Substituted alkylphenols as HCN1 antagonists

Abstract

Provided herein are compounds (e.g., compounds of Formula (I) and Formula (II) that modulate HCN channels, intermembrane proteins that function as non-selective voltage-gated cation channels in the plasma membranes of heart and brain cells. Also provided are pharmaceutical compositions and kits comprising the compounds, as well as methods of treating HCN-related disorders (e.g., pain) with the compounds in a subject by administering the compounds and / or compositions described herein.

Description

[0001] Related Applications

[0002] This application claims priority under 35 USC §119(e) to U.S. Provisional Applications USSN 62 / 690,778, filed on June 27, 2018, and USSN 62 / 803,109, filed on February 8, 2019, each of which is incorporated herein by reference in its entirety. Background Art

[0003] Chronic pain is pain that persists for more than three months. Such pain significantly impairs an individual's quality of life, is prevalent, and has significant economic costs.

[0004] Neuropathic pain is a form of chronic pain caused by lesions or damage to the somatic sensory nervous system. Neuropathic pain is a pathological feature of a variety of conditions, including postherpetic neuralgia, trigeminal neuralgia, sensory (painful) radiculopathy, painful diabetic neuropathy, peripheral nerve damage, stroke, multiple sclerosis, and cancer treatment with antineoplastic drugs. About 7% to 10% of the world's population suffers from chronic pain with neuropathic characteristics. The main symptoms of neuropathic pain are spontaneous pain, abnormal reactions to non-painful (abnormal pain) or painful (hyperalgesia) stimuli, dysesthesia (unpleasant / strange sensations-"tingling" "numbing") and enhanced thermal sensitivity; many pharmacological approaches to relieve neuropathic pain have been proposed, but their efficacy is limited, resulting in a large number of unmet needs in providing meaningful pain relief.

[0005] Recent studies have shown that neuropathic pain may be better understood at a mechanical level based on sensory analysis rather than disease etiology. The hyperexcitability of primary afferent neurons is considered to be an early cause of the development and maintenance of peripheral neuropathic pain. Many ion channel families contribute to this hyperexcitability, including the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel family. In animal models of neuropathic pain, non-selective HCN blockade with ZD7288 or clinically available drug ivabradine produces significant anti-hyperalgesic effects. The expression of HCN isoform HCN1 is upregulated in a variety of neuropathic pain animal models, including those in which the initial injury is mechanical damage, chemotherapy administration or diabetes. Antihypertensive activity is retained in HCN1 selective molecules, indicating that HCN1 selective blockade has therapeutic potential. Summary of the invention

[0006] The present disclosure arises from the recognition that novel compounds, compositions, uses and methods for treating pain are provided by targeting and modulating (e.g., inhibiting) peripheral sensory neuron channels (e.g., HCN1). For example, in a neuropathic pain model, inhibition of HCN1 is anti-hyperalgesic. In particular, 2,6-diisopropylphenol and its non-anesthetic homologue 2,6-di-tert-butylphenol inhibit HCN1 channels by stabilizing the closed state of HCN1, thereby producing a therapeutic effect. Thus, the present disclosure provides a means for locating a plasma membrane permeable pharmacophore tethered to the outer surface of the plasma membrane by generating a molecule with a polar anchoring moiety that confines the pharmacophore to the periphery of the cell, thereby promoting contact with therapeutically important peripheral sensory neuron targets (e.g., HCN1).

[0007] The present disclosure provides novel compounds that modulate HCN channels, and in particular, selective modulators (eg, inhibitors) of HCN1, thereby providing therapeutic agents for the treatment and / or management of pain (eg, chronic pain).

[0008] In one aspect, a compound of formula (I) is provided:

[0009]

[0010] or a pharmaceutically acceptable salt thereof;

[0011] in:

[0012] R 1 is an unsubstituted alkyl group;

[0013] R 2 is an unsubstituted alkyl group;

[0014] X is halogen, -OP, -N(R A )2.-NR A N(R A )2, -SP or -NCO;

[0015] P is hydrogen, an oxygen protecting group, a sulfur protecting group, or a substituted or unsubstituted heterocyclic group;

[0016] L 1 is -(C=O)-, -CH2-, -CH=CH-, -C≡C-, -O-, -S- or -NR A -;

[0017] L 2 is substituted or unsubstituted alkylene, substituted or unsubstituted alkenylene, substituted or unsubstituted alkynylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted heteroalkenylene, or substituted or unsubstituted heteroalkynylene, wherein L 2 Contains from its and L1 a chain of at least 8 consecutive non-hydrogen atoms from its point of attachment to T;

[0018] T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OR C 、-N(R A )2. -SR A 、-CO2H、Halogen、-OS(O2)R B 、-O(C=O)R C 、-(C=O)OR C 、-O(C=O)OR C 、-(C=O)N(R A )2, -O(C=O)N(R A )2.-NR A (C=O)N(R A )2, -CN, -CHO, -N3, -N=C=S,

[0019] Q is S or O;

[0020] Each R A are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted acyl, substituted or unsubstituted heterocyclyl, a nitrogen protecting group or a sulfur protecting group;

[0021] R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group;

[0022] R C is a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted heteroaryl group or an oxygen protecting group;

[0023] R D Is halogen or -OS(O2)R B ;

[0024] R E is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or an oxygen protecting group; and

[0025] R F is hydrogen, substituted or unsubstituted alkyl, or a nitrogen protecting group;

[0026] Provided that the compound is not:

[0027]

[0028] In certain embodiments, the compound of Formula (I) has Formula (Ia):

[0029]

[0030] or a pharmaceutically acceptable salt thereof.

[0031] In certain embodiments, the compound of Formula (I) has Formula (Ib):

[0032]

[0033] or a pharmaceutically acceptable salt thereof.

[0034] In certain embodiments, the compound of Formula (I) has Formula (Ic):

[0035]

[0036] or a pharmaceutically acceptable salt thereof.

[0037] In certain embodiments, the compound of Formula (I) has Formula (Id):

[0038]

[0039] or a pharmaceutically acceptable salt thereof.

[0040] In certain embodiments, the compound of Formula (I) has Formula (Ie):

[0041]

[0042] or a pharmaceutically acceptable salt thereof.

[0043] In another aspect, a compound of formula (II) is provided:

[0044]

[0045] or a pharmaceutically acceptable salt and / or solvate thereof, wherein:

[0046] R 1 It is -CH(OH)CH2(OH), -CH(X 1 )CH3, CH2CH3, -CH=CH2, -CH=CH(R 2 ) or -CH2CH2(R 3 );

[0047] R 2 Yes - (CH2) m CH(OH)CH2(OH), -(CH2)p CH(X 2 )CH3、-(CH2) t CH=CH2 or -(CH2) v CH2CH3;

[0048] R 3 Yes - (CH2) w CH(OH)CH2(OH), -(CH2) x CH(X 3 )CH3、-(CH2) y CH=CH2 or -(CH2) z CH2CH3;

[0049] X 1 , X 2 and X 3 Each independently is Cl or F;

[0050] n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15;

[0051] m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15;

[0052] p is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15;

[0053] t is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15;

[0054] v is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15;

[0055] w is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15;

[0056] x is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15;

[0057] y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15; and

[0058] z is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15.

[0059] In another aspect, a pharmaceutical composition is provided, comprising a compound of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0060] In another aspect, a method for treating pain (e.g., chronic pain) in a subject in need thereof is provided, the method comprising administering to the subject an effective amount of a compound of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof, or administering to the subject a pharmaceutical composition thereof.

[0061] In another aspect, a method of modulating (e.g., inhibiting) HCN channel gating (e.g., HCN1 channel gating) is provided, the method comprising contacting the HCN channel with an effective amount of a compound of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0062] In another aspect, a method of inhibiting HCN1 channels without enhancing γ-aminobutyric acid-A (GABA-A) receptors is provided, the method comprising contacting the HCN1 channel with an effective amount of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0063] In another aspect, a method for inhibiting the activity of HCN1 channels without modulating GABA-A receptors is provided, the method comprising contacting the HCN1 channel with an effective amount of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0064] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the following detailed description. Other features, objects, and advantages of the invention will be apparent from the embodiments and claims.

[0065] definition

[0066] Chemical Definition

[0067] The definition of specific functional groups and chemical terms will be described in more detail below. Chemical elements are determined according to the periodic table of the elements on the inside cover, CAS version, "Handbook of Chemistry and Physics", 75th Ed, and specific functional groups are generally defined as described therein. In addition, the general principles of organic chemistry and specific functional parts and reactivity are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March, "March's Advanced Organic Chemistry", 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, "Comprehensive Organic Transformations", VCH Publishers, Inc., New York, 1989; and Carruthers, "Some Modern Methods of Organic Synthesis", 3rd Edition, Cambridge University Press, Cambridge, 1987.

[0068] The compounds described herein may contain one or more stereocenters and may therefore exist as stereoisomers, e.g., enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from a mixture by methods known to those skilled in the art, including the formation and crystallization of chiral high pressure liquid chromatography (HPLC) and chiral salts; or preferred isomers may be prepared by asymmetric synthesis. See, e.g., Jacques et al., "Enantiomers, Racemates and Resolutions" (Wiley Interscience, New York, 1981); Wilen et al., "Tetrahedron" 33:2725 (1977); Eliel, EL "Stereochemistry of Carbon Compounds" (McGraw-Hill, NY, 1962); and Wilen, SH "Tables of Resolving Agents and Optical Resolutions" p. 268 (EL. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). A compound may exist as an individual isomer substantially free of other isomers, and alternatively as a mixture of various isomers.

[0069] When a range of values ​​is listed, it is intended to include every value and subrange within the range. For example, "C1-6 alkyl" is intended to include C1, C2, C3, C4, C5, C6, C 1-6 , C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-5 , C 2-4 , C 2-3 , C 3-6 , C 3-5 , C 3-4 , C 4-6 , C 4-5 and C 5-6 alkyl.

[0070] As used herein, "alkyl" refers to a straight or branched chain saturated hydrocarbon radical having 1 to 10 carbon atoms ("C 1-10 In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C1-9 In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C 1-8 In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C 1-7 In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C 1-6 In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C 1-5 In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C 1-4 In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C 1-3 In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C 1-2 In some embodiments, the alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, the alkyl group has 2 to 6 carbon atoms (“C 2-6 C 1-6 Examples of alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). Other examples of alkyl groups include n-heptyl (C7), n-octyl (C8), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an "unsubstituted alkyl group") or substituted (a "substituted alkyl group") with one or more substituents. In certain embodiments, an alkyl group is an unsubstituted C 1-10 In certain embodiments, the alkyl group is a substituted C 1-10 alkyl.

[0071] The term "heteroalkyl" refers to an alkyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur, and the heteroatoms are located within (i.e., inserted between adjacent carbon atoms) and / or are placed at one or more terminal positions of the parent chain. In certain embodiments, heteroalkyl refers to a saturated group ("heteroalkyl"). 1-10 In some embodiments, a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms in the parent chain (“heteroalkyl”). 1-9 In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms in the parent chain (“heteroalkyl”). 1-8In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms in the parent chain (“heteroalkyl”). 1-7 In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and 1 or more heteroatoms in the parent chain (“heteroalkyl”). 1-6 In some embodiments, a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms in the parent chain (“heteroalkyl”). 1-5 In some embodiments, a heteroalkyl group is a saturated group having 1 to 4 carbon atoms and 1 or 2 heteroatoms in the parent chain (“heteroalkyl”). 1-4 In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom in the parent chain (“heteroalkyl”). 1-3 In some embodiments, a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom in the parent chain (“heteroalkyl”). 1-2 In some embodiments, a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (a "heteroC1 alkyl"). In some embodiments, a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms in the parent chain (a "heteroC1 alkyl"). 2-6 Unless otherwise specified, each instance of heteroalkyl is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents. In certain embodiments, heteroalkyl is an unsubstituted heteroalkyl. 1-10 In certain embodiments, heteroalkyl is a substituted heteroC 1-10 alkyl.

[0072] The term "alkenyl" refers to a straight or branched chain hydrocarbon radical having 2 to 10 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds). In some embodiments, the alkenyl group has 2 to 9 carbon atoms ("C 2-9 In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C 2-8 In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C 2-7 In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C 2-6 In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C 2-3In some embodiments, an alkenyl group has 2 carbon atoms ("C2 alkenyl"). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). C 2-4 Examples of alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. 2-6 Examples of the alkenyl group include the above-mentioned C 2-4 In some embodiments, alkenyl is an unsubstituted C 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1 2-10 In certain embodiments, alkenyl is a substituted C 2-10 In alkenyl, there is no C=C double bond with a defined stereochemistry (e.g., -CH=CHCH3 or ) can be an (E)- or (Z)-double bond.

[0073] The term "heteroalkenyl" refers to an alkenyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur, and the heteroatoms are located within (i.e., inserted between adjacent carbon atoms) and / or are placed at one or more terminal positions of the parent chain. In certain embodiments, heteroalkenyl refers to a group having 2 to 10 carbon atoms, at least one double bond, and 1 or more heteroatoms in the parent chain ("heteroalkenyl"). 2-10 In some embodiments, a heteroalkenyl group has 2 to 9 carbon atoms, at least one double bond, and 1 or more heteroatoms (“heteroC 2-9 In some embodiments, a heteroalkenyl group has 2 to 8 carbon atoms, at least one double bond, and 1 or more heteroatoms (“heteroC 2-8 In some embodiments, a heteroalkenyl group has 2 to 7 carbon atoms, at least one double bond, and 1 or more heteroatoms (“heteroC 2-7 In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or more heteroatoms (“heteroC 2-6 In some embodiments, a heteroalkenyl group has 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“heteroC 2-5 In some embodiments, a heteroalkenyl group has 2 to 4 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“heteroC2-4 In some embodiments, a heteroalkenyl group has 2 to 3 carbon atoms, at least one double bond, and 1 heteroatom in the parent chain (“heteroC 2-3 In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or 2 heteroatoms (“heteroC 2-6 Unless otherwise specified, each instance of a heteroalkenyl group is independently unsubstituted (an “unsubstituted heteroalkenyl”) or substituted (a “substituted heteroalkenyl”) with one or more substituents. In certain embodiments, a heteroalkenyl group is an unsubstituted heteroC 2-10 In certain embodiments, heteroalkenyl is a substituted heteroC 2-10 Alkenyl.

[0074] The term "alkynyl" refers to a straight or branched chain hydrocarbon radical ("C 2-10 In some embodiments, an alkynyl group has 2 to 9 carbon atoms (“C 2-9 In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C 2-8 In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C 2-7 In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C 2-6 In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C 2-3 In some embodiments, an alkynyl group has 2 carbon atoms (a "C2 alkynyl"). The one or more carbon-carbon triple bonds may be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). 2-4 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. 2-6 Examples of the alkenyl group include the above-mentioned C 2-4 Alkynyl and pentynyl (C5), hexynyl (C6), etc. Other examples of alkynyl include heptynyl (C7), octynyl (C8), etc. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (an “unsubstituted alkynyl group”) or substituted (a “substituted alkynyl group”) with one or more substituents. In certain embodiments, an alkynyl group is an unsubstituted C 2-10 In certain embodiments, the alkynyl group is a substituted C 2-10Alkynyl.

[0075] The term "heteroalkynyl" refers to an alkynyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur, and the heteroatoms are located within (i.e., inserted between adjacent carbon atoms) and / or are placed at one or more terminal positions of the parent chain. In certain embodiments, heteroalkynyl refers to a group having 2 to 10 carbon atoms, at least one triple bond, and 1 or more heteroatoms in the parent chain ("heteroalkynyl"). 2-10 In some embodiments, heteroalkynyl groups have 2 to 9 carbon atoms, at least one triple bond, and 1 or more heteroatoms (“hetero C 2-9 In some embodiments, heteroalkynyl groups have 2 to 8 carbon atoms, at least one triple bond, and 1 or more heteroatoms (“hetero C 2-8 In some embodiments, heteroalkynyl groups have 2 to 7 carbon atoms, at least one triple bond, and 1 or more heteroatoms (“hetero C 2-7 In some embodiments, heteroalkynyl groups have 2 to 6 carbon atoms, at least one triple bond, and 1 or more heteroatoms (“hetero C 2-6 In some embodiments, a heteroalkynyl group has 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“heteroC 2-5 In some embodiments, a heteroalkynyl group has 2 to 4 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“hetero C 2-4 In some embodiments, a heteroalkynyl group has 2 to 3 carbon atoms, at least one triple bond, and 1 heteroatom in the parent chain (“heteroC 2-3 In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms (“heteroC 2-6 Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an “unsubstituted heteroalkynyl”) or substituted (a “substituted heteroalkynyl”) with one or more substituents. In certain embodiments, a heteroalkynyl group is an unsubstituted heteroalkynyl group. 2-10 In certain embodiments, heteroalkynyl is a substituted heteroC 2-10 Alkynyl.

[0076] The term "carbocyclyl" or "carbocycle" refers to a non-aromatic ring system having 3 to 14 ring carbon atoms ("C 3-14 In some embodiments, the carbocyclyl group has 3 to 10 ring carbon atoms ("C 3-10 In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C 3-8In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms (“C 3-7 In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms (“C 4-6 In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C 5-6 In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C 5-10 (Carbocyclyl)) Exemplary C 3-6 Carbocyclic groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. 3-a Carbocyclic groups include but are not limited to the above C 3-6 Carbocyclic groups and cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo [2.2.1] heptyl (C7), bicyclo [2.2.2] octyl (C8), etc. Exemplary C 3-10 Carbocyclic groups include but are not limited to the above C 3-a Carbocyclic groups and cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C9), decahydronaphthyl (C 10 ), spiro[4.5]decyl (C 10 ) and the like. As shown in the foregoing examples, in certain embodiments, the carbocyclyl is monocyclic ("monocyclic carbocyclyl") or polycyclic (e.g., containing a fused, bridged or spiro ring system, such as a bicyclic system ("bicyclic carbocyclyl") or a tricyclic system ("tricyclic carbocyclyl"), and may be saturated or may contain one or more carbon-carbon double or triple bonds. "Carbocyclyl" also includes ring systems in which the carbocyclyl ring as defined above is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the carbocyclyl ring, and in this case, the carbon number continues to refer to the number of carbons in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl is independently unsubstituted ("unsubstituted carbocyclyl") or substituted ("substituted carbocyclyl") with one or more substituents. In certain embodiments, a carbocyclyl is an unsubstituted C 3-14 In certain embodiments, the carbocyclyl group is a substituted C 3-14 Carbocyclic group.

[0077] In some embodiments, a "carbocyclyl" is a monocyclic saturated carbocyclyl ("C 3-14 In some embodiments, a cycloalkyl group has 3 to 10 ring carbon atoms (“C 3-10 In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C 3-8 In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms (“C 4-6 In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C 5-6 In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C 5-10 (cycloalkyl) 5-6 Examples of cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). 3-6 Examples of the cycloalkyl group include the above-mentioned C 5-6 Cycloalkyl and cyclopropyl (C3) and cyclobutyl (C4). 3-8 Examples of the cycloalkyl group include the above-mentioned C 3-6 Cycloalkyl and cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of cycloalkyl is independently unsubstituted (an “unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”) with one or more substituents. In certain embodiments, cycloalkyl is an unsubstituted C 3-14 In certain embodiments, the cycloalkyl group is a substituted C 3-14Cycloalkyl. "Heterocyclyl" or "heterocycle" as used herein refers to a radical of a 3 to 14-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("3 to 14-membered heterocyclyl"). In heterocyclyls containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, where valence permits. Heterocyclyls may be monocyclic ("monocyclic heterocyclyl") or polycyclic (e.g., fused, bridged or spirocyclic systems, such as bicyclic systems ("bicyclic heterocyclyl") or tricyclic systems ("tricyclic heterocyclyl")), and may be saturated or may contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems may include one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocyclyl ring as defined above is fused to one or more carbocyclyls (where the point of attachment is on the carbocyclyl or heterocyclyl ring) or a heterocyclyl ring as defined above is fused to one or more aryl or heteroaryl groups (where the point of attachment is on the heterocyclyl ring), and in such cases, the number of ring members continues to refer to the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently unsubstituted ("unsubstituted heterocyclyl") or substituted ("substituted heterocyclyl") with one or more substituents. In certain embodiments, heterocyclyl is an unsubstituted 3- to 14-membered heterocyclyl. In certain embodiments, heterocyclyl is a substituted 3- to 14-membered heterocyclyl.

[0078] In some embodiments, the heterocyclyl group is a 5-10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5-10-membered heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-8-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5-8-membered heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-6-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5-6-membered heterocyclyl"). In some embodiments, the 5-6-membered heterocyclyl group has 1 to 3 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, the 5-6-membered heterocyclyl group has 1 to 2 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, the 5-6-membered heterocyclyl group has 1 ring heteroatom selected from nitrogen, oxygen and sulfur.

[0079] Exemplary 3-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azirdinyl, oxiranyl, and thiorenyl. Exemplary 4-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azetidinyl, oxetanyl, and thiidine. Exemplary 5-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrophenylthio, dihydrophenylthio, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing 2 heteroatoms include, but are not limited to, dioxolane, oxathiolanyl, and dithiolanyl. Exemplary 5-membered heterocyclic groups containing 3 heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thienyl. Exemplary 6-membered heterocyclic groups containing 2 heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclic groups containing 3 heteroatoms include, but are not limited to, triazinyl. Exemplary 7-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azepanyl, oxepane and thiepanyl. Exemplary 8-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azocanyl, oxepane and thiecanyl. Exemplary bicyclic heterocyclyls include, but are not limited to, dihydroindolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolinyl, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl, decahydroisoquinolyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, dihydroindolinyl, phthalimidyl, naphthylimidyl, chromanyl, chromenyl, 1H-benzo[e][1,4]diazepine, 1,4,5,7-tetrahydro-pyrano[3,4-b]pyrrolyl, 5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydro-thieno[3,2-b]pyridinyl, 1,2,3,4-tetrahydro-1,6-naphthyridinyl and the like.

[0080] As used herein, "aryl" refers to a radical that provides a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., with 6, 10, or 14 π electrons shared in the cyclic array) having 6 to 14 ring carbon atoms and zero heteroatoms in the aromatic ring system ("C6-14 aryl"). In some embodiments, the aryl has 6 ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl has 10 ring carbon atoms ("C10 aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl has 14 ring carbon atoms ("C14 aryl"; e.g., anthracenyl). "Aryl" also includes a ring system in which an aryl ring as defined above is fused to one or more carbocyclic or heterocyclic groups, wherein the radical or point of attachment is on the aryl ring, and in this case, the number of carbon atoms continues to refer to the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted (an "unsubstituted aryl group") or substituted (a "substituted aryl group") with one or more substituents. In certain embodiments, an aryl group is an unsubstituted C6-14 aryl group. In certain embodiments, an aryl group is a substituted C6-14 aryl group.

[0081] "Aralkyl" is a subset of "alkyl" and refers to an alkyl group as described herein substituted with an aryl group as described herein, wherein the point of attachment is on the alkyl portion.

[0082] As used herein, "heteroaryl" refers to a 5- to 14-membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., with 6, 10, or 14 π electrons shared in the cyclic array) having ring carbon atoms and 1 to 4 ring heteroatoms in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5- to 14-membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, where valence permits. Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring as defined above is fused to one or more carbocyclic or heterocyclic groups, wherein the point of attachment is on the heteroaryl ring, and in this case, the number of ring members continues to represent the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring as defined above is fused to one or more aryl groups, wherein the point of attachment is on the aryl or heteroaryl ring, and in this case, the number of ring members refers to the number of ring members in the fused polycyclic (aryl / heteroaryl) ring system. For polycyclic heteroaryls, one of the rings contains no heteroatoms (e.g., indolyl, quinolinyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., the ring with the heteroatom (e.g., 2-indolyl) or on the ring without the heteroatom (e.g., 5-indolyl).

[0083] In some embodiments, heteroaryl is a 5 to 10 membered aromatic ring system provided with ring carbon atoms and 1 to 4 ring heteroatoms in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5 to 10 membered heteroaryl"). In some embodiments, heteroaryl is a 5 to 8 membered aromatic ring system provided with ring carbon atoms and 1 to 4 ring heteroatoms in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5 to 8 membered heteroaryl"). In some embodiments, heteroaryl is a 5 to 6 membered aromatic ring system provided with ring carbon atoms and 1 to 4 ring heteroatoms in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5 to 6 membered heteroaryl"). In some embodiments, 5 to 6 membered heteroaryl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, 5 to 6 membered heteroaryl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, 5 to 6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (an "unsubstituted heteroaryl") or substituted (a "substituted heteroaryl") with one or more substituents. In certain embodiments, a heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl group. In certain embodiments, a heteroaryl group is a substituted 5- to 14-membered heteroaryl group.

[0084] Exemplary 5-membered heteroaryls containing 1 heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Exemplary 5-membered heteroaryls containing 2 heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryls containing 3 heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryls containing 4 heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryls containing 1 heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryls containing 2 heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryls containing 3 or 4 heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryls containing 1 heteroatom include, but are not limited to, azacycloheptatrienyl, oxacycloheptatrienyl, and thiacycloheptatrienyl. Exemplary 5,6-bicyclic heteroaryls include but are not limited to indolyl, isoindolyl, indazolyl, benzotriazolyl, benzophenylthio, isobenzophenylthio, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl and purinyl. Exemplary 6,6-bicyclic heteroaryls include but are not limited to naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl and quinazolinyl. Exemplary tricyclic heteroaryls include but are not limited to phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl and phenazinyl.

[0085] "Heteroaralkyl" is a subset of "alkyl" and refers to an alkyl group as described herein substituted with a heteroaryl group as described herein, wherein the point of attachment is on the alkyl portion.

[0086] The term "partially unsaturated" as used herein refers to a ring portion that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aromatic groups (e.g., aryl or heteroaryl moieties) as defined herein.

[0087] As used herein, the term "saturated" refers to a ring moiety that contains no double or triple bonds, ie, the ring contains all single bonds.

[0088] Appending the suffix "-ene" to a group indicates that the group is a divalent moiety, for example, alkylene is a divalent moiety of alkyl, alkenylene is a divalent moiety of alkenyl, alkynylene is a divalent moiety of alkynyl, heteroalkylene is a divalent moiety of heteroalkyl, heteroalkenylene is a divalent moiety of heteroalkenyl, heteroalkynylene is a divalent moiety of heteroalkynyl, carbocyclylene is a divalent moiety of carbocyclyl, heterocyclylene is a divalent moiety of heterocyclyl, arylene is a divalent moiety of aryl, and heteroarylene is a divalent moiety of heteroaryl.

[0089] As understood above, in certain embodiments, the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl described herein are optionally substituted. Optionally substituted refers to a group that can be substituted or unsubstituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or "unsubstituted" alkynyl, "substituted" or "unsubstituted" heteroalkyl, "substituted" or "unsubstituted" heteroalkenyl, "substituted" or "unsubstituted" heteroalkynyl, "substituted" or "unsubstituted" carbocyclyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl, or "substituted" or "unsubstituted" heteroaryl). In general, the term "substituted", whether preceded by the term "optionally" or not, means that at least one hydrogen present on a group (e.g., a carbon or nitrogen atom) is replaced with a permissible substituent, e.g., a substituent that results in a stable compound (e.g., a compound that does not spontaneously transform, such as by rearrangement, cyclization, elimination, or other reaction) upon substitution. Unless otherwise specified, a "substituted" group has substituents at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituents are the same or different at each position. The term "substituted" is contemplated to include substitution with all permissible substituents of organic compounds, i.e., any substituents described herein that result in the formation of stable compounds. The present invention contemplates any and all such combinations in order to result in stable compounds. For purposes of the present invention, heteroatoms (such as nitrogen) may have hydrogen substituents and / or any suitable substituents described herein that satisfy the valences of the heteroatoms and result in the formation of a stable moiety.

[0090] Exemplary carbon atom substituents include, but are not limited to, halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR aa 、-ON(R bb )2、-N(R bb )2、-N(R bb )3 + X - 、-N(OR cc )R bb , -SH, -SR aa 、-SSR c c. -C(=O)R aa 、-CO2H、-CHO、-C(OR cc 3. -CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(Rbb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)R aa 、-P(=O)(R aa )2、-P(=O)(OR cc )2、-OP(=O)(R aa )2、-OP(=O)(OR cc )2、-P(=O)(N(R bb )2)2、-OP(=O)(N(R bb)2)2、-NR bb P(=O)(R aa )2.-NR bb P(=O)(OR cc )2.-NR bb P(=O)(N(R bb )2)2、-P(R cc )2、-P(OR cc )2.-P(R cc )3 + X - 、-P(OR cc )3 + X - 、-P(R cc )4.-P(OR cc )4.-OP(R cc )2.-OP(R cc )3 + X - 、-OP(OR cc )2.-OP(OR cc )3 + X - 、-OP(R cc )4.-OP(OR cc )4, -B(R aa )2、-B(OR cc )2. -BR aa (OR cc ), C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd wherein X - is the counter ion;

[0091] Or the two geminal hydrogen atoms on the carbon atom are replaced by groups =O, =S, =NN (R bb )2, =NNR bb C(=O)R aa , =NNR bb C(=O)OR aa , =NNR bb S(=O)2Raa , =NR bb or = NOR cc replace;

[0092] R aa Each instance of is independently selected from C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, or two R aa The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution;

[0093] R bb Each instance of is independently selected from hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc 2. -CO2R aa 、-SO2R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc 2. -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)(R aa )2.-P(=O)(OR cc )2、-P(=O)(N(R cc )2)2、C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, or two R bb The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution; wherein X - is the counter ion;

[0094] R cc Each instance of is independently selected from hydrogen, C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, or two R cc The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0095] R dd Each instance of is independently selected from halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee 、-ON(R ff )2、-N(R ff )2、-N(R ff )3 + X - 、-N(OR ee )R ff , -SH, -SR ee 、-SSR ee 、-C(=O)R ee 、-CO2H、-CO2R ee 、-OC(=O)R ee 、-OCO2R ee 、-C(=O)N(R ff )2, -OC(=O)N(R ff )2.-NR ff C(=O)R ee 、-NR ff CO2R ee、-NR ff C(=O)N(R ff )2, -C(=NR ff )OR ee 、-OC(=NR ff )R ee 、-OC(=NR ff )OR ee 、-C(=NR ff )N(R ff )2, -OC(=NR ff )N(R ff )2.-NR ff C(=NR ff )N(R ff )2.-NR ff S02R ee 、-SO2N(R ff 2. -SO2R ee 、-SO2OR ee 、-OSO2R ee 、-S(=O)R ee 、-Si(R ee )3.-OSi(R ee )3. -C(=S)N(R ff )2, -C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)(OR ee )2. -P(=O)(R ee )2.-OP(=O)(R ee )2.-OP(=O)(OR ee 2. C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, 3 to 10 membered heterocyclic group, C 6-10 aryl and 5- to 10-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg group substituted, or two geminal R dd Substituents may be linked to form =O or =S; where X - is the counter ion;

[0096] R eeEach instance of is independently selected from C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, C 6-10 aryl, 3- to 10-membered heterocyclyl, and 3- to 10-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted by 0, 1, 2, 3, 4, or 5 R gg Group substitution;

[0097] R ff Each instance of is independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, 3 to 10 membered heterocyclic group, C 6-10 aryl and 5- to 10-membered heteroaryl, or two R ff The groups are connected to form a 3- to 10-membered heterocyclyl or 5- to 10-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg group substitution; and

[0098] R gg Each instance of is independently selected from halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1-6 Alkyl, -ON(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)3 + X - 、-NH(C 1-6 Alkyl)2 + X - 、-NH2(C 1-6 alkyl) + X - 、-NH3 + X - 、-N(OC 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6Alkyl), -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 alkyl), -C(=O)(C 1-6 alkyl), -CO2H, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 Alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2, -OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 Alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 Alkyl)2, -NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1-6 alkyl), -OC(=NH)(C 1-6 alkyl), -OC(=NH)OC 1-6 Alkyl, -C(=NH)N(C 1-6 Alkyl)2, -C(=NH)NH(C 1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1-6 Alkyl)2, -OC(=NH)NH(CC 1-6 alkyl), -OC(=NH)NH2, -NHC(=NH)N(C 1-6 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 Alkyl), -SO2N(C 1-6 Alkyl)2, -SO2NH(C 1-6 alkyl), -SO2NH2, -SO2(C 1-6 Alkyl), -SO2O(C 1-6 Alkyl), -OSO2(C 1-6 Alkyl), -SO(C 1-6 Alkyl), -Si(C 1-6 Alkyl)3, -OSi(C 1-6 alkyl)3-C(=S)N(C 1-6 alkyl)2, C(=S)NH(C 1-6 alkyl), C(=S)NH2, -C(=O)S(C 1-6 alkyl), -C(=S)SC 1-6 Alkyl, -SC(=S)SC1-6 Alkyl, -P(=O)(OC 1-6 Alkyl)2, -P(=O)(C 1-6 Alkyl)2, -OP(=O)(C 1-6 Alkyl)2, -OP(=O)(OC 1-6 Alkyl)2, C 1-6 Alkyl, C 1-6 Perhaloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, hetero C 1-6 Alkyl, hetero C 2-6 Alkenyl, hetero C 2-6 Alkynyl, C 3-10 Carbocyclic group, C 6-10 aryl, 3- to 10-membered heterocyclyl, 5- to 10-membered heteroaryl; or two geminal R gg Substituents may be linked to form =O or =S; where X - It is a counter ion.

[0099] As used herein, the term "halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -C1), bromine (bromo, -Br), or iodine (iodo, -I).

[0100] The term "acyl" refers to an acyl group having the general formula -C(=O)R X1 、-C(=O)OR X1 、-C(=O)-OC(=O)R X1 、-C(=O)SR X1 、-C(=O)N(R X1 )2, -C(=S)R X1 、-C(=S)N(R X1 )2、-C(=S)O(R X1 )、-C(=S)S(R X1 ),-C(=NR X1 )R X1 、-C(=NR X1 )OR X1 、-C(=NR X1 )SR X1 and -C(=NR X1 )N(R X1 )2, wherein R X1is hydrogen; halogen; substituted or unsubstituted hydroxy; substituted or unsubstituted thiol; substituted or unsubstituted amino; substituted or unsubstituted acyl; cyclic or non-cyclic substituted or unsubstituted branched or unbranched aliphatic; cyclic or non-cyclic substituted or unsubstituted branched or unbranched heteroaliphatic; cyclic or non-cyclic substituted or unsubstituted branched or unbranched alkyl; cyclic or non-cyclic substituted or unsubstituted branched or unbranched alkenyl; substituted or unsubstituted substituted alkynyl; substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, aliphaticoxy, heteroaliphaticoxy, alkoxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthiooxy, heteroaliphaticthiooxy, alkylthiooxy, heteroalkylthiooxy, arylthiooxy, heteroarylthiooxy, mono- or di-aliphatic amino, mono- or di-heteroaliphatic amino, mono- or di-alkylamino, mono- or di-heteroalkylamino, mono- or di-arylamino, or mono- or di-heteroarylamino; or two R X1 The groups are combined to form a 5-6 membered heterocyclic ring. Exemplary acyl groups include aldehydes (-CHO), carboxylic acids (-CO2H), ketones, acyl halides, esters, amides, imines, carbonates, carbamates and ureas. Acyl substituents include, but are not limited to, any substituent described herein that results in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thioxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphatic amino, heteroaliphatic amino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphatic oxy, heteroaliphatic oxy, alkoxy, heteroalkoxy, aryloxy, heteroaryloxy, aliphatic thiooxy, heteroaliphatic thiooxy, alkylthiooxy, heteroalkylthiooxy, arylthiooxy, heteroarylthiooxy, acyloxy, etc., each of which may be further substituted or unsubstituted).

[0101] To maintain electron neutrality, a "counterion" or "anionic counterion" is a negatively charged group associated with a positively charged group. Anionic counterions can be monovalent (i.e., include one formal negative charge). Anionic counterions can also be multivalent (i.e., include more than one formal negative charge), such as divalent or trivalent. Exemplary counterions include halide ions (e.g., F - , Cl - Br - ,I - )、NO3 - 、ClO4 - OH - 、H2PO4 - 、HCO3 - 、HSO4 -, sulfonate ions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphorsulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethyl-1-sulfonic acid-2-sulfonate, etc.), carboxylate ions (e.g., acetate, propionate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, etc.), BF4 - PF4 - PF6 - 、AsF6 - 、SbF6 - 、B[3,5-(CF3)2C6H3]4] - 、B(C6F5)4 - ,BPh4 - 、Al(OC(CF3)3)4 - and carborane anions (e.g., CB 11 H 12 - or (HCB 11 Me5Br6) - ). Exemplary counterions that may be multivalent include CO3 2- 、HPO4 2- PO4 3- 、B4O7 2- 、SO4 2- 、S2O3 2- , carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelaate, sebacate, salicylate, phthalate, aspartate, glutamate, etc.), and carborane.

[0102] The term "leaving group" has its common meaning in the field of synthetic organic chemistry, and refers to an atom or group that can be substituted by a nucleophilic reagent. For example, see Smith, "March's Advanced Organic Chemistry" 6th ed. (501-502). Examples of suitable leaving groups include, but are not limited to, halogen (such as F, Cl, Br or I (iodine)), alkoxycarbonyloxy, aryloxycarbonyloxy, alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (e.g., acetoxy), arylcarbonyloxy, aryloxy, methoxy, N, O-dimethylhydroxyamino, pixyl and haloformate. In some cases, the leaving group is a sulfonate, such as toluenesulfonate (tosylate, -OTs), methanesulfonate (mesylate, -OMs), bromobenzenesulfonyloxy (bromobenzenesulfonate, -OBs), -OS(=O)2(CF2)3CF3 (perfluorobutylsulfonate, -ONf) or trifluoromethanesulfonate (trifluoromethanesulfonate, -OTf). In some cases, the leaving group is bromobenzenesulfonate, such as bromobenzenesulfonyloxy. In some cases, the leaving group is a nitrobenzenesulfonate, such as 2-nitrobenzenesulfonyloxy. The leaving group can also be a phosphine oxide (e.g., formed during the Mitsunobu reaction) or an internal leaving group (such as an epoxide or a cyclic sulfate). Other non-limiting examples of leaving groups are water, ammonia, alcohols, ether moieties, thioether moieties, zinc halides, magnesium moieties, diazonium salts, and copper moieties. Further exemplary leaving groups include, but are not limited to, halogenated (e.g., chloro, bromo, iodo) and activated substituted hydroxyl groups (e.g., -OC(=O)SR aa 、-OC(=O)R aa 、-OCO2R aa 、-OC(=O)N(R bb )2, -OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-OC(=NR bb )N(R bb )2. -OS(=O)R aa 、-OSO2R aa 、-OP(R cc )2.-OP(R cc )3, -OP(=O)2R aa ,-OP(=O)(R aa )2.-OP(=O)(OR cc)2、-OP(=O)2N(R bb )2 and -OP(=O)(NR bb )2, where R aa , R bb and R cc as defined herein).

[0103] As used herein, the term "hydroxyl" or "hydroxyl" refers to the group -OH. The term "substituted hydroxyl" or "substituted hydroxyl" refers by extension to a hydroxyl group in which the oxygen atom directly attached to the parent molecule is replaced by a group other than hydrogen, and includes groups selected from: -OR aa 、-ON(R bb )2, -OC(=O)SR aa 、-OC(=O)R aa 、-OCO2R aa 、-OC(=O)N(R bb )2, -OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-OC(=NR bb )N(R bb )2. -OS(=O)R aa 、-OSO2R aa 、-OSi(R aa )3.-OP(R cc )2.-OP(R cc )3 + X - 、-OP(OR cc )2.-OP(OR cc )3 + X - ,-OP(=O)(R aa )2.-OP(=O)(OR cc )2 and -OP(=O)(N(R bb )2)2, where X - , R aa , R bb and R cc As defined herein.

[0104] As used herein, the term "thiol" or "thio" refers to the group -SH. The term "substituted thiol" or "substituted thio" refers by extension to thiol groups in which the sulfur atom directly attached to the parent molecule is replaced by a group other than hydrogen, and includes thiol groups selected from -SR aa 、-S=SR cc 、-SC(=S)SRaa 、-SC(=O)SR aa 、-SC(=O)OR aa and-SC(=O)R aa A group in which R aa and R cc As defined herein.

[0105] As used herein, the term "amino" refers to the group -NH2. The term "substituted amino" refers by extension to a monosubstituted amino, a disubstituted amino, or a trisubstituted amino as described herein. In certain embodiments, a "substituted amino" is a monosubstituted amino or a disubstituted amino.

[0106] As used herein, "monosubstituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is substituted by one hydrogen atom and one group other than a hydrogen atom, and includes amino groups selected from -NH(R bb )、-NHC(=O)R aa 、-NHCO2R aa 、-NHC(=O)N(R bb )2, -NHC(=NR bb )N(R bb )2. -NHSO2R aa 、-NHP(=O)(OR cc )2 and -NHP(=O)(N(R bb )2)2, wherein R aa , R bb and R cc As defined herein, and wherein the group -NH(R bb ) bb Not hydrogen.

[0107] The term "disubstituted amino" as used herein refers to an amino group in which the nitrogen atom directly attached to the parent molecule is substituted by two groups other than hydrogen, and includes amino groups selected from -N(R bb )2.-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2.-NR bb C(=NR bb )N(R bb )2.-NR bb S02R aa 、-NR bb p(=O)(OR cc )2 and -NR bb P(=O)(N(Rbb )2)2, wherein R aa , R bb and R cc As defined herein, the proviso is that the nitrogen atom directly attached to the parent molecule is not replaced with a hydrogen.

[0108] The term "trisubstituted amino" as used herein refers to an amino group in which the nitrogen atom directly attached to the parent molecule is substituted by three groups, and includes amino groups selected from -N(R bb )3 and -N(R bb )3 + X - A group in which R bb and X - As defined herein.

[0109] As used herein, the term "oxo" refers to the group =0, and the term "thiooxo" refers to the group =S.

[0110] The nitrogen atoms may be substituted or unsubstituted where valence permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents include, but are not limited to, hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc 2. -CO2R aa 、-SO2R aa 、-C(=NR bb )R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc 2. -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)(OR cc )2. -P(=O)(R aa )2、-P(=O)(N(R cc )2)2、C 1-10 Alkyl, C 1-10 Perhaloalkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 Aryl and 5- to 14-membered heteroaryl, or two R groups attached to an N atom cc The groups are connected to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd and wherein R aa , R bb , R cc and R dd As defined herein.

[0111] In certain embodiments, the substituent present on the nitrogen atom is a nitrogen protecting group (also referred to herein as an "amino protecting group"). Nitrogen protecting groups include, but are not limited to, -OH, -OR aa 、-N(R cc )2, -C(=O)R aa 、-C(=O)N(R cc 2. -CO2R aa 、-SO2R aa 、-C(=NR cc )R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc 2. -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc , C 1-10 Alkyl (e.g., aralkyl, heteroaralkyl), C 2-10 Alkenyl, C 2-10 Alkynyl, hetero C 1-10 Alkyl, hetero C 2-10 Alkenyl, hetero C 2-10 Alkynyl, C 3-10 Carbocyclic group, 3 to 14 membered heterocyclic group, C 6-14 aryl and 5- to 14-membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aralkyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd group substituted, and wherein R aa , Rbb , R cc and R dd As defined herein. Nitrogen protecting groups are well known in the art and are included in "Protecting Groups in Organic Synthesis", TW Greene and PGM Wuts, 3 rd Those described in detail in , edition, John Wiley & Sons, 1999.

[0112] For example, a nitrogen protecting group (such as an amide group (e.g., -C(=O)R aa )) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propionamide, 3-(o-nitrophenyl)propionamide, 2-methyl-2-(o-nitrophenoxy)propionamide, 2-methyl-2-(o-phenylazophenoxy)propionamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamamide, N-acetylmethionine derivatives, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0113] A nitrogen protecting group (such as a carbamate group (e.g., -C(=O)OR aa)) includes but is not limited to methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-tert-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthenyl)]carbamate (DBD-Tmoc), 4-methoxyphenylacylcarbamate (Phenoc), 2,2,2-trichloroethylcarbamate (Troc), 2-trimethylsilylethylcarbamate (Teoc), 2-phenylethylcarbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethylcarbamate , 1,1-dimethyl-2,2-dibromoethylcarbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethylcarbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethylcarbamate (Bpoc), 1-(3,5-di-tert-butylphenyl)-1-methylethylcarbamate (t-Bumeoc), 2-(2′- and 4′-pyridyl)ethylcarbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, tert-butylcarbamate (BOC or Boc), 1-adamantyl Carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropyl allyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate , 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethylcarbamate, diphenylmethylcarbamate, 2-methylthioethylcarbamate, 2-methylsulfonylethylcarbamate, 2-(p-toluenesulfonyl)ethylcarbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylphenylthiocarbamate (Mtpc), 2,4-dimethylphenylthiocarbamate (Bmpc), 2-phosphoniumethylcarbamate (Peoc), 2-triphenylphosphoniumisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl (o-nitro) phenyl) methyl carbamate, tert-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropyl methyl carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacyl vinyl carbamate, o-(N,N-dimethylformamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylformamido)propylcarbamate, 1,1-dimethylpropynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furylmethylcarbamate, 2-iodoethylcarbamate, isobornylcarbamate, isobutylcarbamate, isonicotinoylcarbamate, p-(p'-methoxyphenylazo)benzylcarbamate, 1-methylcyclobutylcarbamate, 1-methylcyclohexylcarbamate, 1-methyl-1-cyclopropylmethylcarbamate, 1-methyl -1-(3,5-dimethoxyphenyl)ethylcarbamate, 1-methyl-1-(p-phenylazophenyl)ethylcarbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethylcarbamate, phenylcarbamate, p-(phenylazo)benzylcarbamate, 2,4,6-tri-tert-butylphenylcarbamate, 4-(trimethylammonium)benzylcarbamate and 2,4,6-trimethylbenzylcarbamate. ,

[0114] A nitrogen protecting group (such as a sulfonamide group (e.g., -S(=O)2R aa )) include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchromanyl-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide and benzoylsulfonamide.

[0115] Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivatives, N'-toluenesulfonylaminoacyl derivatives, N'-phenylaminosulfonyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiosuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilazide adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5 -substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenamine (PhF), N-2, 7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamine (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)trimethylphenyl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylamine, N-(5,5-dimethyl-3-oxo-1-cyclohexene

[0063] The invention also includes but is not limited to the following: (i) 2-nitrophenyl sulfonamide, (ii) 2-nitrophenyl sulfonamide, (iii) 2-nitrophenyl sulfonamide, (iv) 2-nitrophenyl sulfonamide, (v ...In certain embodiments, the nitrogen protecting group is benzyl (Bn), tert-butoxycarbonyl (BOC), carbobenzyloxy (Cbz), 9-fluorenylmethyloxycarbonyl (Fmoc), trifluoroacetyl, triphenylmethyl, acetyl (Ac), benzoyl (Bz), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), 2,2,2-trichloroethyloxycarbonyl (Troc), triphenylmethyl (Tr), tosyl (Ts), bromosulfonyl (Bs), nitrosulfonyl (Ns), methanesulfonyl (Ms), trifluoromethanesulfonyl (Tf), or dansyl (Ds).

[0116] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also referred to herein as a "hydroxy protecting group"). Oxygen protecting groups include, but are not limited to, -R aa 、-N(R bb )2, -C(=O)SR aa ,,-C(=O)R aa 、-CO2R aa 、-C(=O)N(R bb )2, -C(=NR bb )R aa 、-C(=NR bb )OR aa 、-C(=NR bb )N(R bb )2, -S(=O)R aa 、-SO2R aa 、-Si(R aa )3. -P(R cc )2.-P(R cc )3 + X - 、-P(OR cc )2、-P(OR cc )3 + X - ,-P(=O)(R aa )2.-P(=O)(OR cc )2 and -P(=O)(N(R bb )2)2, where X - , R aa , R bb and R cc As defined herein. Oxygen protecting groups are well known in the art and are included in "Protecting Groups in Organic Synthesis", TW Greene and P.G.M. Wuts, 3 rdThose described in detail in , edition, John Wiley & Sons, 1999.

[0117] Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxymethyl (MOM), methylthiomethyl (MTM), tert-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), tert-butoxymethyl, 4-pentenyloxymethyl (POM), silyloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl ( THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S, S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxane-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanebenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyl 2-(phenyloxyseleno)ethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenyloxyseleno)ethyl, tert-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolinyl, 4-picolinyl, 3-methyl-2-picolinyl N-oxide, diphenylmethyl, p,p'-dinitrodiphenylmethyl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenyl methyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromobenzoyloxyphenyl)diphenylmethyl, 4,4',4"-tri(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tri(levulinyloxyphenyl)methyl, 4,4',4"-tri(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltert-hexylsilyl, tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), tert-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinic acid ester), 4,4-(ethylenedithio)pentanoate (levulinic acid acetal), pivalate, adamantanoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (lecithin), methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), ethyl carbonate, 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonium)ethyl carbonate ( Peoc), isobutyl carbonate, ethylene carbonate, allyl carbonate, tert-butyl carbonate (BOC or Boc), p-nitrophenyl carbonate, benzyl carbonate, p-methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, o-nitrobenzyl carbonate, p-nitrobenzyl carbonate, S-benzylthiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylvalerate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylbenzene In some embodiments, the oxygen protecting group is silyl. In some embodiments, the oxygen protecting group is methyl styrene. In some embodiments, the oxygen protecting group is methyl styrene. In some embodiments, the oxygen protecting group is methyl styrene.The oxygen protecting group is tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS), triphenylsilyl (TPS), triethylsilyl (TES), trimethylsilyl (TMS), triisopropylsiloxymethyl (TOM), acetyl (Ac), benzoyl (Bz), allyl carbonate, 2,2,2-trichloroethyl carbonate (Troc), 2-trimethylsilylethyl carbonate, methoxymethyl (MOM), 1-ethoxyethyl (EE ), 2-methoxy-2-propyl (MOP), 2,2,2-trichloroethoxyethyl, 2-methoxyethoxymethyl (MEM), 2-trimethylsilylethoxymethyl (SEM), methylthiomethyl (MTM), tetrahydropyranyl (THP), tetrahydrofuranyl (THF), p-methoxyphenyl (PMP), triphenylmethyl (Tr), methoxytrityl (MMT), dimethoxytrityl (DMT), allyl, p-methoxybenzyl (PMB), tert-butyl, benzyl (Bn), allyl or pivaloyl (Piv). ,

[0118] In certain embodiments, the substituent present on the sulfur atom is a sulfur protecting group (also referred to herein as a "thiol protecting group"). Sulfur protecting groups include, but are not limited to, -R aa 、-N(R bb )2, -C(=O)SR aa 、-C(=O)R aa 、-CO2R aa 、-C(=O)N(R bb )2, -C(=NR bb )R aa 、-C(=NR bb )OR aa 、-C(=NR bb )N(R bb )2, -S(=O)R aa 、-SO2R aa 、-Si(R aa )3. -P(R cc )2.-P(R CC )3 + X - 、-P(OR cc )2、-P(OR CC )3 + X - 、-P(=O)(R aa )2.-P(=O)(OR cc )2 and -P(=O)(N(R bb )2)2, where R aa , R bb and Rcc As defined herein. Sulfur protecting groups are well known in the art and are included in "Protecting Groups in Organic Synthesis", TW Greene and P.G.M. Wuts, 3 rd edition, John Wiley & Sons, 1999. In certain embodiments, the sulfur protecting group is acetamidomethyl, tert-butyl, 3-nitro-2-pyridinesulfinyl, 2-pyridinesulfinyl, or triphenylmethyl.

[0119] These and other exemplary substituents are described in more detail in the detailed description, examples, figures, and claims. The present invention is not intended to be limited in any way by the foregoing list of exemplary substituents.

[0120] Other definitions

[0121] The term "antagonist" refers to an agent that (i) reduces or inhibits one or more effects of another agent; and / or (ii) reduces or inhibits one or more biological events. In some embodiments, an antagonist can reduce the level and / or activity of one or more agents that it targets. An antagonist can be direct (in which case it has an effect directly on its target) or indirect (in which case it has an effect by means other than binding to its target; for example, by interacting with a modulator of the target, such that the level or activity of the target is altered). In some embodiments, an antagonist can be a receptor antagonist, such as a receptor ligand or drug, that blocks or inhibits a biological response by binding to and blocking the receptor rather than activating the receptor like an agonist. In certain embodiments, a compound of formula (I) is an antagonist of HCN (e.g., HCN1). In certain embodiments, a compound of formula (II) is an antagonist of HCN (e.g., HCN1). In certain embodiments, a compound of formula (I) inhibits HCN (e.g., HCN1) channel gating. In certain embodiments, compounds of Formula (II) inhibit HCN (eg, HCN1) channel gating.

[0122] The term "agonist" refers to an agent that (i) increases or induces one or more effects of another agent; and / or (ii) increases or induces one or more biological events. In some embodiments, an agonist can increase the level and / or activity of one or more agents that it targets. In various embodiments, an agonist can be or include agents of various chemical classes, including, for example, small molecules, polypeptides, nucleic acids, carbohydrates, lipids, metals, and / or other entities that display relevant agonistic activity. An agonist can be direct (in which case it directly affects its target) or indirect (in which case it affects by means other than binding to its target; for example, by interacting with a modulator of the target, such that the level or activity of the target is altered). A partial agonist can act as a competitive antagonist in the presence of a full agonist because it competes with the full agonist and interacts with its target and / or its modulator, thereby producing a reduction in (i) one or more effects of another agent, and / or (ii) a reduction in one or more biological events compared to that observed with the full agonist alone.

[0123] The term "inverse agonist" refers to an agent that binds to the same receptor as an agonist but induces a pharmacological response opposite to that of the agonist. A neutral antagonist has no activity in the absence of an agonist or inverse agonist, but can block either activity. The action of an inverse agonist is opposite to that of an agonist, but an antagonist can block the action of both. An agonist increases the activity of a receptor above its basal level, while an inverse agonist reduces the activity below the basal level. In certain embodiments, the compound of formula (I) is an inverse agonist of HCN (e.g., HCN1). In certain embodiments, the compound of formula (II) is an inverse agonist of HCN (e.g., HCN1).

[0124] The term "inhibit" or "inhibition" in the context of a regulated level (e.g., expression and / or activity) of a target (e.g., HCN1) is not limited to complete inhibition. Thus, in some embodiments, partial inhibition or relative reduction is included within the scope of the term "inhibition". In some embodiments, the term refers to a reduction in the level (e.g., expression and / or activity) of a target (e.g., HCN1) to a level that is reproducible and / or statistically significantly lower than an initial or other appropriate reference level, which may be, for example, a baseline level of the target. In some embodiments, the term refers to a decrease in the level (e.g., expression and / or activity) of a target to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of the initial level, which initial level can be, for example, a baseline level of the target.

[0125] As used herein, the term "inhibitor" refers to an agent whose presence or level is associated with a decrease in the level or activity of a target to be regulated. In some embodiments, an inhibitor may act directly (in which case it has an effect on its target directly, such as by binding to the target); in some embodiments, an inhibitor may act indirectly (in which case it has an effect by interacting with and / or otherwise altering a modulator of the target, thereby reducing the level and / or activity of the target). In some embodiments, an inhibitor is an inhibitor whose presence or level is associated with a decrease in the level or activity of a target relative to a specific reference level or activity (e.g., observed under appropriate reference conditions such as the presence of a known inhibitor or the absence of an inhibitor disclosed herein). In certain embodiments, a compound of formula (I) is an inhibitor of HCN (e.g., HCN1). In certain embodiments, a compound of formula (II) is an inhibitor of HCN (e.g., HCN1).

[0126] The terms "salt" and "pharmaceutically acceptable salt" refer to those salts which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and which are commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are amino salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or amino salts formed with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or amino salts formed by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, gluconate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N + (C 1-4 Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates, when necessary.

[0127] "Subjects" contemplated for administration include, but are not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young, middle-aged, or elderly)) and / or other non-human animals, such as mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as rodents (e.g., mice, rats), guinea pigs, cows, pigs, horses, sheep, goats, cats, and / or dogs. Non-human animals can be male or female and at any stage of development. Non-human animals can be transgenic animals.

[0128] As used herein, "chronic pain" refers to pain that persists beyond the normal pain process for a specific injury. Chronic pain intensity may vary from mild to severe. Chronic pain includes neuropathic pain, which refers to chronic pain caused by nerves.

[0129] "Disease," "disorder," and "condition" are used interchangeably herein.

[0130] As used herein and unless otherwise indicated, the terms "treat," "treating," and "treatment" contemplate effects that occur while a subject has a particular disease, disorder, or condition and that reduce the severity of the disease, disorder, or condition, or that delay or slow the progression of the disease, disorder, or condition ("therapeutic treatment" or "therapeutically treating"), and also contemplate effects that occur before a subject begins to have a particular disease, disorder, or condition and that inhibit or reduce the severity of the disease, disorder, or condition ("prophylactic treatment" or "prophylactically treating").

[0131] In general, an "effective amount" of a compound refers to an amount sufficient to elicit a desired biological response. As will be appreciated by one of ordinary skill in the art, the effective amount of a compound of the invention may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and condition of the subject. For example, an effective amount of a compound having antiproliferative activity is an amount that results in a concentration sufficient to inhibit cell proliferation. An effective amount encompasses both therapeutic and prophylactic treatments.

[0132] As used herein and unless otherwise indicated, a "therapeutically effective amount" of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the disease, disorder, or condition. A therapeutically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment of a disease, disorder, or condition. The term "therapeutically effective amount" can encompass an amount that improves overall treatment, reduces or avoids symptoms or causes of a disease, disorder, or condition, or enhances the therapeutic effect of another therapeutic agent.

[0133] As used herein and unless otherwise indicated, a "prophylactically effective amount" of a compound is an amount sufficient to prevent a disease, disorder, or condition, or one or more symptoms associated with a disease, disorder, or condition, or to prevent its recurrence. A prophylactically effective amount of a compound means the amount of a therapeutic agent, alone or in combination with other agents, that provides a prophylactic benefit in the treatment of a disease, disorder, or condition. The term "prophylactically effective amount" may encompass an amount that improves overall prevention or enhances the prophylactic efficacy of another prophylactic agent.

[0134] As used herein, use of the phrase "at least one instance" refers to 1, 2, 3, 4 or more instances, but also inclusively encompasses ranges of, for example, 1 to 4, 1 to 3, 1 to 2, 2 to 4, 2 to 3, or 3 to 4 instances. BRIEF DESCRIPTION OF THE DRAWINGS

[0135] Figure 1 shows the structure of HCN1 and the alkylphenol binding site and lipid ordering in the ion channel. Figure 1A and Figure 1B : Parallel to the membrane ( Figure 1A ) or from the outside of the cell ( Figure 1B ) observed, the cryo-EM structure of the HCN1 tetramer in the ligand-free state. The “non-exchangeable” four-fold symmetric arrangement of the subunits is Figure 1B One of the HCN1 subunits is clearly visible in the figure, with a dashed circle around it. Figure 1B In the figure, the sequences of the voltage-sensing blade motifs that form the radial array are labeled S1-S4 in the lower right subunit. Figure 1A The horizontal grey bars in the figure indicate the approximate boundaries of the membrane bilayer. “Outside” and “Inside” indicate the orientation of the tetramer channel in the bilayer.

[0136] Figure 1C and Figure 1D : Structure of a crystallized Kv channel in the presence of lipids, viewed from the extracellular surface (left) and from the side (right). For clarity, on the right, only a single S1-S4 voltage sensing domain and its interface with the lipid-facing shell of the pore are shown (right). Figure 1C and Figure 1D In both cases, the co-crystallized lipids were dissociated and in the gaps between adjacent voltage sensing leaves (in Figure 1C in the middle) and near the membrane-facing surfaces of the leaves and pores (in Figure 1C and Figure 1D The arrows identify the external S4 Arg residues that protrude toward the lipid membrane.

[0137] Figure 1E and Figure 1F : The molecular surface of one of the five general anesthetic subunit internal cavities identified in GLIC (Myxobacterium ligand-gated ion channel, a bacterial homologue of a pentamer ligand-gated ion channel that is a target of alkylphenols; Bocquet et al., Nature 2007 445, 116-119; Nury et al., Nature 2011, 469, 428-431) is shown in black, with the positions of all five radially symmetric cavities shown by arrows. The unlabeled light grey molecular surface volume marks the position of the adjacent intersubunit cavities, and on the lower right, a communication tunnel ( Figure 1E ). Schematic diagram of the helical arrangement of a single subunit of a pentameric ligand-gated channel, with the alkylphenol docked in the subunit's internal cavity (viewed from the outside) ( Figure 1F ).

[0138] Figure 1G and Figure 1H : The predicted structure of the minimal alkylphenol-sensitive HCN1 channel HCN1-ΔNvΔC. Figure 1A As shown, Figure 1G The horizontal grey bars in represent the approximate boundaries of the membrane bilayer. Figure 1G The scale bar on the left indicates the distance from the extracellular apex of the channel to the membrane surface. This distance is approx. . Figure 1G The scale bar on the right represents the distance from the extracellular surface of the membrane to the innermost aspect of the coupling loop connecting the transmembrane helices. This distance is approx. and represents the maximum theoretical orthogonal range required to tether the pharmacophore, and therefore also represents the maximum theoretical orthogonal range of the tether. Figure 1H , the spheres inserted between each voltage-sensing blade represent the hypothetical locations of the anchors (not drawn to scale), and the tether and pharmacophore are inserted in the “targeting groove” between the S1 to S4 blades and enter the page perpendicularly.

[0139] Figure 2 shows the sequence alignment of HCN1-4 ( Figure 2A ) and the cryo-EM structure of the sequence corresponding to the minimal alkylbenzene-sensitive HCN1 channel core HCN1-ΔNvΔC ( Figure 2B ). Figure 2A: In this core structure, there are 34 residues that are unique to mHCN1 relative to mHCN2, hHCN3, and mHCN4 (indicated by residues highlighted in black in white text), and 50 residues that differ from at least one other isoform (indicated by residues highlighted in gray). Residues not highlighted show four-fold conservation among the indicated isoforms. Mouse (m) HCN1 (O88704.1), mHCN2 (O88703.1), human (h) HCN3 (Q9P1Z3.2), and mHCN4 (O70507.2) were aligned using Clustal Omega (1.2.4) (www.ebi.ac.uk / Tools / msa / clustalo / ). Figure 2B : Top view of the core channel tetramer predicted from the cryo-EM structure of the hHCN1 tetramer (left panel) and side view with the anterior and posterior subunits removed for clarity (right panel). Note that the mouse and human HCN1 sequences are identical in the region defined by mHCN1-ΔNvΔC. HCN1 unique residues are shown as spheres in the upper row; residues shared by HCN1 and at least one other HCN1 sequence (but not all) are shown as spheres in the lower row. The structure is based on the atomic coordinates of HCN1 in the cAMP-unbound state from Lee & MacKinnon 2017 (Cell, 168(1-2)111-120) and is deposited in the Protein Data Bank (uniprot.org) under accession number 5U6O.

[0140] Figure 3 Schematic diagram showing the structures of a conceptualized anchor-tethered 2,6-di-tert-butylphenol (2,6-DTBP) therapeutic agent and two tethered derivatives of 2,6-DTBP, BP4C-1 1:0:1 and BP4L-10:0:1. BP4L-10:0:1 is bound to a diol "anchor" (surrounded by a dashed box). BP4C-1 1:0:1 is not anchored.

[0141] Figure 4 Shown are recordings from three individual cells expressing HCN1-ΔNvΔCv, each exposed to the indicated concentrations of BP4C-1 1:0:1, BP4L-10:0:1 or DMSO mediator alone. The recordings on the left are currents activated at hyperpolarizing step potentials, the recordings on the right are tail currents at 0 mV.

[0142] Figure 5 show Figure 4 Tail current activation curves fitted with a Boltzmann function for the three cells are shown.

[0143] Figure 6 Showing V as a function of butylphenol concentration 1 / 2 displacement.

[0144] Figure 7 The effects of propofol and 2,6-DTBP on the V of HCN1-4 channels are shown. 1 / 2 *Indicates a response that is statistically different from that of the control group.

[0145] Figure 8 Showing the V of 2,6- (left) or 2,4- (right) dibutylphenol on HCN1 channel 1 / 2 impact.

[0146] Fig. 9 Display according to EC 50 Estimated rank order of HCN1 antagonism.

[0147] Fig.10 The rates of Tl+-mediated fluorescence quenching are shown in large unilamellar vesicles, demonstrating that these compounds have minimal membrane effects.

[0148] Fig.11 Showing the structures of three 2,6-alkylphenols and their use as general anesthetics (GA) and GABA A GABA receptor A -R) function.

[0149] Fig.12 Display P w，IPSI and P W，CONTRA (probability of withdrawal of the paw ipsilateral or contralateral to the nerve ligation) and the effect of the indicated cumulative ip doses of 2,6-DTBP as a function of the intensity of stimulated fibers.

[0150] Fig.13 A and Fig.13 B shows HPWL (hind paw withdrawal latency) as a function of cumulative ip 2,6-DTBP dose and stimulus intensity. Heat source intensities of 15% and 30% (maximum 150 W) elicited withdrawal responses between the fastest and slowest detection thresholds (2-3 and 30 seconds, respectively), with the lower setting being optimal for examining thermal hyperalgesia and the higher setting examining the majority of nociceptive responses. 2,6-DTBP improved thermal hyperalgesia and had no significant effect on thermal nociception.

[0151] Fig.14 The structures of BP4L-18:1:1 and BP4L-10:0:1 are shown.

[0152] Fig.15Current recordings from individual cells expressing HCN1 are shown. Each cell was pre-incubated for 20 min in recording solution containing 20 mM 2-hydroxypropyl-β-cyclodextrin (HPβCD) + 10 mM DMSO alone or with 30 μM BP4L-18:1:1 as indicated. Black scans are those recorded with an activation potential of -65 mV.

[0153] Fig.16 The inhibition of channel gating by BP4L-18:1:1 was shown to be concentration-dependent.

[0154] Fig.17 BP4L-18:1:1 was shown to effectively inhibit channel gating with a potency and efficacy indistinguishable from free 2,6-DTBP (compare black and grey Hill equation fit lines), whereas BP4L-10:0:1 (pharmacophore that restricts access to the binding site) was largely ineffective.

[0155] Fig.18 Shown are the interactions of alkylphenols and alkylcyclohexanols with HCN1 channel gating, modeled as inverse agonist and partial inverse agonist coupling to channel opening, respectively.

[0156] Fig.19A and Fig.19C Yes Display Fig.19B Graph of the current-voltage relationship for the dialkylbenzene compound structure in FIG. The graph shows that the hydrogen bonding functionality at position 1 is useful for HCN1-gated dialkylbenzene inhibition. The black scan is the scan recorded at an activation potential of -75 mV ( Fig.19A ).

[0157] Fig. 20 The inhibition of 2,6-dialkylbenzene derivatives is shown to reveal hydrogen bonding potentials; the identity of the alkyl side chain and the presence or absence of π electrons contribute differently to the energetics of back-agonism for this class of molecules. In each panel, the thick black line is the Hill fit to 2,6-DTBP. The thin grey line is the Hill fit to 2,6-DTBP in the presence of 50 Shifted on the concentration axis; the thick grey line is the fit of the Hill equation to the concentration response in the plate plot; and the dashed grey lines in the 2,6-DIPP and 2,6-DTBP plate plots are the fits of the Hill equation to the 2,6-DIPCH and 2,6-DTBCH data, with only the maximum response of the fit changing.

[0158] Fig.21A The inverse agonist potency (EC) of 2,6-dialkylbenzene analogs is shown as a function of molecular volume. 50 ) in the table below. Fig. 21Bis a graph showing the potency predicted by the partition coefficient cP corrected for differential accumulation into the lipid bilayer. Fig. 21C show Fig.21A and Fig. 21B Composite structure of the diagram.

[0159] Fig. 22 Alkylphenols and alkylcyclohexanols are shown to associate with the HCN1 channel through four common, radially distributed sites. Fig. 22 A and Fig. 22 B is the ΔV shown as a function of the concentration of isopropyl and tert-butyl phenol and cyclohexanol 1 / 2 of charts. Fig. 22 C. Fig. 22 D and Fig. 22 E is a graph showing ΔV as a function of 2,6-DIPCH, 2,6-DTBCH, and 2,6-DIPP concentrations in the presence of 3 μM 2,6-DTBP. 1 / 2 of charts. Fig. 22 F is a graph showing the common site model based on N=4 (based on Fig. 22 A to Fig. 22 E) The association constant determined by the fit of the model was converted into a free energy term according to -(RT)LnK.

[0160] Fig.23 A is a graph showing ΔV1 / 2 as a function of 2,6-di-sec-butylphenol (2,6-DSBP) concentration in the absence of 3 μM 2,6-DTBP. Fig.23 B is a graph showing ΔV1 / 2 as a function of 2,6-DSBP concentration in the presence of 3 μM 2,6-DTBP. These graphs demonstrate that 2,6-DSBP is a low potency, high efficacy inverse agonist.

[0161] Figure 24 shows that 2,6-DTBP improves mechanical and thermal hyperalgesia in the chronic constriction injury model of peripheral neuropathic pain. Bolus administration of 2,6-DTBP caused approximately 45% reversal of mechanical hypersensitivity in a manner independent of spinal glycine receptors ( FIG. 24A to FIG. 24B ) and complete reversal of thermal hypersensitivity ( FIG. 24C to FIG. 24D ). Data from et al., 2016 J. Clin. Invest 126(7), 2547-60.

[0162] Figure 25 shows that anti-hyperalgesic doses of 2,6-DTBP do not disrupt normal nociception, nor interfere with locomotor activity, motor coordination, and muscle strength. Fig.25A ), autonomous activities in open areas ( Fig.25B ), coordination of movement during accelerated rotation ( Fig.25C ) and horizontal wire test to assess muscle strength ( Fig.25D ). Data from et al., 2016 J. Clin. Invest 126(7), 2547-60.

[0163] Fig.26 HCN1 antagonism of anchor-tethered 2,6-DTBP shown as a function of tether length: The smoothed line is a hypothetical plot of efficacy as a function of tether length generated using a Boltzmann distribution, where the slope incorporates the molecular length distribution due to bond vibrations and the tendency of the pharmacophore to pivot in non-orthogonal directions, shortening the pharmacophore relative to its theoretical maximum extent.

[0164] Fig. 27 Stills showing molecular dynamics simulations of (1) free 2,6-DTBP and (2) its first generation anchor-tethered derivatives BP4L-10:0:1 and BP4L-18:1:1.

[0165] Fig.28 The antagonism of HCN1 by 4-substituted 2,6-DIPP was shown. The behavior of 4-substituted diisopropylphenols was evaluated, where the substitution introduced 4-amino (4-AP), tert-butyl (4-TBP), or 4-trimethyl-amino (4-TMAP). 4-TBP was completely effective. In contrast, 4-TMAP was ineffective. 4-AP was effective, demonstrating that the presence of the nitrogen atom itself was not the cause of the failure.

[0166] Fig.29 Graph showing the concentration of BP4L-18:1:1 in whole blood after administration to rats. Eight adult rats were gavaged with 0.5 mL peanut oil (n=4) or 0.5 mL peanut oil containing 0.58 mmol / kg BP4L-18:1:1 (n=4). Blood samples were collected at designated times. After extraction, precipitation and filtration using an equal volume of acetonitrile, the blood concentration of BP4L-18:1:1 was determined using LC-MS / MS. The black dashed line represents the concentration of BP4L-18:1:1 in individual animals receiving the drug, and the black solid line represents the mean ± SEM of the drug-dosed group. The dashed gray line and the solid gray line are the concentration of BP4L-18:1:1 detected in animals consuming peanut oil alone and their mean ± SEM, respectively. The gray line is superimposed on the time axis and cannot be distinguished. DETAILED DESCRIPTION

[0167] HCN1 channels are assembled as tetramers in a cross-shaped arrangement, with four voltage-sensing "blades" (formed by S1 to S4) arranged around a central complex (formed by the S5 to S6 sequence) (Figure 1). The ion-conducting pore is located at the center of the symmetry axis. Although there are interesting differences between HCN channels and Kv channels, this may be related to the inverse voltage dependence of gating (HCN subunits are four spokes around an axis - see Figure 1B The coding in the lower panel, while in the Kv channel, the voltage sensor of one subunit is located on S5 to S6 of its neighbor - not shown), but in both molecules there is a large gap between each voltage sensor. These gaps expose one aspect of each S5 to S6 motif to the membrane lipids ( Figure 1B , Figure 1C In fact, at high enough resolution, lipids in these “gaps” are visible in the dissolved structure ( Figure 1C and Figure 1D zigzag grey line in the figure).

[0168] Crystallography shows propofol docking at GABA A -R in the cavity ( Figure 1E and Figure 1F ), where it may replace native acyl chains or water. Mass labeling with photoactivated alkylphenols combined with molecular dynamics modeling revealed that alkylphenols are embedded in the α1-β3 subunits of these receptors. Deletion analysis demonstrated that only the core of HCN1 is required for the channel to exhibit full alkylphenol sensitivity; many cytoplasmic nitrogen termini and all cytoplasmic carbon termini can be removed without compromising drug function. The minimal alkylphenol-sensitive channel omits approximately 64% of the full-length human HCN1 sequence ( Figure 1G and Figure 1H ). Figure 2A An alignment of HCN1-4 is shown spanning the region corresponding to the alkylphenol sensitive core. Figure 2B The isotype divergence mapped onto the cryo-EM structure is shown.

[0169] Figure 1G and Figure 1H The approximate position of the smallest channel (HCN1-ΔNvΔC) as well as the membrane surface (horizontal grey bars) and scale bars are shown to illustrate the approximate distance from the apex of the extracellular ring to the outer surface of the membrane (vertical grey bars) and from there to the innermost aspects of the ring between transmembrane structures (vertical black bars).

[0170] Kinetic modeling and other studies suggest that 2,6-DTBP alters HCN1 gating by discriminating between the closed and open configurations of the pore and does so by interacting with the site through which it enters via the lipid phase. There are perpendicular lipid-facing voids along the length of the channel, one aspect of which is the exterior aspect of the S5-S6 pore motif (relative to the central ion conduction path), consistent with the location of the alkylphenol site somewhere along this surface. This description of a prototypical alkylphenol binding site is consistent with the putative location identified in HCN1 based on the GABAA receptor.

[0171] Lipid-filled vertical voids extend along the length of the channel from the outer surface to the cytoplasm, providing evidence that a physical channel (the “targeting groove”) exists for tethered pharmacophore delivery and that its geometry is consistent with pharmacophore delivery regardless of the distance between the outer surface and the alkylphenol site.

[0172] Compound

[0173] Provided herein are novel compounds (e.g., compounds of Formula (I) and Formula (II)) that modulate HCN channels, intermembrane proteins that act as non-selective voltage-gated cation channels in the plasma membrane of excitable cells including excitable cells of the heart, central nervous system (CNS), and peripheral nervous system (PNS). In certain embodiments, the compounds are selective antagonists and / or inhibitors of the HCN isoform HCN1 and can be used to treat pain (e.g., chronic pain). The compounds can be provided as compounds or pharmaceutically acceptable salts for use in any composition, kit, or method described herein.

[0174] In certain embodiments, the compound has the structure (eg, Figure 3 ), which comprises an anchor portion ("T") that may not traverse the lipid bilayer of the cell membrane, thus restricting the anchor end of the compound to the extracellular space. A linker ("L") that is attached to the anchor at one end and to the pharmacophore at the other end 1 -L 2 ”) is used to orient a compound across a cell membrane so that the pharmacophore reaches its effector site in an orientation that allows functionally efficient binding (e.g., inhibitory gating with HCN1).

[0175] Provided are compounds of formula (I):

[0176]

[0177] and pharmaceutically acceptable salts thereof;

[0178] in:

[0179] R 1 is an unsubstituted alkyl group;

[0180] R 2 is an unsubstituted alkyl group;

[0181] X is halogen, -OP, -N(R A )2.-NR A N(R A )2, -SP or -NCO;

[0182] P is hydrogen, an oxygen protecting group, a sulfur protecting group, or a substituted or unsubstituted heterocyclic group;

[0183] L 1 is -(C=O)-, -CH2-, -CH=CH-, -C≡C-, -O-, -S- or -NR A -;

[0184] L 2 is substituted or unsubstituted alkylene, substituted or unsubstituted alkenylene, substituted or unsubstituted alkynylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted heteroalkenylene, or substituted or unsubstituted heteroalkynylene, wherein L 2 Contains from its and L 1 a chain of at least 8 consecutive non-hydrogen atoms from its point of attachment to T;

[0185] T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OR C 、-N(R A )2. -SR A 、-CO2H、Halogen、-OS(O2)R B 、-O(C=O)R C 、-(C=O)OR C 、-O(C=O)OR C 、-(C=O)N(R A )2, -O(C=O)N(R A )2.-NR A (C=O)N(R A )2, -CN, -CHO, -N3, -N=C=S,

[0186] Q is S or O;

[0187] Each R A are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted acyl, substituted or unsubstituted heterocyclyl, a nitrogen protecting group or a sulfur protecting group;

[0188] R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group;

[0189] R C is a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted heteroaryl group or an oxygen protecting group;

[0190] R D Is halogen or -OS(O2)R B ;

[0191] R E is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or an oxygen protecting group; and

[0192] R F is hydrogen, substituted or unsubstituted alkyl, or a nitrogen protecting group;

[0193] Provided that the compound is not:

[0194]

[0195] Group R 1 and R 2

[0196] As described in this article, R 1 is unsubstituted alkyl. 1 is unsubstituted C 1-6 In certain embodiments, R 1 is unsubstituted C 1-4 In certain embodiments, R 1 is neopentyl, tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl. 1 is tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl. 1 is tert-butyl, isobutyl, isopropyl, ethyl or methyl. 1 is tert-butyl, isobutyl, isopropyl, ethyl or methyl. 1 is tert-butyl, isobutyl, isopropyl or ethyl. 1 is tert-butyl, isobutyl or isopropyl. 1 is tert-butyl or isopropyl. 1 In certain embodiments, R1 It is tert-butyl.

[0197] As described in this article, R 2 is unsubstituted alkyl. 2 is unsubstituted C 1-6 In certain embodiments, R 2 is unsubstituted C 1-4 In certain embodiments, R 2 is neopentyl, tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl. 2 is tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl. 2 is tert-butyl, isobutyl, isopropyl, ethyl or methyl. 2 is tert-butyl, isobutyl, isopropyl, ethyl or methyl. 2 is tert-butyl, isobutyl, isopropyl or ethyl. 2 is tert-butyl, isobutyl or isopropyl. 2 is tert-butyl or isopropyl. 2 In certain embodiments, R 2 It is tert-butyl.

[0198] In certain embodiments, R 1 is unsubstituted C 1-6 Alkyl, and R 2 is unsubstituted C 1-6 In certain embodiments, R 1 is unsubstituted C 1-4 Alkyl, and R 2 is an unsubstituted C1-4 alkyl. 1 is neopentyl, tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl; and R 2 is neopentyl, tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl. 1 is tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl; and R 2 is tert-butyl, sec-butyl, isobutyl, n-butyl, isopropyl, n-propyl, ethyl or methyl. 1 is tert-butyl, isobutyl, isopropyl, ethyl or methyl; and R 2 is tert-butyl, isobutyl, isopropyl, ethyl or methyl. 1is tert-butyl, isobutyl, isopropyl, ethyl or methyl; and R 2 is tert-butyl, isobutyl, isopropyl, ethyl or methyl. 1 is tert-butyl, isobutyl, isopropyl or ethyl; and R 2 is tert-butyl, isobutyl, isopropyl or ethyl. 1 is tert-butyl, isobutyl or isopropyl; and R 2 is tert-butyl, isobutyl or isopropyl. 1 is tert-butyl or isopropyl; and R 2 is tert-butyl or isopropyl. 1 is isopropyl; and R 2 In certain embodiments, R 1 is isopropyl; and R 2 In certain embodiments, R 1 is tert-butyl; and R 2 In certain embodiments, R 1 is tert-butyl, and R 2 It is tert-butyl.

[0199] Group X

[0200] As used herein, X is halogen, -OP, -N(R A )2.-NR A N(R A )2, -SP or -NCO; P is hydrogen, an oxygen protecting group, a sulfur protecting group or a substituted or unsubstituted heterocyclic group; and each R A are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted acyl, substituted or unsubstituted heterocyclyl, a nitrogen protecting group, or a sulfur protecting group. In certain embodiments, X is halogen, -OP, -N(R A )2, -SP or -NCO; P is hydrogen, oxygen protecting group or sulfur protecting group; and each R AIn certain embodiments, X is halogen, -OP, -SP or -NCO; and P is hydrogen, oxygen protecting group or sulfur protecting group. In certain embodiments, X is halogen, -OH, -SH or -NCO. In certain embodiments, X is -F, -Cl, -Br, -I, -OH, -SH or -NCO. In certain embodiments, X is -F, -Cl, -OH, -SH or -NCO. In certain embodiments, X is -F, -Cl, -OH, -SH or -NCO. In certain embodiments, X is -F, -OH, -SH or -NCO. In certain embodiments, X is -F, -OH, -SH or -NCO. In certain embodiments, X is -F, -OH or -SH. In certain embodiments, X is -OH or -SH. In certain embodiments, X is -F or -OH. In certain embodiments, X is -F. In certain embodiments, X is -Cl. In certain embodiments, X is -Br. In certain embodiments, X is -I. In certain embodiments, X is -OH. In certain embodiments, X is -OP, wherein P is an oxygen protecting group. In certain embodiments, X is -SH. In certain embodiments, X is -SP, wherein P is a sulfur protecting group. In certain embodiments, X is -NCO.

[0201] Group L 1

[0202] As described in this article, L 1 is -(C=O)-, -CH2-, -CH=CH-, -C≡C-, -O-, -S- or -NR A -. In certain embodiments, L 1 is -(C=O)-, -CH=CH- or -C≡C-. In certain embodiments, L 1 is -CH2-. In certain embodiments, L 1 is -CH=CH-. In certain embodiments, L 1 is -C≡C-. In certain embodiments, L 1 In certain embodiments, L 1 In certain embodiments, L 1 Yes - NR A -, where R A L is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or a nitrogen protecting group. 1 It is -(C=O)-.

[0203] Group L 2

[0204] As described in this article, L 2is substituted or unsubstituted alkylene, substituted or unsubstituted alkenylene, substituted or unsubstituted alkynylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted heteroalkenylene, or substituted or unsubstituted heteroalkynylene, wherein L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 is substituted or unsubstituted alkylene, substituted or unsubstituted alkenylene, substituted or unsubstituted heteroalkylene, or substituted or unsubstituted heteroalkenylene, wherein L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 is substituted or unsubstituted C 8-30 Alkylene; substituted or unsubstituted C 8-30 Alkenylene; substituted or unsubstituted C 4-30 Heteroalkylene; or substituted or unsubstituted C 4-30 In certain embodiments, L 2 is substituted or unsubstituted C 8-20 Alkylene; substituted or unsubstituted C 8-20 Alkenylene; substituted or unsubstituted C 4-20 Heteroalkylene; or substituted or unsubstituted C 4-20 In certain embodiments, L 2 is substituted or unsubstituted C 15-20 Alkylene; substituted or unsubstituted C 15-20 Alkenylene; substituted or unsubstituted C 8-20 Heteroalkylene; or substituted or unsubstituted C 8-20 In certain embodiments, L 2 is substituted or unsubstituted C 8-16 Alkylene; substituted or unsubstituted C 8-16 Alkenylene; substituted or unsubstituted C 4-16 Heteroalkylene; or substituted or unsubstituted C 4-16 In certain embodiments, L 2 is substituted or unsubstituted C a-12 Alkylene; substituted or unsubstituted C 8-12 Alkenylene; substituted or unsubstituted C 4-12 Heteroalkylene; or substituted or unsubstituted C 4-12 Heteroalkenylene.

[0205] In certain embodiments, L 2is substituted or unsubstituted C 8-30 Alkylene; or substituted or unsubstituted C 8-30 In certain embodiments, L 2 is substituted C 8-30 Alkylene; or substituted C 8-30 In certain embodiments, L 2 is unsubstituted C 8-30 Alkylene; or unsubstituted C 8-30 In certain embodiments, L 2 is substituted C 8-20 Alkylene; or substituted C a-20 In certain embodiments, L 2 is unsubstituted C a-20 Alkylene; or unsubstituted C a-20 In certain embodiments, L 2 is substituted C 8-16 Alkylene; or substituted C 8-16 In certain embodiments, L 2 is unsubstituted C 8-16 Alkylene; or unsubstituted C 8-16 In certain embodiments, L 2 is substituted C 8-12 Alkylene; or substituted C 8-12 In certain embodiments, L 2 is unsubstituted C 8-12 Alkylene; or unsubstituted C 8-12 Alkenylene.

[0206] In certain embodiments, L 2 is substituted or unsubstituted C 8-30 Alkylene; or substituted or unsubstituted C 4-30 In certain embodiments, L 2 is substituted C 8-30 Alkylene; or substituted C 4-30 In certain embodiments, L 2 is unsubstituted C 8-30 Alkylene; or unsubstituted C 4-30 In certain embodiments, L 2 is substituted C 8-20 Alkylene; or substituted C 4-20 In certain embodiments, L 2 is unsubstituted C 8-20 Alkylene; or unsubstituted C 4-20In certain embodiments, L 2 is substituted C 8-16 Alkylene; or substituted C 4-16 In certain embodiments, L 2 is unsubstituted C 8-16 Alkylene; or unsubstituted C 4-16 In certain embodiments, L 2 is substituted C 8-12 Alkylene; or substituted C 4-12 In certain embodiments, L 2 is unsubstituted C 8-12 Alkylene; or unsubstituted C 4-12 Heteroalkylene.

[0207] In certain embodiments, L 2 is a substituted or unsubstituted alkylene group. 2 is substituted or unsubstituted C a-30 In certain embodiments, L 2 is substituted C 8-30 In certain embodiments, L 2 is unsubstituted C 8-30 In certain embodiments, L 2 is substituted C 8-20 In certain embodiments, L 2 is unsubstituted C a-20 In certain embodiments, L 2 is substituted C 8-16 In certain embodiments, L 2 is unsubstituted C 8-16 In certain embodiments, L 2 is substituted C a-12 In certain embodiments, L 2 is unsubstituted C a-12 Alkylene.

[0208] In certain embodiments, L 2 is a substituted or unsubstituted alkenylene. 2 is substituted or unsubstituted C a-30 In certain embodiments, L 2 is substituted C 8-30 In certain embodiments, L 2 is unsubstituted C a-30 In certain embodiments, L 2 is substituted C8-20 In certain embodiments, L 2 is unsubstituted C a-20 In certain embodiments, L 2 is substituted C 8-16 In certain embodiments, L 2 is unsubstituted C 8-16 In certain embodiments, L 2 is substituted C 8-12 In certain embodiments, L 2 is unsubstituted C a-12 Alkenylene.

[0209] In certain embodiments, L 2 is a substituted or unsubstituted heteroalkylene. 2 is substituted or unsubstituted C 4-30 In certain embodiments, L 2 is substituted C 4-30 In certain embodiments, L 2 is unsubstituted C 4-30 In certain embodiments, L 2 is substituted C 4-20 In certain embodiments, L 2 is unsubstituted C 4-20 In certain embodiments, L 2 is substituted C 4-16 In certain embodiments, L 2 is unsubstituted C 4-16 In certain embodiments, L 2 is substituted C 4-12 In certain embodiments, L 2 is unsubstituted C 4-12 Heteroalkylene.

[0210] In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 In certain embodiments, L comprises a chain of at least 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive non-hydrogen atoms to its point of attachment to T. 2 Contains from its and L 1to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T. In certain embodiments, L 2 Contains from its and L 1 to its point of attachment to T.

[0211] Group T

[0212] Anchoring moieties ("T") have different functions. In some embodiments, anchoring moieties confine compounds to the periphery and extracellular space. In some embodiments, anchors are used to target pharmacophores to peripheral sensory neurons (e.g., peripheral HCN1 channels). In some embodiments, anchoring moieties are reactive groups (e.g., electrophilic) that can act to promote conversion of an anchoring moiety to another anchoring moiety.

[0213] As described herein, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OR C 、-N(R A )2. -SR A 、-CO2H、Halogen、-OS(O2)R B、-O(C=O)R C 、-(C=O)OR C 、-O(C=O)OR C 、-(C=O)N(R A )2, -O(C=O)N(R A )2.-NR A (C=O)N(R A )2, -CN, -CHO, -N3, -N=C=S, Q is S or O; each R A R is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted acyl, substituted or unsubstituted heterocyclyl, a nitrogen protecting group or a sulfur protecting group; B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group; R C is a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted heteroaryl group or an oxygen protecting group; R D Is halogen or -OS(O2)R B ; R E is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or an oxygen protecting group; and R F is hydrogen, substituted or unsubstituted alkyl, or a nitrogen protecting group.

[0214] In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, -OS(O2)R B 、-O(C=O)R C 、-(C=O)OR C 、-O(C=O)OR C or R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group; and R C is a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heterocyclyl group, or a substituted or unsubstituted heteroaryl group.

[0215] In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, or -OS(O2)R B ; and RB is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group.

[0216] In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, or -OS(O2)R B ; and R B It is methyl, trifluoromethyl, toluoyl or p-nitrophenyl.

[0217] In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, or halogen.

[0218] In certain embodiments, T is substituted or unsubstituted alkenyl, substituted alkyl, substituted or unsubstituted heteroalkyl, -OH, halogen, or -(C=O)OR C ; and R C is a substituted or unsubstituted heterocyclyl. In certain embodiments, T is a substituted or unsubstituted alkenyl, a substituted alkyl, -OH, a halogen, or -(C=O)OR C ; and R c is a substituted or unsubstituted heterocyclic group.

[0219] In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, or -Cl.

[0220] In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted heteroalkyl, -OH, or halogen. In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, -OH, or halogen. In certain embodiments, T is substituted alkyl, substituted or unsubstituted alkenyl, -OH, or -Cl.

[0221] In certain embodiments, T is -Cl, -OH, and n is an integer from 0 to 8. In certain embodiments, T is -Cl, -OH, and n is an integer from 0 to 8. In certain embodiments, T is -Cl. In certain embodiments, T is -OH. In certain embodiments, T is and n is an integer from 0 to 8. In certain embodiments, T is and n is an integer from 0 to 8. In certain embodiments, T is and n is 8. In certain embodiments, T is and n is an integer from 0 to 8. In certain embodiments, T is and n is 8. In certain embodiments, T is In certain embodiments, T is In certain embodiments, T is In certain embodiments, T is

[0222] Certain embodiments

[0223] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C a-30 Alkylene, substituted or unsubstituted C 8-30 Alkenylene, substituted or unsubstituted C 4-30 Heteroalkylene, or substituted or unsubstituted C 4-30 heteroalkenylene; and T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, -OS(O2)R B OR -(C=O)OR C ; R C is a substituted or unsubstituted heterocyclic group; and R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group.

[0224] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-20 Alkylene, substituted or unsubstituted C 8-20 Alkenylene, substituted or unsubstituted C 4-20 Heteroalkylene, or substituted or unsubstituted C 4-20 heteroalkenylene; and T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, -OS(O2)R B OR -(C=O)OR C ; R C is a substituted or unsubstituted heterocyclic group; and R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group.

[0225] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-16 Alkylene, substituted or unsubstituted C 8-16 Alkenylene, substituted or unsubstituted C 4-16 Heteroalkylene, or substituted or unsubstituted C 4-16 heteroalkenylene; and T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, -OS(O2)R B OR -(C=O)OR C ; R C is a substituted or unsubstituted heterocyclic group; and R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group.

[0226] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-12 Alkylene, substituted or unsubstituted C 8-12 Alkenylene, substituted or unsubstituted C 4-12 Heteroalkylene, or substituted or unsubstituted C 4-12 heteroalkenylene; and T is substituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, -OH, -NH2, -SH, -CO2H, halogen, -OS(O2)R B OR -(C=O)OR C ; R C is a substituted or unsubstituted heterocyclic group; and R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group.

[0227] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-12 Alkylene or substituted or unsubstituted C 8-12 alkenylene; and T is substituted alkyl, substituted or unsubstituted alkenyl, -OH, -NH2, -SH, -CO2H, halogen, -OS(O2)R B OR -(C=O)ORC ; R C is a substituted or unsubstituted heterocyclic group; and R B is a substituted or unsubstituted alkyl group or a substituted or unsubstituted aryl group.

[0228] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-12 Alkylene or substituted or unsubstituted C 8-12 alkenylene; and T is substituted alkyl, substituted or unsubstituted alkenyl, -OH or -Cl.

[0229] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-30 Alkylene or substituted or unsubstituted C 8-30 Alkenylene; T is -Cl, -OH, And n is an integer from 0 to 8.

[0230] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-12 Alkylene or substituted or unsubstituted C 8-12 Alkenylene; T is -Cl, -OH, And n is an integer from 0 to 8.

[0231] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-12 Alkylene or substituted or unsubstituted C 8-12 Alkenylene; T is -Cl, -OH, And n is an integer from 0 to 8.

[0232] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-12 Alkylene or substituted or unsubstituted C8-12 Alkenylene; T is And n is an integer from 0 to 8.

[0233] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C 8-30 Alkylene, substituted or unsubstituted C 8-30 Alkenylene, substituted or unsubstituted C 4-30 heteroalkylene; and T is

[0234] In certain embodiments, -L 1 -L 2 -T is Where L 2 is substituted or unsubstituted C a-12 Alkylene; substituted or unsubstituted C a-12 Alkenylene; substituted or unsubstituted C 4-12 Heteroalkylene; or substituted or unsubstituted C 4-12 heteroalkenylene; and n is an integer from 0 to 8.

[0235] In certain embodiments, the compound of Formula (I) has Formula (Ia):

[0236]

[0237] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , L 1 , L 2 and T are as defined herein.

[0238] In certain embodiments, the compound of Formula (I) has Formula (Ib):

[0239]

[0240] or a pharmaceutically acceptable salt thereof, wherein L 1 , L 2 and T are as defined herein.

[0241] In certain embodiments, the compound of formula (I) has formula (I-b1):

[0242]

[0243] or a pharmaceutically acceptable salt thereof, wherein L 1 , L 2 and T are as defined herein.

[0244] In certain embodiments, the compound of formula (I) has formula (I-b2):

[0245]

[0246] or a pharmaceutically acceptable salt thereof, wherein L 1 , L 2 and T are as defined herein.

[0247] In certain embodiments, the compound of Formula (I) has Formula (Ic):

[0248]

[0249] or a pharmaceutically acceptable salt thereof, wherein L 1 , L 2 and T are as defined herein.

[0250] In certain embodiments, the compound of formula (I) has formula (I-c1):

[0251]

[0252] or a pharmaceutically acceptable salt thereof, wherein L 1 , L 2 and T are as defined herein.

[0253] In certain embodiments, the compound of formula (I) has formula (I-c2):

[0254]

[0255] or a pharmaceutically acceptable salt thereof, wherein L 1 , L 2 and T are as defined herein.

[0256] In certain embodiments, the compound of Formula (I) has Formula (Id):

[0257]

[0258] or a pharmaceutically acceptable salt thereof, wherein T is as defined herein.

[0259] In certain embodiments, the compound of formula (I) has formula (I-d1):

[0260]

[0261] or a pharmaceutically acceptable salt thereof, wherein T is as defined herein.

[0262] In certain embodiments, the compound of formula (I) has formula (I-d2):

[0263]

[0264] or a pharmaceutically acceptable salt thereof, wherein T is as defined herein.

[0265] In certain embodiments, the compound of Formula (I) has Formula (Ie):

[0266]

[0267] or a pharmaceutically acceptable salt thereof, wherein T is as defined herein.

[0268] In certain embodiments, the compound of formula (I) has formula (I-e1):

[0269]

[0270] or a pharmaceutically acceptable salt thereof, wherein T is as defined herein.

[0271] In certain embodiments, the compound of formula (I) has formula (I-e2):

[0272]

[0273] or a pharmaceutically acceptable salt thereof, wherein T is as defined herein.

[0274] In certain embodiments, the compound of formula (I) has the following formula:

[0275]

[0276]

[0277] Treatment methods and uses.

[0278] The present disclosure provides a method for treating pain (e.g., chronic pain) in a subject in need thereof, the method comprising administering an effective amount of any compound of formula (I), formula (II), or a pharmaceutically acceptable salt thereof. In certain embodiments, the chronic pain is neuropathic pain, characterized by one or more symptoms selected from the group consisting of persistent negative sensory perception, hyperalgesia, allodynia, burning sensation, and allodynia descriptors.

[0279] The present disclosure provides methods for regulating (e.g., inhibiting or increasing) the activity (e.g., abnormal activity or undesirable activity, such as increased or decreased activity) of HCN channel gating (e.g., HCN1 channel gating). The present disclosure provides methods for regulating (e.g., inhibiting or increasing) the activity (e.g., abnormal activity, such as increased or decreased activity) of HCN channel gating (e.g., HCN1 channel gating) in a subject. In certain embodiments, the method inhibits HCN (e.g., HCN1) channel gating. The present disclosure also provides methods for treating various diseases (e.g., diseases associated with abnormal activity (e.g., increased activity)) of HCN channel gating (e.g., pain (e.g., chronic pain)) in a subject.

[0280] The present disclosure further provides methods of using the compounds described herein (e.g., as biological probes for studying the inhibition of activity of HCN channel gating (e.g., HCN1 channel gating)) and as therapeutic agents (e.g., in the treatment of diseases associated with overexpression and / or abnormal activity of HCN channel gating (e.g., HCN1 channel gating). In certain embodiments, the compound inhibits HCN channel gating (e.g., HCN1 channel gating) without enhancing the activity of gamma-aminobutyric acid-A (GABA-A) receptors (e.g., increasing their activity). In certain embodiments, the compound inhibits HCN channel gating (e.g., HCN1 channel gating) without modulating the activity of GABA-A receptors. In certain embodiments, the diseases treated and / or prevented include, but are not limited to, pain (e.g., chronic pain) in a subject. In certain embodiments, pain is associated with abnormal activity of HCN channel gating (e.g., HCN1 channel gating). The present disclosure also provides pharmaceutical compositions, kits, methods and uses of the compounds of Formula (I) or Formula (II) described herein.

[0281] Certain compounds described herein bind, covalently modify, antagonize, and / or inhibit HCN channel gating (e.g., HCN1 channel gating). In certain embodiments, the compounds described herein modulate the activity of HCN channel gating (e.g., HCN1 channel gating). In certain embodiments, the compounds described herein inhibit the activity of HCN channel gating (e.g., HCN1 channel gating).

[0282] It is expected that the compounds described herein can be used to treat and / or prevent diseases associated with abnormal activity (e.g., increased activity, undesirable activity, abnormal activity) of HCN channel gating (e.g., HCN1 channel gating). It is known in the art that HCN channel gating is implicated in a variety of diseases and conditions in subjects (such as pain (e.g., chronic pain)). Therefore, the compounds described herein are expected to be used to treat and / or prevent diseases (e.g., pain (e.g., chronic pain)).

[0283] The present disclosure also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating a disease in a subject, such as pain (e.g., chronic pain). The present disclosure also provides a compound of formula (II) or a pharmaceutically acceptable salt thereof for use in treating a disease in a subject, such as pain (e.g., chronic pain).

[0284] The present disclosure also provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicine for treating a disease, such as pain (e.g., chronic pain). The present disclosure also provides the use of a compound of formula (II) or a pharmaceutically acceptable salt thereof in the manufacture of a medicine for treating a disease, such as pain (e.g., chronic pain).

[0285] In certain embodiments, methods are provided for reducing the activity of HCN channel gating (e.g., HCN1 channel gating) in a subject (e.g., cell, tissue) by at least about 1%, at least about 3%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% by the methods described herein. In certain embodiments, the activity of HCN channel gating (e.g., HCN1 channel gating) in a subject or cell is reduced by at least about 1%, at least about 3%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% by the methods described herein. In some embodiments, the activity of HCN channel gating (e.g., HCN1 channel gating) in a subject is selectively inhibited by the methods. In some embodiments, the activity of HCN channel gating (eg, HCN1 channel gating) in a subject or cell is selectively reduced by the methods.

[0286] In another aspect, the disclosure provides methods of inhibiting the activity of HCN channel gating (eg, HCN1 channel gating) in a cell, the method comprising contacting the cell with an effective amount of a compound described herein or a pharmaceutical composition thereof.

[0287] In certain embodiments, the subject being treated is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human mammal. In certain embodiments, the subject is a domestic animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a companion animal, such as a dog or cat. In certain embodiments, the subject is a livestock, such as a cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal, such as a rodent, dog, or non-human primate. In certain embodiments, the subject is a non-human transgenic animal, such as a transgenic mouse or transgenic pig. In certain embodiments, the subject is a fish or a reptile. In certain embodiments, the subject is an animal. Such an animal can be of any sex and can also be at any stage of development. In certain embodiments, the subject described herein is a human. In certain embodiments, the subject is a non-human animal. In certain embodiments, the subject is a mammal.

[0288] In certain embodiments, the cell contacted with a compound or composition described herein is in vitro. In certain embodiments, the cell contacted with a compound or composition described herein is in vivo.

[0289] In certain embodiments, the disease (e.g., pain) to be treated or prevented using the compounds described herein may be associated with overexpression of HCN channel gating (e.g., HCN1 channel gating). The disease (e.g., pain) may be associated with abnormal activity of HCN channel gating (e.g., HCN1 channel gating). The abnormal activity of HCN channel gating (e.g., HCN1 channel gating) may be an increase and / or inappropriate and / or undesirable activity of the HCN channel. The compounds described herein and their pharmaceutically acceptable salts, solvates can inhibit the activity of HCN channel gating (e.g., HCN1 channel gating) and can be used to treat and / or prevent diseases (e.g., pain). The compounds described herein and their pharmaceutically acceptable salts can inhibit the activity of HCN channel gating (e.g., HCN1 channel gating) and can be used to treat and / or prevent diseases (e.g., pain). The compounds described herein and their pharmaceutically acceptable salts can inhibit the activity of HCN channel gating (e.g., HCN1 channel gating) and can be used to treat and / or prevent diseases (e.g., pain). The compounds described herein and their pharmaceutically acceptable salts can inhibit the activity of HCN channel gating (e.g., HCN1 channel gating) and can be used to treat and / or prevent diseases (e.g., pain).

[0290] Pharmaceutical compositions, kits and administration

[0291] The present disclosure also provides pharmaceutical compositions comprising the compounds described herein and optional pharmaceutically acceptable excipients. In certain embodiments, the compounds described herein are compounds of formula (I), formula (II) or pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients.

[0292] In certain embodiments, the compounds described herein are provided in an effective amount in a pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a preventive effective amount. In certain embodiments, the therapeutically effective amount is an amount that effectively inhibits the abnormal activity of HCN channel gating (e.g., HCN1 channel gating). In certain embodiments, the therapeutically effective amount is an amount that effectively treats a disease in a subject (e.g., a disease associated with abnormal activity of HCN channel gating (e.g., pain)). In certain embodiments, the preventive effective amount is an amount that effectively inhibits the abnormal activity of HCN channel gating (e.g., HCN1 channel gating). In certain embodiments, the preventive effective amount is an amount that effectively inhibits the abnormal activity of HCN channel gating (e.g., HCN1 channel gating).

[0293] In certain embodiments, an effective amount is an amount that effectively inhibits the activity of HCN channel gating (e.g., HCN1 channel gating) by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98%. In certain embodiments, an effective amount is an amount that effectively inhibits the activity of HCN channel gating (e.g., HCN1 channel gating) by no more than 10%, no more than 20%, no more than 30%, no more than 40%, no more than 50%, no more than 60%, no more than 70%, no more than 80%, no more than 90%, no more than 95%, or no more than 98%.

[0294] Another aspect of the present disclosure relates to methods of inhibiting the activity of HCN channel gating (e.g., HCN1 channel gating) in a subject. In certain embodiments, the methods described herein include administering to the subject an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0295] In certain embodiments, the compound or pharmaceutical composition is a solid. In certain embodiments, the compound or pharmaceutical composition is a powder. In certain embodiments, the compound or pharmaceutical composition can be dissolved in a liquid to make a solution. In certain embodiments, the compound or pharmaceutical composition is dissolved in water to make an aqueous solution. In certain embodiments, the pharmaceutical composition is a liquid for parenteral injection. In certain embodiments, the pharmaceutical composition is a liquid for oral administration (e.g., intake). In certain embodiments, the pharmaceutical composition is a liquid (e.g., an aqueous solution) for intravenous injection. In certain embodiments, the pharmaceutical composition is a liquid (e.g., an aqueous solution) for subcutaneous injection.

[0296] After being formulated with suitable pharmaceutically acceptable excipients at desired doses, the pharmaceutical compositions of the present disclosure can be administered to humans and other animals orally, parenterally, intracisternal, intraperitoneally, topically, buccally, etc., depending on the disease or condition to be treated.

[0297] In certain embodiments, the pharmaceutical composition comprising the compound of Formula I is administered orally or parenterally with each pharmaceutical composition being sufficient to deliver a dosage level of about 0.001 mg / kg to about 200 mg / kg in one or more doses, for one or more days (depending on the mode of administration). In certain embodiments, the effective amount of each dose is about 0.001 mg / kg to about 200 mg / kg, about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 50 mg / kg, preferably about 0.1 mg / kg to about 40 mg / kg, preferably about 0.5 mg / kg to about 30 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg of the subject's body weight per day, once or more in the range of obtaining the desired therapeutic effect and / or preventive effect. In certain embodiments, the compounds described herein may be in a dosage level sufficient to deliver about 0.001 mg / kg to about 200 mg / kg, about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 50 mg / kg, preferably about 0.1 mg / kg to about 40 mg / kg, preferably about 0.5 mg / kg to about 30 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg, and more preferably about 1 mg / kg to about 25 mg / kg of the subject's body weight per day, once or more a day to obtain the desired therapeutic effect and / or preventive effect. The desired dose can be delivered three times a day, twice a day, once a day, every other day, every three days, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen or more administrations) can be used to deliver the desired dose. In certain embodiments, a composition described herein is administered at a dose below that at which the agent causes non-specific effects.

[0298] In certain embodiments, the pharmaceutical composition is administered at a dosage of about 0.001 mg to about 1000 mg per unit dose. In certain embodiments, the pharmaceutical composition is administered at a dosage of about 0.01 mg to about 200 mg per unit dose. In certain embodiments, the pharmaceutical composition is administered at a dosage of about 0.01 mg to about 100 mg per unit dose. In certain embodiments, the pharmaceutical composition is administered at a dosage of about 0.01 mg to about 50 mg per unit dose. In certain embodiments, the pharmaceutical composition is administered at a dosage of about 0.01 mg to about 10 mg per unit dose. In certain embodiments, the pharmaceutical composition is administered at a dosage of about 0.1 mg to about 10 mg per unit dose.

[0299] The pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. Generally, these preparation methods include the following steps: associating a composition comprising a compound of Formula I with a carrier and / or one or more other auxiliary ingredients, and then, if necessary and / or desired, shaping and / or packaging the product into a desired single-dose or multi-dose unit.

[0300] Pharmaceutical compositions can be prepared, packaged and / or sold in bulk as a single unit dose and / or multiple single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to a subject and / or a convenient fraction of this dose, such as one-half or one-third of this dose.

[0301] In the pharmaceutical compositions of the present invention, the relative amounts of active ingredient, pharmaceutically acceptable excipient and / or any additional ingredients will vary, depending on the identity, size and / or condition of the subject being treated, and further on the route of administration of the composition. For example, the composition may contain between 0.1% and 100% (w / w) active ingredient.

[0302] Pharmaceutically acceptable excipients used to make the provided pharmaceutical compositions include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. Excipients such as cocoa butter and suppository waxes, colorants, coating agents, sweeteners, flavoring agents and perfuming agents may also be present in the compositions.

[0303] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, dibasic calcium phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and mixtures thereof.

[0304] Exemplary granulating and / or dispersing agents include potato starch, corn starch, tapioca starch, sodium glycolate starch, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium glycolate starch), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (magnesium aluminosilicate), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0305] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, waxes, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and magnesium aluminosilicate (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glycerol monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60), polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate (Span 40), sorbitan monostearate (Span 60), sorbitan tristearate (Span 65), glycerol monooleate, sorbitan monooleate (Span 80)), polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45), polyoxyethylene hydrogenated castor oil, polyoxyethylene castor oil, polyoxymethylene stearate and Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor TM ), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether (Brij 30)), poly(vinyl pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic F-68, poloxamer-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium and / or mixtures thereof.

[0306] Exemplary binders include starch (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., gum arabic, sodium alginate, Irish moss extract, panwar gum, ghatti gum, isapol husk pulp, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl pyrrolidone), magnesium aluminum silicate (magnesium aluminosilicate), and larch arabinogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohols, and / or mixtures thereof.

[0307] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives and other preservatives. In certain embodiments, the subject is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0308] Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0309] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and its salts and hydrates (e.g., sodium EDTA, disodium EDTA, trisodium EDTA, disodium EDTA calcium, dipotassium EDTA, etc.), citric acid and its salts and hydrates (e.g., citric acid monohydrate), fumaric acid and its salts and hydrates, malic acid and its salts and hydrates, phosphoric acid and its salts and hydrates, and tartaric acid and its salts and hydrates. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerol, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0310] Exemplary antifungal preservatives include butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0311] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and phenethyl alcohol.

[0312] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

[0313] Other preservatives include tocopherol, tocopheryl acetate, deferoxamine mesylate, cetrimonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, and Euxyl.

[0314] Exemplary buffers include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glucuronate, calcium glucoheptonate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium levulinate, valeric acid, dibasic calcium phosphate, phosphoric acid, tricalcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium hydrogen phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and mixtures thereof.

[0315] Exemplary lubricants include magnesium stearate, calcium stearate, stearic acid, silicon dioxide, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0316] Exemplary natural oils include almond oil, apricot kernel oil, avocado oil, babassu oil, bergamot oil, black currant oil, borage oil, juniper oil, chamomile oil, canola oil, caraway oil, Brazilian palm oil, castor oil, cinnamon oil, cocoa butter oil, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, evening primrose oil, fish oil, linseed oil, geraniol oil, gourd oil, grapeseed oil, hazelnut oil, hyssop oil, isopropyl myristate oil, jojoba oil, macadamia nut oil, lavender oil, lavender oil. , lemon oil, litsea cubeba oil, macadamia oil, mallow oil, mango seed oil, meadowfoam seed oil, mink oil, nutmeg oil, olive oil, citrus oil, orange roughy oil, palm tree oil, palm kernel oil, peach kernel oil, peanut oil, poppy seed oil, pumpkin seed oil, rapeseed oil, rice bran oil, rosemary oil, safflower oil, sandalwood oil, camellia (sasquana) oil, savory oil, sea buckthorn oil, sesame oil, shea butter, silicone oil, soybean oil, sunflower oil, tea tree oil, thistle oil, toon oil, vetiver oil, walnut oil and wheat germ oil. Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil and mixtures thereof.

[0317] Liquid dosage forms for oral and parenteral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active agent, the liquid dosage form can contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof. In addition to the inert diluent, oral compositions can also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and flavoring agents. In certain embodiments of parenteral administration, the medicament of the present invention is mixed with a solubilizer, such as polyoxyethylene castor oil (polyethoxylated castor oil), alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers and combinations thereof.

[0318] Injectable preparations (e.g., sterile injectable aqueous or oily suspensions) can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in nontoxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable carriers and solvents that can be employed are water, Ringer's solution, USP, and isotonic sodium chloride solution. In addition, sterile fixed oils are typically used as solvents or suspension media. For this reason, any gentle fixed oil can be employed, including synthetic monoglycerides or diglycerides. In addition, fatty acids (such as oleic acid) are used to prepare injections.

[0319] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0320] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active agent is mixed with at least one inert, pharmaceutically acceptable excipient or carrier (such as sodium citrate or dicalcium phosphate) and / or the following: a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; b) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic; c) wetting agents, such as glycerol; d) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; e) solution retardants, such as paraffin; f) absorption promoters, such as quaternary ammonium compounds; g) wetting agents, such as, for example, cetyl alcohol and glyceryl monostearate; h) absorbents, such as kaolin and bentonite; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0321] Solid compositions of similar types can also be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells (such as other coatings well-known in the field of enteric coatings and pharmaceutical formulations). They can optionally contain opacifiers and can also be compositions that release active ingredients only or preferentially in a delayed manner at a certain position in the intestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of similar types can also be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0322] The active agent can also be in the form of a microcapsule with one or more of the above-mentioned excipients. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells (such as enteric coatings, controlled release coatings and other coatings well-known in the field of drug formulation). In this solid dosage form, the active agent can be mixed with at least one inert diluent (such as sucrose, lactose or starch). This dosage form can also generally include other substances that are normally practical except the inert diluent, such as tableting lubricants and other tableting aids, (such as magnesium stearate and microcrystalline cellulose). In the case of capsules, tablets and pills, the dosage form can also include a buffer. They can optionally contain an opacifier, and can also be the following composition, which only or preferentially optionally releases the active ingredient in a delayed manner in a certain part of the intestinal tract. Examples of usable embedding compositions include polymeric substances and waxes.

[0323] The formulation suitable for topical application includes liquid or semi-liquid preparations, such as liniments, lotions, gels, applications, oil-in-water or water-in-oil emulsions, such as creams, ointments or pastes; or solutions or suspensions, such as drops. The formulation for topical application to the skin surface can be prepared by dispersing the drug with a dermatologically acceptable carrier (such as a lotion, cream, ointment or soap). Useful carriers can form a film or layer on the skin for topical application and inhibition of removal. For topical application to internal tissue surfaces, the medicament can be dispersed in a liquid tissue adhesive or other known substances that enhance tissue surface adsorption. For example, hydroxypropylcellulose or fibrinogen / thrombin solutions can be advantageously used. Alternatively, tissue coating solutions, such as pectin-containing formulations, can be used. Ophthalmic formulations, ear drops and eye drops are also considered to be within the scope of the present invention. In addition, the present disclosure contemplates the use of transdermal patches, which have the additional advantage of providing controlled delivery of medicaments to the body. This dosage form can be prepared by dissolving or distributing the medicament in a suitable medium. Absorption enhancers can also be used to increase the flux of the agent across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the agent in a polymer matrix or gel.

[0324] In addition, the carrier of the topical formulation can be in the form of a hydroalcoholic system (e.g., liquids and gels), an anhydrous oil or silicone system, or an emulsion system (including but not limited to oil-in-water, water-in-oil, water-in-oil-in-water, and water-in-oil-in-silicone emulsions). The emulsions can cover a wide range of consistencies, including thin lotions (which may also be suitable for spray or aerosol delivery), ointment lotions, light creams, thick creams, etc. Emulsions can also include microemulsion systems. Other suitable topical carriers include anhydrous solids and semisolids (such as gels and sticks); and water-based mousse systems.

[0325] In certain embodiments, the kits described herein include a first container comprising a compound or pharmaceutical composition described herein. In certain embodiments, the kits described herein can be used to treat and / or prevent diseases in subjects, such as pain (e.g., chronic pain). In certain embodiments, the kits described herein are used to inhibit the activity of HCN channel gating (e.g., HCN1 channel gating) in subjects. In certain embodiments, the kits described herein also include instructions for using the compounds or pharmaceutical compositions included in the kits. The kits described herein may also include information required by regulatory agencies such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information included in the kits is prescription information. In certain embodiments, the kits and instructions provide for treating diseases in subjects (e.g., pain (e.g., chronic pain)), inhibiting the activity of HCN channel gating (e.g., HCN1 channel gating). The kits described herein may include one or more additional agents described herein as separate compositions.

[0326] Example

[0327] In order that the present disclosure may be more fully understood, the following examples are set forth.The synthetic and biological examples described in this application are intended to illustrate the compounds, pharmaceutical compositions and methods provided herein and should not be construed in any way as limiting the scope thereof.

[0328] Example 1. Exemplary compounds are potent antagonists of HCN1 channels when applied externally in two-electrode voltage clamp (TEVC)

[0329] Figure 3 BP4C-11:0:1 and BP4L-10:0:1 are shown relative to a hypothetical anchor-tethered 2,6-DTBP. Figure 4 Recordings from three individual cells expressing HCN1-ΔNvΔCv are shown. Each cell was preincubated for 20 min alone or with 10 μM BP4C-11:0:1 or BP4L-10:0:1 as indicated in recording solution containing 0.04% DMSO. Current-voltage relationships (IV) were collected with 3- and 5-second activation steps (5-s scans are shown in each case). V reported by both protocols 1 / 2 There were no systematic differences, indicating that activation in both conditions was balanced at each voltage in all drug and vehicle conditions (not shown). Scans and tails acquired in response to activation at -65 mV are highlighted in black. Figure 4 ( Figure 5 ) The tail current activation curves of the three cells shown are fitted with a Boltzmann function. The data for BP4C-11∶0∶1 are from 9 cells (3 cells expressing wtHCN1 at 10μM, 3 cells expressing HCN1-ΔNvΔCv at 10μM; 2 cells expressing wtHCN1 at 100μM, and 1 cell expressing HCN1-ΔNvΔCv at 100μM). The data for BP4L-10∶0∶1 are from 6 cells (3 cells expressing HCN1-ΔNvΔCv at 10μM, and 3 cells expressing wtHCN1 at 100μM). The smooth line is the fit of the Hill function to the 2,6-DTBP data and its right-shifted version shown.

[0330] BP4C-11:0:1 is a tethered but unanchored 2,6-DTBP. That is, BP4C-11:0:1 is a molecule that should be relatively freely distributed into the membrane. In fact, the new data greatly expand this point, as they demonstrate that 4-adduct tolerance is retained when the more bulky 2,6-DTBP pharmacophore is used in place of the diisopropylphenol head group. It was previously assumed that a long tether could be attached to the 4 position.

[0331] BP4L-10:0:1 is a tethered and anchored 2,6-DTBP, with the diol group strongly preferring to remain in the water compartment. This suggests that pharmacophore penetration of the membrane is still required when the more bulky 2,6-DTBP pharmacophore is used in place of diisopropylphenol. The alkylphenol sites on HCN1 are important and perhaps measurable (although still unknown) depths of penetration into the membrane.

[0332] BP4C-11:0:1 and BP4L-10:0:1 are easily soluble in DMSO (dimethyl sulfoxide) and disperse fairly easily in aqueous buffers.

[0333] Data were collected from wild-type HCN1 (wtHCN1) and truncated HCN1 channels heterologously expressed in Xenopus oocytes in which the variable N and C domains were removed but the cyclic intact nucleotide binding domain (CNBD) was retained (HCN1-ΔNvΔCv). The HCN1-ΔNvΔCv gate is essentially the same as wtHCN1, but has the advantage that it is amenable to inside-out patch clamp (IOPC) recording techniques, whereas wtHCN1 is extremely difficult to record in IOPCs due to its tendency to cluster. The data recordings presented here were acquired using two-electrode voltage clamp.

[0334] The data is presented as V 1 / 2 relative to the displacement of the appropriate solvent mediator (ΔV 1 / 2 ), where V 1 / 2 is the midpoint of voltage activation. Since cells were not exposed to more than one condition, the population mean of Mediator was subtracted from each individual plus drug measurement. The drug effects were indistinguishable for wtHCN1 and HCN1-ΔNvΔCv. Therefore, the ΔV values ​​for wtHCN1 and HCN1-ΔNvΔCv were subtracted from the population mean of Mediator. 1 / 2 The values ​​were combined for this preliminary data set and the data are referred to simply as HCN1 unless they were from a specific cell.

[0335] Example 2. 2,6-DTBP is a potent HCN1 selective inverse agonist.

[0336] Effects of propofol and 2,6-DTBP on the V of HCN1-4 channels 1 / 2 *Indicates a response that is statistically different from the control group ( Figure 7 ). 2,6-(left) or 2,4-(right) dibutylphenol on the V of HCN1 channel 1 / 2 The solid line and the indicated parameter values ​​are from the Hill function fit to the 2,6-DTBP relationship; the dashed lines are the fits shifted 2, 15, and 23 times for 2,4-DTBP, 2,6-DSBP, and 2,4-DSBP, 2,6-DTBP, respectively. 1 / 2The shift was significant at ≥1 μM for DTBP and significant at 20 μM for DSBP ( Figure 8 ).

[0337] According to EC 50 The rank order of estimated HCN1 antagonism is as follows Fig. 9 The rate of Tl+-mediated fluorescence quenching in large unilamellar vesicles demonstrates that these compounds have minimal membrane effects ( Fig.10 Structures of three 2,6-alkylphenols and their efficacy as general anesthetics (GA) and positive modulators of GABAA receptor (GABAA-R) function ( Fig.11 ).

[0338] The novel compounds of the present technology are therefore demonstrated to be useful as anti-hyperalgesic agents for the treatment of peripheral neuropathic pain. The data demonstrate that: (1) 2,6- and 2,4-di-tert-butylphenol (2,6- and 2,4-DTBP) are more potent inverse agonists of HCN1 than the intravenous general anesthetic propofol (2,6-diisopropylphenol), (2) 2,6- and 2,4-di-sec-butylphenol (2,6- and 2,4-DSBP) are less potent, and (3) 2,6-DTBP retains the selectivity of propofol for HCN1 versus HCN2-4.

[0339] Example 3. 2,6-DTBP selectively inhibits mechanical and thermal hyperalgesia associated with mechanical and thermal nociception.

[0340] Fig.12 Display P w，IPSI and P W，CONTRA (probability of withdrawal of the paw ipsilateral or contralateral to the nerve ligation) and the effect of the indicated cumulative ip doses of 2,6-DTBP as a function of the intensity of stimulated fibers ( Fig.12 )). Fig.13 A and Fig.13 B shows HPWL (hind paw withdrawal latency) as a function of cumulative ip 2,6-DTBP dose at low stimulation intensity (15%) and high stimulation intensity (30%).

[0341] Notably, DTBP is ineffective as a general anesthetic due to lack of efficacy as a functional agonist of GABA-A receptors; additional data suggest that alkylphenols impair HCN1 gating through a spatially confined site where a network of hydrogen bonds contributes to the initial binding energy while coupling energy is rarely involved.

[0342] Example 4. Access of the pharmacophore to its site of action depends on the tether length.

[0343] New chemical entities (NCEs) were synthesized that retained 2,6-DTBP as the pharmacophore and modified it with an emphasis on the "tether anchor" in order to maintain selective inhibition of HCN1 while preventing passage across lipid membranes (and ultimately to the CNS). One of the compounds, BP4L-18:1:1, possessed many desirable properties.

[0344] Current recordings from individual cells expressing HCN1. Each cell was pre-incubated for 20 min in a recording solution containing 20 mM 2-hydroxypropyl-β-cyclodextrin (HPβCD) + 10 mM DMSO alone or with 30 μM BP4L-18:1:1 as indicated ( Fig.15 ). The black scans are those recorded with an activation potential of -65 mV. Inhibition of channel gating is concentration dependent ( Fig.16 BP4L-18:1:1 effectively inhibited channel gating with a potency and efficacy indistinguishable from free 2,6-DTBP (compare Fig.17 ), while BP4L-10∶0∶1 (which restricts access to the pharmacophore in the binding site) was largely ineffective ( Fig.17 ).

[0345] Example 5. Interaction of alkylphenols and alkylcyclohexanols with the HCN1 channel gating model.

[0346] Fig.18 CR, CA, and O are shown, which represent closed-resting, closed-activated, and open states; KV and L are equilibrium constants describing activation and openness; I and H are inverse agonists and partial inverse agonists; p and q represent the number of binding events for I and H; KC and KO represent the association constants of I and H with closed and open channels, with I and H labeled with appropriate superscripts. For simplicity, only activation and open transitions between uncoordinated states are shown, but all CR-CA and CA-O transitions are allowed. When H and I are associated with a common site, the model contains 3 states per plane, 9 states in total (N=1), and 15 states per plane, 45 states in total (N=4). When H and I are associated with different sites, the model contains 4 states per plane, 12 states in total (N=1), and 25 states per plane, 75 states in total (N=4). Importantly, since the equilibrium constant is the only free parameter and the number of equilibrium constants is independent of the spread in the number of states, if these models are equally able to describe the data, then each of them should be equally well determined.

[0347] Example 6. Hydrogen bonding functionality at position 1 is useful for HCN1-gated alkylbenzene inhibition.

[0348] Fig.19ARepresentative voltage clamp recordings of HCN1 currents (left) are shown after 20 min incubation in the absence or presence of 10 μM of the indicated reagents. The tail currents (right) are shown on an expanded time scale. In each case, the black trace is the current recorded at an activation potential of -75 mV. All recordings were obtained on the same day from different oocytes from one donor frog. Scale bars are 2 μA and 1 s (left) and 200 nA and 50 ms (right). Fig.19B The chemical representation of 2,6-diisopropylbenzene is shown. The substitution at the 1-position (according to the legend) describes its effect on Fig.19A and Fig.19C The molecules reported in . Fig.19C Displayed by Fig.19A Normalized steady-state activation curve constructed from the recordings shown. The smoothed line is the fit of the Boltzmann function. Symbols represent Fig.19B The molecules described.

[0349] Example 7. Inhibitory effects of 2,6-dialkylbenzene derivatives reveal the different contributions of hydrogen bonding potential, alkyl side chain identity, and the presence or absence of π electrons to drug function.

[0350] Fig. 20 Displays V as a function of each specified ligand concentration 1 / 2 In each panel, the thick black line is the Hill fit to 2,6-DTBP. The thin grey line is the Hill fit to 2,6-DTBP in the presence of 50 Shifted on the concentration axis; the thick gray line is the Hill equation fit to the concentration response in the plate; and the dashed gray lines in the 2,6-DIPP and 2,6-DTBP plate plots are the Hill equation fits to the 2,6-DIPCH and 2,6-DTBCH data, with only the maximum response of the fit changing so that the cyclohexanol fit line varies according to the ratio ΔV 1 / 2 Maximum phenol / ΔV 1 / 2 The maximum cyclohexanol was scaled, and all other terms of the cyclohexanol fit were held constant. All data are mean ± SEM, with 6 or more determinations per point.

[0351] Example 8. Inverse agonist potency of 2,6-dialkylbenzene analogs as a function of molecular volume.

[0352] Fig.21A The observed aqueous EC 50 Plotted as a function of the calculated molecular volume. For fully (2,6-DIPP, 2,6-DTBP) and partially (2,6-DEP, 2,6-DEIC, 2,6-DIPCH, 2,6-DTBCH, 2,6-DSBP) potent ligands, EC 50 According to the Hill equation Fig. 20For ligands with poorly defined maximal potency to allow Hill function fitting (2,6-DMBT, 2,6-DIPBT, 2,6-DIPFB, 2,6-DECH), the Hill fit of 2,6-DTBP was determined based on the shifted Fig. 20 The thin grey line in the figure estimates the EC 50 When Fig. 20 When no inflection point is observed in the concentration-response curve, the corrected EC 50 is set equal to 100 mM. Fig.21A and Fig. 21B In both cases, the dashed ellipse encloses the data for the isopropyl family of reagents. Fig. 21B The cumulative corrected EC is plotted as a function of the calculated molecular volume. 50 (Aqueous EC determined as described in A 50 For clarity, 2,6-DMP (▲), 2,6-DMIC (▲), 2,6-DMCH (▲), 2,6-DECH (■), and 2,6-DSBCH are omitted. ).

[0353] Example 9. Alkylphenols and alkylcyclohexanols are associated with the HCN1 channel through four common, radially distributed sites.

[0354] Fig. 22 A and Fig. 22 B is shown as isopropyl ( Fig. 22 A) and tert-butyl ( Fig. 22 B) ΔV as a function of phenol and cyclohexanol concentrations 1 / 2 Data from Fig. 20 The solid black line is the common site model (N = 4) for the data in these two plates and the plate Fig. 22 Simultaneous fitting of data in CE. Fig. 22 CE showed that in the presence of 3 μM 2,6-DTBP, as 2,6-DIPCH ( Fig. 22 C), 2,6-DTBCH ( Fig. 22 D) or 2,6-DIPP ( Fig. 22 E) ΔV as a function of concentration 1 / 2 . Black solid line as shown in the figure Fig. 22 A and Fig. 22 B. The black dashed lines are the predicted behavior of the different site models using the association constants determined from the fit of the common site model when N = 4. The gray lines report the ΔV elicited by 3 μM 2,6-DTBP in the paired controls under each additivity condition. 1 / 2 Mean (solid lines) and SEM (dashed lines). Fig. 22 F shows the common site model based on N=4 (based on Fig. 22 A to Fig. 22 E) The association constant determined by the fit of the model was converted into a free energy term according to -(RT)LnK. The reported value of the open equilibrium constant (L) is from this same fit.

[0355] Example 10. Reduced inverse agonist potency of large 2,6-dialkylbenzene analogs indicates a clear upper limit to drug cavity volume.

[0356] Fig.23 Shown in the absence of 3 μM 2,6-DTBP ( Fig.23 A) and existence ( Fig.23 B) ΔV1 / 2 as a function of 2,6-DSBP concentration. The solid black line is a simultaneous fit of the common site model (N=4) to the data in these two panels, with all constants except JoI and JcI kept constant. Fig.23 The JoI and JcI determined from these fits were 881 ± 1,801 and 3,696 ± 2,948, respectively. Fig.23 The black dashed line in B is the predicted behavior of the different site model with N = 4 parameters as described above. When N equals 1 and 4, the discrete fit of the four models produces χ2 values ​​of 600 and 128 (common sites) and 165 and 167 (different sites), respectively. Fig.23 The symbols and gray lines in A are for Fig. 20 Plate map of 2,6-DSBP replicates of 2,6-DSBP and EC 50 Adjusted 2,6-DTBP fitting line (EC 50 was set to 370 μM) and the ΔV 1 / 2 .exist Fig.23 In B, the gray line reports the ΔV elicited by 3 μM 2,6-DTBP in paired controls. 1 / 2 Mean (solid lines) and SEM (dashed lines).

[0357] Example 11. Simulation of the interaction of exemplary compounds with cell membrane lipid bilayer

[0358] 2,6-DTBP is highly mobile and explores the entire volume of the hydrophobic core of the lipid bilayer and adopts completely random orientations ( Fig. 27). Furthermore, the diol-anchored molecules do not behave like free 2,6-DTBP. Despite being more hydrophobic overall, the diol-anchored molecules are not freely distributed into the lipid phase nor can they move freely within the lipid phase. Visual inspection of the simulation trajectories suggests that the anchors generally function as designed. Thus, in both molecules (BP4L-18:1:1 and BP4L-10:0:1), the diol exhibits a strong preference for polar environments and the diol settles into the hydrated headgroup phase and spends little time in the hydrophobic core or away from entering the aqueous phase.

[0359] Long and short molecules (BP4L-18:1:1 and BP4L-10:0:1, respectively) tend to stay in a perpendicular orientation in a manner that is typically by design. This behavior may be a reflection of two design constraints: The diols are preferentially retained at the hydrophobic-polar interface. The stacking interactions of the acyl tethers embedded within the hydrophobic core. The tails of the membrane lipids present a dynamic, but relatively ordered space that would impose an energetic cost on the tethers to adopt any orientation other than perpendicular. This cost would likely be the result of a combination of tether C-C bond rotations and ocean-coupled displacements of the membrane lipid (ordered) tails. While bond rotations present an intrinsically low thermodynamic cost, displacements of the membrane tails would represent a high entropic cost.

[0360] Due to the flexibility of the tether, the time-averaged depth of the pharmacophore appears to be smaller than the calculated average linear length for a static molecule, but this is expected. On the other hand, since the diol is located in the polar headgroup phase, rather than outside / above it as expected for a fully efficient anchor, the pharmacophores of both BP4L-10:0:1 and BP4L-18:1:1 extend deep into the membrane. Both can reach the middle of the bilayer. This rarely happens for BP4L-10:0:1; for BP4L-18:1:1, this may be the preferred arrangement. In fact, BP4L-18:1:1 can exceed the midpoint of the bilayer without the diol anchor separating from the headgroup phase.

[0361] Regardless of the overall perpendicular orientation of the tether, both BP4L-10:0:1 and BP4L-18:1:1 exhibit considerable flexibility along the long axis of the tether, allowing the pharmacophore to explore different depths and orientations associated with the channel protein, which would be (largely) orthogonal to the bilayer. The presentation of the alkyl-hydroxyl face of the pharmacophore orthogonal to the lipid tail appears to require rotation of the molecule about a series of bonds to bring the pharmacophore toward the surface.

[0362] In the initial 300 ns runs, neither molecule showed any obvious flipping. However, BP4L-18:1:1 apparently made several (brief) "diving" sojourns to the inner leaflet. In this arrangement, the phenol was temporarily bound to the headgroup of the inner leaflet, while the diol was in the lipid phase.

[0363] Example 12. Compound Synthesis

[0364] Synthesis of BP4C-11∶0∶1

[0365]

[0366] At 0 ℃, under inert atmosphere, titanium tetrachloride (107 μ L, 0.96 mmol) is added dropwise to a solution of 2,6-di-tert-butylphenol (200 mg, 0.96 mmol) and 10-undecenoyl chloride (206 μ L, 0.96 mmol) in dichloromethane (3.0 mL). The reaction mixture is stirred at 0 ℃ for 30 minutes. Add saturated ammonium chloride solution (2.0 mL) and water (10 mL) to quench the reaction. Then, extract the aqueous layer with ethyl acetate (2x10 mL). Wash the combined ethyl acetate layer with saline (10 mL), dry with anhydrous sodium sulfate, filter and concentrate. The crude residue was purified by silica gel column chromatography using EtOAc:hexanes (1:25) to provide BP4C-11:0:1 (290 mg, 74% yield) as an oil; Rf = 0.36 using 3% ethyl acetate in hexanes as the developing solvent on silica TLC plates. 1 H NMR (500 MHz, CDCl3): 8 = 1 H NMR (500MHz, CDC13): 8=7.85 (s, 2H), 5.69 (s, 1H), 4.04-4.00 (m, 1H), 2.90 (t, J=7 .4Hz, 2H), 1.74-1.68(m, 4H), 1.51-1.49(m, 3H), 1.47(s, 18H), 1.4-1.29(m, 10H).

[0367] Synthesis of BP4K-11∶0∶1

[0368]

[0369] 2,6-di-tert-butylphenol (294.6 mg, 1.42 mmol) was added to a mixture of trifluoroacetic anhydride (199 μL, 1.42 mmol) and 10-undecenoic acid (263 mg, 1.42 mmol) at 0°C under an inert atmosphere. The reaction mixture was stirred at room temperature for 3 hours and 30 minutes. The reaction mixture was then diluted to ethyl acetate (30 mL), and the resulting organic layer was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography using 3% EtOAc in hexane to provide the corresponding unsaturated ketone intermediate BP4K-11:0:1 (388 mg, 73% yield) in the form of an oil; Rf=0.36 using 3% ethyl acetate in hexane as a developing solvent on a silica TLC plate. 1 HNMR (500MHz, CDCl3): 8=7.85 (s, 2H), 5.85-7.76 (m, 1H), 5.70 (s, 1H), 5.00-4.91 (m, 2H), 2. 90(t, J=7.4Hz, 2H), 2.06-2.01(m, 2H), 1.17-1.69(m, 2H), 1.47(s, 18H), 1.37-1.30(m, 10H).

[0370] Synthesis of BP4L-10∶0∶1

[0371]

[0372] At room temperature, 4-methylmorpholino N-oxide (51 mg, 0.37 mmol) was added to the unsaturated ketone BP4K-11: 0: 1 (35 mg, 0.09 mmol) dissolved in a mixture of acetone (0.5 mL) and tert-butanol (100 μL). Then, water (300 μL) and a catalytic amount of osmium tetroxide were added, and the reaction was stirred for 40 minutes. Then, the reaction was diluted with ethyl acetate (10 mL), water (5 mL), and quenched with sodium bisulfite (87 mg, 0.43 mmol) dissolved in water (2.0 mL). The reaction mixture was stirred for 10 minutes. Then, the aqueous mixture was extracted with ethyl acetate (2x10 mL). The combined ethyl acetate layer was washed with saturated sodium bicarbonate solution (10 mL), water (15 mL), and saline (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product obtained was purified by silica gel column chromatography using EtOAc:hexane (9:1) to provide BP4L-10:0:1 (31 mg, 81% yield) as a yellow oil; Rf=0.38 using 30% ethyl acetate in hexane as the developing solvent on silica TLC plates. 1H NMR (500MHz, CDCl3): 8=7.84 (s, 2H), 5.70 (s, 1H), 3.72-3.69 (m, 1H), 3.66-3.63 (m, 1H), 3.45-3.41 (m, 1H) , 2.91-2.88(m, 2H), 2.21(br, 2H), 1.72-1.70(m, 2H), 1.46(s, 18H), 1.43-1.42(m, 3H), 1.33-1.31(m, 9H). 13 C NMR (125MHz, CDCl3): 8=200.2, 158.2, 135.7, 128.9, 125.8, 72.3, 66.9, 38.19, 34.3, 33.2, 29.5, 29.4, 29.3, 25.5, 24.8

[0373] General strategy for the synthesis of 4-(oxo)-substituted 2,6-dialkylphenols

[0374]

[0375] Exemplary Synthesis of 4-(Oxo)-Substituted 2,6-Dialkylphenols

[0376]

[0377] Oxacyclononadecane-2,19-dione: To a stirred solution of octadecane dioic acid (300 mg, 875 μmol) in dichloromethane (2 mL) was added acetic anhydride (860 μL, 8.76 mmol). The reaction mixture was stirred at ambient temperature for 3 h and then concentrated in vacuo to give the title compound (220 mg, 677 μmol, 77%) as a clear, colorless oil that foamed under vacuum. This material was then used without further purification. 1 H NMR (500MHz, DMSO-d6) δ2.22 (m, 4H), 1.55 (m, J=7.1Hz, 4H), 1.25 (br, 28H).

[0378]

[0379] 18-(3,5-di-tert-butyl-4-hydroxyphenyl)-18-oxooctadecanoic acid, 1: 2,6-di-tert-butylphenol (1.0 g, 4.85 mmol), oxacyclopentadecane-2,19-dione (1.44 g, 4.85 mmol) and dichloromethane (DCM, 100 mL) were added to an oven-dried flask. The resulting mixture was stirred magnetically until all reactants dissolved, and then aluminum chloride (646 mg, 4.85 mmol) was added in batches. The reaction was heated to 45 ° C and stirred overnight. The reaction was quenched by adding water (100 mL) and extracted with DCM (3x100 mL). The combined organic layer was dried (Na2SO4) and concentrated. Automatic flash chromatography (50% MeOH in DCM) was used to purify to obtain compound 1 (1.77 g, 3.34 mmol, 68%) in the form of a yellow oil. 1 H NMR (500MHz, CDCl3) δ7.84 (s, 2H), 5.72 (s, 1H) 3.49 (s, 2H), 2.55 (s, 3H), 2.22 (s, 31H), 1.47 (s, 20H), 1.39-1.14 (br, 21H), 0.93-0.81 (brm, 13H).

[0380]

[0381] 2,5-Dioxopyrrolidin-1-yl 18-(3,5-di-tert-butyl-4-hydroxyphenyl)-18-oxooctadecanoate, 2: Compound 1 (52 mg, 98 μmol), EDC (37 mg, 196 μmol), NHS (23 mg, 196 μmol) and DMF (1 ml) were added to an oven-dried flask. The mixture was stirred magnetically and triethylamine (27 μL, 196 μmol) was added. The reaction mixture was stirred overnight at ambient temperature, then the solvent was removed by rotary evaporation, and the crude mixture was purified using mass-guided LCMS. The fractions containing compound 2 were combined and lyophilized to give a slightly yellowish gel-like material. 1 H NMR (500MHz, CHCl3) δ7.86 (s, 3H), 5.74 (s, 1H), 2.58 (s, 4H), 2.18 (d, J = 16.2Hz, 2H), 1.94-1.55 (m, 3H), 1.49 (br, 25H), 1.43-0.77 (brm, 13H).

[0382]

[0383] BSA conjugate: BSA (66.5 kDa) was dissolved in 0.05 M phosphate buffered saline (pH 7.4) in a glass vial at a concentration of 2.5 mg / mL. 500 μL of compound 2 was added as a 55 mg / mL solution in DMF / acetonitrile (1:1 v / v). The mixture was incubated at 37°C for 12 hours, at which time LCMS reaction analysis showed no signs of compound 1 (m / z = 529.5 for [MH]) or compound 2 ([M+H] + The protein was precipitated by diluting 10 μL of the reaction mixture with 0.5 ml of chloroform, followed by centrifugation and sampling of the supernatant for analysis. The crude reaction mixture was freeze-dried and used without further purification.

[0384] Alternative strategies for the synthesis of 4-(oxo)-substituted 2,6-dialkylphenols

[0385]

[0386] Exemplary Synthesis of 4-(Oxo)-Substituted 2,6-Dialkylphenols with Trans-Alkenyl Moieties

[0387]

[0388] 1-(3,5-di-tert-butyl-4-hydroxyphenyl)undec-10-ene-1-one, 4: Anhydrous MeCN (10 mL) was added to an oven-dried pressure tube containing potassium iodide (1.64 g, 9.87 mmol) and 2,6-di-tert-butylphenol (1.02 g, 4.93 mmol), followed by addition of undec-10-enoyl chloride (2 g, 9.87 mmol). The tube was then sealed, and the reaction mixture was heated to 82 ° C, stirred for 24 hours, and then cooled to ambient temperature. The solution was quenched with water (10 mL) and extracted with DCM (3x50 mL). The combined organic layers were washed with saturated sodium thiosulfate solution, then dried (Na2SO4), and concentrated in vacuo. Purified by column chromatography (1% EtOAc in hexane) to obtain compound 4 (3.2 g, 10.2 mmol, 87%) in the form of a yellow oil. 1HNMR (500MHz, CDCl3) δ7.87 (s, 2H), 7.33 (d, J=7.9Hz, 2H), 7.15 (q, J=8.3Hz, 1H), 5.83 (m, 1H), 5.08-4.87 (m, 3H), 2.92 (m, 2H) , 2.70-2.57(m, 2H), 2.13-1.97(m, 3H), 1.97-1.90(m, 2H), 1.76(m, 5H), 1.69-1.54(m, 7H), 1.49(s, 20H), 1.42-1.28(br, 39H). C 25 H 40 ESIMS of O2; [MH] - Theoretical m / z = 371.3, measured m / z of [MH]- = 371.5

[0389]

[0390] (E) -1- (3,5-di-tert-butyl-4-hydroxyphenyl) -17- (oxirane-2-yl) heptadeca-10-ene-1-one, 5: Under an argon atmosphere, Grubbs second generation catalyst (114 mg, 1.34 mmol) was added to a stirred suspension of CuI (26 mg, 134 μmol) in a solution of compound 4 (500 mg, 1.34 mmol) and 2- (octa-7-ene-1-yl) oxirane (207 mg, 1.34 mmol) in dry DCM (6 mL). The resulting mixture was stirred at ambient temperature for 15 minutes, then heated to reflux and stirred for 4 hours. At this point, the reaction mixture was cooled to ambient temperature, and three drops of DMSO were added. The resulting solution was stirred overnight and then concentrated in vacuo. The resulting residue was purified by automated flash chromatography (EtOAc / pet ether, 15:85) to afford compound 5 (475 mg, 0.95 mmol, 71%) as a light brown liquid. 1 H NMR (500MHz, CDCl3) δ7.86 (s, 2H), 5.71 (s, 1H), 5.50-5.29 (m, 2H), 2.92 (t, J=7.3Hz, 3H ), 2.77(m, 1H), 2.48(m, 1H), 2.11-1.91(m, 5H), 1.81-1.65(m, 3H), 1.60-1.21(br, 43H).

[0391]

[0392] (E)-1-(3,5-di-tert-butyl-4-hydroxyphenyl)-18,19-dihydroxynonadeca-10-ene-1-one, 6 (BP4L-18:1:1): Compound 5 (600 mg, 1.20 mmol) was dissolved in a mixture of dioxane / water / H2SO4 (2:1:1 v / v., 7 mL) and heated under reflux for 18 hours. The reaction mixture was then cooled to ambient temperature, extracted with water and dichloromethane, and then concentrated in vacuo. The resulting residue was purified by automated flash chromatography (20% EtOAc in DCM) to give compound 6 (450 mg, 0.87 mmol, 72%) as a dark brown liquid. m / z = 514.4 for [MH]-, m / z = 514.5 for [MH]-, 1 HNMR (500MHz, CDCl3) δ7.86 (s, 2H), 5.72 (s, 1H), 5.50-5.30 (m, 2H), 3.78-3.63 (m, 3H), 3.46 (dd, J=11.0, 7.7Hz, 1H), 2.92 (t, J=7.4Hz, 2H), 1.98 (d, J=7.4Hz, 5H), 1.52-1.19 (br, 46H); 13 C NMR (126MHz, CDCl3) δ200.3, 158.3, 135.7, 128.9, 125.8, 114.2, 72.3, 66.8, 53.4, 38.2, 34.4, 33.7, 33.2, 33.2, 32.6, 32.5, 30.2, 29.6, 29.5, 29.5, 29.4, 29.1, 28.8, 25.5, 25.4, 24.8, 24.8. C 33 H 56 O4 ESIMS: [MH] - The theoretical m / z = 514.4, the actual value is 514.5.

[0393]

[0394] 1-(3,5-di-tert-butyl-4-hydroxyphenyl) heneicosine-20-ene-1-one, 7: Anhydrous MeCN (10 mL) was added to an oven-dried pressure tube containing potassium iodide (871 mg, 5.25 mmol) and 2,6-di-tert-butylphenol (1.2 g, 5.25 mmol), followed by the addition of heneicosine-20-ene acid chloride (1.8 g, 5.25 mmol). The tube was then sealed, and the reaction mixture was heated to 82 ° C, stirred for 24 hours, and then cooled to ambient temperature. The solution was quenched with water (10 mL) and extracted with DCM (3x50 mL). The combined organic layer was washed with saturated sodium thiosulfate solution, then dried (Na2SO4), and concentrated in vacuo. Purified by column chromatography (1% EtOAc in hexane) to obtain compound 7 (1.4 g, 2.69 mmol, 52%) in the form of a yellow oil. 1 H NMR (500MHz, CDCl3) δ7.20 (d, J=7.8Hz, 2H), 6.85 (t, J=7.8Hz, 1H), 5.23 (s, 1H), 3.46-3.31 (m, 4H), 1.91-1.78 (m, 4H), 1.55-1.21 (br, 36H).

[0395] LCMS / MS was performed on an Agilent 1290 Infinity II UPLC system and an Agilent 6495 triple quadrupole mass spectrometer. Separation was achieved at 45°C using a BEH C18, 1.7 μm, 2.1 x 50 mm column (Waters) and a 7-minute gradient consisting of 10% B for 0.5 minutes, then 10% to 95% B in 3 minutes, then 95% B for the remainder of the separation; mobile phase A consisted of 0.1% formic acid in water; mobile phase B consisted of 0.1% formic acid in acetonitrile. Each injection was followed by a 3-minute equilibration at 10% B concentration. The flow rate was 0.5 mL / min; 2 μL of sample was introduced into the column via an Agilent 1290 Infinity II multisampler. The retention time of BP4L-18.1.1 was 5.93 minutes. Quantitative analysis was performed using multiple reaction monitoring in positive electrospray ionization mode, using transitions 517.4→233.1 and collision energies of 42. Source specific parameters were: gas temperature, 120°C; gas flow, 11 L / min; nebulizer, 22 psi; sheath gas temperature, 400°C; sheath gas flow, 12 L / min; capillary voltage, 2500 V; nozzle voltage 0 V; high-voltage RF, 150 V; low-voltage RF, 110 V.

[0396] Sample preparation

[0397] 200 μL of LCMS grade acetonitrile was added to 200 μL of blood in a 1 mL Eppendorf tube and vortexed for 10 seconds, then centrifuged at 2400 x g for 15 minutes. 200 μL of the supernatant was removed, placed in a fresh 1 mL Eppendorf tube, and centrifuged at 5300 x g for 15 minutes. Finally, 100 μL of the supernatant was removed and passed through a 0.2 μm modified nylon centrifugal filter to remove any protein residues. The resulting supernatant was used for LCMS analysis without further modification.

[0398]

[0399] Other compounds can be synthesized according to the following schemes.

[0400]

[0401] Example 14. Pharmacokinetics

[0402] The pharmacokinetic properties of the exemplary compound BP4L-18:1:1 (Compound 6) were evaluated in rats ( Fig.29 In particular, the concentration of BP4L-18:1:1 was determined in whole blood samples within 30 hours after oral administration. The results indicate that the compound is absorbed, thereby exhibiting bioavailability.

[0403] Equivalent forms and scope

[0404] In the claims, articles such as "a," "an," and "the" may mean one or more than one, unless indicated to the contrary or otherwise obvious from the context. Claims or descriptions that include "or" between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, used in, or otherwise related to a given product or process, unless indicated to the contrary or otherwise obvious from the context. The present disclosure includes embodiments in which exactly one member of the group is present in, used in, or related to a given product or process. The present disclosure includes embodiments in which more than one or all of the group members of the group are present in, used in, or related to a given product or process.

[0405] In addition, the present disclosure encompasses all variations, combinations and permutations in which one or more limitations, elements, clauses and descriptive terms from one or more listed claims are introduced into another claim. For example, any claim that depends on another claim can be modified to include one or more limitations found in any other claim that depends on the same basic claim. Where an element is presented in the form of a list (e.g., in a Markush group format), each subgroup of the element is also disclosed, and any element can be removed from the group. It should be understood that, in general, when the present disclosure or the aspects described herein are referred to as including specific elements and / or features, certain embodiments described herein or the aspects described herein are composed of or substantially composed of these elements and / or features. For simplicity, these embodiments are not specifically described herein. It should also be noted that the terms "comprising" and "containing" are intended to be open-ended and allow the inclusion of additional elements or steps. In the case of a given range, endpoints are included. In addition, unless otherwise specified or otherwise apparent from the context and the understanding of a person of ordinary skill in the art, the value expressed as a range can present any specific value or subrange within the range described in the different embodiments described herein, up to one tenth of the lower limit of the range, unless the context clearly specifies otherwise.

[0406] This application relates to various issued patents, published patent applications, journal articles and other publications, all of which are incorporated herein by reference. If there is a conflict between any incorporated reference and this specification, this specification shall prevail. In addition, any specific embodiment of the present disclosure that falls into the prior art may be explicitly excluded from any one or more claims. Because these embodiments are considered to be known to those of ordinary skill in the art, they may be excluded even if they are not explicitly stated herein. Any specific embodiment described herein may be excluded from any claim for any reason, whether or not it is related to the existence of the prior art.

[0407] Those skilled in the art will recognize, or be able to ascertain using only routine experimentation, many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above specific embodiments, but as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications may be made to this description without departing from the spirit or scope of the present disclosure as defined in the following claims.

Claims

1. A compound of the following formula or a pharmaceutically acceptable salt thereof:

2. The compound according to claim 1, wherein the compound has the following formula: or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1, wherein the compound has the following formula: or a pharmaceutically acceptable salt thereof.

4. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 4 in the preparation of a medicament for treating pain.

6. The use according to claim 5, wherein the pain is chronic pain.

7. The use according to claim 6, wherein the chronic pain is neuropathic pain characterized by one or more symptoms selected from the group consisting of persistent negative sensory perception, hyperalgesia, allodynia, burning sensation and allodynia descriptors.

8. Use of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 4 in the preparation of a medicament for regulating HCN channel gating.

9. The use according to claim 8, comprising inhibiting HCN channel gating.

10. The use according to claim 8 or 9, wherein the HCN channel is an HCN1 channel.

11. Use of a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 4 in the preparation of a medicament for inhibiting HCN1 channels without enhancing γ-aminobutyric acid-A (GABA-A) receptors.

Images