”, and their grammatical equivalents, as used herein, encompass administration of two or more agents to an animal, including humans, so that both agents and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which both agents are present.

[0063] “Pharmaceutically acceptable” refers to compounds, salts, compositions, dosage forms and other materials which are useful in preparing a pharmaceutical composition that is suitable for veterinary or human pharmaceutical use.

[0064] “Pharmaceutically acceptable salt” includes both acid and base addition salts.

[0065] “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor- 10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid,Attorney Docket No. 01270-0017-00PCT ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2- oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene- 1,5-disulfonic acid, naphthal ene-2-sulfonic acid, 1 -hydroxy -2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, -toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like.

[0066] “Pharmaceutically acceptable base addition salt” refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, 7V-ethylpiperidine, polyamine resins and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0067] In some embodiments, pharmaceutically acceptable salts include quaternary ammonium salts such as quaternary amine alkyl halide salts (e.g., methyl bromide).

[0068] As used herein, “therapeutic agent” refers to a biological, pharmaceutical, or chemical compound or other moiety. Non-limiting examples include a simple or complex organic or inorganic molecule, a peptide, a protein, an oligonucleotide, an antibody, an antibody derivative, antibody fragment, a vitamin derivative, a carbohydrate, a toxin, or a chemotherapeutic compound. Various compounds can be synthesized, for example, small molecules and oligomers (e.g., oligopeptides and oligonucleotides), and synthetic organic compounds based onAttorney Docket No. 01270-0017-00PCT various core structures. In addition, various natural sources can provide compounds for screening, such as plant or animal extracts, and the like.

[0069] The term “zzz vivo" refers to an event that takes place in a subject’s body.

[0070] Embodiments of the disclosure are also meant to encompass all pharmaceutically acceptable compounds of Formula (I), Formula (II) or Formula (III) being isotopically-labelled by having one or more atoms replaced by an atom having a different atomic mass or mass number (z.e., an “isotopic form” or “isotope” of a compound of Formula (I), Formula (II) or Formula (III)). Examples of isotopes that can be incorporated into the compounds of Formula (I), Formula (II) or Formula (III) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as2H,3H,nC,13C,14C,13N,15N,15O,17O,18O,31P,32P,35S,18F,36C1,123I, and125I, respectively. These radiolabeled compounds could be useful to help determine or measure the effectiveness of the compounds, by characterizing, for example, the site or mode of action, or binding affinity to pharmacologically important site of action. Certain isotopically-labeled compounds of Formula (I), Formula (II) or Formula (III), for example, those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The radioactive isotopes tritium, z.e.,3H, and carbon-14, z.e.,14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection.

[0071] Substitution with heavier isotopes such as deuterium, z.e.,2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence are preferred in some circumstances.

[0072] Substitution with positron emitting isotopes, such asnC,18F,15O and13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds of Formula (I), Formula (II) or Formula (III) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Examples as set out below using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

[0073] Certain embodiments are also meant to encompass the in vivo metabolic products of the disclosed compounds. Such products may result from, for example, the oxidation, reduction, hydrolysis, amidation, esterification, and the like of the administered compound, primarily due to enzymatic processes. Accordingly, the embodiments include compounds produced by a process comprising administering a compound of this disclosure to a mammal for a period of time sufficient to yield a metabolic product thereof. Such products are typically identified by administering a radiolabeled compound of the disclosure in a detectable dose to an animal, suchAttorney Docket No. 01270-0017-00PCT as rat, mouse, guinea pig, monkey, or to human, allowing sufficient time for metabolism to occur, and isolating its conversion products from the urine, blood or other biological samples.

[0074] “ Stable compound” and “stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0075] Often crystallizations produce a solvate of the compound of the disclosure. As used herein, the term “solvate” refers to an aggregate that comprises one or more molecules of a compound of Formula (I), Formula (II) or Formula (III) with one or more molecules of solvent. In some embodiments, the solvent is water, in which case the solvate is a hydrate. Alternatively, in other embodiments, the solvent is an organic solvent. Thus, the compounds of Formula (I), Formula (II) or Formula (III) may exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. In some aspects, the compound of Formula (I), Formula (II) or Formula (III) is a true solvate, while in other cases, the compound of the disclosure merely retains adventitious water or is a mixture of water plus some adventitious solvent.

[0076] “Optional” or “optionally” means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. For example, “optionally substituted aryl” means that the aryl radical may or may not be substituted and that the description includes both substituted aryl radicals and aryl radicals having no substitution. Polymers or similar indefinite structures arrived at by defining substituents with further substituents appended ad infinitum (e.g., a substituted aryl having a substituted alkyl which is itself substituted with a substituted aryl group, which is further substituted by a substituted heteroalkyl group, etc.) are not intended for inclusion herein. Similarly, the above definitions are not intended to include impermissible substitution patterns (e.g., methyl substituted with 5 fluorines or heteroaryl groups having two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to the skilled artisan.

[0077] A “pharmaceutical composition” or “pharmaceutically acceptable composition” refers to a formulation of a compound of the disclosure and a medium generally accepted in the art for the delivery of the biologically active compound to mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents, or excipients therefor.

[0078] “Pharmaceutically acceptable carrier, diluent or excipient” includes, without limitation, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonicAttorney Docket No. 01270-0017-00PCT agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.

[0079] The compounds of Formula (I), Formula (II) or Formula (III) or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, may contain one or more centers giving rise to geometric asymmetry and may thus provide enantiomers, diastereomers, and other stereoisomeric forms that are defined, in terms of absolute stereochemistry, as (R)- or (5)- or, as (D)- or (L)- for amino acids. Embodiments thus include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (5)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0080] A “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0081] “Diastereoisomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.

[0082] A “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule. Embodiments thus include tautomers of the disclosed compounds.

[0083] The chemical naming protocol and structure diagrams used herein are a modified form of the I.U.P.A.C. nomenclature system, using the ACD / Name Version 9.07 software program and / or ChemDraw Ultra Version 11.0.1 software naming program (CambridgeSoft). For complex chemical names employed herein, a substituent group is typically named before the group to which it attaches. For example, cyclopropylethyl comprises an ethyl backbone with a cyclopropyl substituent. Except as described below, all bonds are identified in the chemical structure diagrams herein, except for all bonds on some carbon atoms, which are assumed to be bonded to sufficient hydrogen atoms to complete the valency.Attorney Docket No. 01270-0017-00PCT

[0084] Although various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, although the invention may be described herein in the context of separate embodiments for clarity, the invention may also be implemented in a single embodiment.Compounds

[0085] In an aspect, provided herein is a compound of structural Formula (I):or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein:L is absent, O, S, or CH2; wherein: when L is absent, structure A is a 6- to 10-membered, N-linked spiroheterocyclyl, N- linked fused heterocyclyl, or N-linked bridged heterocyclyl; or when L is O, S, or CH2, structure A is a 6- to 10-membered spirocyclyl, fused cyclyl, or bridged cyclyl, or 3- to 6-membered, monocyclyl;X1, X2, and X3are each independently CR60or N; each R60is independently H, halo, -(Ci-C3)alkyl, or -O(Ci-C3)alkyl;X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), orC(R50)(N(R51)C(O)N(R5)2), or X4is absent; each R5is independently H, (Ci-Cs)alkyl, or (C3-Cs)cycloalkyl; each R50is independently H, halo, or (Ci-Cs)alkyl;R51is H or (Ci-Cs)alkyl; andL' is absent, methylene, or ethylene;R1is -C(O)NR10Rn;R10is H or (Ci-Cs)alkyl;R11is H or (Ci-Cs)alkyl;R2is H or halo;Attorney Docket No. 01270-0017-00PCTR3is phenyl, (C3-C6)cycloalkyl, or 6-membered heteroaryl or heterocyclyl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30; wherein: when R3is (C3-Ce)cycloalkyl, then L' is absent; each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci- C3)alkyl,-O-halo(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl; each R4is independently halo, -OH, or (Ci-Cs)alkyl; and m is 0, 1, 2, 3, or 4.

[0086] In another aspect, provided herein is a compound of structural formula (II):or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein:L is absent, O, S, or CH2; wherein: when L is absent, structure B is a 6- to 10-membered, N-linked spiroheterocyclyl, N- linked fused heterocyclyl, or N-linked bridged heterocyclyl, or 3- to 6-membered, N-linked monocyclic heterocyclyl; or when L is O, S, or CH2, structure B is a 6- to 10-membered spirocyclyl, fused cyclyl, or bridged cyclyl, or 3- to 6-membered monocyclyl;X1, X2, and X3are each independently CR60or N; each R60is independently H, halo, -(Ci-C3)alkyl, or -O(Ci-C3)alkyl;X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), orC(R50)(N(R51)C(O)N(R5)2), or X4is absent; each R5is independently H, (Ci-Cs)alkyl, or (C3-Cs)cycloalkyl; each R50is independently H, halo, or (Ci-Cs)alkyl;R51is H or (Ci-Cs)alkyl;L' is absent, methylene, or ethylene;Attorney Docket No. 01270-0017-00PCTR2is H or halo;R3is phenyl, (C3-C6)cycloalkyl, or 6-membered heteroaryl or heterocyclyl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30; wherein: when R3is (C3-Ce)cycloalkyl, then L' is absent; each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci- C3)alkyl,-O-halo(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl; each R4is independently halo, -OH, or (Ci-Cs)alkyl;R6is H or methyl;R7is H or methyl; and m is 0, 1, 2, 3, or 4.

[0087] In yet another aspect, provided herein is a compound of structural formula (III):or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein:L is absent, O, S, or CH2; wherein: when L is absent, structure C is a 6- to 10-membered, N-linked spiroheterocyclyl orN-linked bridged heterocyclyl, provided structure C is not orwhen L is O, S, or CH2, structure C is a 6- to 10-membered spirocyclyl, fused cyclyl, or bridged cyclyl, or 3- to 6-membered monocyclyl;X1, X2, and X3are each independently CR60or N; each R60is independently H, halo, -(Ci-C3)alkyl, or -O(Ci-C3)alkyl;Attorney Docket No. 01270-0017-00PCTX4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), or C(R50)(N(R51)C(O)N(R5)2), or X4is absent; each R5is independently H, (Ci-Cs)alkyl, or (C3-Cs)cycloalkyl; each R50is independently H, halo, or (Ci-Cs)alkyl;R51is H or (Ci-Cs)alkyl;R2is H or halo;R3is phenyl, (C3-Ce)cycloalkyl, or 6-membered heteroaryl or heterocyclyl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30; each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci- C3)alkyl,-O-halo(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl; each R4is independently halo, -OH, or (Ci-Cs)alkyl;R8is H or (Ci-Cs)alkyl; and m is 0, 1, 2, 3, or 4.

[0088] In some embodiments, R10and R11are each independently H or methyl. In some embodiments, R10and R11are each H. In some embodiments, R10and R11are each methyl. In some embodiments, R10is H, and R11is methyl.

[0089] In some embodiments, when L is absent, structure A is a 6- to 10-membered, N-linked spiroheterocyclyl or N-linked bridged heterocyclyl. In some embodiments, when L is absent, structure A is a 6- to 10-membered, N-linked spiroheterocyclyl. In some embodiments, when L is absent, structure A is a 6- to 10-membered, N-linked bridged heterocyclyl. In some embodiments, when L is absent, structure A is a 6- to 10-membered, N-linked fused heterocyclyl.

[0090] In some embodiments, when L is absent, structure A is:Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTome embodiments, when L is absent, structure A is:

[0091] In some embodiments, when L is O, S, or CH2, structure A is a 6- to 10-membered, C- linked spirocyclyl, C-linked fused cyclyl, or C-linked bridged cyclyl, or 3- to 6-membered, C- linked monocyclyl.

[0092] In some embodiments, when L is O, S, or CH2, structure A is a 6- to 10-membered, C- linked spirocyclyl or 3- to 6-membered, C-linked monocyclyl. In some embodiments, when L is O, S, or CH2, structure A is a 6- to 10-membered, C-linked spirocyclyl. In some embodiments, when L is O, S, or CH2, structure A is a 6- to 10-membered, C-linked spirocyclyl or 3- to 6- membered, C-linked monocyclic cycloalkyl.Attorney Docket No. 01270-0017-00PCT

[0094] In some embodiments, R6is H and R7is H. In some embodiments, R6is methyl and R7is H.In some embodiments, R6is methyl and R7is methyl.

[0095] In some embodiments, when L is absent, structure B is a 6- to 10-membered, N-linked spiroheterocyclyl or N-linked bridged heterocyclyl. In some embodiments, when L is absent, structure B is a 6- to 10-membered, N-linked spiroheterocyclyl. In some embodiments, when L is absent, structure B is a 6- to 10-membered, N-linked bridged heterocyclyl. In some embodiments, when L is absent, structure B is a 6- to 10-membered, N-linked fused heterocyclyl.

[0096] In some embodiments, when L is absent, structure B is:Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0097] In some embodiments, when L is O, S, or CH2, structure B is a 6- to 10-membered, C- linked spirocyclyl, C-linked fused cyclyl, C-linked bridged cyclyl, or 3- to 6-membered, C-Attorney Docket No. 01270-0017-00PCT linked monocyclyl. In some embodiments, when L is O, S, or CH2, structure B is a 6- to 10- membered, C-linked spirocyclyl or 3- to 6-membered, C-linked monocyclyl. n some embodiments, when L is O, S, or CH2, structure B is a 6- to 10-membered, C-linked spirocyclyl.

[0099] In some embodiments, R8is H or (C2-Cs)alkyl. In some embodiments, R8is H or ethyl. In some embodiments, R8is H.

[0100] In some embodiments, when L is absent, structure C is a 6- to 10-membered, N-linked spiroheterocyclyl, provided structure C is notembodiments, when L is absent, structure C is a 6- to 10-membered, N-linked bridged heterocyclyl.

[0101] In some embodiments, when L is absent, structure C is:Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0102] In some embodiments, when L is O, S, or CH2, structure C is a 6- to 10-membered, C- linked spirocyclyl, C-linked fused cyclyl, or C-linked bridged cyclyl, or 3- to 6-membered, C- linked monocyclyl. In some embodiments, when L is O, S, or CH2, structure C is a 6- to 10- membered, C-linked spirocyclyl or 3- to 6-membered, C-linked monocyclyl. In some embodiments, when L is O, S, or CH2, structure C is a 6- to 10-membered, C-linked spirocyclyl.

[0104] In some embodiments, L is absent. In alternative embodiments, L is O, S, or Clfc.In some embodiments, L is O or S. In some embodiments, L is O. In some embodiments, L is S. In some embodiments, L is CH2.

[0105] In some embodiments, X1is C(R60); X2is C(R60); and X3is N; or X1is N; X2is C(R60); and X3is C(R60); or X1is C(R60); X2is N; and X3is C(R60); or X1is N; X2is C(R60); and X3is N. In some embodiments, X1is N; X2is C(R60); and X3is C(R60). In some embodiments, X1is C(R60); X2is N; and X3is C(R60). In some embodiments, X1is C(R60); X2is C(R60); and X3is N. In some embodiments, X1is N; X2is C(R60); and X3is N. In some embodiments, X1is C(R60); X2is N; and X3is N. In some embodiments, X1is C(R60); X2is C(R60); and X3is C(R60).

[0106] In some embodiments, X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), or C(R50)(N(R51)C(O)N(R5)2). In some embodiments, X4is C(R50)(N(R5)2), N(R5), or C(R50)(C(R50)2N(R5)2). In some embodiments, X4is C(R50)(N(R5)2). In some embodiments, X4is C(H)(NH2), C(H)(N(H)CH3), C(H)(N(CH3)2), C(H)(N(H)CH(CH3)2), C(H)(N(H)cyclopropyl), C(H)(OH), C(H)2, N(H), N(CH3), N(CH(CH3)2), N(cyclopropyl), C(H)(CH2NH2), C(H)(N(H)C(O)OC(CH3)3) or C(H)(N(H)C(O)N(H)C(CH3)3).

[0107] In some embodiments, each R5is independently H, methyl, isopropyl, tert-butyl, or cyclopropyl.Attorney Docket No. 01270-0017-00PCT

[0108] In some embodiments, each R50is independently H, fluoro, or methyl.

[0109] In some embodiments, each R50is H.

[0110] In some embodiments, R51is H or methyl. In some embodiments, R51is H. In some embodiments, R51is methyl.[OHl] In some embodiments, R2is H.

[0112] In some embodiments, R3is phenyl or 6-membered heteroaryl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30. In some embodiments, R3is phenyl or pyridinyl, and is optionally substituted with one or two R30. In some embodiments, R3is phenyl, and is optionally substituted with one or two R30.

[0113] In some embodiments, each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci- Cs)alkyl, -O(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl. In some embodiments, each R30is independently fluoro, chloro, -CN, methyl, trifluoromethyl, methoxy, difluoromethoxy, or cyclopropyl. In some embodiments, each R30is independently fluoro, chloro, -CN, methyl, trifluoromethyl, methoxy, or cyclopropyl. In some embodiments, each R30is fluoro.Attorney Docket No. 01270-0017-00PCT

[0115] In some embodiments, each R4is independently fluoro, -OH, or methyl. In some embodiments, m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0.

[0116] In some embodiments, each R60is independently H or fluoro. In some embodiments, each R60is H.

[0117] In some embodiments, there are at least two (e.g., at least three, at least four, two, three, four, or five) intervening atoms between L and X4.

[0118] In some embodiments, L' is absent. In some embodiments, L' is methylene or ethylene.In some embodiments, L' is methylene.

[0119] In some embodiments,

[0120] In the descriptions herein, it is understood that every description, variation, embodiment, or aspect of a moiety may be combined with every description, variation, embodiment, or aspect of other moieties the same as if each and every combination of descriptions is specifically andAttorney Docket No. 01270-0017-00PCT individually listed. For example, every description, variation, embodiment, or aspect provided herein with respect to L of structural Formula (I) may be combined with every description, variation, embodiment, or aspect of the remaining variables (e.g., structure A, L', X1, X2, X3, X4, R1, R2, R3, R4, and m) the same as if each and every combination were specifically and individually listed. It is also understood that, except when indicated otherwise, all descriptions, variations, embodiments, or aspects of structural Formula (I), where applicable, apply equally to other formulae detailed herein, and are equally described, the same as if each and every description, variation, embodiment, or aspect were separately and individually listed for all formulae. For example, all descriptions, variations, embodiments, or aspects of structural Formula (I), where applicable, apply equally to any of the formulae as detailed herein, such as structural Formulas (II) and (III), and are equally described, the same as if each and every description, variation, embodiment, or aspect were separately and individually listed for all formulae.

[0121] Another aspect is a compound of Table 1, or a pharmaceutically acceptable salt or solvate thereof. In some embodiments, the compound is a compound of Table 1, or a pharmaceutically acceptable salt thereof. Although certain compounds described in the present disclosure, including in Table 1, are presented as specific stereoisomers and / or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms, such as isotopic forms, of any of the compounds of the present disclosure, including in Table 1, are herein described. Thus, in some embodiments, provided is a compound selected from the compounds in Table 1, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof.Table 1. CompoundsAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0122] In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is Compound No. 1, 2, 3, 4, 5, 11, 13, 17, 19, 23, 34, 68, 70, 77, or 87, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is Compound No. 1, 2, 3, 4, 5, 11, 13, 17, 19, 23, 34, 68, 70, 77, 87, 111, 112, 113, 120, 122, 124, 125, 126, 127, 128, 129, 132, 133, 134, 136, 137, 141, 142, 143, 149, 154, 156 or 157, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is Compound No: 1, 2, 3, 4, 5, 6, 10, 11, 12, 13, 16, 17, 18, 19, 20, 22, 23, 24, 25, 27, 33, 34, 36, 37, 39, 42, 45, 46, 54, 56, 59, 68, 70, 77, 87, 88, 92, 94, 95, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 132, 133, 134, 136, 137, 141, 142, 143, 144, 145, 146, 148, 149, 150, 154, 155, 156 or 157 or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is Compound No: 7, 8, 9, 15, 21, 28, 30, 35, 40, 41, 43, 47, 49, 53, 66, 80, 84, 85, 89, 93, 97, 99, 101, 104, 107, or 110, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof.

[0123] It is understood that in the present description, combinations of substituents and / or variables of the depicted formulae are permissible only if such contributions result in stable compounds.

[0124] Furthermore, all compounds of the structural formulas depicted herein which exist in free base or free acid form can be converted to their salts (e.g., their pharmaceutically acceptable salts) by treatment with the appropriate inorganic or organic base, or inorganic or organic acid by methods known to one skilled in the art. Salts (e.g., pharmaceutically acceptable salts) of the compounds of the structural formulas depicted herein can be converted to their free base or freeAttorney Docket No. 01270-0017-00PCT acid form by standard techniques. Some embodiments provide a compound of a structural formula depicted herein in free base form. Some embodiments provide a compound of a structural formula depicted herein in free acid form.Methods of Synthesis

[0125] Compounds of the structural formulas depicted herein, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, can be prepared by using organic chemistry synthesis methods known in the art. In general, starting components may be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, etc. or synthesized according to methods known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)) or prepared as described herein.General Reaction Scheme.

[0126] Compounds of the general structure shown in the General Reaction Scheme where X1and X2are N or C(H) and X2is N, C(H) or C(F) can be synthesized by the route shown therein.Attorney Docket No. 01270-0017-00PCTAn acid chloride of the appropriate aromatic acid can be prepared e.g., with SOCI2), and then condensed with ethyl (E)-3-(dimethylamino)acrylate. The condensation product can be reacted with 2,4-difluoroanaline and the resultant product treated with a base, such as potassium carbonate, to provide a fused bicyclic compound. An appropriate amine can be added to the fused bicyclic ring system by direct aromatic nucleophilic substitution of the halogen (R = Br or Cl) or by substitution using Buchwald conditions. The resulting esters can be converted to the corresponding carboxylic acid by hydrolysis, e.g., with LiOH. Alternatively, the resulting esters can be converted directly to the corresponding amide by heating with ammonia in methanol. Carboxylic acids can be converted to corresponding amides using HATU and ammonia. Final compounds can be produced by deprotection, e.g., of a Boc-protected amino group, e.g., with TFA.

[0127] It should be noted that various alternative strategies for preparation of the compounds disclosed herein are available to those of ordinary skill in the art. For example, other compounds of the structural formulas depicted herein can be prepared according to analogous methods using the appropriate starting material.

[0128] It will also be appreciated by those skilled in the art that in the processes for preparing the compounds described herein the functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups may include hydroxy, amino, and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (for example, Z-butyldimethylsilyl, Z-butyldiphenylsilyl or trimethyl silyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino and amidino include / -butoxycarbonyl (Boc), benzyloxycarbonyl, and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl, or arylalkyl esters. Protecting groups are optionally added or removed in accordance with standard techniques, which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As one of skill in the art would appreciate, the protecting group may also be a polymer resin such as a Wang resin, Rink resin or a 2-chlorotrityl-chloride resin.Pharmaceutical Compositions and Formulations

[0129] In a further aspect, provided herein are pharmaceutical compositions. The pharmaceutical composition comprises any one (or more) of the foregoing compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition isAttorney Docket No. 01270-0017-00PCT formulated for injection. In still more embodiments, the pharmaceutical compositions comprise any of the foregoing compounds (e.g., of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and an additional therapeutic agent. Non-limiting examples of such therapeutic agents are described herein below.

[0130] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, otic, nasal, and topical administration. In addition, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injections.

[0131] In certain embodiments, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered in a local rather than systemic manner, for example, via injection of the compound directly into an organ, often in a depot preparation or sustained release formulation. In specific embodiments, long acting formulations are administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Furthermore, in other embodiments, the drug is delivered in a targeted drug delivery system, for example, in a liposome coated with organ-specific antibody. In such embodiments, the liposomes are targeted to and taken up selectively by the organ. In yet other embodiments, the compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is provided in the form of a rapid release formulation, in the form of an extended release formulation, or in the form of an intermediate release formulation. In yet other embodiments, the compound described herein is administered topically.

[0132] The compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are effective over a wide dosage range. For example, in the treatment of adult humans, dosages from 0.01 to 1000 mg, from 0.01 to 300 mg, or from 0.5 to 200 mg per day. The exact dosage will depend upon the route of administration, the form in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, and the preference and experience of the attending physician.

[0133] In some embodiments, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered in a single dose. Typically, such administration will be by injection, e.g., intravenous injection, in order to introduce the agent quickly. However, other routes are used asAttorney Docket No. 01270-0017-00PCT appropriate. A single dose of a compound of the disclosure may also be used for treatment of an acute condition (e.g., peripheral inflammation).

[0134] In some embodiments, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered in multiple doses. In some embodiments, dosing is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, dosing is about once a month, once every two weeks, once a week, or once every other day. In another embodiment a compound of Formula (I), Formula (II), or Formula (III) or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and another therapeutic agent are administered together about once per day to about 6 times per day. In another embodiment the administration of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and a therapeutic agent continues for less than about 7 days. In yet another embodiment the administration continues for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous dosing is achieved and maintained as long as necessary.

[0135] Administration of the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, may continue as long as necessary. In some embodiments, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered chronically on an ongoing basis, e.g., for the treatment of chronic effects (e.g., peripheral inflammation).

[0136] In some embodiments, the compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is administered in dosages. It is known in the art that due to intersubject variability in compound pharmacokinetics, individualization of dosing regimen is necessary for optimal therapy. Dosing for a compound may be found by routine experimentation in light of the instant disclosure.

[0137] In some embodiments, the compounds Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated into pharmaceutical compositions. In specific embodiments, pharmaceuticalAttorney Docket No. 01270-0017-00PCT compositions are formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Any pharmaceutically acceptable techniques, carriers, and excipients are used as suitable to formulate the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkinsl999).

[0138] Provided herein are pharmaceutical compositions comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s). Also provided herein are methods for administering a pharmaceutical composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s).

[0139] In certain embodiments, the compounds are administered as pharmaceutical compositions in which compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are mixed with other therapeutic agents, as in combination therapy. Encompassed herein are all combinations of active ingredients set forth in the methods section below and throughout this disclosure. In specific embodiments, the pharmaceutical compositions include one or more compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof.

[0140] A pharmaceutical composition, as used herein, refers to a mixture of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments, practicing the methods of treatment or use provided herein, therapeutically effective amounts of compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, orAttorney Docket No. 01270-0017-00PCT stereoisomer thereof, provided herein are administered in a pharmaceutical composition to a mammal having a disease, disorder or medical condition to be treated. In specific embodiments, the mammal is a human. In certain embodiments, therapeutically effective amounts vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used and other factors. The compounds described herein are used singly or in combination with one or more therapeutic agents as components of mixtures.

[0141] In one embodiment, one or more compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated in an aqueous solution. In specific embodiments, the aqueous solution is selected from, by way of example only, a physiologically compatible buffer, such as Hank’s solution, Ringer’s solution, or physiological saline buffer. In other embodiments, one or more compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated for transmucosal administration. In specific embodiments, transmucosal formulations include penetrants that are appropriate to the barrier to be permeated (e.g., the blood-brain barrier). In still other embodiments wherein the compounds described herein are formulated for other parenteral injections, appropriate formulations include aqueous or non-aqueous solutions. In specific embodiments, such solutions include physiologically compatible buffers and / or excipients.

[0142] In another embodiment, compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated for oral administration. Compounds are formulated by combining the active compounds with, e.g., pharmaceutically acceptable carriers or excipients. In various embodiments, the compounds Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated in oral dosage forms that include, by way of example only, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, and the like.

[0143] In certain embodiments, pharmaceutical preparations for oral use are obtained by mixing one or more solid excipient with one or more of the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as: for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose,Attorney Docket No. 01270-0017-00PCT sodium carboxymethylcellulose; or others such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. In specific embodiments, disintegrating agents are optionally added. Disintegrating agents include, by way of example only, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0144] In one embodiment, dosage forms, such as dragee cores and tablets, are provided with one or more suitable coating. In specific embodiments, concentrated sugar solutions are used for coating the dosage form. The sugar solutions, optionally contain additional components, such as by way of example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs and / or pigments are also optionally added to the coatings for identification purposes. Additionally, the dyestuffs and / or pigments are optionally utilized to characterize different combinations of active compound doses.

[0145] In certain embodiments, therapeutically effective amounts of at least one of the compounds Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. In specific embodiments, push-fit capsules contain the active ingredients in admixture with one or more filler. Fillers include, by way of example only, lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In other embodiments, soft capsules contain one or more active compound that is dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

[0146] In other embodiments, therapeutically effective amounts of at least one of the compounds of Formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, described herein are formulated for buccal or sublingual administration. Formulations suitable for buccal or sublingual administration include, by way of example only, tablets, lozenges, or gels. In still other embodiments, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated for parental injection, including formulations suitable for bolus injection or continuous infusion. In specific embodiments, formulations for injection are presented in unit dosage form (e.g., in ampoules) or in multi-dose containers. Preservatives are, optionally, added to the injection formulations. In still other embodiments, the pharmaceutical compositions are formulated in a form suitable for parenteral injection as sterile suspensions, solutions orAttorney Docket No. 01270-0017-00PCT emulsions in oily or aqueous vehicles. Parenteral injection formulations optionally contain formulatory agents such as suspending, stabilizing and / or dispersing agents. In specific embodiments, pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. In additional embodiments, a suspension of an active compound or compounds (e.g., compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. In certain specific embodiments, aqueous injection suspensions contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension contains suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, in other embodiments, the active ingredient is in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0147] In still other embodiments, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are administered topically. The compounds are formulated into a variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams or ointments. Such pharmaceutical compositions optionally contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.

[0148] In yet other embodiments, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated for transdermal administration. In specific embodiments, transdermal formulations employ transdermal delivery devices and transdermal delivery patches and can be lipophilic emulsions or buffered, aqueous solutions, dissolved and / or dispersed in a polymer or an adhesive. In various embodiments, such patches are constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents. In additional embodiments, the transdermal delivery of the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, is accomplished by means of iontophoretic patches and the like. In certain embodiments, transdermal patches provide controlled delivery of the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In specific embodiments, the rate of absorption is slowed by using rate-controlling membranes or byAttorney Docket No. 01270-0017-00PCT trapping the compound within a polymer matrix or gel. In alternative embodiments, absorption enhancers are used to increase absorption. Absorption enhancers or carriers include absorbable pharmaceutically acceptable solvents that assist passage through the skin. For example, in one embodiment, transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound to the skin of the host at a controlled and predetermined rate over a prolonged period of time and means to secure the device to the skin.

[0149] In other embodiments, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated for administration by inhalation. Various forms suitable for administration by inhalation include, but are not limited to, aerosols, mists or powders. Pharmaceutical compositions of any of compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas). In specific embodiments, the dosage unit of a pressurized aerosol is determined by providing a valve to deliver a metered amount. In certain embodiments, capsules and cartridges of, such as, by way of example only, gelatin for use in an inhaler or insufflator is formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0150] In still other embodiments, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are formulated in rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides, as well as synthetic polymers such as polyvinylpyrrolidone, PEG, and the like. In suppository forms of the compositions, a low- melting wax such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with cocoa butter is first melted.

[0151] In certain embodiments, pharmaceutical compositions are formulated in any conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Any pharmaceutically acceptable techniques, carriers, and excipients are optionally used as suitable. Pharmaceutical compositions comprising a compound of FormulaAttorney Docket No. 01270-0017-00PCT(I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are manufactured in a conventional manner, such as, by way of example only, by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or compression processes.

[0152] Pharmaceutical compositions include at least one pharmaceutically acceptable carrier, diluent or excipient and at least one compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, described herein as an active ingredient. The active ingredient is in free-acid or free-base form, or in a pharmaceutically acceptable salt form. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), as well as active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein. Additionally, the compounds Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, encompass unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. The solvated forms of the compounds presented herein are also considered to be disclosed herein. In addition, the pharmaceutical compositions optionally include other medicinal or pharmaceutical agents, carriers, adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure, buffers, and / or other therapeutically valuable substances.

[0153] Methods for the preparation of compositions comprising the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, include formulating the compounds with one or more inert, pharmaceutically acceptable excipients or carriers to form a solid, semi-solid or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which a compound is dissolved, emulsions comprising a compound, or a solution containing liposomes, micelles, or nanoparticles comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, or stereoisomer thereof. Semi-solid compositions include, but are not limited to, gels, suspensions and creams. The form of the pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for solution or suspension in a liquid prior to use, or as emulsions. These compositions also optionally contain minor amounts of nontoxic, auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and so forth.Attorney Docket No. 01270-0017-00PCT

[0154] In some embodiments, pharmaceutical composition comprising at least one compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, illustratively takes the form of a liquid where the agents are present in solution, in suspension or both. Typically, when the composition is administered as a solution or suspension a first portion of the agent is present in solution and a second portion of the agent is present in particulate form, in suspension in a liquid matrix. In some embodiments, a liquid composition includes a gel formulation. In other embodiments, the liquid composition is aqueous.

[0155] In certain embodiments, useful aqueous suspensions contain one or more polymers as suspending agents. Useful polymers include water-soluble polymers such as cellulosic polymers, e.g., hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Certain pharmaceutical compositions described herein comprise a mucoadhesive polymer, selected for example from carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate and dextran.

[0156] Useful pharmaceutical compositions also, optionally, include solubilizing agents to aid in the solubility of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. The term "solubilizing agent" generally includes agents that result in formation of a micellar solution or a true solution of the agent. Certain acceptable nonionic surfactants, for example polysorbate 80, are useful as solubilizing agents, as can ophthalmically acceptable glycols, polyglycols, e.g., polyethylene glycol 400, and glycol ethers.

[0157] Furthermore, useful pharmaceutical compositions optionally include one or more pH adjusting agents or buffering agents, including acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; and buffers such as citrate / dextrose, sodium bicarbonate and ammonium chloride. Such acids, bases and buffers are included in an amount required to maintain pH of the composition in an acceptable range.

[0158] Additionally, useful compositions also, optionally, include one or more salts in an amount required to bring osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions; suitable salts includeAttorney Docket No. 01270-0017-00PCT sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0159] Other useful pharmaceutical compositions optionally include one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0160] Still other useful compositions include one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10, or octoxynol 40.

[0161] Still other useful compositions include one or more antioxidants to enhance chemical stability where required. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0162] In certain embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multiple-dose reclosable containers are used, in which case it is typical to include a preservative in the composition.

[0163] In alternative embodiments, other delivery systems for hydrophobic pharmaceutical compounds are employed. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also employed. In additional embodiments, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials are useful herein. In some embodiments, sustained-release capsules release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization are employed.

[0164] In certain embodiments, the formulations described herein comprise one or more antioxidants, metal chelating agents, thiol containing compounds and / or other general stabilizing agents. Examples of such stabilizing agents, include, but are not limited to: (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% w / v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / v ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v. polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrins, (1) pentosanAttorney Docket No. 01270-0017-00PCT polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc; or (n) combinations thereof.Kits / Articles of Manufacture

[0165] For use in the therapeutic applications described herein, kits and articles of manufacture are also provided. In some embodiments, such kits comprise a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers are formed from a variety of materials such as glass or plastic.

[0166] The articles of manufacture provided herein contain packaging materials. Packaging materials for use in packaging pharmaceutical products include those found in, e.g., U.S. Pat. Nos. 5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment. For example, the container(s) includes one or more compounds described herein, optionally in a composition or in combination with another agent as disclosed herein. The container(s) optionally have a sterile access port (for example the container is an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). Such kits optionally comprise a compound with an identifying description or label or instructions relating to its use in the methods described herein.

[0167] For example, a kit typically includes one or more additional containers, each with one or more of various materials (such as reagents, optionally in concentrated form, and / or devices) desirable from a commercial and user standpoint for use of a compound disclosed herein (e.g., a compound of structural Formula (I), (II), or (III)), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. Non-limiting examples of such materials include, but not limited to, buffers, diluents, filters, needles, syringes, carrier, package, container, vial, and / or tube labels listing contents and / or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included. A label is optionally on or associated with the container. For example, a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In addition, a label is used to indicate that the contents are to be used for a specific therapeutic application. In addition, the label indicates directions for use ofAttorney Docket No. 01270-0017-00PCT the contents, such as in the methods described herein. In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack for example contains metal or plastic foil, such as a blister pack. Or, the pack or dispenser device is accompanied by instructions for administration. Or, the pack or dispenser is accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the US Food and Drug Administration for prescription drugs, or the approved product insert. In some embodiments, compositions containing a compound disclosed herein (e.g., a compound of structural Formula (I), (II), or (III)), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, formulated in a compatible pharmaceutical carrier are prepared, placed in an appropriate container, and labeled for treatment of an indicated disease or condition.Methods of Use / Treatments

[0168] Embodiments of the present disclosure provide a method for modulating (e.g., inhibiting) pannexin-1 (Pannex-1) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof). In some embodiments, the Pannexin-1 is human Pannexin-1 (e.g., Access No. NP 056183.2). In some embodiments, a compound described herein blocks passage of large molecules through the Pannexin-1 channel. In some embodiments, a compound described herein reduces release of IL-lbeta from immune cells. In some embodiments, a compound described herein reduces Pannexin-1 -mediated calcium release. In some embodiments, a compound described herein reduces Pannexin-1 -mediated calcium release and IL-lbeta release. Modulation (e.g., inhibition of pannexin-1, calcium release, and IL- lbeta release) can be assessed and demonstrated by a wide variety of ways known in the art. Kits and commercially available assays can be utilized for determining whether and to what degree pannexin-1 been modulated (e.g., inhibited) and / or calcium and / or IL-lbeta has been released.

[0169] In some embodiments, provided herein are compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, for use in modulating (e.g., inhibiting) pannexin-1 in a subject in need thereof. In someAttorney Docket No. 01270-0017-00PCT embodiments, provided herein are compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for modulating (e.g., inhibiting) pannexin-1 in a subject in need thereof.

[0170] Applicant has discovered that the compounds of Formula (I), Formula (II), or Formula (III) show promising activity related to certain diseases of interest. Accordingly, in one aspect, provided herein is a method for treating an inflammatory condition in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof.

[0171] By way of non-limiting examples, inflammatory conditions include sepsis, arthritis, acute kidney injury, and the like. In some embodiments, the inflammatory condition is a systemic inflammatory condition and / or peripheral inflammatory condition. In some embodiments, the inflammatory condition is sepsis. In some embodiments, the inflammatory condition is arthritis. In some embodiments, the inflammatory condition is acute kidney injury.

[0172] In one aspect, provided herein is a method for treating epilepsy in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some embodiments, the epilepsy is treatmentresistant epilepsy, generalized seizures, partial seizures, pediatric epilepsy (e.g., pediatric epilepsy including Dravet syndrome), or temporal lobe epilepsy. In some embodiments, the epilepsy is pediatric epilepsy. In some embodiments, the method is for treating epileptic seizure. In some embodiments, the seizure is a generalized seizure or partial seizure. In some embodiments, the seizure is associated with a pediatric epileptic condition. In some embodiments, the pediatric epilepsy or pediatric epileptic condition is Dravet syndrome or Lennox-Gastaut syndrome. In some embodiments, the epilepsy is temporal lobe epilepsy. In some embodiments, the epilepsy is treatment-resistant.

[0173] In one aspect, provided herein is a method for treating seizure or seizures in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some embodiments, the seizure or seizures are associated with epilepsy. In some embodiments, the seizure or seizures are generalized seizures or partial seizures. In some embodiments, the seizure or seizures are pediatric epileptic seizures. In some embodiments, the seizure or seizures are associated with a pediatric epilepticAttorney Docket No. 01270-0017-00PCT condition. In some embodiments, the pediatric epileptic condition is Dravet syndrome or Lennox-Gastaut syndrome. In some embodiments, the seizure or seizures are treatment-resistant.

[0174] In one aspect, provided herein is a method for treating pain in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some embodiments, the pain is neuropathic pain or visceral pain. In some embodiments, the pain is neuropathic pain. In some embodiments, the pain is visceral pain.

[0175] In one aspect, provided herein is a method of treating a disease, disorder, or condition mediated by pannexin-1 activity, comprising administering to a subject having the disease, disorder or condition, a compound or salt thereof of Formula (I), Formula (II) or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In some aspects, the disease, disorder, or condition is epilepsy. In some embodiments, the disease, disorder, or condition is a seizure or seizures. In some embodiments, the disease, disorder, or condition is a seizure or seizures associated with epilepsy. In some embodiments, the seizure or seizures are generalized seizures or partial seizures. In some embodiments, the seizure or seizures are associated with a pediatric epileptic condition. In some embodiments, the pediatric epileptic condition is Dravet syndrome or Lennox-Gastaut syndrome. In some embodiments, the epilepsy is temporal lobe epilepsy.

[0176] In some aspects, the disease, disorder, or condition is pain. In some embodiments, the pain is neuropathic pain or visceral pain. In some embodiments, the pain is neuropathic pain. In some embodiments, the pain is visceral pain.

[0177] In some embodiments, the disclosure provides methods of modulating (e.g., inhibiting) pannexin-1 activity in a cell, comprising contacting the cell with an amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, sufficient to modulate (e.g., inhibit) pannexin-1. In some embodiments, the disclosure provides methods of modulating (e.g., inhibiting) pannexin-1 activity in a tissue, comprising contacting the tissue with an amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, sufficient to modulate (e.g., inhibit) the activity of pannexin-1 in the tissue. In some embodiments, the disclosure provides methods of modulating (e.g., inhibiting) Pannexin-1 activity in an organism, comprising contacting the organism with an amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, sufficient to modulate (e.g.,Attorney Docket No. 01270-0017-00PCT inhibit) pannexin-1 activity in the organism. In some embodiments, the disclosure provides methods of modulating (e.g., inhibiting) pannexin-1 activity in an animal, comprising contacting the animal with an amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, sufficient to modulate (e.g., inhibit) pannexin-1 in the animal. In some embodiments, the disclosure provides methods of modulating (e.g., inhibiting) pannexin-1 activity in a mammal, comprising contacting the mammal with an amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, sufficient to modulate (e.g., inhibit) pannexin-1 in the mammal. In some embodiments, the disclosure provides methods of modulating (e.g., inhibiting) pannexin-1 activity in a human, comprising contacting the human with an amount of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, sufficient to modulate (e.g., inhibit) pannexin-1 activity in the human. In other embodiments, the present disclosure provides methods of treating a disease mediated by pannexin-1 activity in a subject in need of such treatment.

[0178] Other embodiments provide methods for combination therapies in which a therapeutic agent known to modulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes are used in combination with a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof. In one aspect, such therapy includes but is not limited to the combination of one or more compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, with therapeutic agents, therapeutic antibodies, and other forms of treatment, to provide a synergistic or additive therapeutic effect.

[0179] Hence, in some embodiments the one or more compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, will be co-administered with other therapeutic agents. When used in combination therapy, the compounds of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are administered with the second therapeutic agent simultaneously or separately. This administration in combination can include simultaneous administration in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and any of the therapeutic agents described above canAttorney Docket No. 01270-0017-00PCT be formulated together in the same dosage form and administered simultaneously. Alternatively, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and any of the therapeutic agents described above can be simultaneously administered, wherein both are present in separate formulations. In another alternative, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, can be administered just followed by and any of the therapeutic agents described above, or vice versa. In some embodiments of the separate administration protocol, a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and any of the therapeutic agents described above are administered a few minutes apart, or a few hours apart, or a few days apart.

[0180] The examples and preparations provided below further illustrate and exemplify the compounds of Formula (I), Formula (II) or Formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, and methods of preparing such compounds. It is to be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples and preparations. In the following examples, and throughout the specification and claims, molecules with a single stereocenter, unless otherwise noted, exist as a racemic mixture. Those molecules with two or more stereocenters, unless otherwise noted, exist as a racemic mixture of diastereomers. Single enantiomers / diastereomers may be obtained by methods known to those skilled in the art.EXAMPLES

[0181] Methods for preparation of compounds of the structural formulas depicted herein, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, are provided herein or can be derived by one of ordinary skill in the art.

[0182] The examples and preparations provided below further illustrate and exemplify the compounds of the present disclosure and methods for testing such compounds. It is to be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples.

[0183] The chemical reactions in the Examples described can be readily adapted to prepare a number of other compounds disclosed herein, and alternative methods for preparing the compounds of this disclosure are deemed to be within the scope of this disclosure. For example, the synthesis of non-exemplified compounds according to the present disclosure can be performed by modifications apparent to those skilled in the art, for example by appropriatelyAttorney Docket No. 01270-0017-00PCT protecting interfering groups, by utilizing other suitable reagents known in the art other than those described, or by making routine modification of reaction conditions, reagents, and starting materials. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present disclosure.

[0184] Unless indicated otherwise in the following Examples, the compounds are isolated as a racemic mixture.

[0185] The following abbreviations may be relevant for the application.AbbreviationsAcOH: acetic acidACN: acetonitrileBOC: tert-butyloxy carbonylCyPF-tBu: (A)-l-[(SP)-2-(Dicyclohexylphosphino)ferrocenyl]ethyldi-tert-butylphosphineDCM: dichloromethaneDIPEA: N,N-diisopropylethylamineDMF: dimethylformamideDMSO: dimethyl sulfoxideEq or eq: equivalentsEtOAc: ethyl acetateH or h: hour(s)HATU: (l-[bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphateLC / MS: liquid chromatography -mass spectrometryMe: methylMeOH: methanolN: NormalPrep HPLC: preparative high performance liquid chromatographyRT or rt: room temperatureTEA: TriethanolamineTFA: trifluoroacetic acidTHF: tetrahydrofuranTLC: thin layer chromatographyVol. or vol.: volume(s)Attorney Docket No. 01270-0017-00PCTSynthetic Examples

[0186] Example SI: General Procedure 1 - 1,8-naphthyridine scaffoldsScheme 1“la”, “lb”, and “1c” refer to Synthetic Procedures la, lb, and 1c, respectively.

[0187] Synthetic Procedure la: A solution of ethyl 7-chloro-l-(2,4-difluorophenyl)-6-fluoro-4- oxo- l,4-dihydro-l,8-naphthyridine-3 -carboxylate (1 eq), TEA (3 eq) and the appropriate Boc protected diamine or Boc protected aminoalcohol (1.1 eq) in ACN (10 Vol) were stirred at 80°C for 3h. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT and then diluted with water and EtOAc. The organic layer was separated, washed with brine solution, and dried over Na2SO4. The mixture was filtered and then the solvent was removed in vacuo to obtain the desired product.

[0188] Synthetic Procedure lb: A solution of ethyl 7-chloro-l-(2,4-difluorophenyl)-6-fluoro-4- oxo- l,4-dihydro-l,8-naphthyridine-3 -carboxylate (1 eq), cesium carbonate (3 eq) and the appropriate amine or alcohol (1.1 eq) in DMF (10 Vol) were heated for 5h at 80°C. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT and then diluted with water and EtOAc. The organic layer was separated, washed with brine solution, and dried over Na2SO4. The mixture was filtered and then the solvent was removed in vacuo to obtain the desired products.

[0189] Synthetic Procedure 1c: To a stirred solution of the Boc protected products produced in Synthetic Procedure la or lb (1 eq) in DCM (10 Vol) was added trifluoroacetic acid (5 eq) and the solution was stirred for 2h at 25°C. The reaction was monitored by TLC. After completion, the reaction mixture was concentrated under vacuum to obtain a gummy solid, which was washed and triturated using diethyl ether to obtain the crude material. The crude material was purified by prep-HPLC to afford the desired products.Attorney Docket No. 01270-0017-00PCT

[0190] Synthetic Procedure Id: Some intermediates produced by Synthetic Procedure la or lb were monomethylated prior to conversion to final compounds. To a stirred solution of compounds produced by Synthetic Procedure la or lb (1 eq) in THF (10 vol) was added sodium hydride (60% in mineral oil) (1.5 eq) at 0°C and the mixture stirred for 30 mins at 25°C. Methyl iodide (5.0 eq) was added, and the mixture was stirred for 12 at 25°C. After TLC indicated complete reaction, the reaction mixture was diluted with water and EtOAc. The organic layer was separated, washed with brine solution and dried with Na2SO4. The solvent was concentrated in vacuo. The crude was purified by Comb-flash to afford the desired products.Scheme le

[0191] Synthetic Procedure le: Some intermediates produced by Synthetic Procedure 1c were dimethylated prior to conversion to final compounds. To the stirred solution of compound (1 eq) in MeOH (10 vol) were added formaldehyde (40% in water) (5 eq) and acetic acid (0.1 eq) at room temperature and then the mixture was stirred for 30 mins at 25°C. Then the mixture was added to sodium cyanoborohydride (2.0 eq) and the mixture was stirred for 12 at 25°C. The reaction was monitored by TLC. After completion, the reaction mixture was diluted with water and EtOAc. The organic layer was separated, washed with brine solution and dried with Na2SO4. The solvent was concentrated in vacuo. The crude material was purified by Comb-flash to afford the desired product.Attorney Docket No. 01270-0017-00PCTScheme If

[0192] Synthetic Procedure If:

[0193] Step 1 : Some mono-alkyl intermediates were produced by an alternate synthetic route. A solution of ethyl 7-chloro-l-(2,4-difluorophenyl)-6-fluoro-4-oxo-l,4-dihydro-l,8-naphthyridine- 3-carboxylate (1 eq), TEA (3 eq) and 2-azaspiro [3.3]heptan-6-one (1.1 eq) in ACN (10 Vol) was stirred at 80°C for 5h. After TLC indicated reaction completion, the reaction mixture was cooled to RT diluted with water and EtOAc. The organic layer was separated, washed with brine solution, dried with Na2SO4, and then solvent was concentrated in vacuo to obtain the desired product.

[0194] Step 2 : To the stirred solution of the product from Step 1 (1 eq) in MeOH (10 vol) were added isopropylamine or cyclopropylamine (2 eq) and acetic acid (0.1 eq) at room temperature and then the mixture was stirred for 30 mins at 25°C. Then sodium cyanoborohydride (2.0 eq) was added to it at 0°C and the mixture stirred for 12 at 25°C. The reaction was monitored by TLC. After completion, the reaction mixture was diluted with water and EtOAc. The organic layer was separated, washed with brine solution and dried with Na2SO4; the solvent was concentrated in vacuo. The crude was purified by Comb-flash to afford the desired product.Attorney Docket No. 01270-0017-00PCTGeneral Procedure 2. Ester Hydrolysiscyclopropyl-NH or OH cyclopropyl-NHScheme 2

[0195] Synthetic Procedure 2a: To a stirred solution of a compound produced by SyntheticProcedure la, lb, or Id (1 eq) in THF:MeOH:H2O (3: 1 : 1) (10 Vol) was added LiOH.H2O (5 eq)Attorney Docket No. 01270-0017-00PCT and the mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion, the reaction mixture was concentrated, acidified with 2N HC1 solution (pH=6.5) and filtered. The filtered solid was washed with water and dried under vacuum to afford the desired product.

[0196] Synthetic Procedure 2b: A solution of a compound produced by Synthetic Procedure la or lb (1 eq) was dissolved in AcOH:conc. HC1 (3: 1) (5 Vol) and heated for 3h at 100°C. The reaction was monitored by TLC. After completion, the reaction mixture was concentrated under vacuum and the solid obtained was washed with water, then filtered and dried to afford the desired product.

[0197] Synthetic Procedure 2c: Compounds with a Boc protected amine were deprotected using the following procedure: To a stirred solution of a compound produced by Synthetic Procedure 2a or 2b (1 eq) in DCM (10 Vol) was added trifluoroacetic acid (5 eq) and the solution was stirred for approximately 2h at 25°C. The reaction was monitored by TLC. After completion, the reaction mixture was concentrated under vacuum to obtain a gummy solid. The solid was triturated using diethyl ether to obtain the crude material. The crude material was purified by prep-HPLC to afford the desired product.

[0198] Synthetic Procedure 2d: Some acid compounds were converted to sodium salts by the following procedure. To a stirred solution of a compound produced by Synthetic Procedure 2a, 2b or 2c (1 eq) in water was added 2M NaOH solution (1 Vol) and the mixture was stirred for 2h at 25°C. The reaction was monitored by TLC. After completion, the reaction mixture was concentrated under vacuum to obtain a solid, which was washed and triturated using diethyl ether. The crude obtained was lyophilized to afford the desired product.Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTGeneral Procedure 3. Amidationcyclopropyl-NH or OH cyclopropyl-NHScheme 3

[0199] Synthetic Procedure 3a: To a stirred solution of a compound produced by Synthetic Procedure 2a or 2b (1 eq) in DMF (10 vol) were added HATU (1.5 eq), DIPEA (5 eq) and ammonia (1 eq) at room temperature and then the mixture was stirred for 12 h at 50°C. The reaction was monitored by TLC. After completion, the reaction mixture was diluted with water and EtOAc. The organic layer was separated, washed with brine solution, dried over Na2SO4, filtered and then the filtrate was concentrated in vacuo. The crude material was purified by Comb-flash to afford the desired product.Attorney Docket No. 01270-0017-00PCT

[0200] Synthetic Procedure 3b: A solution of a compound produced by Synthetic Procedure 2a or 2b (1 eq) was dissolved in 7N ammonia in MeOH (10 Vol) and heated for 12 h at 75°C. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to RT, then the solvent was concentrated under vacuum and the solid obtained was washed using water, filtered and dried to obtain the desired product.

[0201] Synthetic Procedure 3c: Compounds with a Boc protected amine were deprotected using the following procedure: To a stirred solution of a compound produced by Synthetic Procedure 3a or 3b (1 eq) in DCM (10 Vol) was added trifluoroacetic acid (5 eq) and the mixture was stirred for 2h at 25°C. The reaction was monitored by TLC. After completion, the reaction mixture was concentrated under vacuum to obtain a gummy solid, which was washed and triturated using diethyl ether to obtain the crude material. The crude material was purified by prep-HPLC to afford the desired product.Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTGeneral Procedure 4

[0202] Compounds having a quinoline scaffold were synthesized using procedures described from the corresponding commercially available amino acid by protecting the amino group with a Boc protecting group, forming the amide using Synthetic procedure 3a and removing the Boc protecting group using Synthetic procedure 3c.Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0203] Example S2: General Procedure 5 - 1,6-Naphthyridine ScaffoldsScheme 5

[0204] Synthetic procedure 5a: Ethyl (Z)-2-(4,6-dichloronicotinoyl)-3- (dimethylamino)acrylate). To a stirred solution of 4,6-dichloronicotinic acid (1 eq) in toluene (10 Vol) was added SOCh (3 eq) and the mixture was refluxed for 2 hr. After 2h, the reaction mixture was concentrated in vacuo under an inert atmosphere to obtain a pale brown solid. In a 250 mL round bottom flask was added ethyl (E)-3-(dimethylamino)acrylate (1 eq), TEA (5 eq) and THF (5 Vol). The obtained pale brown solid was dissolved in THF (5 Vol) and was added to the reaction mass dropwise. Then the reaction was heated to reflux for 8h. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT. The organic layer was separated, washed with 10% bicarbonate solution and brine solution and then dried with Na2SO4. The solvent was concentrated in vacuo. The crude was purified by Combi- flash to afford ethyl (Z)-2-(4,6-dichloronicotinoyl)-3-(dimethylamino)acrylate as a brown oil. LC-MS: 316.90 [M+H]+.

[0205] Synthetic procedure 5b: Ethyl 7-chloro-l-(2,4-difluorophenyl)-4-oxo-l,4-dihydro-l,6- naphthyridine-3-carboxylate. A solution of ethyl (Z)-2-(4,6-dichloronicotinoyl)-3- (dimethylamino)acrylate (1 eq), DIPEA (1 eq) and 2,4-difluoroaniline (2 eq) in THF (10 Vol) was heated for 12h at 60°C. The reaction was monitored by TLC. After completion, the reaction mass was cooled to RT, and then extracted using water and EtOAc. The organic layer was separated, washed with a brine solution and dried with Na2SO4. The solvent was concentrated inAttorney Docket No. 01270-0017-00PCT vacuo. The crude was purified by Combi-flash to afford ethyl 7-chloro-l-(2,4-difluorophenyl)-4- oxo-1, 4-dihydro-l,6-naphthyridine-3-carboxylate as a pale yellow solid. LC-MS: 365.20[M+H]+.

[0206] Synthetic Procedure 5c: Ethyl 7-chloro-l-(2,4-difluorophenyl)-4-oxo-l,4-dihydro-l,6- naphthyridine-3 -carboxylate (1 eq) was dissolved in 1,4-dioxane (10 Vol). The appropriate amine or alcohol (1 eq) and cesium carbonate (3 eq) were added and the vessel was purged with nitrogen for 15 min. X-phos-Pd-G4 (0.1 eq) was added and then the mixture was heated to 100°C and stirred for 12 hr. The reaction was monitored by TLC. After the reaction was complete, the mixture was poured into crushed ice, stirred for 15 min and extracted with EtOAc. The organic layer was separated, washed with brine solution, dried over Na2SO4 and filtered. The solution was concentrated in vacuo. The crude was purified by Comb-flash to afford the desired product.

[0207] Synthetic Procedure 5d: Ethyl 7-chloro-l-(2,4-difluorophenyl)-4-oxo-l,4-dihydro-l,6- naphthyridine-3 -carboxylate (1 eq) was dissolved in 1,4-dioxane (10 Vol). The appropriate amine or alcohol (1 eq) and cesium carbonate (3 eq) were added and the vessel was purged with nitrogen for 15 min. Allylpalladium(II) chloride dimer (0.05 eq) and CyPF-tBu (0.1 eq) were added and then the mixture was heated to 100°C and stirred for 12 hr. The reaction was monitored by TLC. After the reaction was complete, the mixture was poured into crushed ice, stirred for 15 min and extracted with EtOAc. The organic layer was separated, washed with brine solution, dried over Na2SO4 and filtered. The solution was concentrated in vacuo. The crude was purified by Comb-flash to afford the desired product.Scheme 5e

[0208] Synthetic Procedure 5e: Synthetic procedure 5e is similar to Synthetic Procedure 2a.

[0209] Synthetic Procedure 5f: Synthetic Procedure 5f is similar to synthetic procedure 3a.

[0210] Synthetic Procedure 5g: Synthetic Procedure 5g is similar to synthetic procedure 3b.

[0211] Synthetic Procedure 5h: Synthetic Procedure 5h is similar to Synthetic Procedure 3cAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0212] Example S3: General Procedure 6 - 1,5-Naphthyridine ScaffoldsScheme 6

[0213] Synthetic Procedure 6a: Ethyl (Z)-2-(5-bromo-3-chloropicolinoyl)-3- (dimethylamino)acrylate. To a stirred solution of 5-bromo-3-chloropicolinic acid (1 eq) in toluene (10 Vol) was added SOCh (2 eq) and the solution was refluxed for 2 hr. After 2h, the reaction mixture was concentrated in vacuo under an inert atmosphere to obtain a pale brown solid. Ethyl (E)-3 -(dimethylamino) acrylate (1 eq) was dissolved in TEA (3 eq) and THF (5 Vol) was added. The obtained pale brown solid was dissolved in THF (5 Vol) and it was added to the reaction mass dropwise. Then the reaction was refluxed for approximately 8 h. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT and washed with 10% bicarbonate solution, brine solution and dried over Na2SO4. The mixture was filtered and the solvent was concentrated in vacuo. The crude material was purified by Combflash to afford ethyl (Z)-2-(5-bromo-3-chloropicolinoyl)-3-(dimethylamino) acrylate as a brown oil.

[0214] Synthetic Procedure 6b: Ethyl (Z)-2-(5-bromo-3-chloropicolinoyl)-3-((2,4- difluorophenyl)amino)acrylate. A solution of ethyl (Z)-2-(5-bromo-3-chloropicolinoyl)-3- (dimethylamino)acrylate (1 eq), DIPEA (1 eq) and 2,4-difluoroaniline (1 eq) in THF (10 Vol) was heated for approximately 12h at 50°C. The reaction was monitored by TLC. After completion, the reaction mass was allowed to cool to RT, and then extracted using water andAttorney Docket No. 01270-0017-00PCTEtOAc. The organic layer was separated, washed with brine solution and dried over Na2SO4. The mixture was filtered, and the solvent was concentrated in vacuo. The crude material was purified by Comb-flash to afford ethyl (Z)-2-(5-bromo-3-chloropicolinoyl)-3-((2,4- difluorophenyl)amino)acrylate as a pale yellow solid. LC-MS: 446.8 [M+H]+.

[0215] Synthetic Procedure 6c: Ethyl 7-bromo-l-(2,4-difluorophenyl)-4-oxo-l,4-dihydro-l,5- naphthyridine-3-carboxylate. To a stirred solution of ethyl (Z)-2-(5-bromo-3-chloropicolinoyl)- 3-((2,4-difluorophenyl) amino)acrylate (1 eq) in DMF (5 vol) was added K2CO3 (3 eq). Then the reaction was heated for approximately 12 hr at 70°C. The reaction was monitored by TLC. After completion, the reaction mass was allowed to cool to RT. The reaction mixture was added to ice cold water, and the solid which precipitated which was washed with water, filtered and dried under vacuo to afford ethyl 7-bromo-l-(2,4-difluorophenyl)-4-oxo-l,4-dihydro-l,5- naphthyridine-3 -carboxylate as a yellow solid.

[0216] Synthetic Procedure 6d: To a stirred solution of the compound from Synthetic Procedure 6c (1 eq) in 1,4-dioxane (10 Vol), the appropriate diamine or amino alcohol (1 eq) and cesium carbonate (3 eq) were added and the solution was purged under nitrogen for 15 min.Then X-phos-Pd-G4 (0.1 eq) was added and the mixture was stirred at 100°C for approximately 12 hr. The reaction was monitored by TLC. After completion, the reaction mixture was poured into crushed ice, stirred for 15 min and extracted with EtOAc. The organic layer was separated, washed with brine solution and dried over Na2SO4 and filtered. The solvent was removed in vacuo. The crude material was purified by Comb-flash to afford the desired product.

[0217] Synthetic Procedure 6e: To a stirred solution of the compound from Synthetic Procedure 6d (1 eq) in toluene (10 Vol), the appropriate diamine or amino alcohol and cesium carbonate (3 eq) were added and the solution was purged under nitrogen for 15 min. Then allylpalladium(II) chloride dimer (0.05 eq) and CyPF-tBu (0.1 eq) were added and the mixture was stirred at 100°C for 12 hr. The reaction was monitored by TLC. After completion, the reaction mixture was poured into crushed ice, stirred for 15 min and extracted with EtOAc. The organic layer was separated, washed with brine solution and dried with Na2SO4. The solvent was concentrated in vacuo. The crude was purified by Comb-flash to afford the desired product.Attorney Docket No. 01270-0017-00PCTScheme 6f

[0218] Synthetic Procedure 6f: Synthetic procedure 6f is similar to Synthetic Procedure 2a.

[0219] Synthetic Procedure 6g: Synthetic procedure 6g is similar to Synthetic Procedure 3a.

[0220] Synthetic Procedure 6h: Synthetic procedure 6h is similar to Synthetic Procedure 3b.

[0221] Synthetic procedure 6i: Synthetic procedure 6i is similar to Synthetic Procedure 3c.Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0222] Example S4: General Procedure 7 - Pyrazine ScaffoldsScheme 7

[0223] Synthetic Procedure 7a: Ethyl (Z)-2-(3,5-dichloropyrazine-2-carbonyl)-3-(dimethylamino)acrylate. To a stirred solution of 3,5-dichloropyrazine-2-carboxylic acid (1 eq) in DCM (10 Vol) was added oxalyl chloride (3 eq) and the mixture was refluxed for 2 hr. AfterAttorney Docket No. 01270-0017-00PCT2h, the reaction mixture was concentrated in vacuo under an inert atmosphere to obtain a pale brown solid. In a 250 mL round bottom flask was added ethyl (E)-3-(dimethylamino) acrylate (1.1 eq), TEA (3 eq) and toluene (5 Vol). The obtained pale brown solid was dissolved in toluene (5 Vol) and it was added to the reaction mixture dropwise. Then the reaction was heated to reflux for approximately 8h. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT and washed with 10% bicarbonate solution and then brine solution, dried over solid Na2SO4 and filtered. The solvent was removed in vacuo. The crude material was purified by Comb-flash to afford ethyl (Z)-2-(3,5-dichloropyrazine-2-carbonyl)-3- (dimethylamino) acrylate. LC-MS: 317.90 [M+H]+.

[0224] Synthetic procedure 7b: Ethyl 3-chloro-5-(2,4-difluorophenyl)-8-oxo-5,8- dihydropyrido[2,3-b]pyrazine-7-carboxylate. A solution of ethyl (Z)-2-(3,5-dichloropyrazine-2- carbonyl)-3-(dimethylamino) acrylate (1 eq), acetic acid (2 eq) and 2,4-difluoroaniline (1,5 eq) in THF (10 Vol) was stirred for approximately 12h at room temperature. The reaction was monitored by TLC. After completion, the reaction mass was neutralized using bicarbonate solution and extracted using EtOAc. The organic layer was separated, washed with brine solution, dried over Na2SO4 and filtered. The solvent was removed in vacuo to obtain the crude material. The crude material (1 eq) was taken up in 1,4-Dioxane (10 Vol) and K2CO3 (1.5 eq) was added to the stirred solution. Then the reaction was heated to 60°C for approximately 8 hrs. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT. The organic layer was separated, washed with brine solution, dried over Na2SO4 and filtered. The solvent was concentrated in vacuo. The crude material was purified by Comb-flash to afford ethyl 3-chloro-5-(2,4-difluorophenyl)-8-oxo-5,8-dihydropyrido[2,3-b]pyrazine-7- carboxylate as a pale brown solid. LC-MS: 365.95 [M+H]+.

[0225] Synthetic procedure 7c: A solution of ethyl 3-chloro-5-(2,4-difluorophenyl)-8-oxo-5,8- dihydropyrido[2,3-b]pyrazine-7-carboxylate (1 eq), TEA (3 eq) and tert-butyl (2- azaspiro[3.3]heptan-6-yl)carbamate (1.1 eq) in ACN (10 Vol) was stirred at 80°C for 3h. The reaction was monitored by TLC. After completion, the reaction mixture was allowed to cool to RT and then diluted with water and EtOAc. The organic layer was separated, washed with brine solution, dried over Na2SO4 and filtered. The solvent was concentrated in vacuo to obtain the desired product.Attorney Docket No. 01270-0017-00PCTScheme 7d

[0226] Synthetic procedure 7d: Synthetic procedure 7d is similar to Synthetic Procedure 2a.

[0227] Synthetic procedure 7e: Synthetic procedure 7e is similar to Synthetic Procedure 3b.

[0228] Synthetic procedure 7f: Synthetic procedure 7f is similar to Synthetic Procedure 3c.Attorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCTBiological Examples

[0229] Example Bl: Dye Uptake AssayAttorney Docket No. 01270-0017-00PCT

[0230] The activity of selected compounds was determined using a YO-PRO® dye uptake assay. Pannexin-1 assembles into hexameric hemichannels (Boassa et al., J Biol Chem. 2007,282:31733-31743; Bao et al., FEBS Lett. 2004; 572:65-68) that are permeable to relatively large organic molecules (Pelegrin P, Surprenant A. EMBO J. 2006;25:5071-5082; Locovei S et al., FEBS Lett. 2007; 581 :483-488) like ATP, the YO-PRO®-1 dye and ethidium bromide (Bao L, FEBS Lett. 2004; 572:65-68; Huang YJ, Proc Natl Acad Sci USA. 2007, 104:6436-6441), and ions like Ca2+(Vanden Abeele F et al., J Cell Biol. 2006,174:535-546).

[0231] Briefly, a human embryonic kidney (HEK) cell line stably expressing human Pannexin-1 when induced with doxycycline (DOX) (HEK-Pannexin-1 cells) was used. The expression of Pannexin-1 is under the control of the TET-On inducible expression system wherein upon induction with DOX, the Pannexin-1 channel is expressed in the cells in an open conformation, allowing uptake of large molecular dyes like YO-PRO®.

[0232] Oxazole Yellow, also known as YO-PRO® dye, is a green-fluorescent, cell-impermeant, high-affinity carbocyanine monomeric nucleic acid stain. It only fluoresces upon entry into the cell. In the absence of Pannexin-1 antagonists, YO-PRO® dye enters the cell through the open Pannexin-1 channel, and the resulting fluorescence signal can be quantified on a plate reader. Pannexin-1 antagonists can inhibit the uptake of YO-PRO® dye in a concentration-dependent manner; hence, the potency of compounds can be identified in this assay.

[0233] In this study, the HEK-Pannexin-1 cells were plated in a 96-well format at 50,000 cells / well and treated with DOX for 72-96 hours (about 4 days) to induce and activate Pannexin- 1 in the cells. On the day of the study, select compounds were added across a range of concentrations, and YO-PRO® dye was simultaneously added to all treatment wells.Fluorescence readouts were captured over a 210-minute period and were used to determine compound (ICsos). The results are shown in Table 10, wherein “****” indicates an ICso <10 nM; “***” indicates an ICso of >10 nM to 100 nM; “**” indicates an IC50 of > 100 nM to 1 micromolar.; indicates an IC50 of > 1 micromolar.Table 10. Dye Uptake AssayAttorney Docket No. 01270-0017-00PCTAttorney Docket No. 01270-0017-00PCT

[0234] Example B2: IL-ip Release Assay

[0235] PANXI is known to upregulate inflammatory signaling. Certain PANXI compounds disclosed herein were tested for their ability to reduce inflammasome activation in immortalized immune cell culture. An inflammasome activation assay was conducted using a protocol adapted from Pelegrin and Surprenant (EMBO J., 2006, 25(21): 5071-5082). A THP-1 macrophage cell line (ATCC) was utilized for IL-ip release studies on selected compounds. Briefly, 50,000 / well THP-1 cells were seeded in a 96-well format. Differentiation of non-adherent monocytes into adherent macrophages was achieved by treating with 100 ng / mL PMA (phorbol 12-myristate 13- acetate) for 72 h. Subsequently, adherent macrophages were washed once with warmed media (RPMI plus 10% FBS) before exposure to 2 ng / ml LPS (lipopolysaccharides) for 3 hours in the first phase of NLRP3 inflammasome activation. During this priming phase, the cells produce increased levels of pro-ILip, the immature form of IL-ip. Following LPS treatment, cells were treated with various concentrations of selected compounds for 30 min.Attorney Docket No. 01270-0017-00PCT

[0236] For IL-ip release studies, the second phase of NLRP3 inflammasome activation was induced by incubating cells with 3 mM ATP for 30 min. The addition of extracellular ATP activates an ATP receptor, P2X7R, which together with Pannexin-1, promotes a caspase- mediated proteolytic cleavage of pro-IL-lBeta to mature IL-ip which is finally released into the extracellular environment. The extracellular supernatant is subsequently analyzed for mature human IL-ip levels using a homogenous time resolved fluorescence (HTRF) assay. Maximum percent (Max %) inhibition of IL-ip released upon treatment with select compounds are listed in Table 10. Max % inhibition was calculated as a percentage decrease relative to IL-ip levels secreted by the LPS + ATP treatment group.Table 11: IL-ip Release Assay

[0237] Example B3: LDH release assay

[0238] Programmed cell death triggered by infection and inflammation, pyroptosis, is a key driver of inflammatory damage in multiple cell types. Pyroptosis is primarily driven by a series of large-pore membrane complexes, the gasdermins. Gasdermins can be activated by the activity of caspase and inflammasome proteins, both of which are downstream of PANXI activation, which represent a key regulator of programmed cell death (Zhou et al., Sci Signal. 2022; 15). In experimental model hematopoietic cell lines, including THP-1 monocytes, pyroptosis signaling can be stimulated by the chemotherapeutic agent etoposide, and this study model was used to interrogate the ability of PANXI inhibitors to interrupt the process of programmed cell death.Attorney Docket No. 01270-0017-00PCTEtoposide-stimulated PANXI activity drives activation of the inflammasome and gasdermins, which leads to cell lysis and release of intracellular protein contents. The magnitude of this pyroptotic cell lysis can be measured by the presence of intracellular lactate dehydrogenase (LDH) in the cell culture media.

[0239] Briefly, 50,0000 / well THP-1 cells were seeded in a 96-well format and cultured for 24 hours. Etoposide (50 uM) and test PANXI inhibitors were applied to cells simultaneously and treatments continued for 24 hours. At the end of the treatment period, culture media was removed for LDH quantification via CyQuant LDH kit (Invitrogen). Data were normalized between untreated cultures (0%) and etoposide-treated cultures (100%). Reduction of LDH release in PANXI inhibitor-treated wells indicated a reduction in etoposide-dependent pyroptosis. Statistical analysis consisted of one-way ANOVA with Dunnett’s multiple comparisons post-test with comparison against the etoposide treatment.

[0240] The results of the LDH release assay are reported in Table 12.Table 12: Etoposide-stimulated LDH release assayAttorney Docket No. 01270-0017-00PCT“+” indicates a significant reduction in LDH release at the indicated concentration compared to etoposide-alone treatment by one-way ANOVA with Dunnett’s post test, p<0.05. indicates no significant change in LDH release at the indicated concentration.

[0241] Example B4: Multi-Electrode Array Studies

[0242] PANXI activity is thought to contribute to seizure-like neuron firing activity in epilepsy. To evaluate the anti-seizure potential of test compounds, seizure-like firing in hippocampal neurons was evaluated by stimulation of neuron cultures with pyridine-4-amine (4-AP) and recording of neuron firing activity in multi-electrode array (MEA) culture plates. MEA systems use a grid of tightly spaced microscopic electrodes embedded in the bottom of each well in a multi -well MEA plate. Neurons, which are electrically active, can be cultured over the electrodes creating a cohesive network. The functional behavior or electrical activity of this network can be recorded as extracellular field potentials.

[0243] To study the activity of selected compounds on epileptiform activity, hippocampus tissue was harvested from embryonic day 18 chrNMRJ mouse embryos (Charles River Laboratories). Pregnant mice were sacrificed by cervical dislocation according to the German Animal Protection Act §4 and Animal Welfare Laboratory Animal Ordinance §2 Section 2, Appendix 2.Attorney Docket No. 01270-0017-00PCTThe tissue was dissociated by enzymatic digestion and mechanical trituration. The resulting suspension was cell-counted, vitality-controlled, set to 7.5 million cells / ml, and plated in a 20 pl drop of DMEM containing laminin (10 pg / ml), 10% fetal bovine serum, and 10% horse serum on 48-well MEA plates. Cultures in the MEA apparatus were incubated at 37 °C in a 10% CO2 atmosphere for 27-29 days in vitro prior to study. Culture media were replenished two times per week with DMEM containing 10% horse serum. The developing co-cultures were treated with the mitosis inhibitor 5-fluoro-2'-deoxyuridine and uridine between days 3 and 5 after seeding to prevent further glia proliferation.

[0244] For the recording, a multichannel recording system, MAESTRO™ from Axion Biosystems Inc. (USA), was used. For extracellular recording, 48-well MEA plates were placed into the recording station and maintained at 37 °C. Recordings were made in the original medium. The pH was maintained at 7.4 with a continuous stream of filtered, humidified airflow with appropriate CO2 levels. The action potentials, or "spikes", were recorded.

[0245] After recording the native baseline activity, 4-aminopyridin (4-AP) was added at 100 pM to induce epileptiform activity. This activity was followed for 60 minutes. After 60 minutes, test compounds were added. The neuronal activity was recorded for 1 hour, and the last 30 minutes of activity were analyzed. Twenty-four hours after compound application, MEAs were recorded again for 60 minutes.

[0246] Analysis: Epileptiform activity in neuron cultures was characterized by spike rate, with greater spikes per second indicating greater seizure-like firing activity. Within the 4-AP treated group, the effects of the test compounds were compared to vehicle (DMSO). The data of the 4- AP episodes of all experiments were pooled to characterize its effects on the cells.

[0247] For the compound effects, data were normalized to the 4-AP activity. Compound effects were compared to the vehicle effects and the 4-AP control “water / DMSO”. Spikes were quantitatively described via direct spike train analysis using the program NPWaveX™ (NeuroProof GmbH, Rostock, Germany).

[0248] Statistical significance was determined by paired Student’s t-test versus 4-AP effect. The data, which are reported in Table 13, show that the compounds tested significantly reduced seizure-like spiking behavior after 24 hours of incubation.Attorney Docket No. 01270-0017-00PCTTable 13: Results of Exemplary Compounds in MEA study characterizing reduction of 4- AP-stimulated epileptiform activityResults of hippocampal neuron epileptiform spiking behavior characterization. Table indicates statistical difference between indicated compound incubated at the indicated concentration for the indicated time interval vs stimulatory 4-AP treatment. Statistical analysis: Student's t-test versus 4-AP control. NS: not significant, +: p < 0.05, ++: p < 0.01.

[0249] Although the present invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are expressly incorporated herein in their entirety by reference.

Claims

Attorney Docket No. 01270-0017-00PCTCLAIMS1. A compound of the following structural formula:or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein:L is absent, O, S, or CH2; wherein: when L is absent, structure A is a 6- to 10-membered, N-linked spiroheterocyclyl, N- linked fused heterocyclyl, or N-linked bridged heterocyclyl; or when L is O, S, or CH2, structure A is a 6- to 10-membered spirocyclyl, fused cyclyl, or bridged cyclyl, or 3- to 6-membered, monocyclyl;X1, X2, and X3are each independently CR60or N; each R60is independently H, halo, -(Ci-C3)alkyl, or -O(Ci-C3)alkyl;X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), or C(R50)(N(R51)C(O)N(R5)2), or X4is absent; each R5is independently H, (Ci-Cs)alkyl, or (C3-Cs)cycloalkyl; each R50is independently H, halo, or (Ci-Cs)alkyl;R51is H or (Ci-Cs)alkyl; andL' is absent, methylene, or ethylene;R1is -C(O)NR10Rn;R10is H or (Ci-Cs)alkyl;R11is H or (Ci-Cs)alkyl;R2is H or halo;R3is phenyl, (C3-Ce)cycloalkyl, or 6-membered heteroaryl or heterocyclyl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30; wherein: when R3is (C3-Ce)cycloalkyl, then L' is absent;Attorney Docket No. 01270-0017-00PCT each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci-C3)alkyl,-O-halo(Ci-C3)alkyl,-C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl; each R4is independently halo, -OH, or (Ci-Cs)alkyl; and m is 0, 1, 2, 3, or 4.

2. The compound of claim 1, wherein R10and R11are each independently H or methyl.3 The compound of claim 1 or 2, wherein when L is absent, structure A is a 6- to 10- membered, N-linked spiroheterocyclyl or N-linked bridged heterocyclyl.4 The compound of claim 1 or 2, wherein when L is absent, structure A is:Attorney Docket No. 01270-0017-00PCTThe compound of any one of claims 1-2, wherein when L is O, S, or CH2, structure A is a 6- to 10-membered, C-linked spirocyclyl, C-linked fused cyclyl, or C-linked bridged cyclyl, or 3- to 6-membered, C-linked monocyclyl.The compound of claim 5, wherein when L is O, S, or CH2, structure A is a 6- to 10- membered, C-linked spirocyclyl or 3- to 6-membered, C-linked monocyclyl.The compound of claim 6, wherein structure A is:A compound of the following structural formula:or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein:L is absent, O, S, or CH2; wherein: when L is absent, structure B is a 6- to 10-membered, N-linked spiroheterocyclyl, N- linked fused heterocyclyl, or N-linked bridged heterocyclyl, or 3- to 6-membered, N-linked monocyclic heterocyclyl; or when L is O, S, or CH2, structure B is a 6- to 10-membered spirocyclyl, fused cyclyl, or bridged cyclyl, or 3- to 6-membered monocyclyl;X1, X2, and X3are each independently CR60or N;Attorney Docket No. 01270-0017-00PCT each R60is independently H, halo, -(Ci-C3)alkyl, or -O(Ci-C3)alkyl;X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), or C(R50)(N(R51)C(O)N(R5)2), or X4is absent; each R5is independently H, (Ci-Cs)alkyl, or (C3-Cs)cycloalkyl; each R50is independently H, halo, or (Ci-Cs)alkyl;R51is H or (Ci-Cs)alkyl;L' is absent, methylene, or ethylene;R2is H or halo;R3is phenyl, (C3-Ce)cycloalkyl, or 6-membered heteroaryl or heterocyclyl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30; wherein: when R3is (C3-Ce)cycloalkyl, then L' is absent; each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci- C3)alkyl,-O-halo(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl; each R4is independently halo, -OH, or (Ci-Cs)alkyl;R6is H or methyl;R7is H or methyl; and m is 0, 1, 2, 3, or 4. The compound of claim 8, wherein R6is H and R7is H. The compound of claim 8, wherein R6is methyl and R7is H. The compound of claim 8, wherein R6is methyl and R7is methyl. The compound of any one of claims 8-11, wherein when L is absent, structure B is a 6- to10-membered, N-linked spiroheterocyclyl or N-linked bridged heterocyclyl. The compound of any one of claims 8-11, wherein when L is absent, structure B is:Attorney Docket No. 01270-0017-00PCT14. The compound of any one of claims 8-11, wherein when L is O, S, or CH2, structure B is a 6- to 10-membered, C-linked spirocyclyl, C-linked fused cyclyl, or C-linked bridged cyclyl, or 3- to 6-membered, C-linked monocyclyl.

15. The compound of claim 14, wherein when L is O, S, or CH2, structure B is a 6- to 10- membered, C-linked spirocyclyl or 3- to 6-membered, C-linked monocyclyl.16.Attorney Docket No. 01270-0017-00PCT17. A compound of the following structural formula:or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, wherein:L is absent, O, S, or CH2; wherein: when L is absent, structure C is a 6- to 10-membered, N-linked spiroheterocyclyl orN-linked bridged heterocyclyl, provided structure C is not orwhen L is O, S, or CH2, structure C is a 6- to 10-membered spirocyclyl, fused cyclyl, or bridged cyclyl, or 3- to 6-membered monocyclyl;X1, X2, and X3are each independently CR60or N; each R60is independently H, halo, -(Ci-C3)alkyl, or -O(Ci-C3)alkyl;X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), or C(R50)(N(R51)C(O)N(R5)2), or X4is absent; each R5is independently H, (Ci-Cs)alkyl, or (C3-Cs)cycloalkyl; each R50is independently H, halo, or (Ci-Cs)alkyl;R51is H or (Ci-Cs)alkyl;R2is H or halo;R3is phenyl, (C3-Ce)cycloalkyl, or 6-membered heteroaryl or heterocyclyl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30; each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci- C3)alkyl, -O-halo(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3-C5)cycloalkyl; each R4is independently halo, -OH, or (Ci-Cs)alkyl;Attorney Docket No. 01270-0017-00PCTR8is H or (Ci-Cs)alkyl; and m is 0, 1, 2, 3, or 4.

18. The compound of claim 17, wherein R8is H or (C2-Cs)alkyl.

19. The compound of claim 18, wherein R8is H.

20. The compound of any one of claims 17-19, wherein when L is absent, structure C is:

21. The compound of any one of claims 17-19, wherein when L is O, S, or CH2, structure C is a 6- to 10-membered, C-linked spirocyclyl, C-linked fused cyclyl, or C-linked bridged cyclyl, or 3- to 6-membered, C-linked monocyclyl.

22. The compound of claim 21, wherein when L is O, S, or CH2, structure C is a 6- to 10- membered, C-linked spirocyclyl or 3- to 6-membered, C-linked monocyclyl.Attorney Docket No. 01270-0017-00PCT24. The compound of any one of claims 1-23, wherein L is absent.

25. The compound of any one of claims 1-23, wherein L is O, S, or CH2.

26. The compound of claim 25, wherein L is O or S.

27. The compound of claim 26, wherein L is O.

28. The compound of any one of claims 1-27, wherein X1is C(R60); X2is C(R60); and X3is N; or X1is N; X2is C(R60); and X3is C(R60); or X1is C(R60); X2is N; and X3is C(R60); or X1is N; X2is C(R60); and X3is N.

29. The compound of any one of claims 1-28, wherein X1is N; X2is C(R60); and X3is C(R60).

30. The compound of any one of claims 1-28, wherein X1is C(R60); X2is N; and X3is C(R60).

31. The compound of any one of claims 1-28, wherein X1is C(R60); X2is C(R60); and X3is N.

32. The compound of any one of claims 1-28, wherein X1is N; X2is C(R60); and X3is N.

33. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, and 24-32, wherein X4is C(R50)(N(R5)2), N(R5), C(R50)(C(R50)2N(R5)2), C(R50)(N(R51)C(O)OR5), or C(R50)(N(R51)C(O)N(R5)2).

34. The compound of claim 33, wherein X4is C(R50)(N(R5)2), N(R5), or C(R50)(C(R50)2N(R5)2).

35. The compound of claim 34, wherein X4is C(R50)(N(R5)2).

36. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, and 24-35, wherein each R5is independently H, methyl, isopropyl, tert-butyl, or cyclopropyl.

37. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, and 24-36, wherein each R50is independently H, fluoro, or methyl.

38. The compound of claim 37, wherein each R50is H.

39. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, 24-33, and 36- 38, wherein R51is H or methyl.

40. The compound of claim 39, wherein R51is H.Attorney Docket No. 01270-0017-00PCT41. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, and 24-32, wherein X4is C(H)(NH2), C(H)(N(H)CH3), C(H)(N(CH3)2), C(H)(N(H)CH(CH3)2), C(H)(N(H)cyclopropyl), C(H)(OH), C(H)2, N(H), N(CH3), N(CH(CH3)2), N(cyclopropyl), C(H)(CH2NH2), C(H)(N(H)C(O)OC(CH3)3) or C(H)(N(H)C(O)N(H)C(CH3)3).

42. The compound of any one of claims 1-41, wherein R2is H.

43. The compound of any one of claims 1-42, wherein R3is phenyl or 6-membered heteroaryl containing one or two nitrogen atoms, and is optionally substituted with one, two, or three R30.

44. The compound of claim 43, wherein R3is phenyl or pyridinyl, and is optionally substituted with one or two R30.

45. The compound of any one of claims 1-44, wherein each R30is independently halo, -CN, -(Ci-C3)alkyl, -halo(Ci-C3)alkyl, -O(Ci-C3)alkyl, -C(O)O(Ci-C3)alkyl, or (C3- C5)cycloalkyl.

46. The compound of any one of claims 1-44, wherein each R30is independently fluoro, chloro,-CN, methyl, trifluoromethyl, methoxy, difluoromethoxy, or cyclopropyl.

47. The compound of claim 45 or 46, wherein each R30is fluoro.

48. The compound of any one of claims 1-42, whereinAttorney Docket No. 01270-0017-00PCT49. The compound of claim 46, wherein50. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, and 24-49, wherein each R4is independently fluoro, -OH, or methyl.

51. The compound of any one of claims 1-49, wherein m is 0, 1, or 2.

52. The compound of claim 51, wherein m is 0.

53. The compound of any one of claims 1-52, wherein each R60is independently H or fluoro.

54. The compound of any one of claims 1-3, 5, 6, 8-12, 14, 15, 17-19, 21, 22, and 24-53, wherein there are at least two intervening atoms between L and X4.

55. The compound of any one of claims 1-16 and 24-54, wherein L' is absent.

56. The compound of any one of claims 1-16 and 24-54, wherein L' is methylene or ethylene.

57. A compound of Table 1, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof.

58. The compound of claim 57, selected from: a. Compound No.: 1, 2, 3, 4, 5, 11, 13, 17, 19, 23, 34, 68, 70, 77, or 87; b. Compound No.: 1, 2, 3, 4, 5, 11, 13, 17, 19, 23, 34, 68, 70, 77, 87, 111, 112, 113, 120, 122, 124, 125, 126, 127, 128, 129, 132, 133, 134, 136, 137, 141, 142, 143,149, 154, 156 or 157; c. Compound No.: 1, 2, 3, 4, 5, 6, 10, 11, 12, 13, 16, 17, 18, 19, 20, 22, 23, 24, 25, 27, 33, 34, 36, 37, 39, 42, 45, 46, 54, 56, 59, 68, 70, 77, 87, 88, 92, 94, or 95; d. Compound No.: 1, 2, 3, 4, 5, 6, 10, 11, 12, 13, 16, 17, 18, 19, 20, 22, 23, 24, 25, 27, 33, 34, 36, 37, 39, 42, 45, 46, 54, 56, 59, 68, 70, 77, 87, 88, 92, 94, 95, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 132, 133, 134, 136, 137, 141, 142, 143, 144, 145, 146, 148, 149,150, 154, 155, 156 or 157; or e. Compound No.: 7, 8, 9, 15, 21, 28, 30, 35, 40, 41, 43, 47, 49, 53, 66, 80, 84, 85, 89, 93, 97, 99, 101, 104, 107, or 110, or a pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof.

59. A pharmaceutical composition comprising a compound, or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, of any one of claims 1-58, and a pharmaceutically acceptable carrier.Attorney Docket No. 01270-0017-00PCT60. A method of inhibiting pannexin-1 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound, or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, of any one of claims 1-58, or pharmaceutical composition of claim 59.

61. The method of claim 60, wherein the compound or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, or the pharmaceutical composition reduces release of IL-lbeta by immune cells.

62. The method of claim 60 or 61, wherein the compound or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, or the pharmaceutical composition reduces Pannexin-1 -mediated calcium release.

63. A method of treating an inflammatory condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound, or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, of any one of claims 1-58, or pharmaceutical composition of claim 59.

64. The method of claim 63, wherein the inflammatory condition is a systemic and / or peripheral inflammatory condition.

65. The method of claim 63 or 64, wherein the inflammatory condition is sepsis, arthritis, or acute kidney injury.

66. The method of claim 65, wherein the inflammatory condition is sepsis.

67. A method of treating epilepsy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound, or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, of any one of claims 1-58, or pharmaceutical composition of claim 59.

68. The method of claim 67, wherein the epilepsy is pediatric epilepsy.

69. The method of claim 67 or 68, wherein the epilepsy is Dravet syndrome or Lennox-Gastaut syndrome.

70. The method of claim 67 or 68, wherein the epilepsy is temporal lobe epilepsy.

71. A method of treating seizure or seizures in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound, or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, of any one of claims 1-58, or pharmaceutical composition of claim 59.

72. The method of claim 71, wherein the seizure or seizures is an epileptic seizure or seizures.Attorney Docket No. 01270-0017-00PCT73. The method of claim 71 or 72, wherein the seizure or seizures is a pediatric epileptic seizure or seizures.

74. The method of any one of claims 71-73, wherein the seizure or seizures are generalized seizure or seizures or partial seizure or seizures.

75. The method of any one of claims 67-74, wherein the epilepsy or seizure or seizures is treatment-resi stant.

76. A method of treating pain in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound, or pharmaceutically acceptable salt, solvate, tautomer, isotope, or stereoisomer thereof, of any one of claims 1-58, or pharmaceutical composition of claim 59.

77. The method of claim 75, wherein the pain is neuropathic pain or visceral pain.

78. The method of claim 76, wherein the pain is neuropathic pain.

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