A3 adenosine receptor agonists for use in medicine

Abstract

This disclosure provides adenosine analogues for treating diseases such as pain and inflammatory conditions.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 066,757, filed August 17, 2020; U.S. Provisional Patent Application No. 63 / 171,949, filed April 7, 2021; and U.S. Provisional Patent Application No. 63 / 172,570, filed April 8, 2021, the entire contents of each of which are incorporated herein by reference. Background Technology

[0003] Extracellular concentrations of adenosine, a purine nucleoside, increase significantly when cells are stressed or damaged, or when neurotransmitters are released from neurons. This occurs in most (if not all) organs, including the nervous system. Cellular responses to adenosine are categorized as A1, A2, ... 2A A 2B Adenosine receptors (A3) are mediated by four G protein-coupled receptors. These adenosine receptor subtypes are found in many different cell types in most (if not all) tissues, and their expression is known to increase in the presence of lesions or diseases in at least some cases. Although adenosine activates all four receptor subtypes, various compounds and drugs have varying abilities to activate one subtype more than the others. Therapeutically, the use of selective adenosine receptor agonists may have the advantage of avoiding potentially harmful effects attributable to activation of one or more other subtypes.

[0004] Acting through adenosine receptors, increased extracellular adenosine concentrations modulate responses from the innate immune system, the inappropriate activation of which has been implicated in numerous diseases and conditions. Adenosine receptor activation can also alter the properties of astrocytes, microglia, and neurons in the nervous system. Through one mechanism, activation of the A3 adenosine receptor (A3AR) can inhibit the formation of the NLRP3 (containing the NOD-like receptor family heat-containing protein domain 3) inflammasome, an intracellular polyprotein complex that leads to the production of interleukin-1β (IL-1β). IL-1β is a key mediator of both acute and chronic inflammatory responses. Activation of A3AR with agonists has been shown to suppress inflammasome activity, thereby alleviating inflammation and pain induced by IL-1β and other pro-inflammatory cytokines. Furthermore, inflammasome-mediated chronic inflammation is frequently associated with cellular degeneration, such as the loss of central nervous system cells in neurodegenerative conditions like Alzheimer's and Parkinson's diseases.

[0005] Chronic inflammatory diseases affecting various tissue types are also known to involve mitochondrial dysfunction in cells, which provide the energy needed to drive all cellular processes. Insufficient energy enhances cellular stress and, when severe, leads to cell degeneration. Mitochondrial dysfunction can lead to inflammation, and inflammation can lead to mitochondrial dysfunction. A3 adenosine receptor (AR) agonists are known to prevent mitochondrial damage and inhibit cell degeneration.

[0006] Activation of A3AR is known to promote multiple cellular signaling pathways. A3AR inhibits adenylate cyclase activity via Gi G proteins and stimulates the phospholipase C / inositol triphosphate / diacylglycerol pathway via Gβγ G proteins. These receptors are also coupled to mitogen-activated protein kinases (MAPKs), including ERK1 and ERK2. Gβγ proteins also mediate the regulation of voltage-gated calcium channels and G protein-gated inward rectifier potassium channels (GIRK and Kir6) to inhibit overexcitability. Furthermore, A3AR activation is coupled with translocation of β-repressor proteins. Previous studies have shown that some A3AR agonists with a bridged methylene carba motif rather than a ribocyclic motif exhibit biased agonistic activity, meaning that the compounds show different potencies and effects on pathways triggered by ligand binding (Baltos et al., Mol. Pharmacol. 90:12 (2016)). The efficacy and safety of A3AR agonists depend on the activity spectrum and profile required for anti-inflammatory or analgesic activity, and previous studies with structurally dissimilar A3AR agonists could not predict the activity spectrum of the novel compounds described in this application. As indicated, biased agonist activity has been observed in compounds with a bridged methylene carbazin motif, and the compounds reported herein contain modifications in the bridged methylene carbazin substituent that result in different patterns of biased agonist activity.

[0007] Inflammation of tissues is often accompanied by pain. Pain attributable to damage to peripheral nerves and certain areas of the central nervous system is called neuropathic pain. Multiple pieces of evidence indicate that some chronic inflammatory pain conditions, including but not limited to chronic neuropathic pain and chronic mixed pain conditions (which combine elements of common inflammatory pain and neuropathic pain), involve inflammasome activity. Selective A3AR agonists are analgesics used in chronic inflammatory pain, neuropathic pain, and mixed pain conditions. A3AR-mediated analgesia is accompanied by mitochondrial protection and a decrease in the levels of IL-1β and other pro-inflammatory cytokines.

[0008] In the central nervous system, chronic inflammasome activity leading to increased levels of IL-1β and other pro-inflammatory cytokines induces widespread cognitive impairment. For example, patients exposed to certain chemotherapy drugs exhibit a persistent condition (lasting months to years) known as chemotherapy-induced cognitive impairment (“chemo-brain”; “chemo-fog”). Patients recovering from head trauma exhibit a similar syndrome (traumatic brain injury-induced cognitive impairment or “post-concussion syndrome”). Cognitive impairment syndromes are also seen after surgery, especially after cardiopulmonary bypass surgery, and particularly in the elderly (postoperative cognitive impairment). Selective A3AR agonists can treat and prevent cognitive impairment syndromes.

[0009] The innate immune system responds to various toxins, including certain plant alkaloids such as morphine and its synthetic homologs (collectively known as opioids). Exposure to analgesic levels of opioids leads to inflammasome formation, increased IL-1β levels, and a pro-inflammatory cascade, promoting many undesirable side effects of opioids, including but not limited to physiological dependence (a contributing factor to addiction) and analgesic tolerance (the phenomenon that the analgesic effect of repeated doses of opioids gradually diminishes, thus requiring escalating doses to maintain adequate analgesia). Selective A3AR agonists can attenuate opioid-induced inflammasome activation, dependence / addiction, and tolerance.

[0010] Inhibition of voltage-gated calcium channels in nociceptors is a clinically proven approach to treating neuropathic pain, as demonstrated in clinical studies using gabapentin and the peptide Prialt. Activation of adenosine receptors in nociceptors inhibits the activation of CaV2.1 and CaV2.2 channels, mimicking the direct inhibitory effects of these channels and thereby suppressing painful nociceptive sensation. In some studies, the effect of adenosine on voltage-gated calcium channels has been recapitulated by selective activation of A3AR, indicating the importance of this pathway for analgesic activity.

[0011] Typically, disease, trauma, or other lesions can lead to the upregulation of A3AR on cells, thus providing opportunities for selective A3AR agonist therapy or management of a wide range of diseases and conditions affecting humans and other animals.

[0012] Adenosine receptor agonists are still needed to treat the disease. Summary of the Invention

[0013] In some respects, this disclosure provides a compound represented by formula (I):

[0014]

[0015] Or its pharmaceutically acceptable salt, wherein:

[0016] R 1 Selected from -CD3, C2 fluoroalkyl, C4-C6 fluoroalkyl and C2-C6 alkyl substituted with phenyl, wherein the phenyl is substituted with one or more fluorine atoms;

[0017] R 2 Selected from hydrogen, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl are optionally selected independently by one or more of R. * Substituents; and

[0018] R 3 Selected from hydrogen, C1-C6 alkyl, and C1-C6 haloalkyl; and

[0019] Each R * Independently selected from fluorine-OR 20 -N(R) 20 )2、-N(R 20 )2、-C(O)R 20 -C(O)OR 20 -OC(O)R 20 -NO2 and -CN; and C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 20 -N(R) 20 2, -NO2, -CN and C 1-3 Alkyl; and

[0020] R 20 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0021] In some respects, this disclosure provides a compound represented by formula (II'):

[0022]

[0023] Or its pharmaceutically acceptable salt, wherein:

[0024] R 4’The group is selected from hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C3-C8 carbon rings, and optionally substituted 3- to 8-membered heterocycles, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R. 7 Furthermore, the optional substituents on the C3-C8 carbon rings and 3- to 8-membered heterocycles are independently selected from R. 8 ;

[0025] R 5’ Selected from hydrogen, -NO2, -CN, -NH2, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl are optionally selected independently by one or more of R 9 Substituents of the substituents;

[0026] R 6’ Selected from hydrogen, -C(O)(NR) 50 2) and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl groups are independently selected from R 10 ;

[0027] Each R 7 R 9 and R 10 Selected independently each time it appears:

[0028] Fluorine-OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30 -OC(O)R 30 -NO2 and -CN;

[0029] C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 alkyl;

[0030] And when R 6’ It is -C(O)(NHMe), and R 9 When R is a 3- to 6-membered heterocycle that is optionally substituted, 9 Optionally substituted by one or more substituents selected from one or more of the following substituents independently selected: fluorine, bromine, -OR 30 -N(R) 30 2, -NO2, -CN and C 1-3 alkyl;

[0031] Each R 8 Selected independently each time it appears:

[0032] Halogen, -OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30 -OC(O)R 30 -NO2 and -CN;

[0033] C10 ... 1-3 Alkyl groups: halogens, -OR 30 -N(R) 30 )2、-NO2、-CN、C 3-6 Carbon rings and 3- to 6-membered heterocycles;

[0034] Each R 30 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3- to 6-membered heterocycles;

[0035] Each R 50 Each time it appears, it is independently selected from hydrogen and C. 1-6 Alkyl; and

[0036] Each R 31 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles, wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocyclic ring are optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, -NO2, -NH2, -OC. 1-6 Alkyl and -C 1-6 Alkyl; and

[0037] R 12’ Selected from hydrogen, halogen, hydroxyl, -NO2, -CN, -NH2, -OC 1-6 Alkyl and C 1-6 Alkyl group, wherein the -OC 1-6 The alkyl moiety and C 1-6The alkyl group is optionally substituted by one or more substituents selected from the following: halogen, -OH, -NH2, -NO2, -CN, -OC. 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles; wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents selected from the following: halogen, hydroxyl, -NO2, -CN, -NH2, -OC. 1-6 Alkyl and C 1-6 alkyl.

[0038] In some respects, this disclosure provides a compound represented by formula (III):

[0039]

[0040] Or its pharmaceutically acceptable salt, wherein:

[0041] R 21 Selected from C3-C6 cycloalkyl and 3-6 membered heterocyclic alkyl, wherein the C3-C6 cycloalkyl and 3-6 membered heterocyclic alkyl are optionally selected independently by one or more of R 29 Substituents of the substituents;

[0042] R 22 The substituents are selected from optionally substituted C1-C6 alkyl groups, optionally substituted C3-C8 carbon rings, and optionally substituted 3- to 8-membered heterocycles, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R each time they appear. 27 Furthermore, the optional substituents on the C3-C8 carbon rings and 3- to 8-membered heterocycles are independently selected from R. 28 ;

[0043] R 23 Selected from hydrogen and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl groups are independently selected from R. 40 ;

[0044] Each R 27 R 29 and R 40 Selected independently each time it appears:

[0045] Fluorine-OR 50 -N(R) 50 )2、-N(R 50 )2、-C(O)R 50 -C(O)OR 50 -OC(O)R 50 -NO2 and -CN;

[0046] C 3-6The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 50 -N(R) 50 2, -NO2, -CN and C 1-3 alkyl;

[0047] Each R 28 Selected independently each time it appears:

[0048] Halogen, -OR 50 -N(R) 50 )2、-N(R 50 )2、-C(O)R 50 -C(O)OR 50 -OC(O)R 50 -NO2 and -CN;

[0049] C10 ... 1-3 Alkyl groups: halogens, -OR 50 -N(R) 50 )2、-NO2、-CN、C 3-6 Carbon rings and 3- to 6-membered heterocycles; and

[0050] R 50 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0051] In some respects, this disclosure provides a compound represented by formula (IV):

[0052]

[0053] Or its pharmaceutically acceptable salt, wherein:

[0054] R 31 Selected from hydrogen, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl are optionally selected independently by one or more of R. 39 Substituents of the substituents;

[0055] R 32 Selected from C1-C6 alkyl groups;

[0056] R 33 Selected from C1-C3 haloalkyl groups;

[0057] Each R 39 Selected independently each time it appears:

[0058] Fluorine, -OR 60 -N(R) 60 )2、-N(R 60 )2、-C(O)R 60 -C(O)OR 60 -OC(O)R 60 -NO2 and -CN;

[0059] C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 60 -N(R) 60 2, -NO2, -CN and C 1-3 alkyl;

[0060] Each R 60 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0061] In some aspects, this disclosure provides a pharmaceutical composition comprising a compound described herein or a salt of any of the compounds described herein, and a pharmaceutically acceptable excipient.

[0062] In some aspects, this disclosure provides a method for activating A3 adenosine receptors, the method comprising administering the compounds or salts described herein to a subject with a desired condition. In some embodiments, relative to A1, A 2A and A 2B The agonistic effect of the receptor is ten times greater; compounds or salts have an agonistic effect on A3 adenosine receptors.

[0063] In some embodiments, this disclosure provides a method for treating conditions selected from vascular inflammation, arthritis, allergies, asthma, wound healing, stroke, heart failure, acute spinal cord injury, acute head injury or trauma, seizures, neonatal hypoxia, cerebral palsy, chronic hypoxia attributable to arteriovenous malformations and occlusive cerebral artery disease, ischemia and reperfusion injury in skeletal muscle, severe neurological disorders associated with excitotoxicity, Parkinson's disease, Huntington's chorea, CNS diseases, heart disease, kidney disease, glaucoma, cancer, neuropathic pain, transient ischemic attack, myeloprotection, dry eye syndrome, osteoarthritis, rheumatoid arthritis, loss of skin pigmentation, inflammatory bowel disease, pulmonary inflammation, uveitis, and septic shock.

[0064] In some embodiments, this disclosure provides a method for treating conditions selected from chemotherapy-induced peripheral neuropathy, diabetic peripheral neuropathy, neurodegeneration, drug-induced ototoxicity, spinocerebellar degeneration, symptoms associated with traumatic brain injury, chemotherapy-induced cognitive impairment, pain and discomfort of irritable bowel syndrome, and neuropathic pain.

[0065] Additional aspects and advantages of this disclosure will become apparent to those skilled in the art from the following detailed description, in which only illustrative embodiments of the disclosure are shown and described. It will be appreciated that the disclosure is capable of other and different embodiments, and that several details thereof can be modified in various obvious respects without departing from the disclosure. Therefore, the drawings and descriptions should be considered illustrative in nature and not restrictive.

[0066] Incorporation

[0067] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference as if each individual publication, patent, or patent application had been specifically and individually indicated to be incorporated herein by reference. If any publication, patent, or patent application incorporated herein by reference conflicts with any disclosure contained in this specification, the specification is intended to supersede and / or give precedence to any such conflicting material. Detailed Implementation

[0068] Although various embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous changes, variations, and substitutions may be conceived by those skilled in the art without departing from the invention. Various alternatives to the embodiments of the invention described herein should be understood.

[0069] A. Definition

[0070] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. All patents and publications mentioned herein are incorporated herein by reference.

[0071] "Alkyl" refers to a straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, without unsaturation, and preferably having one to fifteen carbon atoms (i.e., C1-C1). 15 Alkyl groups. In some embodiments, the alkyl group comprises one to thirteen carbon atoms (i.e., C1-C1). 13 Alkyl group. In some embodiments, the alkyl group comprises one to eight carbon atoms (i.e., C1-C8 alkyl). In other embodiments, the alkyl group comprises one to five carbon atoms (i.e., C1-C5 alkyl). In other embodiments, the alkyl group comprises one to four carbon atoms (i.e., C1-C4 alkyl). In other embodiments, the alkyl group comprises one to three carbon atoms (i.e., C1-C3 alkyl). In other embodiments, the alkyl group comprises one to two carbon atoms (i.e., C1-C2 alkyl). In other embodiments, the alkyl group comprises one carbon atom (i.e., C1 alkyl). In other embodiments, the alkyl group comprises five to fifteen carbon atoms (i.e., C5-C6 alkyl). 15 Alkyl group. In other embodiments, the alkyl group comprises five to eight carbon atoms (i.e., C5-C8 alkyl). In other embodiments, the alkyl group comprises two to five carbon atoms (i.e., C2-C5 alkyl). In other embodiments, the alkyl group comprises three to five carbon atoms (i.e., C3-C5 alkyl). In some embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). The alkyl group is attached to the rest of the molecule by a single bond.

[0072] Term "C" x-y "When used in conjunction with chemical motifs such as alkyl, alkenyl, or alkynyl, it is intended to include groups containing x to y carbons in the chain. For example, the term 'C'..." 1-6 "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group containing 1 to 6 carbon atoms, including straight-chain alkyl and branched-chain alkyl groups. Term –C x-y Alkylene refers to an alkylene chain, whether substituted or unsubstituted, having x to y carbons. For example, –C 1-6 Alkylene – can be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any of which may be optionally substituted.

[0073] "Alkoxy" refers to a group bonded by an oxygen atom of the formula –O-alkyl, where the alkyl is an alkyl chain as defined above.

[0074] "Alkenyl" refers to a straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having two to twelve carbon atoms (i.e., C2-C). 12 Alkenyl group. In some embodiments, the alkenyl group comprises two to eight carbon atoms (i.e., C2-C8 alkenyl). In some embodiments, the alkenyl group comprises two to six carbon atoms (i.e., C2-C6 alkenyl). In other embodiments, the alkenyl group comprises two to four carbon atoms (i.e., C2-C4 alkenyl). The alkenyl group is attached to the rest of the molecule by a single bond, such as ethenyl (i.e., vinyl), propenyl (i.e., allyl), butenyl, pentenyl, pent-1,4-dienyl, etc.

[0075] "Alkyne" refers to a straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and preferably having two to twelve carbon atoms (i.e., C2-C). 12 The alkynyl group (C2-C8 alkynyl) is present in some embodiments. In other embodiments, the alkynyl group contains two to eight carbon atoms (C2-C6 alkynyl). In still other embodiments, the alkynyl group contains two to four carbon atoms (C2-C4 alkynyl). The alkynyl group is attached to the rest of the molecule by a single bond, and is an example of ethynyl, propynyl, butynyl, pentylyl, hexynyl, etc.

[0076] Term "C" x-y "Alkenyl" and "C" x-y "Alkyne" refers to an alkyl group, substituted or unsubstituted, that is similar in length and possible substitutions to the alkyl groups described above, but contains at least one double or triple bond of an unsaturated aliphatic group. Term –C x-y An alkenyl group refers to an alkenyl chain, whether substituted or unsubstituted, having x to y carbons. For example, –C 2-6 Alkenyl group – optionally selected from vinylene, propenene, butenene, pentenene, and hexenene, any of which may be optionally substituted. The alkenyl chain may have one or more double bonds. Term – C x-y Alynyl groups refer to substituted or unsubstituted alynyl chains having x to y carbons in the alynyl chain. For example, –C 2-6 The alkenyl group can be selected from alkenylene, propynylene, butynylene, pentylyne, and hexynylene, any of which may be optionally substituted. The alkenylene chain may have one or more triple bonds.

[0077] "alkylene" or "alkylene chain" refers to a straight-chain or branched divalent hydrocarbon chain, consisting only of carbon and hydrogen, without unsaturation, and preferably having one to twelve carbon atoms, with the remainder of the molecule attached to a group. Examples include methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is attached to the remainder of the molecule and to the group via single bonds. The connection points between the alkylene chain and the remainder of the molecule, as well as with the group, can be achieved by any two carbon atoms within the chain. In some embodiments, the alkylene comprises one to ten carbon atoms (i.e., C1-C8 alkylene). In some embodiments, the alkylene comprises one to eight carbon atoms (i.e., C1-C8 alkylene). In other embodiments, the alkylene comprises one to five carbon atoms (i.e., C1-C5 alkylene). In other embodiments, the alkylene comprises one to four carbon atoms (i.e., C1-C4 alkylene). In other embodiments, the alkylene comprises one to three carbon atoms (i.e., C1-C3 alkylene). In other embodiments, the alkylene group comprises one to two carbon atoms (i.e., C1-C2 alkylene). In other embodiments, the alkylene group comprises one carbon atom (i.e., C1 alkylene). In other embodiments, the alkylene group comprises five to eight carbon atoms (i.e., C5-C8 alkylene). In other embodiments, the alkylene group comprises two to five carbon atoms (i.e., C2-C5 alkylene). In other embodiments, the alkylene group comprises three to five carbon atoms (i.e., C3-C5 alkylene).

[0078] "Alkenyl" or "alkenyl chain" refers to a straight-chain or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing at least one carbon-carbon double bond, and preferably having two to twelve carbon atoms, with the remainder of the molecule attached to a group. The alkenyl chain is attached to the remainder of the molecule and to the group via single bonds. The connection points between the alkenyl chain and the remainder of the molecule, as well as with the group, can be achieved by any two carbon atoms within the chain. In some embodiments, the alkenyl group comprises two to ten carbon atoms (i.e., C2-C2). 10 (Alkenyl group). In some embodiments, the alkenyl group comprises two to eight carbon atoms (i.e., C2-C8 alkenyl). In other embodiments, the alkenyl group comprises two to five carbon atoms (i.e., C2-C5 alkenyl). In other embodiments, the alkenyl group comprises two to four carbon atoms (i.e., C2-C4 alkenyl). In other embodiments, the alkenyl group comprises two to three carbon atoms (i.e., C2-C3 alkenyl). In other embodiments, the alkenyl group comprises two carbon atoms (i.e., C2 alkenyl). In other embodiments, the alkenyl group comprises five to eight carbon atoms (i.e., C5-C8 alkenyl). In other embodiments, the alkenyl group comprises three to five carbon atoms (i.e., C3-C5 alkenyl).

[0079] "Imyynyl" or "Imyynyl chain" refers to a straight-chain or branched divalent hydrocarbon chain in which the rest of the molecule is attached to a group, consisting only of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having two to twelve carbon atoms. The ynylyl chain is attached to the rest of the molecule and to the group via single bonds. The connection points between the ynylyl chain and the rest of the molecule, as well as with the group, can be achieved by any two carbons within the chain. In some embodiments, the ynylyl group contains two to ten carbon atoms (i.e., C2-C). 10 The ynyl group (C2-C8 ynyl group) comprises two to eight carbon atoms in some embodiments. In other embodiments, the ynyl group comprises two to five carbon atoms (C2-C5 ynyl group). In other embodiments, the ynyl group comprises two to four carbon atoms (C2-C4 ynyl group). In other embodiments, the ynyl group comprises two to three carbon atoms (C2-C3 ynyl group). In other embodiments, the ynyl group comprises two carbon atoms (C2 ynyl group). In other embodiments, the ynyl group comprises five to eight carbon atoms (C5-C8 ynyl group). In other embodiments, the ynyl group comprises three to five carbon atoms (C3-C5 ynyl group).

[0080] "Aryl" refers to a group derived from an aromatic monocyclic or polycyclic aromatic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. An aromatic monocyclic or polycyclic aromatic hydrocarbon ring system contains only hydrogen and carbon, with five to eighteen carbon atoms, wherein at least one ring in the ring system is aromatic, meaning it contains a ring-shaped delocalized (4n+2)π electron system conforming to Hückel's theory. Ring systems derived from aryl groups include, but are not limited to, groups such as benzene, fluorene, indene, indene, tetrahydronaphthalene, and naphthalene.

[0081] "Arane" refers to a group having the formula -Rc-aryl, where Rc is an alkylene chain as defined above, such as methylene, ethylene, etc.

[0082] "Arylene" refers to a group having the formula –Rd-aryl, where Rd is an alkenyl chain as defined above. "Arylynyl" refers to a group having the formula –Re-aryl, where Re is an alkenyl chain as defined above.

[0083] A “carbocyclic ring” refers to a saturated, unsaturated, or aromatic ring in which each atom of the ring is carbon. Carbocyclic rings can include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic carbocyclic ring can be selected from a saturated, unsaturated, or aromatic ring. Aromatic rings, such as phenyl, can be fused to saturated or unsaturated rings such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of a carbocyclic ring, where valence allows. Exemplary carbocyclic rings include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl.

[0084] “Cycloalkyl” refers to a fully saturated monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, including fused ring systems or bridging ring systems, and preferably having three to twelve carbon atoms. In some embodiments, the cycloalkyl group contains three to ten carbon atoms. In other embodiments, the cycloalkyl group contains five to seven carbon atoms. The cycloalkyl group can be linked to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl groups include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptyl), norbornenyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, etc.

[0085] "Cycloalkenyl" refers to an unsaturated non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, including fused ring systems or bridging ring systems, preferably having three to twelve carbon atoms, and containing at least one double bond. In some embodiments, the cycloalkenyl group contains three to ten carbon atoms. In other embodiments, the cycloalkenyl group contains five to seven carbon atoms. The cycloalkenyl group can be linked to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyl groups include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0086] "Cycloalkylalkyl" refers to a group having the formula –Rc-cycloalkyl, where Rc is an alkylene chain as described above.

[0087] "Cycloalkylalkoxy" refers to a group bonded by the oxygen atom of the formula –O-Rc-cycloalkyl, where Rc is an alkylene chain as described above.

[0088] "Halogen" or "halogen" refers to halogen substituents, such as bromine, chlorine, fluorine, and iodine substituents.

[0089] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl group as defined above that has been substituted with one or more halogen groups, such as trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl group may optionally be further substituted. Examples of halogen-substituted alkanes (“haloalkanes”) include halomethanes (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), dihalomethanes and trihalomethanes (e.g., chloroform, tribromomethane, trifluoromethane, triiodomethane), 1-haloethane, 2-haloethane, 1,2-dihaloethane, 1-halopropane, 2-halopropane, 3-halopropane, 1,2-dihalopropane, 1,3-dihalopropane, 2,3-dihalopropane, 1,2,3-trihalopropane, and any other suitable combination of alkanes (or substituted alkanes) and halogens (e.g., Cl, Br, F, I, etc.). When the alkyl group is substituted by more than one halogen group, each halogen can be chosen independently, for example, 1-chloro,2-fluoroethane.

[0090] "Fluoroalkyl" refers to an alkyl group as defined above that is substituted with one or more fluorine groups, such as trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc.

[0091] A "heterocycle" is a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring in a bicyclic heterocycle may be a self-saturated, unsaturated, or aromatic ring. A "hypocyclic group" is a divalent heterocycle to which the remainder of the molecule is attached.

[0092] "Heteroaryl" or "aromatic heterocycle" refers to a group derived from a heteroaromatic ring group comprising one to eleven carbon atoms and at least one heteroatom, wherein each heteroatom may be selected from N, O, and S. As used herein, the heteroaryl ring may be selected from monocyclic or bicyclic and fused or bridged ring systems, wherein at least one ring in the ring system is aromatic, i.e., it contains a ring-delocalized (4n+2)π electron system conforming to Hückel's theory. The heteroatom in the heteroaryl may optionally be oxidized. If present, one or more nitrogen atoms may optionally be quaternized. Where valence permits, the heteroaryl may be attached to the remainder of the molecule by any atom of the heteroaryl, such as the carbon or nitrogen atom of the heteroaryl. Examples of heteroaryls include, but are not limited to, pyridine, pyrimidine, oxazole, furan, thiophene, benzothiazole, and imidazopyridine. "X-aryl" refers to the number of intracyclic atoms in the ring, i.e., X. For example, a 5-membered heteroaryl ring or a 5-membered aromatic heterocycle has 5 internal ring atoms, such as triazole, oxazole, thiophene, etc.

[0093] "Heterocyclic alkyl" refers to a 3- to 12-membered non-aromatic cyclic group comprising two to twelve carbon atoms and at least one heteroatom, wherein each heteroatom may be selected from N, O, Si, P, B, and S atoms. Heterocyclic alkyl groups may be monocyclic or bicyclic and fused or bridged ring systems. The heteroatom in the heterocyclic alkyl group may optionally be oxidized. If present, one or more nitrogen atoms may optionally be quaternized. The heterocyclic alkyl group may be partially or fully saturated. Where valence permits, the heterocyclic alkyl group is attached to the remainder of the molecule by any atom of the heterocyclic alkyl group, such as any carbon or nitrogen atom of the heterocyclic alkyl group. Examples of heterocyclic alkyl groups include, but are not limited to, dioxacyclopentyl, thienyl[1,3]dithiohexacyclohexyl, decahydroisoquinolinyl, imidazolinyl, imidazoalkyl, isothiazolinyl, isoxazolinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopiperidyl, oxazolinyl, piperidinyl, piperazinyl, 4-piperidinoneyl, pyrrolyl, pyrazolyl, quininecycloyl, thiazoalkyl, tetrahydrofuranyl, trithiohexacyclohexyl, tetrahydropyranyl, thiomorpholinyl, thiomorpholinyl, 1-oxo-thiomorpholinyl and 1,1-dioxo-thiomorpholinyl.

[0094] The term “substituted” refers to a substituent on one or more carbon atoms or substituted heteroatoms, such as NH, that partially have a substituted structure. It should be understood that “substitution” or “substituted by” includes the implicit condition that such substitution conforms to the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, i.e., a compound that does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc. In some embodiments, substituted means having a substituent that partially replaces two hydrogen atoms on the same carbon atom, such as replacing two hydrogen atoms on a single carbon atom with an oxo group, imino group, or thio group. As used herein, the term “substituted” is considered to include all permissible substituents of an organic compound. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. For a suitable organic compound, permissible substituents may be one or more, and may be the same or different. For the purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents conforming to the valence of the heteroatom in the organic compound described herein.

[0095] In some embodiments, the substituents may include any substituents described herein, such as: halogen, hydroxyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=N-OH), hydrazine (=N-NH2), -R b -OR a -R b -OC(O)-R a -R b-OC(O)-OR a -R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a -R b -C(O)OR a -R b -C(O)N(R a )2、-R b -OR c -C(O)N(R a )2、-R b -N(R a )C(O)OR a -R b -N(R a )C(O)R a -R b -N(R a S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, areneyl, arynyl, cycloalkyl, cycloalkylalkyl and heterocyclic, any of which may optionally be substituted by: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=N-OH), hydrazine (=N-NH2), -R b -OR a -R b -OC(O)-R a -R b -OC(O)-OR a -R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a -R b-C(O)OR a -R b -C(O)N(R a )2、-R b -OR c -C(O)N(R a )2、-R b -N(R a )C(O)OR a -R b -N(R a )C(O)R a -R b -N(R a S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2); where each R a Independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, and heterocyclic, wherein each R a Subject to the permissible oxidation state, the following groups may be optionally substituted: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=N-OH), hydrazine (=N-NH2), -R b -OR a -R b -OC(O)-R a -R b -OC(O)-OR a -R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a -R b -C(O)OR a -R b -C(O)N(R a )2、-R b -OR c -C(O)N(R a )2、-Rb -N(R a )C(O)OR a -R b -N(R a )C(O)R a -R b -N(R a S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2); and where each R b Independently selected from direct-chain or straight-chain or branched alkylene, alkenyl, or ynylene chains, and each R c It is a straight-chain or branched alkylene, alkenyl, or ynylene chain.

[0096] Unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” as used in the specification and claims include a plurality of indicators.

[0097] The term "salt" or "pharmaceutically acceptable salt" refers to a salt derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed from inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed from inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, etc., particularly isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts.

[0098] As used herein, the phrases “parenteral administration” and “administered via a parenteral route” refer to administration methods that are normally administered by injection, other than enteral and topical administration, and include, but are not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intra-bursal, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions.

[0099] The phrase “pharmaceutically acceptable” is used herein to refer to compounds, materials, compositions, and / or dosage forms that, to the extent of reasonable medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0100] As used herein, the phrase “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with other components of the formulation and is harmless to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil and cottonseed oil. (10) Safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (11) Diols, such as propylene glycol; (12) Polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Atherless water; (18) Isotonic saline; (19) Ringer's solution; (20) Ethanol; (21) Phosphate buffer solution; and (22) Other non-toxic and compatible substances used in pharmaceutical preparations.

[0101] In some implementations, the term “preventing,” as used in relation to a disease or condition, may refer to, in a statistical sample, a compound reducing the occurrence of a disease or condition in a treated sample relative to an untreated control sample, or delaying the onset of one or more symptoms of a disease or condition or reducing the severity of said one or more symptoms relative to an untreated control sample.

[0102] As used herein, the term "treatment" may include alleviating, reducing, or improving symptoms of a disease or condition; preventing additional symptoms; improving or preventing the underlying cause of symptoms; suppressing a disease or condition, such as halting its progression; reducing a disease or condition; causing a disease or condition to subside; alleviating the condition caused by a disease or condition; or preventing and / or therapeutically stopping the symptoms of a disease or condition.

[0103] B. Compounds disclosed herein

[0104] In some embodiments, this disclosure provides a compound represented by formula (I):

[0105]

[0106] Or its pharmaceutically acceptable salt, wherein:

[0107] R 1 Selected from -CD3, C2 fluoroalkyl, C4-C6 fluoroalkyl and C2-C6 alkyl substituted with phenyl, wherein the phenyl is substituted with one or more fluorine atoms;

[0108] R 2 Selected from hydrogen, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl are optionally selected independently by one or more of R. * Substituents; and

[0109] R 3 Selected from hydrogen, C1-C6 alkyl, and C1-C6 haloalkyl; and

[0110] Each R * Independently selected from fluorine-OR 20 -N(R) 20 )2、-N(R 20 )2、-C(O)R 20 -C(O)OR 20 -OC(O)R 20 -NO2 and -CN; and C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 20 -N(R) 20 2, -NO2, -CN and C 1-3 Alkyl; and

[0111] R 20 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following.1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0112] In some embodiments, the compound of formula (I) is represented by formula (IA) or a pharmaceutically acceptable salt thereof:

[0113]

[0114] In some embodiments, for compounds or salts of formula (I) or (IA), R1 is selected from C2 fluoroalkyl, C4-C6 fluoroalkyl and C2-C6 alkyl substituted with phenyl, wherein the phenyl is substituted with one or more fluorine atoms.

[0115] In some embodiments, for compounds or salts of formula (I) or (IA), R1 is selected from C1-C6 alkyl groups, wherein the alkyl group is substituted with at least one deuterium atom. In some cases, R1 is -CD3.

[0116] In some implementations, for compounds or salts of formula (I) or formula (IA), R 1 Selected from CH2CFH2, CH2CF2H, and CH2CF3. In some cases, R 1 It is CH2CF2H.

[0117] In some implementations, for compounds or salts of formula (I) or (IA), when R 2 When selected from hydrogen, fluorine, bromine, iodine, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl are optionally selected independently by one or more of R * Substituents of R 1 Further selected from C3 fluoroalkyl and C1 alkyl with phenyl substituted atoms, wherein the phenyl is substituted with one or more fluorine atoms. In some cases, when R 2 When it is not Cl, R 1 Further selected from C3 fluoroalkyl and C1 alkyl substituted with phenyl, wherein the phenyl is substituted with one or more fluorine atoms. In some embodiments, when R 2 When it is not Cl, R 1 It is fluorobenzyl. In some embodiments, when R 2 When it is not Cl, R 1 It is a C3 fluoroalkyl group.

[0118] In some embodiments, for compounds or salts of formula (I) or (IA), R2 is selected from C3-C6 cycloalkyl groups. In some embodiments, R2 is...

[0119] In some implementations, for compounds or salts of formula (I) or formula (IA), R 2 Selected from hydrogen, CH3, CH2CH3, and Cl. In some cases, R 2 Selected from Cl, CH3, In some cases, R 2 It is Cl. In some cases, R 2 yes In some cases, R 2 Selected from Cl and CH3. In some cases, R 2 Selected from Cl, CH3 and

[0120] In some embodiments, for compounds or salts of formula (I) or (IA), R3 is selected from hydrogen, CH3, CH2F, CHF2, and CF3. In some cases, R3 is hydrogen.

[0121] On the other hand, this disclosure provides a compound represented by formula (II'):

[0122]

[0123] Or its pharmaceutically acceptable salt, wherein:

[0124] R 4’ The group is selected from hydrogen, optionally substituted C1-C6 alkyl groups, optionally substituted C3-C8 carbon rings, and optionally substituted 3- to 8-membered heterocycles, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R. 7 Furthermore, the optional substituents on the C3-C8 carbon rings and 3- to 8-membered heterocycles are independently selected from R. 8 ;

[0125] R 5’ Selected from hydrogen, -NO2, -CN, -NH2, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl are optionally selected independently by one or more of R 9 Substituents of the substituents;

[0126] R 6’ Selected from hydrogen, -C(O)(NR) 50 2) and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl groups are independently selected from R 10 ;

[0127] Each R 7 R 9 and R10 Selected independently each time it appears:

[0128] Fluorine-OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30 -OC(O)R 30 ,

[0129] -NO2 and -CN;

[0130] C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 alkyl;

[0131] And when R 6’ It is -C(O)(NHMe), and R 9 When R is a 3- to 6-membered heterocycle that is optionally substituted, 9 Optionally substituted by one or more substituents selected from one or more of the following substituents independently selected: fluorine, bromine, -OR 30 -N(R) 30 2, -NO2, -CN and C 1-3 alkyl;

[0132] Each R 8 Selected independently each time it appears:

[0133] Halogen, -OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30 -OC(O)R 30 -NO2 and -CN;

[0134] C10 ... 1-3 Alkyl groups: halogens, -OR 30 -N(R) 30 )2、-NO2、-CN、C 3-6 Carbon rings and 3- to 6-membered heterocycles;

[0135] Each R 30 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3- to 6-membered heterocycles;

[0136] Each R 50 Each time it appears, it is independently selected from hydrogen and C. 1-6 Alkyl; and

[0137] Each R 31 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles, wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocyclic ring are optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, -NO2, -NH2, -OC. 1-6 Alkyl and -C 1-6 alkyl;

[0138] R 12’ Selected from hydrogen, halogen, hydroxyl, -NO2, -CN, -NH2, -OC 1-6 Alkyl and C 1-6 Alkyl group, wherein the -OC 1-6 The alkyl moiety and C 1-6 The alkyl group is optionally substituted by one or more substituents selected from the following: halogen, -OH, -NH2, -NO2, -CN, -OC. 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles; wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents selected from the following: halogen, hydroxyl, -NO2, -CN, -NH2, -OC. 1-6 Alkyl and C 1-6 alkyl.

[0139] In some embodiments, compounds or salts of formula (II') are represented by formula (IIA'):

[0140]

[0141] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 4’ The substituents are selected from optionally substituted C1-C6 alkyl groups, optionally substituted C3-C8 carbon rings, and optionally substituted 3- to 8-membered heterocycles, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R each time they appear.7 Furthermore, the optional substituents on the C3-C8 carbon rings and the 3 to 8-membered heterocycles are independently selected from R8.

[0142] In some embodiments, for compounds or salts of formula (II') or (IIA'), R4' is selected from optionally substituted C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted by one or more substituents selected independently of the following each time they appear: fluorine; C 3-6 The carbon ring and 3 to 6-membered heterocycles are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR31, -N(R31)2, -NO2, -CN and C1-3 alkyl.

[0143] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 4’ Selected from optionally substituted C1-C6 alkyl groups, wherein the C1-C6 alkyl group is optionally substituted by one or more substituents selected independently of the following groups each time it appears: fluorine; C1-C6 alkyl group is optionally substituted by one or more substituents selected independently of the following groups. 3-6 Carbon rings: halogens, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 Alkyl group. In some cases, C 3-6 The carbon ring is optionally substituted by one or more substituents independently selected from the following: halogen, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 Alkyl group. In some cases, C 3-6 The carbon ring is optionally substituted by one or more substituents independently selected from the following: halogen, -OR 31 -N(R) 31 )2 and C 1-3 Alkyl group. In some cases, C 3-6 The carbon ring is optionally substituted by one or more substituents independently selected from the following: halogen, -OR 31 and -N(R) 31 2. In some cases, each R 31 Each occurrence is independently selected from C groups that are optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles, wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocyclic ring are optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, -NO2, -NH2, -OC.1-6 Alkyl and -C 1-6 Alkyl groups. In some cases, each R... 31 Each occurrence is independently selected from C groups that are optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles, wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocyclic ring are optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, -NO2, -NH2, -OC. 1-6 Alkyl and -C 1-6 Alkyl groups. In some cases, each R... 31 Each occurrence is independently selected from C groups that are optionally substituted by one or more substituents independently selected from 3 to 6-membered heterocycles. 1-6 Alkyl group, wherein the 3- to 6-membered heterocycle is optionally selected independently from -OC 1-6 Alkyl and -C 1-6 Alkyl substituents. In some cases, each R... 31 Each occurrence is independently selected from C1646, which is optionally substituted by one or more substituents independently selected from 5-membered heterocycles. 1-6 Alkyl group, wherein the 5-membered heterocycle is optionally selected independently from -C 1-6 Alkyl substituents.

[0144] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 4’ Selected from C1-C6 alkyl groups substituted with one or more fluorine molecules. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with one or more hydroxyl groups. In some cases, R 4’ Selected from the unreplaced C 3-6 Carbocyclic substituted C1-C6 alkyl groups. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with phenyl groups, wherein the phenyl group is substituted with one or more halogens. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with phenyl groups, wherein the phenyl group is substituted with one or more halogens selected from fluorine and chlorine. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with phenyl groups, wherein the phenyl group is substituted with at least one or two chlorine groups. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with phenyl groups, wherein the phenyl group is substituted with at least one or two fluorine molecules. In some cases, R 4’Selected from C1-C6 alkyl groups substituted with phenyl groups, wherein the phenyl group is substituted with two chlorine groups. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with phenyl groups, wherein the phenyl group is substituted with two fluorine groups. In some cases, R 4’ Selected from C1-C6 alkyl groups substituted with phenyl, wherein the phenyl group is converted to a chlorine and -OR 31 Replacement. In some cases, -OR 31 R 31 C is replaced by a 5-membered heterocycle. 1-6 Alkyl group, wherein the 5-membered heterocycle is optionally selected independently from -C 1-6 Alkyl substituents. In some cases, the 5-membered heterocycle has at least two heteroatoms. In some cases, the 5-membered heterocycle has at most two heteroatoms. In some cases, the 5-membered heterocycle has two heteroatoms. In some cases, the heteroatoms of the 5-membered heterocycle are different.

[0145] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 4’ Selected from In some cases, R 4’ yes In some cases, R 4’ yes In some cases, R 4’ yes In some cases, R 4’ yes In some cases, R 4’ yes

[0146] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 6’ Selected from hydrogen and -C(O)(NR) 50 2); R 5’ Selected from hydrogen, -NO2, -CN, -NH2, halogen, C3-C6 cycloalkyl, C1-C6 alkyl; R 12’ Selected from -NO2, -CN, -NH2 and -OC 1-6 Alkyl; and R 4’ Selected from optionally substituted C1-C6 alkyl groups, wherein the C1-C6 alkyl group is optionally substituted by one or more substituents selected independently of the following groups each time it appears: fluorine; C1-C6 alkyl group is optionally substituted by one or more substituents selected independently of the following groups. 3-6 Carbon rings: halogens, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 Alkyl group. In some cases, R 6’It is -C(O)(NR) 50 2). In some cases, R6' is hydrogen.

[0147] In some embodiments, for compounds or salts of formula (II') or (IIA'), R4' is selected from unsubstituted C1-C6 alkyl groups.

[0148] In some embodiments, for compounds or salts of formula (II') or (IIA'), when R6' is -C(O)(NHMe) and R9 is an optionally substituted 3- to 6-membered heterocycle, R9 is optionally substituted with one or more substituents selected independently from: fluorine, bromine, -OR30, -N(R30)2, -NO2, -CN, and C1-3 alkyl. In some cases, when R6' is -C(O)(NHMe), R9 is not an optionally substituted 3- to 6-membered heterocycle. In some cases, when R6' is -C(O)(NHMe), R9 is not an optionally substituted 5-membered heterocycle.

[0149] In some embodiments, for compounds or salts of formula (II') or (IIA'), R4' is hydrogen.

[0150] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 5’ Selected from hydrogen, -NO2, -CN, -NH2, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl are optionally selected independently by one or more of R 9 Substituents are substituted. In some cases, R 5’ Selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, -NO2, -CN, -NH2, and halogens. In some cases, R 5’ Selected from C1-C6 alkyl groups, -NO2, -CN, -NH2, and halogens. In some cases, R 5’ Selected from -CN and halogens.

[0151] In some embodiments, for compounds of formula (II') or formula (IIA'), R 6’ The substituents are selected from hydrogen and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl group of R6' are independently selected from R10.

[0152] In some embodiments, for compounds or salts of formula (II') or (IIA'), R6' is selected from hydrogen, CH3, CH2F, CHF2, and CF3. In some cases, R6' is hydrogen.

[0153] In some embodiments, for compounds of formula (II') or (IIA'), R6' is selected from -C(O)(NR502) and hydrogen. In some cases, R6' is -C(O)(NR502). In some cases, R50 is selected from hydrogen and C1-3 alkyl. In some cases, R50 is selected from C1-3 alkyl. In some cases, R6' is -C(O)(NHMe).

[0154] In some embodiments, for compounds or salts of formula (II') or (IIA'), when R6' is -C(O)(NHMe) and R9 is an optionally substituted 3- to 6-membered heterocycle, R9 is optionally substituted with one or more substituents selected independently from one or more of the following: -OR30, -N(R30)2, -NO2, -CN and C1-3 alkyl.

[0155] In some embodiments, for compounds or salts of formula (II') or (IIA'), when R6' is -C(O)(NHMe) and R9 is an optionally substituted 5-6 membered heterocycle, R9 is optionally substituted with one or more substituents selected from one or more substituents independently selected from: halogen, -OR30, -N(R30)2, -NO2, -CN and C1-3 alkyl.

[0156] In some embodiments, for compounds or salts of formula (II') or (IIA'), R12' is selected from hydrogen, halogens, and unsubstituted C1-6 alkyl groups. In some cases, R12' is selected from fluorine and chlorine. In some cases, R12' is fluorine. In some cases, R12' is hydrogen. In some cases, R12' is selected from unsubstituted C1-3 alkyl groups. In some cases, R12' is methyl. In some cases, R12' is selected from -NO2, -CN, -NH2, and -O-C1-6 alkyl groups. In some cases, R12' is selected from -NO2, -CN, and -NH2. In some cases, R12' is -CN. In some cases, R12' is selected from hydrogen, -CN, and unsubstituted C1-3 alkyl groups.

[0157] In some embodiments, for compounds or salts of formula (II') or (IIA'), R 12’ Selected from hydrogen, halogen, hydroxyl, -NO2, -CN, -NH2, -OC 1-6 Alkyl and C 1-6 Alkyl group, wherein the -OC 1-6 The alkyl moiety and C 1-6 The alkyl group is optionally substituted by one or more substituents selected from the following: halogen, -OH, -NH2, -NO2, -CN, -OC. 1-6 Alkyl, C 3-6Carbon rings, 3- to 6-membered heterocycles; wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents selected from the following: halogen, hydroxyl, -NO2, -CN, -NH2, -OC. 1-6 Alkyl and C 1-6 Alkyl group. In some cases, R 12’ Selected from hydrogen, halogen, hydroxyl, -CN, -OC 1-6 Alkyl and C 1-6 Alkyl group, wherein the -OC 1-6 The alkyl moiety and C 1-6 The alkyl group is optionally substituted by one or more substituents selected from the following: halogen, -OH, -CN, -OC. 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles; wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents selected from the following: halogen, hydroxyl, -NO2, -CN, -NH2, -OC. 1-6 Alkyl and C 1-6 Alkyl group. In some cases, R 12’ yes

[0158] In some embodiments, for compounds of formula (II') or formula (IIA'), R 6’ The substituents are selected from hydrogen and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl groups are independently selected from R10.

[0159] In some embodiments, for compounds or salts of formula (II') or (IIA'), each R7, R9, and R10 is independently selected each time it appears:

[0160] Fluorine, -OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30 -OC(O)R 30

[0161] -NO2 and -CN; and

[0162] C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 30 -N(R) 30 2, -NO2, -CN and C 1-3 alkyl.

[0163] In some embodiments, for compounds or salts of formula (II') or (IIA'), each R 7 R 9 and R 10 Each time it appears, it is selected independently from: fluorine, -OR 30 -N(R) 30 2. -NO2 and -CN; and C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 30 -N(R) 30 2, -NO2, -CN and C 1-3 alkyl.

[0164] In some embodiments, formula (II') is represented by formula (II) or a pharmaceutically acceptable salt thereof:

[0165]

[0166] in:

[0167] R 4’ The substituent is selected from optionally substituted C1-C6 alkyl groups, optionally substituted C3-C8 carbon rings, and optionally substituted 3- to 8-membered heterocycles, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R. 7 Furthermore, the optional substituents on the C3-C8 carbon rings and 3- to 8-membered heterocycles are independently selected from R. 8 ;

[0168] R 5’ Selected from hydrogen, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl, wherein the C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 ynyl are optionally selected independently by one or more of R. 9 Substituents of the substituents;

[0169] R 6’ Selected from hydrogen and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl groups are independently selected from R. 10 ;

[0170] Each R 7 R 9 and R 10 Selected independently each time it appears:

[0171] Fluorine-OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30-OC(O)R 30 -NO2 and -CN;

[0172] C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 30 -N(R) 30 2, -NO2, -CN and C 1-3 alkyl;

[0173] Each R 8 Selected independently each time it appears:

[0174] Halogen, -OR 30 -N(R) 30 )2、-N(R 30 )2、-C(O)R 30 -C(O)OR 30 -OC(O)R 30 -NO2 and -CN;

[0175] C10 ... 1-3 Alkyl groups: halogens, -OR 30 -N(R) 30 )2、-NO2、-CN、C 3-6 Carbon rings and 3- to 6-membered heterocycles; and

[0176] Each R 30 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogens, -OH, -CN, -NO2, -NH2,

[0177] -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0178] In some embodiments, for compounds or salts of formula (II'), formula (II), formula (IIA'), or formula (IIA), R 4’ The components are selected from C3-C6 carbon rings and optionally substituted C1-C6 alkyl groups, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R7 each time they appear.

[0179] In some embodiments, for compounds or salts of formula (II') or (IIA), R7 is selected from fluorine, -NO2, and -CN; and C 3-6 Carbon ring, wherein the C 3-6 The carbon ring is optionally substituted by one or more substituents independently selected from the following: halogen, -NO2, -CN, and C.1-3 alkyl.

[0180] In some embodiments, the compound or salt of formula (II') is represented by formula (IIA):

[0181]

[0182] In some embodiments, for compounds or salts of formula (II'), formula (II), formula (IIA'), or formula (IIA), R 4’ Selected from C which is arbitrarily replaced 1-2 Alkyl groups and optionally substituted C3-C6 carbon rings. In some cases, R 4’ It is an optionally substituted C1 alkyl group. In some cases, R 4’ It is an optionally substituted C2 alkyl group. In some cases, R 4’ It is an ethyl group. In some cases, C... 1-2 The optional substituents of the alkyl group are independently selected from R each time they appear. 7 .

[0183] In some embodiments, for compounds or salts of formula (II'), formula (II), formula (IIA'), or formula (IIA), R 7 It is selected independently from fluorine-OR each time it appears. 30 -N(R) 30 )2、-N(R 30 )2, -NO2, -CN; and C 3-6 The carbon rings are optionally substituted by one or more substituents independently selected from the following: halogens, -OR 30 -N(R) 30 )2、-NO2、-CN、C 1-3 Alkyl group. In some cases, R 7 Selected from fluorine and optionally substituted phenyl groups. In some cases, R 4’ Selected from an R 7 Substituted C1 alkyl, wherein R 7 It is a phenyl group that is optionally substituted; and is substituted with one or more R 7 Substituted C2 alkyl, wherein R 7 It's fluorine. In some cases, R... 4’ It is by one or more R 7 Substituted C2 alkyl, wherein R 7 It's fluorine. In some cases, R... 4’ It is at most three Rs 7 Substituted C2 alkyl, wherein R 7 It's fluorine. In some cases, R... 4’ It was by an R 7Substituted C1 alkyl, wherein R 7 It is an optionally substituted phenyl group. In some cases, R 7 The optional substituents of the phenyl group are independently selected from halogens, -OR, each time they appear. 30 -N(R) 30 )2、-NO2、-CN、C 1-3 Alkyl group. In some cases, R 7 It is a substituted phenyl group. In some cases, R 7 The phenyl group is substituted with one or more fluorine atoms. In some cases, R 7 The phenyl group is replaced by two fluorine atoms. In some cases, R 4’ Selected from CH2CH3, CH2CFH2, CH2CF2H, CH2CF3,

[0184] In some embodiments, for compounds or salts of formula (II'), formula (II), formula (IIA'), or formula (IIA), R 5’ Selected from hydrogen, CH3, CH2CH3, And Cl. In some implementations, R 5’ Selected from hydrogen, CH3, CH2CH3, and Cl. In some cases, R 5’ Selected from hydrogen, CH3, and Cl. In some cases, R 5’ It is Cl. In some cases, R 5’ yes

[0185] In some embodiments, for compounds of formula (II'), formula (II), formula (IIA'), or formula (IIA), R 6’ Selected from hydrogen, CH3, CH2F, CHF2, and CF3. In some cases, R 6’ It is hydrogen.

[0186] In some embodiments, for compounds or salts of formula (II') or formula (IIA'),

[0187] R 4’ Selected from hydrogen and optionally substituted C1-C6 alkyl groups, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R 7 ;

[0188] R 5’ Selected from -CN, halogens, C3-C6 cycloalkyl and C1-C6 alkyl, wherein the C1-C6 alkyl is optionally selected by one or more independently from R. 9 Substituents of the substituents;

[0189] R 6’Selected from hydrogen and -C(O)(NR) 50 2;

[0190] Each R 7 Selected independently each time it appears:

[0191] Fluorine, -OR 30 ;as well as

[0192] C10 ... 3-6 Carbon rings: halogens, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 alkyl;

[0193] Each R 9 Selected independently each time it appears:

[0194] Fluorine, -OR 30 -N(R) 30 2. -NO2 and -CN; and

[0195] C10 ... 3-6 Carbon rings: halogens, -OR 31 -N(R) 31 2, -NO2, -CN and C 1-3 alkyl;

[0196] Each R 30 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3- to 6-membered heterocycles;

[0197] Each R 50 Each time it appears, it is independently selected from hydrogen and C. 1-6 alkyl;

[0198] Each R 31 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings, 3- to 6-membered heterocycles, wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocyclic ring are optionally substituted by one or more substituents independently selected from the following: halogen, -OH, -CN, -NO2, -NH2, -OC. 1-6Alkyl and -C 1-6 Alkyl; and

[0199] R 12’ Selected from hydrogen, halogen, hydroxyl, -NO2, -CN, -NH2, -OC 1-6 Alkyl and C 1-6 Alkyl group, wherein the -OC 1-6 The alkyl moiety and C 1-6 The alkyl group is optionally substituted by one or more substituents selected from the following: halogen, -OH, -NH2, -NO2, -CN, C. 3-6 Carbon rings, 3- to 6-membered heterocycles; wherein the C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents selected from the following: halogen, hydroxyl, -NO2, -CN, -NH2, -OC. 1-6 Alkyl and C 1-6 alkyl.

[0200] On the other hand, this disclosure provides a compound represented by formula (III):

[0201]

[0202] Or its pharmaceutically acceptable salt, wherein:

[0203] R 21 Selected from C3-C6 cycloalkyl and 3-6 membered heterocyclic alkyl, wherein the C3-C6 cycloalkyl and 3-6 membered heterocyclic alkyl are optionally selected independently by one or more of R 29 Substituents of the substituents;

[0204] R 22 The substituents are selected from optionally substituted C1-C6 alkyl groups, optionally substituted C3-C8 carbon rings, and optionally substituted 3- to 8-membered heterocycles, wherein the optional substituents on the C1-C6 alkyl groups are independently selected from R each time they appear. 27 Furthermore, the optional substituents on the C3-C8 carbon rings and 3- to 8-membered heterocycles are independently selected from R. 28 ;

[0205] R 23 Selected from hydrogen and optionally substituted C1-C3 alkyl groups, wherein the optional substituents on the C1-C3 alkyl groups are independently selected from R. 40 ;

[0206] Each R 27 R 29 and R 40 Selected independently each time it appears:

[0207] Fluorine-OR 50 -N(R) 50)2、-N(R 50 )2、-C(O)R 50 -C(O)OR 50 -OC(O)R 50 -NO2 and -CN;

[0208] C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: halogen, -OR 50 -N(R) 50 2, -NO2, -CN and C 1-3 alkyl;

[0209] Each R 28 Selected independently each time it appears:

[0210] Halogen, -OR 50 -N(R) 50 )2、-N(R 50 )2、-C(O)R 50 -C(O)OR 50 -OC(O)R 50 -NO2 and -CN;

[0211] C10 ... 1-3 Alkyl groups: halogens, -OR 50 -N(R) 50 )2、-NO2、-CN、C 3-6 Carbon rings and 3- to 6-membered heterocycles; and

[0212] R 50 Each time it appears, it is independently selected from hydrogen and optionally substituted by one or more substituents independently selected from the following. 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0213] In some embodiments, compounds of formula (III) are represented by formula (IIIA):

[0214]

[0215] In some embodiments, R21 is selected from C3-C6 cycloalkyl groups. In some cases, R21 is selected from C3 cycloalkyl groups.

[0216] In some implementation schemes, R 22 Selected from optionally substituted C1-C6 alkyl groups. In some cases, R 22Selected from unsubstituted C1-C6 alkyl groups.

[0217] In some implementations, each R 27 Independently selected from -OR 50 -N(R) 50 )2、-N(R 50 )2、-C(O)R 50 -C(O)OR 50 -OC(O)R 50 -NO2 and -CN; C 3-6 The carbocyclic ring and the 3- to 6-membered heterocycle are each optionally substituted by one or more substituents independently selected from the following: - OR 50 -N(R) 50 2, -NO2, -CN and C 1-3 Alkyl groups. In some cases, each R... 50 Each time it appears, it is independently selected from hydrogen and C. 1-6 alkyl.

[0218] In some cases, R23 is selected from hydrogen, CH3, CH2F, CHF2, and CF3.

[0219] On the other hand, this disclosure provides a compound represented by formula (IV):

[0220]

[0221] Or its pharmaceutically acceptable salt, wherein:

[0222] R 31 Selected from hydrogen, halogen, C3-C6 cycloalkyl, C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl.

[0223] The C1-C6 alkyl, C2-C6 alkenyl, and C2-C6 alkynyl groups are optionally selected independently from R. 39 Substituents of the substituents;

[0224] R 32 Selected from C1-C6 alkyl groups;

[0225] R 33 Selected from C1-C3 haloalkyl groups;

[0226] Each R 39 Selected independently each time it appears:

[0227] Fluorine, -OR 60 -N(R) 60 )2、-N(R 60 )2、-C(O)R 60 -C(O)OR 60-OC(O)R 60

[0228] -NO2 and -CN;

[0229] C 3-6 The carbon ring and 3 to 6-membered heterocycles are each optionally selected independently from one or more of the following:

[0230] Substituents: halogen, -OR 60 -N(R) 60 2, -NO2, -CN and C 1-3 Alkyl; each R 60 Each time it appears, it is independently selected from hydrogen and optionally selected by one or more independent selections.

[0231] The following substituents replace C 1-6 Alkyl groups: halogen, -OH, -CN, -NO2, -NH2, -OC 1-6 Alkyl, C 3-6 Carbon rings and 3 to 6-membered heterocycles.

[0232] In some embodiments, compounds of formula (IV) are represented by formula (IVA):

[0233]

[0234] In some implementations, for compounds of formula (IV) or formula (IVA), R 31 Selected from hydrogen, CH3, CH2CH3, Cl, and In some implementation schemes, R 31 Selected from hydrogen, CH3, CH2CH3, Cl and In some cases, R 31 Selected from Cl.

[0235] In some implementations, for compounds of formula (IV) or formula (IVA), R32 is ethyl.

[0236] In some implementations, for compounds of formula (IV) or formula (IVA), R 33 Selected from C1-C3 fluoroalkyl groups. In some cases, R 33 yes

[0237] This disclosure includes salts of the compounds described herein, particularly pharmaceutically acceptable salts. Compounds of the present invention having sufficiently acidic functional groups, sufficiently basic functional groups, or both, can react with a variety of inorganic bases and any of inorganic and organic acids to form salts. Alternatively, inherently charged compounds, such as those having quaternary nitrogen, can form salts with suitable counterions such as halide ions such as bromide, chloride, or fluoride ions, particularly bromide ions.

[0238] Chemical entities having carbon-carbon or carbon-nitrogen double bonds can exist in Z or E forms (or cis or trans forms). Furthermore, some chemical entities can exist in various tautomeric forms. Unless otherwise specified, the compounds described herein are intended to include all Z, E, and tautomeric forms.

[0239] A "tautomer" is a molecule in which a proton may move from one atom of the molecule to another atom of the same molecule. In some embodiments, the compounds provided herein exist in tautomer form. Where tautomerism is possible, a chemical equilibrium of tautomerism will exist. The precise ratio of tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeristic equilibria include:

[0240]

[0241] In some embodiments, the compounds disclosed herein are used in different enriched isotopic forms, for example in 2 H, 3 H, 11 C 13 C and / or 14 The content of C is enriched. In one particular embodiment, the compound is deuterated at at least one position. Such a deuterated form can be prepared by the procedures described in U.S. Patent Nos. 5,846,514 and 6,334,997. As described in U.S. Patent Nos. 5,846,514 and 6,334,997, deuteration can improve metabolic stability and / or efficacy, thereby increasing the duration of action of the drug.

[0242] Unless otherwise stated, the compounds described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of the invention, except where hydrogen is replaced by deuterium or tritium, or carbon is replaced by 13C or 14C enriched carbon, are within the scope of this disclosure.

[0243] The compounds disclosed herein optionally contain atomic isotopes in non-natural proportions at one or more atoms constituting such compounds. For example, the compounds may be labeled with isotopes such as deuterium (…). 2 H), tritium (3 H), Iodine-125 ( 125 I) or carbon-14 ( 14 C). Use 2 H, 11 C 13 C 14 C 15 C 12 N、 13 N、 15 N、 16 N、 16 O、 17 O、 14 F, 15 F, 16 F, 17 F, 18 F, 33 S, 34 S, 35 S, 36 S, 35 Cl、 37 Cl、 79 Br、 81 Br and 125 All isotopic substitutions performed by I are taken into consideration. All isotopic variations of the compounds of the present invention, whether or not they are radioactive, are covered within the scope of the present invention.

[0244] In some embodiments, some or all of the compounds disclosed herein 1 H atoms are 2 H atom substitution. Methods for synthesizing deuterium-containing compounds are known in the art, and, by way of example only, include the following synthetic methods.

[0245] Deuterium-substituted compounds were synthesized using various methods such as those described in the following: Dean, Dennis C., ed., Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [Curr., Pharm.Des., 2000; 6(10)] 2000, p. 110; George W.; Varma, Rajender S., The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony., Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.

[0246] Deuteration starting materials are readily available and can be synthesized using the methods described herein to provide the synthesis of deuterium-containing compounds. A wide range of deuterium-containing reagents and building blocks are commercially available from chemical suppliers such as Aldrich Chemical Co.

[0247] The compounds of the present invention also include crystalline and amorphous forms of these compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrous forms), conformational polymorphs and amorphous forms of the compounds, and mixtures thereof.

[0248] In some cases, the compounds described herein may exist as diastereomers, enantiomers, or other stereoisomers. When absolute stereochemistry is not specified, the compounds provided herein include all diastereomers, enantiomers, and epimers, as well as suitable mixtures thereof. Separation of stereoisomers can be performed by chromatography or by forming diastereomers and separating them by recrystallization or chromatography, or any combination thereof. (Jean Jacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John Wiley and Sons, Inc., 1981, incorporated herein by reference). Stereoisomers can also be obtained by stereoselective synthesis.

[0249] The methods and compositions described herein include the use of amorphous and crystalline forms (also known as polymorphs). The compounds described herein may be in pharmaceutically acceptable salt form. Similarly, in some embodiments, active metabolites of these compounds having the same type of activity are included within the scope of this disclosure. Furthermore, the compounds described herein may be in unsolvated form as well as in solvated form with pharmaceutically acceptable solvents such as water, ethanol, etc. The solvated forms of the compounds provided herein are also considered to be disclosed herein.

[0250] In some embodiments, the compound or a salt of a compound may be a prodrug, for example, wherein the hydroxyl group in the parent compound is presented as an ester or carbonate, or the carboxylic acid present in the parent compound is presented as an ester. The term "prodrug" is intended to cover compounds that are converted into the pharmaceutical agents disclosed herein under physiological conditions. One method for preparing a prodrug involves hydrolyzing one or more selected portions of a desired molecule under physiological conditions to expose them. In other embodiments, the prodrug is converted by the enzymatic activity of a host animal, such as specific target cells in the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids, and esters of phosphonates) are preferred prodrugs of this disclosure.

[0251] The scope of the claims includes the prodrug form of the compounds described herein, wherein the prodrug is metabolized in vivo to produce the compounds as set forth herein. In some cases, some of the compounds described herein may be prodrugs of another derivative or active compound.

[0252] Prodrugs are often useful because, in some cases, they can be easier to administer than the parent drug. They can achieve bioavailability, for example, through oral administration, which the parent drug cannot. Prodrugs can help enhance the cellular permeability of compounds compared to the parent drug. Prodrugs may also have improved solubility in pharmaceutical compositions compared to the parent drug. Prodrugs can be engineered as reversible drug derivatives to act as modulators to enhance drug transport to site-specific tissues or increase drug retention within cells.

[0253] In some implementations, the prodrug is designed to increase the lipophilicity of the drug. In some implementations, the prodrug is designed to increase effective water solubility. See, for example, Fedorak et al., Am. J. Physiol., 269: G210-218 (1995); McLoed et al., Gastroenterol, 106: 405-413 (1994); Hochhaus et al., Biomed. Chrom., 6: 283-286 (1992); J. Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J. Larsen et al., Int. J. Pharmaceutics, 47, 103 (1988); Sinkula et al., J. Pharm. Sci., 64: 181-210 (1975); T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Volume 14 of the ACS Symposium Series; and Edward B. Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987 (all incorporated herein by reference in this disclosure). According to another embodiment, this disclosure provides a method for producing compounds as defined above. The compounds can be synthesized using conventional techniques. Advantageously, these compounds are conveniently synthesized from readily available starting materials.

[0254] Synthetic chemical transformations and methods that can be used to synthesize the compounds described herein are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations (1989); T.W. Greene and P.G. M. Wuts, Protective Groups in Organic Synthesis, 2nd edition (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis (1995).

[0255] C. Pharmaceutical Composition

[0256] In some embodiments, this document provides compositions comprising a therapeutically effective amount of any compound or salt of either of formulas (I) and (II) (also referred to herein as a “pharmaceutical”).

[0257] Pharmaceutical compositions may be formulated using one or more physiologically acceptable carriers (including excipients and adjuvants) that facilitate the processing of the pharmaceutical agent into a pharmaceutically usable formulation. The appropriate formulation depends on the chosen route of administration. An overview of pharmaceutical compositions can be found, for example, in the following: Remington: The Science and Practice of Pharmacy, 19th edition (Easton, Pa., Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L. (eds.), Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th edition (Lippincott Williams & Wilkins, 1999).

[0258] The compositions and methods disclosed herein can be used to treat individuals in need. In some embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal such as a human, the composition or agent is preferably administered in the form of a pharmaceutical composition comprising, for example, a pharmaceutical agent and a pharmaceutically acceptable carrier or excipient. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline, or other solvents or mediators such as glycols, glycerols, oils such as olive oil, or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive routes of administration, such as those bypassing transport or diffusion through the epithelial barrier, such as injection or implantation, the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients may be selected, for example, to achieve delayed release of the agent or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition may be in the form of dosage units, such as tablets, capsules, granules, hydrophilic agents for decongestion, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system such as a skin patch. The composition may also be present in solutions suitable for surface application, such as eye drops.

[0259] Pharmaceutically acceptable excipients may contain physiologically acceptable agents, for example, that stabilize compounds such as pharmaceuticals, increase the solubility of compounds such as pharmaceuticals, or increase the absorption of compounds such as pharmaceuticals. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The choice of pharmaceutically acceptable excipients including physiologically acceptable agents depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix, which may contain compounds such as those of the present invention incorporated therein. For example, liposomes containing phospholipids or other lipids are relatively simple to prepare and administer, and are non-toxic, physiologically acceptable, and metabolizable carriers.

[0260] Pharmaceutical compositions (formulations) can be administered to a subject via any of a number of routes of administration, including, for example, oral administration, such as oral enemas, tablets, capsules (including sprinkle capsules and gelatin capsules), pellets, powders, granules, pastes for application to the tongue, as an aqueous or non-aqueous solution or suspension; absorption through the oral mucosa, such as sublingually; anal, rectal, or vaginal administration, such as in the form of vaginal suppositories, creams, or foams; parenteral administration, including intramuscular, intravenous, subcutaneous, or intrathecal administration, in the form of, for example, sterile solutions or suspensions; nasal administration; intraperitoneal administration; subcutaneous administration; transdermal administration, such as in the form of patches applied to the skin; and surface administration, such as in the form of creams, ointments, or sprays applied to the skin, or in the form of eye drops. The compound can also be formulated for inhalation. In some embodiments, the compound may simply be dissolved or suspended in sterile water.

[0261] Pharmaceutical compositions may be sterile aqueous or non-aqueous solutions, suspensions, or emulsions, such as microemulsions. The excipients described herein are examples and are by no means limiting. An effective amount, or therapeutically effective amount, refers to the amount of one or more pharmaceutical agents administered to a subject in a single dose or as part of a series of doses to effectively produce the desired therapeutic effect.

[0262] Treatment efficacy in a subject can be monitored using assays and methods generally suitable for the condition being treated, assays familiar to those skilled in the art and described herein. The pharmacokinetics of the drug or one or more metabolites administered to the subject can be monitored by measuring the levels of the drug or metabolites in biological fluids, such as blood, blood fractions such as serum, and / or urine and / or other biological samples or tissues from the subject. Any method practiced in the art and described herein for detecting agents can be used to measure the levels of the drug or metabolites during treatment.

[0263] The dosage of the pharmaceutical agents described herein for treating a disease or condition may depend on the subject's condition, i.e., the stage of the disease, the severity of the symptoms caused by the disease, overall health status, and age, sex, and weight, as well as other factors obvious to a person skilled in the medical field. The pharmaceutical composition may be administered in a manner suitable for the disease to be treated, as determined by a person skilled in the medical field. In addition to the factors described herein and above related to the use of pharmaceutical agents for treating a disease or condition, the appropriate duration and frequency of administration of the pharmaceutical agent may also be determined or adjusted based on factors such as the patient's condition, the type and severity of the patient's disease, the specific form of the active ingredient, and the method of administration. The optimal dosage of the agent may generally be determined using experimental models and / or clinical trials. The optimal dosage may depend on the subject's body mass, weight, or blood volume. Using the minimum dose sufficient to provide effective treatment is generally preferred. The design and execution of preclinical and clinical studies of the pharmaceutical agents described herein (including when administered for preventative benefit) are entirely within the skill of a person skilled in the relevant field. When two or more pharmaceutical agents are administered to treat a disease or condition, the optimal dosage of each agent may differ, such as being less than when any agent is administered alone as a single-agent therapy. In certain specific embodiments, the two combined agents may act synergistically or additively, and either agent may be used in a smaller amount than when administered alone. The daily dose of the agent, based on body weight, may be, for example, between about 0.01 mg / kg and 100 mg / kg, for example, between about 0.1 and 1 mg / kg, between about 1 and 10 mg / kg, between about 10 and 50 mg / kg, or between about 50 and 100 mg / kg. In other embodiments, the daily dose of the agent, based on body weight, may be between about 0.01 mg / kg and 1000 mg / kg, between about 100 and 500 mg / kg, or between about 500 and 1000 mg / kg. The optimal daily dose or dose per course of treatment may vary depending on the disease or condition to be treated and may also vary with the route of administration and treatment regimen.

[0264] Pharmaceutical compositions containing pharmaceutical agents can be formulated using techniques conventionally practiced in the art in a manner suitable for delivery methods. The compositions can be in the form of solids (e.g., tablets, capsules), semi-solids (e.g., gels), liquids, or gases (e.g., aerosols). In other embodiments, the pharmaceutical composition is administered as a bolus infusion.

[0265] Pharmaceutically acceptable excipients are well known in the pharmaceutical industry and are described, for example, in Rowe et al., Handbook of Pharmaceutical Excipients: A Comprehensive Guide to Uses, Properties, and Safety, 5th Edition, 2006, and Remington: The Science and Practice of Pharmacy (Gennaro, 21st Edition, Mack Pub. Co., Easton, PA (2005)). Exemplary pharmaceutically acceptable excipients include sterile saline and phosphate-buffered saline at physiological pH. Preservatives, stabilizers, dyes, buffers, etc., may be provided in the pharmaceutical composition. In addition, antioxidants and suspending agents may be used. Generally, the type of excipient is selected based on the administration method and the chemical composition of the active ingredient. Alternatively, the compositions described herein may be formulated as lyophilized products. The compositions described herein may be lyophilized or otherwise formulated as lyophilized products, and the pharmaceutical agent of the composition may be dissolved and / or diluted with one or more suitable excipient solutions at the time of administration. In other embodiments, the pharmaceutical agent may be encapsulated in liposomes using techniques known and practiced in the art. In certain specific embodiments, the agent is not formulated in liposomes for application to a stent used to treat highly but not completely occluded arteries. The pharmaceutical composition may be formulated for any suitable administration method described herein and in the art.

[0266] For example, pharmaceutical compositions intended for oral administration or for injection, infusion, subcutaneous delivery, intramuscular delivery, intraperitoneal delivery, or other methods may be in liquid form. Liquid pharmaceutical compositions may include one or more of the following: sterile diluents such as water, saline solution (preferably physiological saline), Ringer's solution, isotonic sodium chloride, non-volatile oils that can act as solvents or suspending media, polyethylene glycol, glycerin, propylene glycol, or other solvents; antibacterial agents; antioxidants; chelating agents; buffers and reagents for adjusting tension, such as sodium chloride or dextran. Parenteral compositions may be packaged in ampoules, disposable syringes, or multi-dose vials made of glass or plastic. The use of physiological saline is preferred, and injectable pharmaceutical compositions are preferably sterile. In another embodiment, for the treatment of ophthalmic conditions or diseases, the liquid pharmaceutical composition may be applied to the eye in the form of eye drops. Liquid pharmaceutical compositions may also be delivered orally.

[0267] For oral formulations, at least one of the pharmaceutical agents described herein may be used alone or in combination with suitable additives to prepare tablets, powders, granules, or capsules, and, if desired, in combination with diluents, buffers, humectants, preservatives, colorants, and flavoring agents. The pharmaceutical agent may be formulated with a buffer to provide protection of the compound from the low pH of the gastric environment and / or with an enteric coating. Pharmaceutical agents included in pharmaceutical compositions may be formulated with a flavoring agent, for example, in liquid, solid, or semi-solid dosage form, and / or with an enteric coating, for oral delivery.

[0268] Pharmaceutical compositions comprising any of the agents described herein may be formulated for sustained or slow release, also known as timed release or controlled release. Such compositions may generally be prepared using well-known techniques and administered, for example, orally, rectally, intradermally, or subcutaneously, or by implantation at a desired target site. Sustained-release formulations may contain compounds dispersed in a carrier matrix and / or contained in a reservoir surrounded by a rate-controlled membrane. Excipients used in such formulations are biocompatible and may also be biodegradable; preferably, the formulation provides a relatively constant level of release of the active ingredient. The amount of agent contained in a sustained-release formulation depends on the implantation site, the rate of release and the expected duration, and the nature of the condition, disease, or ailment to be treated or prevented.

[0269] In some embodiments, the pharmaceutical composition containing the agent is formulated for transdermal, intradermal, or topical application. The composition can be applied using syringes, bandages, transdermal patches, inserts, or syringe-like applicators in the form of powder / talcum powder or other solids, liquids, sprays, aerosols, ointments, foams, creams, gels, or pastes. This is preferably in the form of a controlled-release or sustained-release formulation, which is applied topically or injected, for example, intradermally or subcutaneously, directly into the skin adjacent to or within the area to be treated. The active composition can also be delivered via iontophoresis. Preservatives may be used to prevent the growth of fungi and other microorganisms. Suitable preservatives include, but are not limited to, benzoic acid, butylparaben, ethylparaben, methylparaben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzyl chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethanol, thimerosal, and combinations thereof.

[0270] Pharmaceutical compositions containing agents can be formulated as emulsions for surface application. The emulsion contains a liquid distributed within a bulk of a second liquid. The emulsion can be an oil-in-water emulsion or an oil-in-water emulsion. Either or both of the oil and aqueous phases may contain one or more surfactants, emulsifiers, emulsion stabilizers, buffers, and other excipients. The oil phase may contain other pharmaceutically approved oily excipients. Suitable surfactants include, but are not limited to, anionic surfactants, nonionic surfactants, cationic surfactants, and amphoteric surfactants. Compositions for surface application may also include at least one suitable suspending agent, antioxidant, chelating agent, emollient, or moisturizing agent.

[0271] Ointments and creams can be formulated, for example, with an aqueous or oil-based base, in the addition of suitable thickeners and / or gelling agents. Lotions can be formulated with an aqueous or oil-based base and will typically also contain one or more emulsifiers, stabilizers, dispersants, suspending agents, thickeners, or colorants. Liquid sprays can be delivered from pressurized packaging, such as through specially shaped closures. Oil-in-water emulsions can also be used in compositions, patches, bandages, and articles. These systems are semi-solid emulsions, microemulsions, or foam emulsion systems.

[0272] In some embodiments, the pharmaceutical agents described herein may be formulated as inhalers. Inhalation methods deliver the drug directly to the airways. The agents may be formulated as aerosols, microspheres, liposomes, or nanoparticles. The agents may be formulated with solvents, gases, nitrates, or any combination thereof. The compositions described herein are optionally formulated for delivery as liquid aerosols or inhalable dry powders. Liquid aerosol formulations are optionally primarily atomized to a particle size deliverable to the terminal and respiratory bronchioles. Liquid aerosols and inhalable dry powder formulations are preferably delivered throughout the bronchial tree to the terminal bronchioles and ultimately to the parenchymal tissue.

[0273] The aerosolized formulations described herein are optionally delivered using aerosol forming devices such as injectors, vibrating perforated plates, or ultrasonic nebulizers, preferably selected to allow the formation of aerosol particles having a median mass average diameter primarily between 1 and 5 μm. Furthermore, the formulation preferably has an equilibrium volumetric molality, ionic strength, and chloride ion concentration, as well as a minimum aerosolizable volume capable of delivering an effective dose of the drug. Additionally, the aerosolized formulation preferably does not adversely impair airway function and does not cause unwanted side effects.

[0274] Suitable aerosol devices for administering the aerosol formulations described herein include, for example, jet injectors, vibrating perforated plates, ultrasonic nebulizers, and electrically powered dry powder inhalers, capable of atomizing the formulation into aerosol particles primarily in the size range of 1-5 μm. In this application, "primarily" means that at least 70%, but preferably more than 90%, of all generated aerosol particles are in the 1-5 μm range. Jet injectors operate by means of air pressure to break up liquid solutions into aerosol droplets. Vibrating perforated plate nebulizers operate by means of an acoustic vacuum generated by a rapidly vibrating perforated plate to expel solvent droplets through the perforated plate. Ultrasonic nebulizers operate by means of piezoelectric crystals that shear liquids into small aerosol droplets. A variety of suitable devices are available, including, for example, the AeroNeb™ and AeroDose™ vibrating perforated plate nebulizers (AeroGen, Inc., Sunnyvale, California). Nebulizer (Medic-Aid Ltd., West Sussex, England), Pari and Pari LC Jet atomizers (Pari Respiratory Equipment, Inc., Richmond, Virginia) and Aerosonic™ (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and (Omron Healthcare, Inc., Vernon Hills, Illinois) Ultrasonic nebulizer.

[0275] In some embodiments, the agent may be formulated with an oily matrix or ointment to form a semi-solid composition having the desired shape. In addition to the agent, these semi-solid compositions may also contain dissolving and / or suspending bactericides, preservatives, and / or buffering systems. The petrolatum component that may be included may be any paraffin, in terms of viscosity, ranging from mineral oil containing isobutylene, colloidal silica, or stearates to solid paraffin. The absorbent matrix may be used with the oily system. Additives may include cholesterol, lanolin (lanolin derivatives), beeswax, fatty alcohols, lanolin alcohol, low HLB (hydrophobic-oleophobic balance) emulsifiers, and a wide variety of ionic and nonionic surfactants, alone or in combination.

[0276] Controlled or sustained release of transdermal or surface formulations can be achieved by adding timed-release additives available in the art, such as polymeric structures and matrices. For example, the composition can be applied using a hot-melt extruded article, such as a bioadhesive hot-melt extruded film. The formulation may contain a crosslinked polycarboxylic acid polymer formulation. The crosslinking agent can provide sufficient adhesion to allow the system to remain attached to the surface of the target epithelial or endothelial cells for a sufficient time to allow the desired amount of compound release to be achieved.

[0277] Inserts, transdermal patches, bandages, or articles may contain mixtures or coatings of polymers that provide the release of a drug at a constant rate over an extended period. In some embodiments, the article, transdermal patch, or insert contains a water-soluble pore-forming agent, such as polyethylene glycol (PEG), that can be mixed with water-insoluble polymers to increase the durability of the insert and prolong the release of the active ingredient.

[0278] Transdermal devices (insertions, patches, bandages) may also contain water-insoluble polymers. Rate-controlled polymers can be used for application to sites where pH changes can be used to achieve release. These rate-controlled polymers can be applied using a continuous coating process during spraying and drying with an active compound. In one embodiment, a coating formulation is used to coat granules containing the active ingredient, which are then compressed to form a solid biodegradable insert.

[0279] Polymer formulations can also be used to provide controlled or sustained release. Bioadhesive polymers described in the art can be used. For example, sustained-release gels and compounds can be incorporated into a polymeric matrix such as a hydrophobic polymer matrix. Examples of polymeric matrices include microparticles. Microparticles can be microspheres, and the core can have a material different from the polymeric shell. Alternatively, the polymer can be cast into sheets or films, powders produced by grinding or other standard techniques, or gels such as hydrogels. Polymers can also be in the form of coatings or portions of bandages, stents, catheters, vascular grafts, or other devices to facilitate drug delivery. The matrix can be formed by solvent evaporation, spray drying, solvent extraction, and other methods known to those skilled in the art.

[0280] Kits are provided that contain unit doses of one or more of the pharmaceutical agents described herein, typically in oral or injectable form. Such kits may include a container containing the unit dose, an informational insert describing the use of the drug in treating a disease and associated benefits, and optionally an apparatus or device for delivering the composition.

[0281] D. Treatment methods

[0282] The compounds described herein can be used to prepare medicaments for the prevention or treatment of diseases or conditions. Furthermore, a method for treating any of the diseases or conditions described herein in a subject requiring such treatment involves administering a pharmaceutical composition containing at least one compound described herein, or a pharmaceutically acceptable salt, pharmaceutically acceptable prodrug, or pharmaceutically acceptable solvate thereof, to the subject in a therapeutically effective amount.

[0283] Compositions containing the compounds described herein may be applied for preventative and / or therapeutic treatment. In therapeutic use, the composition is administered to a patient suffering from the disease or condition in an amount sufficient to cure or at least partially suppress the symptoms of the disease or condition. The effective amount for this use will depend on the severity and course of the disease or condition; prior therapy; the patient's health status, weight, and response to the medication; and the judgment of the treating physician.

[0284] In prophylactic use, a composition containing the compounds described herein is administered to a patient who is susceptible to a particular disease, condition, or illness, or otherwise at risk of such a disease, condition, or illness. This amount is defined as the “preventative effective amount or dose.” In this application, the precise amount also depends on the patient’s health status, weight, etc. When used in a patient, the effective amount for this purpose will depend on the severity and duration of the disease, condition, or illness; prior therapy; the patient’s health status and response to the drug; and the judgment of the treating physician.

[0285] In cases where the patient’s condition does not improve, the compound may be administered for an extended period, as determined by a physician, that is, for a prolonged period, including throughout the patient’s life, to improve or otherwise control or limit the symptoms of the patient’s disease or condition.

[0286] Once the patient's condition has improved, a maintenance dose is administered as needed. Subsequently, the dosage or frequency of administration, or both, may be reduced to maintain the level of improvement in the disease, condition, or illness, depending on the symptoms. However, after any recurrence of symptoms, the patient may require long-term intermittent treatment.

[0287] The amount of a given agent corresponding to this quantity will vary depending on factors such as the specific compound, the disease or condition and its severity, and the identity (e.g., weight) of the person or host requiring treatment. However, it can be determined in a manner recognized in the art based on the specific circumstances surrounding the case, including, for example, the specific agent administered, the route of administration, the condition being treated, and the person or host being treated. Generally, however, the dosage for adult treatment will typically range from about 0.02 mg to about 5000 mg daily, and in some embodiments, from about 1 mg to about 1500 mg daily. The required dosage can be conveniently provided as a single dose or in fractionated doses, which are administered simultaneously (or over a short period) or at appropriate intervals, for example, as sub-dose twice, three, four, or more times daily.

[0288] The pharmaceutical compositions described herein may be in unit dosage forms suitable for precise single-dose administration. In a unit dosage form, the formulation is divided into unit doses containing appropriate amounts of one or more compounds. The unit dose may be in the form of a package containing discrete amounts of the formulation. Non-limiting examples are packaged tablets or capsules and powders in vials or ampoules. Aqueous suspension compositions may be packaged in single-dose, non-reclosable containers. Alternatively, multiple-dose, reclosable containers may be used, in which case a preservative is typically included in the composition. By way of example only, formulations for parenteral injection may be provided in unit dosage forms, including but not limited to ampoules, or in multiple-dose containers with added preservatives.

[0289] The toxicity and efficacy of such treatment regimens can be determined in cell cultures or laboratory animals using standard pharmaceutical procedures, including but not limited to determining the LD50. 50 (A dose that is lethal to 50% of the population) and ED 50 (The dose that has therapeutic efficacy in 50% of the population). The dose ratio between toxicity and therapeutic effect is the therapeutic index, and it can be expressed as LD50. 50 With ED 50 The ratio between these values. Compounds exhibiting a high therapeutic index are preferred. Data obtained from cell culture assays and animal studies can be used to determine the dosage range for human use. The dosage of such compounds is preferably within the range of minimal toxicity, including ED. 50 The cyclic concentration range. Depending on the dosage form and route of administration, the dosage may vary within this range.

[0290] In some embodiments, the present invention provides a method for treating or preventing a disease, state, or condition in a patient in need, the method comprising administering to the patient an effective amount of any of the compounds in the embodiments of the present invention. Diseases, states, or conditions may be selected from neuropathic pain, vascular inflammation, arthritis, allergies, asthma, wound healing, stroke, heart failure, acute spinal cord injury, acute head injury or trauma, seizures, neonatal hypoxia, cerebral palsy, chronic hypoxia attributed to arteriovenous malformations and occlusive cerebral artery disease, ischemia and reperfusion injury in skeletal muscle, severe neurosis associated with excitotoxicity, Parkinson's disease, Huntington's disease, CNS disease, heart disease, kidney disease, glaucoma, cancer, neuropathic pain, diabetic neuropathic pain, transient ischemic attack, myeloprotection, dry eye syndrome, osteoarthritis, rheumatoid arthritis, loss of skin pigmentation, inflammatory bowel disease, pulmonary inflammation, uveitis, and septic shock. In a preferred embodiment, the present invention provides a method for treating or preventing neuropathic pain in a patient in need. In another preferred embodiment, the present invention provides a method for treating or preventing postoperative pain in patients in need.

[0291] In some embodiments, this disclosure provides a method for treating conditions selected from chronic inflammatory conditions, chronic neuropathic pain and mixed pain conditions, neurodegenerative conditions, cognitive impairment conditions, unintended side effects of opioid analgesia, congestive heart failure, myocarditis, giant cell arteritis, temporal arteritis, aortic (Takayasu's) arteritis, vasculitis, atherosclerotic vascular disease, chronic bronchitis, chronic pancreatitis, fatty liver disease (including alcoholic and non-alcoholic types), inflammatory bowel disease (including Crohn's disease and ulcerative colitis), inflammatory bowel syndrome, cholangitis, cholecystitis, interstitial cystitis, duodenitis, lymphadenitis, prostatitis, salpingitis, arthritis (including osteoarthritis and rheumatoid arthritis), temporomandibular joint dysfunction, myositis (including polymyositis and dermatomyositis), osteitis (including periostitis and osteomyelitis), jaundice, etc. Macular degeneration (wet and dry types), glaucoma, uveitis, iritis, dry eye syndrome, and ototoxicity induced by drugs (including but not limited to platinum-based chemotherapeutic agents, aminoglycoside antibiotics, and loop diuretics) and noise (deafness, hyperacusis, and vestibular dysfunction), trigeminal neuralgia, post-traumatic painful neuropathy (burning neuralgia and complex regional pain syndrome), postherpetic neuralgia, diabetic neuropathy, small fiber neuropathy, burning mouth syndrome (glossodynia), vulvar pain (including vulvar vestibulitis), chemotherapy-induced peripheral neuropathy (including but not limited to neuropathy caused by vinca alkaloids, taxanes, platinum-based chemotherapeutic agents, and proteasome inhibitors), spinal cord injury pain, chronic low back pain, chronic neck pain, sciatica, discogenic pain, fibromyalgia, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Leber's optic neuropathy. Optical neuropathy, frontotemporal dementia, Lewy body dementia (DLB), spinocerebellar degeneration, multiple sclerosis, diabetic neuropathy, small fiber neuropathy, chemotherapy-induced neuropathy, traumatic brain injury (including concussion), postoperative cognitive impairment, chemotherapy-induced or radiation-induced damage to the oral and gastrointestinal mucosa (mucositis), hepatocellular carcinoma, adverse effects attributable to anticancer drugs, overactive bladder, pelvic pain, prostatodynia, interstitial cystitis, septic shock, erectile dysfunction, acne, dynamic wrinkles, and psoriasis.

[0292] In some embodiments, this disclosure provides a method for treating conditions selected from hyperactivity, hypertension, acute hypoxia, depression, and infertility.

[0293] In some embodiments, this disclosure provides a method for treating conditions selected from inflammatory conditions such as vasculitis and arthritis, allergies, asthma, wound healing, stroke, heart failure, acute spinal cord injury, acute head injury or trauma, seizures, neonatal hypoxia (cerebral palsy; prophylactic treatment involves long-term exposure through placental circulation), chronic hypoxia attributable to arteriovenous malformations and occlusive cerebral artery disease, ischemia and reperfusion injury in skeletal muscle, severe neurological conditions associated with excitotoxicity, Parkinson's disease, Huntington's disease and other CNS diseases, heart disease, kidney disease, and contraception.

[0294] In some embodiments, the compounds of the present invention can also be used to treat pain associated with chemotherapy-induced peripheral neuropathy (CIPN), which is induced by a combination of one or more chemotherapeutic agents as part of a treatment regimen. Non-limiting examples of combinations include CHOPP (cyclophosphamide, doxorubicin, vincristine, prednisone, and procarbazine); CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone); COP (cyclophosphamide, vincristine, and prednisone); CAP-BOP (cyclophosphamide, doxorubicin, procarbazine, bleomycin, vincristine, and prednisone); m-BACOD (methotrexate, bleomycin, doxorubicin, and procarbazine); Cyclophosphamide, vincristine, dexamethasone, and leucovorin; ProMACE-MOPP (prednisone, methotrexate, doxorubicin, cyclophosphamide, etoposide, leucovorin, mechloethamine, vincristine, prednisone, and procarbazine); ProMACE-CytaBOM (prednisone, methotrexate, doxorubicin, cyclophosphamide, etoposide, leucovorin, cytarabine, bleomycin, and vincristine); MACOP- B (methotrexate, doxorubicin, cyclophosphamide, vincristine, prednisone, bleomycin, and leucovorin); MOPP (dichloromethyldiethylamine, vincristine, prednisone, and procarbazine); ABVD (adriamycin / doxorubicin, bleomycin, vinblastine, and dacarbazine); alternating MOPP (dichloromethyldiethylamine, vincristine, prednisone, and procarbazine) with ABV (adriamycin / doxorubicin, bleomycin, and vinblastine); alternating MOPP (dichloromethyldiethylamine, vincristine, prednisone, and procarbazine) with ABVD (adriamycin / doxorubicin, bleomycin, and vinblastine); alternating MOPP (dichloromethyldiethylamine, vincristine, prednisone, and procarbazine) with ABVD (adriamycin / doxorubicin, bleomycin, and vinblastine). Alternating regimens including: doxorubicin, bleomycin, vinblastine, and dacarbazine; ChIVPP (chlorambucil, vinblastine, procarbazine, and prednisone); IMVP-16 (ifosfamide, methotrexate, and etoposide); MIME (methylgag, ifosfamide, methotrexate, and etoposide); DHAP (dexamethasone, high-dose cytarabine, and cisplatin); ESHAP (etoposide, methylprednisolone, high-dose cytarabine, and cisplatin).CEPP(B) (cyclophosphamide, etoposide, procarbazine, prednisone, and bleomycin); CAMP (lomustine, mitoxantrone, cytarabine, and prednisone); CVP-1 (cyclophosphamide, vincristine, and prednisone); ESHOP (etoposide, methylprednisolone, high-dose cytarabine, vincristine, and cisplatin); EPOCH (etoposide, vincristine, and doxorubicin for 96 hours, plus a bolus dose). Cyclophosphamide and oral prednisone), ICE (ifosfamide, cyclophosphamide, and etoposide), CEPP(B) (cyclophosphamide, etoposide, procarbazine, prednisone, and bleomycin), CHOP-B (cyclophosphamide, doxorubicin, vincristine, prednisone, and bleomycin), CEPP-B (cyclophosphamide, etoposide, procarbazine, and bleomycin), and P / DOCE (epirarubicin or doxorubicin, vincristine, cyclophosphamide, and prednisone).

[0295] In some embodiments, the method includes administering to a subject a first amount of a compound or salt described herein combined with a second amount of an analgesic, the first and second amounts together constituting a pharmaceutically effective amount. The first amount, the second amount, or both may be less than the effective amount of each compound administered as a monotherapy. The therapeutically effective amounts of the compound of the invention and the analgesic may be administered to the subject simultaneously or separately in any given order and via the same or different routes of administration. It may be advantageous to begin administration of the compound of the invention first, for example, one or more days or one or more weeks before the start of administration of the analgesic. Furthermore, additional drugs may be administered in combination with the above combination therapy. In some embodiments, this disclosure provides a method for treating or preventing chemotherapy-induced peripheral neuropathy (CIPN) in a subject, the method comprising administering to the subject a compound, salt, or pharmaceutical composition described herein. In some embodiments, CIPN is attributed to anticancer chemotherapy. In some cases, the anticancer chemotherapy is a taxane chemotherapeutic agent, a platinum complex chemotherapeutic agent, a vinca alkaloid chemotherapeutic agent, or a proteasome inhibitor chemotherapeutic agent. In some cases, CIPN is attributed to antiviral chemotherapy. In some cases, the antiviral chemotherapy is anti-HIV chemotherapy.

[0296] In some embodiments, this disclosure provides a method for treating or preventing diabetic peripheral neuropathy in a subject, the method comprising administering to the subject a compound, salt, or pharmaceutical composition as described herein.

[0297] In some embodiments, this disclosure provides a method for treating or preventing neurodegeneration in a subject, the method comprising administering to the subject a compound, salt, or pharmaceutical composition as described herein. In some cases, neurodegeneration is attributed to Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, or Lieber's optic neuropathy.

[0298] In some embodiments, this disclosure provides a method for preventing or treating drug-induced ototoxicity in a subject, the method comprising administering to the subject a compound, salt, or pharmaceutical composition described herein. In some embodiments, drug-induced ototoxicity is hearing loss, tinnitus, or hyperacusis.

[0299] In some embodiments, this disclosure provides a method for treating or preventing spinocerebellar degeneration in a subject, the method comprising administering to the subject a compound, salt, or pharmaceutical composition described herein.

[0300] In some embodiments, this disclosure provides a method for treating or preventing symptoms associated with traumatic brain injury in a subject of need, the method comprising administering to the subject the compounds, salts, or pharmaceutical compositions described herein. In some cases, the method comprises treating one or more symptoms associated with traumatic brain injury. In some cases, one or more symptoms are cognitive impairment. In some cases, cognitive impairment includes at least one of the following: memory loss; impairment of insight, judgment, and thinking; reduced processing speed; inattention and / or deficits in executive functions (such as abstract reasoning, planning, problem-solving, and multitasking). In some cases, the compounds, salts, or pharmaceutical compositions are administered within 24 hours of the traumatic brain injury or within 48 hours of the traumatic brain injury. In some cases, the compounds, salts, or pharmaceutical compositions are administered in multiple doses.

[0301] In some embodiments, this disclosure provides a method for treating or preventing chemotherapy-induced cognitive impairment, the method comprising administering the compounds, salts, or pharmaceutical compositions described herein to a patient who is receiving or will receive cancer chemotherapy. In some cases, the method comprises administering the compounds, salts, or pharmaceutical compositions prior to cancer chemotherapy. In some cases, the method comprises administering the compounds, salts, or pharmaceutical compositions approximately one minute to seven days prior to cancer chemotherapy. In some cases, the method comprises administering the compounds, salts, or pharmaceutical compositions concurrently with cancer chemotherapy. In some cases, the method comprises administering the compounds, salts, or pharmaceutical compositions only on the days on which cancer chemotherapy is administered. In some embodiments, the method comprises administering the compounds, salts, or pharmaceutical compositions on one or more days, including the days on which cancer chemotherapy is administered and those days between successive doses of the chemotherapy agent. In some cases, the method comprises administering the compounds, salts, or pharmaceutical compositions after cancer chemotherapy. In some cases, the method comprises administering the compounds, salts, or pharmaceutical compositions approximately one minute to seven days after cancer chemotherapy. In some cases, cancer chemotherapy is selected from taxanes, platinum complexes, vinca alkaloids, proteasome inhibitors, 5-fluorouracil, methotrexate, doxorubicin, and combinations thereof.

[0302] In some embodiments, this disclosure provides a method for treating pain and discomfort in irritable bowel syndrome, the method comprising administering a compound, salt, or pharmaceutical composition described herein to a patient undergoing or to undergo cancer chemotherapy. In some cases, the method includes administration before the onset of pain or discomfort. In some cases, the method includes administration after the onset of pain or discomfort. In some cases, the method includes administration during the onset of pain or discomfort. In some cases, the pain or discomfort is reduced by at least about 10%, as determined by measuring the abdominal response to colonic or rectal distension. In some cases, the pain or discomfort is reduced by at least about 50%. In some cases, the pain or discomfort is reduced by at least about 90%. In some cases, the pain or discomfort is reduced by at least about 10%, as determined by measuring the visceral motility response to colonic or rectal distension. In some cases, the pain or discomfort is reduced by at least 50%. In some cases, the pain or discomfort is reduced by at least 90%. In some cases, administration is performed at least 5 days after the onset of pain or discomfort.

[0303] In some embodiments, regardless of the cause of the pain, the compounds or salts of this disclosure, when administered together with an analgesic, can be used to alleviate symptoms of neuropathic pain, the causes of which include, but are not limited to, spinal cord injury, multiple sclerosis, stroke, diabetes, herpes zoster infection, HIV-related neuropathy, nutritional deficiencies, toxins, distal manifestations of malignancies, hereditary immune-mediated conditions or physical trauma to nerve trunks, cancer, chemotherapy, radiation damage or surgery (e.g., postoperative pain), vulvar pain, and burning mouth syndrome. In one embodiment, the neuropathic pain is associated with long-term use of opioids.

[0304] Analgesics used in combination with the compounds of the present invention can be selected based on the specific condition being treated. Currently known analgesics include, but are not limited to, opioids, morphine analogues, antidepressants, antiepileptics, NMDA receptor antagonists, fatty acid amine hydrolase inhibitors, anticonvulsants, nonsteroidal anti-inflammatory drugs (NSAIDs), COX-2 inhibitors, NOS inhibitors, acetaminophen, and calcium channel subunit α2δ ligands.

[0305] Examples of opioids include any natural or synthetic opioid analgesics, such as morphine, fentanyl, codeine, thebaine, diacetylmorphine (heroin), dihydrocodeine, hydrocodone, hydromorphone, nicomorphine, oxycodone, oxymorphone, alpha-methylfentanyl, alfentanil, sufentanil, remifentanil, carfentanyl, ohmefentanyl, nocaine, pethidine (meperidine), ketobemidone, MPPP, allylprodine, prodine, PEPAP, propoxyphene, and dextropropoxyphene. The following are listed: tropropoxyphene, dextromoramide, bezitramide, piritramide, methadone, dipipanone, levoalphacetylmethadol (LAAM), loperamide, diphenoxylate, pentazocine, phenazocine, buprenorphine, etorphine, butorphanol, nalbuphine, levorphanol, levomethorphan, dezocine, lefetamine, tilidine, tramadol, propoxyphene, and oxycodone. As anticipated in this article, opioids also include any natural or synthetic narcotic antagonists such as nalmefene, naloxone, or naltrexone, as well as any natural or synthetic mixed opioid agonists / antagonists such as nalbuphine, butorphanol, buprenorphine, and pentazocine.

[0306] Examples of nonsteroidal anti-inflammatory drugs (NSAIDs) include aspirin, ibuprofen, acetaminophen, naproxen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, indomethacin, sulindac, etodolac, ketorolac, diclofenac, nabumetone, piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, isoxicam, mefenamic acid, meclofenamic acid, flufenamic acid, and tolfenamic acid. The following are listed: acid, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, nimesulide, and licofelone.Examples of antidepressants include tricyclic antidepressants such as: amitriptyline, amitriptyline oxide, butriptyline, clomipramine, demetriptiline, desipramine, dibenzepin, dimetacrine, dosulepin, doxepin, imipramine, imipramin oxide, lofepramine, melitracen, and methapramine. The following drugs are listed: etapramine, nitroxazepine, nortriptyline, noxiptiline, pipofezine, propizepine, protriptyline, and quinupramine; amineptine, norepinephrine, iprindole, opipramol, tianeptine, trimipramine, carbamazepine, and flupirtine.

[0307] It is anticipated that the compounds or salts of the present invention may be particularly suitable for the treatment of pain when co-administered with opioids, tricyclic antidepressants, or analgesics believed to bind to the α2δ calcium channel subunit, i.e., calcium channel subunit α2δ ligands. Examples of such ligands include GABA analogues such as gabapentin (2-[1-(aminomethyl)cyclohexyl]acetic acid) and pregabalin ((S)-3-(aminomethyl)-5-methylhexanoic acid).

[0308] In some embodiments, the method includes administering to a subject a first amount of a combination of the compound or salt described herein and a second amount of a dopamine agonist such as carbidopa or levodopa.

[0309] The relative amounts of the compound or its salts can be selected to provide synergistic pain relief. For example, a suitable ratio of the compound of the present invention to gabapentin may be in the range of about 0.1 parts by weight of the compound to about 3 to about 30 parts by weight of gabapentin. A suitable ratio of the compound of the present invention to morphine may be in the range of about 0.1 parts by weight of the compound to about 1 to about 5 parts by weight of morphine. Although these ratios are calculated with respect to the free compound (non-salt form), it should be understood that a pharmaceutically acceptable equivalent ratio of a salt or prodrug of the compound can also be readily determined by using the ratio of the molecular weights of the salts.

[0310] In some embodiments, the co-administration of the compounds of the present invention and the analgesic is achieved by formulating the compounds together in a combination composition. The combination composition may comprise a first pharmaceutically acceptable composition containing a first amount of the compound of the present invention and a second pharmaceutically acceptable composition containing a second amount of the analgesic, wherein the first and second amounts together constitute a pharmaceutically effective amount. The first amount, the second amount, or both may be less than the effective amount of each compound administered as a monotherapy. The combination composition is a pharmaceutically acceptable composition comprising a first amount of the compound or salt of the present invention and a second amount of the analgesic, wherein the first and second amounts together constitute a pharmaceutically effective amount. The first amount, the second amount, or both may be less than the effective amount of each compound administered as a monotherapy.

[0311] In one embodiment, the present invention provides a method for reducing opioid nociceptive tolerance and / or hypersensitivity in a subject receiving opioid therapy, the method comprising administering to the subject an amount of a compound or salt of the present invention sufficient to reduce opioid nociceptive tolerance.

[0312] In another embodiment, a method is provided for preventing or treating opioid dependence, i.e., withdrawal, in a subject receiving an opioid preparation, the method comprising administering to the subject an amount sufficient to treat one or more symptoms of opioid withdrawal of the present invention, either a compound or a salt thereof. The opioid may be morphine, oxycodone, fentanyl, cocaine, heroin, or opium. The compound or salt thereof may be delivered before or after the initiation of withdrawal. The compound or salt thereof may be administered in conjunction with a tapering dose of opioid. The compound or salt thereof may be delivered before the initiation of opioid therapy. The compound or salt thereof may be delivered for a period of time after opioid administration to the subject has ceased. The compound or salt thereof may be delivered for a period of one week, two weeks, three weeks, four weeks, one month, two months, three months, four months, five months, or six months after opioid preparations have ceased administration to the subject. The opioid and / or the compound or salt thereof may be delivered by continuous infusion, such as via an implanted pump.

[0313] One or more symptoms of opioid withdrawal may include agitation, anxiety, muscle pain, increased tearing, insomnia, runny nose, sweating, and yawning, while late-stage withdrawal symptoms include abdominal cramps, diarrhea, dilated pupils, goosebumps, nausea, and / or vomiting. Methods may also include subjecting the subject to a pharmacological treatment procedure, such as methadone or buprenorphine.

[0314] In other embodiments, the compounds or salts of the present invention are administered in combination with agents such as TNF-α inhibitors, IL-1β inhibitors, p38 kinase inhibitors, ERK inhibitors, JNK inhibitors, regulators of transcription factors such as NF-κB, agents regulating glial cell function, agents blocking the expression and / or activity of adenosine kinases, recombinant exonucleases, ENT inhibitors, etc. Non-limiting examples of p38 kinase inhibitors include PH-797804, BIRB 796, VX-702, SB 239063, SB202190, SCIO 469, and BMS 582949. One example of an ERK inhibitor is sorafenib. One example of a JNK inhibitor is AM-111. Non-limiting examples of NF-κB regulators include disulfiram, olmesartan, dithiocarbamate, and anatatabine.

[0315] The following embodiments further illustrate the invention, but should not be construed as limiting its scope in any way.

[0316] Example

[0317] The present invention, which has been generally described, will be more readily understood by referring to the following embodiments, which are included only for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention in any way.

[0318] The following synthetic schemes are provided for illustrative purposes and not for limitation. The following examples illustrate various methods for preparing the compounds described herein. It should be understood that those skilled in the art can prepare these compounds by similar methods or by combinations of other methods known to them. It should also be understood that those skilled in the art will be able to prepare them in a similar manner as described below by using appropriate starting materials and modifying the synthetic route as needed. Generally, starting materials and reagents are available from commercial suppliers, or synthesized from sources known to those skilled in the art, or prepared as described herein.

[0319] Examples 1–11 show exemplary procedures for the preparation of key intermediates and the claimed A3 receptor agonist.

[0320] Example 1. Preparation of 2,6-dichloro-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-9H-purine (5.1)

[0321]

[0322] Step-1

[0323]

[0324] CeCl3 (7.19 g, 1.84 mL, 1.00 equivalent) was added to a solution of compound 1 (4.50 g) in MeOH (225 mL), followed by fractional addition of NaBH4 (1.27 g, 1.15 equivalent) at 0 °C. The mixture was stirred at 0 °C for 1 h. TLC (petroleum ether: ethyl acetate = 5:1) showed that the starting material was consumed and new spots were formed. The mixture was quenched with water (100 mL) and extracted with DCM (50 mL × 3), followed by washing of the combined organic extracts with brine.

[0325] After drying with Na2SO4 and filtering, the solvent was concentrated under vacuum. The mixture was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 50:1 to 5:1) to produce compound 2 (3.20 g) as a yellow liquid. 1 H NMR: 400MHz CDCl3δ (ppm) 5.96-5.86 (m, 2H), 5.02 (d, J = 5.5Hz, 1H), 4.75 (t, J = 5.6Hz ,1H),4.56(dd,J=5.6,9.8Hz,1H),2.71(d,J=9.9Hz,1H),1.44(s,3H),1.41(s,3H).

[0326] Step-2

[0327]

[0328] At 0 °C, a solution of diethylzinc in DCM (1.0 M, 89.6 mL, 4.00 equivalents) was added dropwise to a solution of compound 2 (3.50 g, 1.00 equivalents) in DCM (110 mL). After stirring the mixture at 0 °C for 15 min, CH2I2 (48.0 g, 14.4 mL, 8.00 equivalents) was added, and the mixture was stirred at 20 °C for 16 h. TLC (petroleum ether:ethyl acetate = 2:1) showed that the starting material was consumed. The mixture was quenched with saturated NH4Cl solution (100 mL) and extracted with DCM (50 mL × 3). The organic extract was dried over Na2SO4 and concentrated to produce a crude product, which was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 20:1 to 5:1) to produce compound 3 (1.90 g) as a colorless oil. 1 H NMR: 400MHz CDCl3δ (ppm) 4.89 (t, J = 5.9Hz, 1H), 4.59-4.43 (m, 2H), 2.35 (s, 1H), 1.91-1.80 (m,1H),1.71-1.60(m,1H),1.56(s,3H),1.30(s,3H),1.05-0.88(m,1H),0.69-0.58(m,1H).

[0329] Step 3

[0330]

[0331] Under N2, PPh3 (5.86 g, 2.00 equivalent) and DIAD (4.51 g, 4.34 mL, 2.00 equivalent) were added to a solution of compound 4 (3.16 g, 1.50 equivalent) in THF (40 mL). After stirring the mixture at 20 °C for 15 min, compound 3 (1.90 g, 1.00 equivalent) was added, and the mixture was stirred at 20 °C for 16 h. TLC (petroleum ether:ethyl acetate = 1:1) showed that the starting material was consumed. The reaction mixture was concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 20:1 to 5:1) to produce compound 5.1 (3.50 g) as a white solid. MS: 340.9 (M+H) + . 1 H NMR:400MHz CDCl3δ(ppm)8.16(s,1H),5.43-5.32(m,1H),5.05(s,1H),4.67(d,J=7.2Hz,1H),2 .21-2.11(m,1H),1.72-1.63(m,1H),1.55(s,3H),1.26(s,3H),1.01–0.99(s,2H).

[0332] The following intermediates were prepared similarly according to the method of Example 1:

[0333] • In step-3, 6-chloro-2-iodo-9H-purine was substituted for 4 to produce 6-chloro-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-2-iodo-9H-purine (5.2). MS: 433.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.71(s,1H),5.31-5.24(m,1H),5.01(s,1H),4.73(d,J=7.2Hz,1H),2. 02-1.96(m,1H),1.83-1.76(m,1H),1.44(s,3H),1.17-1.13(m,3H),0.90-0.76(m,2H);

[0334] • In step-3, 5,7-dichloro-3H-imidazo[4,5-b]pyridine was used to replace 4, resulting in 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine (5.3). MS: 339.9 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.69(s,1H),7.66(s,1H),5.28(t,J=6.0Hz,1H),5.03(s,1H),4.69(d,J=6.8 Hz,1H),2.01-2.99(m,1H),1.81-1.78(m,1H),1.44(s,3H),1.16(s,3H),0.88-0.79(m,2H);

[0335] • In step-3, 7-chloro-3H-imidazo[4,5-b]pyridine was used to replace 4, resulting in 7-chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine (5,4). MS: 306.1 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.66(s,1H),8.36(d,J=5.2Hz,1H),7.47(d,J=5.0Hz,1H),5.34-5.26(m,1H),5.10(s,1H),4. 69(d,J=7.2Hz,1H),2.04-1.98(m,1H),1.83-1.76(m,1H),1.44(s,3H),1.14-1.12(m,3H),0.90-0.78(m,2H);

[0336] • In step-3, substitution of 4 with 5-bromo-7-chloro(bromo)-3H-imidazo[4,5-b]pyridine yielded 5-bromo-7-chloro(bromo)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine (5.5). MS: 386.0 (M+H) + and 430.0(M+H) + 7-Chloroisomeric, 1 H NMR:400MHz DMSO-d6δ(ppm)8.74(s,1H),7.82(s,1H),5.34(t,J=6.0Hz,1H),5.09(d,J=5.2Hz,1H),4.76(d,J =6.8Hz,1H),2.07-2.04(m,1H),1.86-1.84(m,1H),1.51(s,3H),1.23(s,3H),0.94-0.85(m,2H). 7-Bromoisomer δ (ppm) 8.74 (s, 1H), 7.93 (s, 1H), 5.34 (t, J = 6.0 Hz, 1H), 5.09 (d, J = 5.2 Hz, 1H), 4.76 (d, J = 6.8 Hz, 1H), 2.07-2.04 (m, 1H), 1.86-1.84 (m, 1H), 1.51 (s, 3H), 1.23 (s, 3H), 0.94-0.85 (m, 2H).

[0337] Table 1. Intermediates prepared by the method of Example 1

[0338]

[0339] Example 2. Preparation of 2-chloro-N-(2,2-difluoroethyl)-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-9H-purine-6-amine (7.1).

[0340]

[0341] A solution of compound 5.1 (150.0 mg, 1.00 equivalent), compound 6 (71.2 mg, 2.00 equivalent), and TEA (222.4 mg, 305.9 μL, 5.00 equivalent) in t-BuOH (2 mL) was stirred for 2 h at 65 °C. LC-MS analysis showed that the starting materials were completely consumed. The reaction mixture was concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (SiO2, EtOAc) to yield compound 7.1 (110.0 mg) as a yellow solid. MS: 386.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.63(s,1H),8.31(s,1H),6.35-6.01(m,1H),5.25(t,J=6.0Hz,1H),4.89(s,1H),4.63(d,J=7.2Hz ,1H),3.95-3.75(m,2H),1.99-1.93(m,1H),1.78-1.69(m,1H),1.44(s,3H),1.18-1.14(m,3H),0.89-0.74(m,2H).

[0342] The following intermediates were prepared similarly using the method described in Example 2:

[0343] Replacing 6 with 2-fluoroethylamine yielded 2-chloro-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl

[0344] )-N-(2-fluoroethyl)-9H-purine-6-amine (7.2). MS: 368.2 (M+H) + . 1 H NMR: 400

[0345] MHz DMSO-d6δ(ppm)8.48-8.42(m,1H),8.27(s,1H),5.25(t,J=

[0346] 5.8Hz,1H),4.80-4.71(m,1H),4.68-4.60(m,2H),4.54(t,J=4.8Hz,1H),3.81-3.67(m,2H),1.98-1.96(m,1H),1.76-1.69(m,1H),1.44(s,

[0347] 3H),1.19(s,3H),0.89-0.74(m,2H);

[0348] Replacing 6 with 2,2,2-trifluoroethylamine produced 2-chloro...

[0349] -9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenyl[3,4]cyclopentadiene

[0350] [1,2-d][1,3]dioxacyclopenten-5-yl)-N-(2,2,2-trifluoroethyl)-9H-purine-6-amine

[0351] (7.3). MS: 404.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.92(s,

[0352] 1H),8.35(s,1H),5.25(t,J=6.4Hz,1H),4.90(s,1H),4.65(d,J=7.2

[0353] Hz,1H),4.26(s,2H),1.98-1.96(m,1H),1.76-1.74(m,1H),1.43(s,

[0354] 3H),1.11(s,3H),0.85-0.77(m,2H).

[0355] Replacing 6 with methyl-d3amine yielded 2-chloro-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-N-methyl-d3-9H-purine-6-amine (7,4). MS 339(M+H) + .

[0356] Table 2. Intermediates prepared by the method of Example 2.

[0357]

[0358] Example 3. Preparation of N-(2,2-difluoroethyl)-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-2-iodo-9H-purine-6-amine (8.1).

[0359]

[0360] TEA (350.8 mg, 482.6 μL, 5.00 equivalent) was added to a solution of compound 5.2 (300.0 mg, 1.00 equivalent) and compound 6 (84.3 mg, 1.50 equivalent) in t-BuOH (5 mL) at 20 °C. The mixture was stirred at 65 °C for 3 h. TLC analysis (EtOAc) showed that the starting material was completely consumed. The mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 mL × 4). The organic extract was dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative TLC (EtOAc) to yield compound 8.1 (250.0 mg) as a yellow solid. 1 H NMR:400MHz DMSO-d6δ(ppm)8.46(s,1H),8.22(s,1H),6.33-6.01(m,1H),5.22-5.29(m,1H),4.88(s,1H),4.62(d,J=6.8H z,1H),3.91-3.72(m,2H),1.98-1.93(m,1H),1.65-1.74(m,1H),1.44(s,3H),1.17(s,3H),0.74-0.88(m,2H).

[0361] Prepared similarly using the method described in Example 3:

[0362] Replacing 6 with ethylamine yielded 9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-2-iodo-9H-purine-6-amine (8.2). MS: 442.0 (M+H) + . 1 H NMR: 400 MHz CDCl3δ (ppm) 7.65 (s, 1H), 5.68 (s, 1H), 5.36-5.33 (m, 1H), 4.94 (s, 1H), 4.64 (d, J = 7.2 Hz,1H),3.65(s,2H),2.10-2.08(m,1H),1.65-1.60(m,1H),1.54(s,3H),1.31-1.25(m,6H),0.97-0.89(m,2H).

[0363] Table 3. Intermediates prepared by the method of Example 3

[0364]

[0365] Example 4. Preparation of 5-chloro-N-(2,2-difluoroethyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine-7-amine (9.1)

[0366]

[0367] DIPEA (569.8 mg, 768.0 μL, 5.0 equivalent) and compound 6 (285.9 mg, 130.5 μL, 4.0 equivalent) were added to a solution of compound 5.3 (300.0 mg, 1.0 equivalent) in NMP (3 mL). The mixture was stirred at 120 °C for 16 h. TLC analysis (ethyl acetate) showed that compound 5.3 / compound 9.1 = 2 / 1. The mixture was extracted with ethyl acetate (10 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (DCM / MeOH = 40 / 1) to obtain compound 9.1 (130.0 mg) as a yellow solid. MS: 385.1 (M+H) + . 1 H NMR:400 MHzDMSO-d6δ(ppm)8.26(s,1H),7.47(t,J=6.8Hz,1H),6.57(s,1H),6.32-6.03(m,1H),5.09(t,J=6.4Hz,1H),4.92(s,1H) ,4.58(d,J=6.8Hz,1H),3.97(brs,2H),1.98-1.95(m,1H),1.72-1.70(m,1H),1.44(s,3H),1.16(s,3H),0.84-0.79(m,2H).

[0368] The following compounds were prepared similarly using the method described in Example 4:

[0369] Replacing 6 with 2-fluoroethylamine yielded 5-chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl

[0370] )-N-(2-fluoroethyl)-3H-imidazo[4,5-b]pyridine-7-amine (9.2). MS: 366.9 (M+H) + .

[0371] 1H NMR:400MHz DMSO-d6δ(ppm)8.21(s,1H),7.37-7.34(m,1H),

[0372] 6.47(s,1H),5.30-5.25(m,1H),4.91(s,1H),4.59-4.52(m,3H),

[0373] 3.81-3.73(m,2H),2.00-1.95(m,1H),1.73-1.68(m,1H),1.44(s,3H),

[0374] 1.19-1.16(m,4H),0.80-0.78(m,1H);

[0375] Replacing 6 with 2,2,2-trifluoroethylamine produced 5-chloro...

[0376] -3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenyl[3,4]cyclopentadiene

[0377] [1,2-d][1,3]dioxacyclopenten-5-yl)-N-(2,2,2-trifluoroethyl)-3H-imidazo[4,5-b]

[0378] Pyridine-7-amine (9.3). MS: 403.2 (M+H) + . 1 H NMR: 400MHz DMSO-d6δ

[0379] (ppm)8.28(s,1H),7.71(t,J=6.8Hz,1H),6.66(s,1H),5.28-5.25(m,

[0380] 1H),4.93(s,1H),4.59(d,J=7.2Hz,1H),4.47(s,2H),1.98-1.97(m,

[0381] 1H),1.73-1.718(m,1H),1.44(s,3H),1.62(s,3H),0.85-0.78(m,2H);

[0382] Replacing 6 with ethylamine yielded 5-chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl

[0383] -3H-imidazo[4,5-b]pyridine-7-amine (9.4). MS: 349.0 (M+H) + . 1 H NMR: 400

[0384] MHz DMSO-d6δ(ppm)8.18(s,1H),7.19(t,J=5.6Hz,1H),6.35(s,1H),

[0385] 5.27(t,J=6.0Hz,1H),4.91(s,1H),4.59(d,J=6.8Hz,1H),3.41(brs,

[0386] 2H),1.99-1.97(m,1H),1.72-1.69(m,1H),1.44(s,3H),1.20-1.17(m,

[0387] 6H), 0.84-0.79(m,2H);

[0388] Replacing 6 with cyclobutylamine produced 5-chloro-N-cyclobutyl

[0389] -3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenyl[3,4]cyclopentadiene

[0390] [1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine-7-amine (9.5). MS:

[0391] 375.2(M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.19(s,1H),7.44(d,J=7.2Hz,1H),6.29(s,1H),5.30-5.23(m,1H),4.90(s,1H),4.57(d,J=7.2Hz,1H),4 .47-4.15(m,1H),2.37-2.27(m,2H),2.09-1.93(m,3H),1.75-1.64(m,3H),1.43(s,3H),1.16(s,3H),0.87-0.75(m,2H);

[0392] Replacing 6 with 2,5-difluorobenzylamine yielded 5-chloro-N-(2,5-difluorobenzyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine-7-amine (9.6) MS: 447.3 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.24(s,1H),7.82-7.79(m,1H),7.28-7.26(m,1H),7.17-7.12(m,2H),6.38(s,1H),5.27-5.24(m,1H),4.91(s, 1H),4.71-4.69(m,2H),4.60-4.58(m,1H),1.99-1.96(m,1H),1.72-1.71(m,1H),1.43(s,3H),1.16(s,3H),0.86-0.78(m,2H).

[0393] Replacing 6 with (5-chloro-2-((3-methylisoxazol-5-yl)methoxy)phenyl)methylamine yielded 5-chloro-N-(5-chloro-2-((3-methylisoxazol-5-yl)methoxy)benzyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridin-7-amine (9.7). LCMS: MS: 556.1 (M+H)+.

[0394] Replacing 6 with (6-methoxypyridin-2-yl)methylamine yielded 5-chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-((6-methoxypyridin-2-yl)methyl)-3H-imidazo[4,5-b]pyridin-7-amine (9.8). LCMS: 442.1 (M+H)+.

[0395] Replacing 6 with propan-1-amine yielded 5-chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-propyl-3H-imidazo[4,5-b]pyridin-7-amine (9.9). LCMS: 363.1 (M+H)+

[0396] Replacing 6 with cyclobutylmethylamine yielded 5-chloro-N-(cyclobutylmethyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine-7-amine (9.10). LCMS: 389.1 (M+H)+.

[0397] Replacing 6 with (2,5-dichlorophenyl)methylamine yielded 5-chloro-N-(2,5-dichlorophenylmethyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine-7-amine (9.11). LCMS: Rt = 0.798, MS: 479.1 (M+H)+.

[0398] Table 4. Intermediates prepared by the method of Example 4

[0399]

[0400]

[0401] Example 5.2-(cyclopropylethynyl)-N-(2,2-difluoroethyl)

[0402] Preparation of 9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-9H-purine-6-amine (11.1)

[0403]

[0404] At 20°C and under N2, TEA (466.5 mg, 641.6 μL, 10.0 equivalent), CuI (17.6 mg, 0.20 equivalent), and KF (80.4 mg, 3.00 equivalent) were added to a solution of compound 8.1 (220.0 mg, 1.00 equivalent) and compound 10 (382.5 mg, 312.8 μL, 6.00 equivalent) in DMF (5 mL). Then, Pd(PPh3)2Cl2 (64.7 mg, 0.20 equivalent) was added at 20°C and under N2.

[0405] The mixture was stirred at 20°C for 12 h. LCMS analysis showed that the starting material was completely consumed. The mixture was diluted with H₂O (20 mL) and extracted with EtOAc (20 mL × 4). The organic extract was dried over Na₂SO₄, filtered, and concentrated to produce a residue, which was purified by preparative TLC (EtOAc) to yield compound 11.1 (160.0 mg) as a red oil. MS 416.2 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.32(s,1H),8.25-8.10(m,1H),6.35-6.05(m,1H),5.24-5.22(m,1H),4.91(s,1H),4.57(d,J=7.2Hz,1H), 3.98-3.76(m,2H),1.77-1.69(m,1H),1.63-1.54(m,1H),1.44(s,3H),1.16(s,3H),0.95-0.89(m,2H),0.86-0.73(m,5H).

[0406] Prepared similarly using the method described in Example 5:

[0407] Replacing 8.1 with 8.2 produces 2-(cyclopropylethynyl)

[0408] )-9-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenyl[3,4]cyclopentadiene

[0409] [1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-9H-purine-6-amine (11.2). MS: 380.2

[0410] (M+H) +1 H NMR:400MHz CDC13δ(ppm)7.77(s,1H),5.32-5.30(m,1H),5.11(s,1H),4.58(d,J=7.2Hz,1H),3.74(s,2H),2.10-2. 07(m,1H),1.69-1.65(m,1H),1.54-1.48(m,4H),1.31(t,J=7.2Hz,3H),1.24(s,3H),0.97-0.90(m,6H);

[0411] Replacing 8.1 with 26.3 yielded 5-(cyclopropylethynyl)-3-((3aR,3bS,4aS,5R,5aS)-3b-(difluoromethyl)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-3H-imidazo[4,5-b]pyridine-7-amine (11.3). MS: 429.3 (M+H) + . 1 H NMR: 400MHzDMSO-d6δ(ppm)8.06(s,1H),6.90(s,1H),6.54-6.22(m,2H),5.35(d,J=7.2Hz,1H),5.01(s,1H),4.67(d,J=6.8Hz,1H),3.43-3.35(m, 2H),1.99(s,3H),1.96-1.90(m,1H),1.61-1.51(m,1H),1.46(s,3H),1.3 2-1.26(m,1H),1.20-1.18(m,4H),0.96-0.88(m,2H),0.79-0.69(m,2H).

[0412] Table 5. Intermediates prepared by the method of Example 5

[0413]

[0414] Example 6. Preparation of 5-(cyclopropylethynyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-3H-imidazo[4,5-b]pyridine-7-amine (14.1)

[0415]

[0416] Step-1

[0417]

[0418] At 20 °C, DIPEA (1.11 g, 1.50 mL, 23.6 equivalents) and ethylamine hydrochloride (148.4 mg, 5.00 equivalents) were added to a solution of compound 5.5 (140.0 mg, crude) in NMP (5 mL). The mixture was stirred at 100 °C for 12 h. TLC analysis (dichloromethane:methanol = 10:1) indicated that compound 5.5 was consumed and a major new spot with high polarity was detected. The reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (10 mL × 3). The combined organic extracts were washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to produce compound 12 (40.0 mg) as a yellow solid. MS: 393.0 (M+H) + ... 1 H NMR:400MHz DMSO-d6δ(ppm)8.16(s,1H),7.18(t,J=5.8Hz,1H),6.47(s,1H),5.27(t,J=6Hz,1H),4.90(s,1H),4.58(d,J=6. 8Hz,1H),3.40(s,2H),1.70-1.68(s,1H),1.44(s,3H),1.28-1.24(m,1H),1.19-1.15(m,6H),0.84-0.78(m,2H).

[0419] Step-2

[0420]

[0421] A solution consisting of compound 12 (120.0 mg, 1.00 equivalent), compound 13 (239.1 mg, 300.0 mL, 11.8 equivalent), Pd(PPh3)2Cl2 (42.8 mg, 0.20 equivalent), CuI (11.6 mg, 0.20 equivalent), TEA (436.2 mg, 600.0 μL, 14.13 equivalent), and DMF (6 mL) was prepared at 20 °C. The resulting mixture was stirred at 80 °C for 12 h. TLC (petroleum ether:ethyl acetate = 10:1) indicated that compound 12 was consumed and a major new spot with high polarity was detected. The reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (10 mL × 3). The combined extracts were washed with brine (18 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to yield compound 14.1 (120.0 mg, crude) as a brown solid. MS: 379.3 (M+H)+ . 1 H NMR:400MHz DMSO-d6δ(ppm)8.18(s,1H),6.81(t,J=5.8Hz,1H),6.40(s,1H),5.25(t,J=6Hz,1H),4.95(s,1H),4.52(d,J=7.2Hz,1H), 3.38(s,2H),1.99-1.96(m,1H),1.71-1.69(m,1H),1.57-1.53(m,1H),1.44(s,3H),1.19-1.15(m,6H),1.92-1.75(m,6H).

[0422] Table 6. Intermediates prepared by the procedure of Example 6

[0423]

[0424] Example 7. Preparation of N-(2,2-difluoroethyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-5-methyl-3H-imidazo[4,5-b]pyridine-7-amine (16.1)

[0425]

[0426] Under N2, Pd(PPh3)4 (69.1 mg, 0.10 equivalent) was added to a solution of compound 9.1 (230.0 mg, 1.00 equivalent), compound 15 (150.1 mg, 167.1 μL, 2.00 equivalent), and K2CO3 (330.4 mg, 4.00 equivalent) in dioxane (3 mL). The mixture was stirred at 100 °C for 12 h. LCMS analysis showed that 9.1 was consumed. The mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 mL × 4). The organic extract was washed with saturated brine (40 mL × 3), dried over Na2SO4, filtered, and concentrated to produce a residue. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 1:5) to produce compound 16.1 (200.0 mg) as a yellow oil. MS: 365.0 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.15-8.10(m,1H),6.97-6.88(m,1H),6.38(s,1H),6.32-6.00(m,1H),5.35-5.24(m,1H),5.03-4.98(m,1H),4.56(d ,J=7.0Hz,1H),4.01-3.85(m,2H),2.40(s,3H),2.00-1.92(m,1H),1.68-1.61(m,1H),1.44(s,3H),1.16(s,3H),0.88-0.77(m,2H).

[0427] The following compounds were prepared similarly according to the method of Example 7:

[0428] Replacing 9.1 with 9.2 yielded 3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-(2-fluoroethyl)-5-methyl-3H-imidazo[4,5-b]pyridine-7-amine (16.2). MS 347(M+H) + ;

[0429] Replacing 9.1 with 9.3 yielded 3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-5-methyl-3H-imidazo[4,5-b]pyridine-7-amine (16.3). MS: 329.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.04(s,1H),6.62-6.53(m,1H),6.21(s,1H),5.33-5.27(m,1H),4.97(s,1H),4.59-4.53(m,1H),3.41 -3.34(m,2H),2.39(s,3H),2.00-1.92(m,1H),1.68-1.61(m,1H),1.44(s,3H),1.20-1.15(m,6H),0.84-0.76(m,2H);

[0430] Replacing 9.1 with 9.6 yielded N-(2,5-difluorobenzyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-5-methyl-3H-imidazo[4,5-b]pyridine-7-amine (16.4). MS 427(M+H) + .

[0431] Table 7. Intermediates prepared by the procedure of Example 7

[0432]

[0433]

[0434] Example 8.2-Chloro-N-(2,2-difluoroethyl)-9-((3aR,3bS,4aS,5R,5aS)-3b-(fluoromethyl)

[0435] )-2,2-Dimethylhexahydrocyclopropen[3,4]cyclopentadien[1,2-d][1,3]dioxacyclopentene-5-

[0436] Preparation of 9H-purine-6-amine (23)

[0437]

[0438] 5 steps - 1

[0439]

[0440] At 20 °C, DIAD (4.28 g, 4.11 mL, 2.00 equivalent) and PPh3 (5.55 g, 2.00 equivalent) were added to a solution of compound 4 (2.00 g, 1.00 equivalent) and compound 17 (5.15 g, 1.10 equivalent) in THF (70 mL). The resulting mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with H2O (200 mL) and extracted with EtOAc (90 mL × 3). The organic extract was washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 10:1 to 0:1) to produce compound 18.1 (4.30 g, crude) as a pale yellow solid. 1 H NMR: 400

[0441] MHz DMSO-d6δ(ppm)8.78(s,1H),7.39-7.24(m,15H),5.20(d,J=7.2Hz,1H),5.05(s,1H),4.83(d,J=6.4Hz,1H ),4.79-4.75(m,1H),4.06-4.01(m,1H),1.83-1.79(m,1H),1.47(s,3H),1.18(s,3H),1.04-1.06(m,2H).

[0442] • Following the procedure of step-1, 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethyl-3b-((triphenylmethyloxy)methyl)hexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine (18.2) was prepared by replacing compound 4 with 5,7-dichloro-3H-imidazo[4,5-b]pyridine. MS: 611.9 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.67(s,1H),7.60(s,1H),7.38-7.24(m,15H),5.25(d,J=7.2Hz,1H),5.05(s,1H),4.79(d,J=7.2H z,1H),3.38-3.28(m,2H),1.75-1.72(m,1H),1.46(s,3H),1.19(s,3H),1.03(t,J=4.8Hz,1H),0.95-0.93(m,1H);

[0443] • Following the procedure of step-1, 5-bromo-,7-bromo / chloro-3H-imidazo[4,5-b]pyridine was similarly prepared by replacing compound 4 with 5-bromo-7-bromo / chloro-3H-imidazo[4,5-b]pyridine to prepare 5-bromo-,7-bromo / chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethyl-3b-((triphenylmethyloxy)methyl)hexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine (18.3). MS: 658.1 (M+H) + and 702.0(M+H) + .

[0444] Step-2

[0445]

[0446] At 30 °C, a mixture of compound 18.1 (4.20 g, crude), AcOH (44.1 g, 42.0 mL, 80.0%, in water, 220.0 equivalents) in ACN (13 mL) was stirred for 12 h. TLC (petroleum ether:ethyl acetate = 0:1) indicated approximately 0% of compound 18.1 remaining, and a major new spot with high polarity was detected. The reaction mixture was alkalized to pH approximately 8 with aqueous NH3·H2O and extracted with EtOAc (50 mL × 3). The combined organic extracts were washed with brine (70 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 50:1 to 0:1) to produce compound 19.1 (840.0 mg) as a white solid. MS: 371.1 (M+H) + . 1 HNMR:400MHz DMSO-d6δ(ppm)8.85(s,1H),5.18(d,J=7.2Hz,1H),4.99(s,1H),4.95(t,J=5.4Hz,1H),4.72(d,J=3.6Hz,1H),3.85- 3.81(m,1H),3.79-3.44(m,1H),1.75-1.72(m,1H),1.45(s,3H),1.16(s,3H),0.99-0.97(m,1H),0.95-0.92(m,1H).

[0447] • Following the procedure in step-2, compound 18.2 was used instead of 18.1 to similarly prepare ((3aR,3bR,4aS,5R,5aS)-5-(5,7-dichloro-3H-imidazo[4,5-b]pyridin-3-yl)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b-yl)methanol (19.2). MS: 370.1 (M+H) + . 1 H NMR: 400MHzDMSO-d6δ(ppm)8.75(s,1H),7.67(s,1H),5.20(d,J=7.2Hz,1H),5.01(s,1H),4.95(t,J=5.2Hz, 1H),4.66-4.65(m,2H),3.47-3.43(m,1H),1.72-1.69(m,1H),1.45(s,3H),1.16(s,3H),1.00-0.92(m,2H).

[0448] • Following the procedure in step-2, compound 18.3 was used instead of 18.1 to similarly prepare ((3aR,3bR,4aS,5R,5aS)-5-(5-bromo-7-bromo / chloro-3H-imidazo[4,5-b]pyridin-3-yl)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b-yl)methanol (19.3). MS: 416.0 (M+H) + and 460.0(M+H) +

[0449] Step 3

[0450]

[0451] Compound 20 (651.0 mg, 378.5 μL, 4.00 equivalents), compound 21 (347.4 mg, 351.2 μL, 4.00 equivalents), and TEA (436.1 mg, 599.9 μL, 8.00 equivalents) were added to a solution of compound 19.1 (200.0 mg, 1.00 equivalents) in 2 mL of THF. The resulting mixture was stirred at 20 °C for 12 h. TLC (petroleum ether:ethyl acetate = 3:1) indicated approximately 0% of compound 19 remaining, and a major new spot with low polarity was detected. The reaction mixture was concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 3:1) to produce compound 22 (0.17 g) as a pale yellow solid. MS: 373.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.71(s,1H),5.26(d,J=7.2Hz,1H),5.08(d,J=3.2Hz,1H),4.99-4.84(m,2H) ,4.46-4.31(m,1H),1.94(t,J=7.6Hz,1H),1.47(s,3H),1.18(s,3H),1.13(d,J=7.6Hz,1H).

[0452] Step 4

[0453]

[0454] TEA (230.4 mg, 317.0 μL, 5.00 equivalent) was added to a solution of compound 22 (0.17 g, 1.00 equivalent) and compound 6 (110.7 mg, 3.00 equivalent) in t-BuOH (10 mL) at 20 °C. The resulting mixture was stirred at 65 °C for 2 h. The reaction mixture was diluted with H₂O (30 mL) and extracted with EtOAc (10 mL × 3). The combined organic extracts were washed with brine (15 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to yield compound 23 (0.17 g, crude) as a pale yellow solid. MS: 418.1 (M + H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.65(s,1H),8.22(s,1H),6.18(t,J=56Hz,1H),5.25(d,J=7.2Hz,1H),4.95(d,J=2.4Hz,1H),4.8 4-4.73(m,2H),4.47-4.32(m,1H),3.85(s,2H),1.83-1.80(m,1H),1.46(s,3H),1.18(s,3H),1.11-1.06(m,2H).

[0455] Table 8. Intermediates prepared by the procedure of Example 8

[0456]

[0457] Example 9.2 Preparation of chloro-9-((3aR,3bS,4aS,5R,5aS)-3b-(difluoromethyl)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-9H-purine-6-amine (26.1)

[0458]

[0459] 5 steps - 1

[0460]

[0461] A solution of compound 19.1 (370.0 mg, 1.00 equivalent) and Dess-Martin periodane (507.3 mg, 0.37 μL, 1.20 equivalent) in DCM (4 mL) was stirred for 0.5 h at 20 °C. LC-MS analysis showed approximately 0% of compound 19.1 remaining. The mixture was filtered through a diatomaceous earth filter, diluted with 5% Na₂SO₃ aqueous solution (20 mL), 5% NaHCO₃ aqueous solution (40 mL), and extracted with DCM (25 mL × 5). The organic extract was washed with brine (60 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to yield compound 24 (360.0 mg, crude) as a pale yellow solid. MS: 369.1 (M+H) + .

[0462] Step-2

[0463]

[0464] A solution of compound 24 (360.0 mg, 1.00 equivalent) in DCM (4 mL) and DAST (738.7 mg, 0.61 μL, 4.70 equivalent) in DCM (0.5 mL) were stirred for 1 h at 20 °C. TLC (petroleum ether:ethyl acetate = 0:1) indicated approximately 0% of compound 24 remaining. The reaction mixture was diluted with saturated aqueous NaHCO3 solution (25 mL) and extracted with DCM (20 mL × 3). The combined organic extracts were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 0:1) to produce compound 25 (260.0 mg) as a pale yellow solid. MS: 391.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.68(s,1H),6.42-6.13(m,1H),5.39(d,J=7.2Hz,1H),5.11(s,1H),4.92(d,J= 6.8Hz,1H),2.16-2.13(m,1H),1.48(s,3H),1.34-1.32(m,1H),1.19(s,3H),1.17-1.15(m,1H).

[0465] Step 3

[0466]

[0467] TEA (194.0 mg, 0.27 μL, 5.00 equivalent) was added to a solution of compound 25 (150.0 mg, 1.00 equivalent) and ethylamine hydrochloride (62.5 mg, 2.00 equivalent) in t-BuOH (3 mL) at 20 °C. The resulting mixture was stirred at 65 °C for 2 h. LC-MS analysis showed approximately 0% of compound 25 remaining. The reaction mixture was concentrated under reduced pressure to yield compound 26.1 (160.0 mg, crude) as a pale yellow solid. MS: 400.2 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.38(s,1H),8.10(s,1H),6.42-6.13(m,1H),5.35(d,J=6.4Hz,1H),4.96(s,1H),4.78(d ,t=3.6Hz,1H),3.47-3.44(m,2H),2.01-1.99(m,1H),1.46(s,3H),1.31-1.28(m,1H),1.22-1.14(m,7H).

[0468] • Using the method of Example 9, 5-chloro-3-((3aR,3bS,4aS,5R,5aS)-3b-(difluoromethyl)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-3H-imidazo[4,5-b]pyridine-7-amine (26.2) was prepared similarly in step-1 by replacing compound 19.2. MS: 399.0 (M+H) + . 1 H NMR: 400MHz

[0469] MeODδ(ppm)8.02(s,1H),6.49-6.20(m,2H),5.47(d,J=7.2Hz,1H),5.00(s,1H),4.79( d,J=6.8Hz,1H),3.41-3.35(m,2H),1.90-1.87(m,1H),1.53(s,3H),1.34-1.25(s,8H);

[0470] • Using the method of Example 9, 5-bromo-3-((3aR,3bS,4aS,5R,5aS)-3b-(difluoromethyl)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-3H-imidazo[4,5-b]pyridine-7-amine (26.3) was prepared similarly in step-1 by replacing compound 19.3. MS: 443.1 (M+H) + .1 H NMR: 400MHzDMSO-d6δ(ppm)8.06(s,1H),7.25(t,J=5.6Hz,1H),6.19-6.18(m,1H),6.53-6.18(m,1H),5.36(d,J=7.2Hz,1H), 4.99(s,1H),4.74(d,J=6.8Hz,1H),3.48-3.35(m,2H),1.98-1.91(m,1H),1.46(s,3H),1.33-1.25(m,2H),1.20-1.16(m,6H).

[0471] Table 9. Intermediates prepared by the procedure of Example 9

[0472]

[0473] Example 10. Preparation of 2-chloro-N-(2,2-difluoroethyl)-9-((3aR,3bR,4aS,5R,5aS)-2,2,3b-trimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-yl)-9H-purine-6-amine (34)

[0474]

[0475] Step-1

[0476]

[0477] At 0 °C, NaH (298.2 mg, 60.0%, in mineral oil, 1.10 equivalent) was added to a solution of compound 17 (3.00 g, 1.00 equivalent) in THF (30 mL), and the reaction mixture was stirred at 0 °C for 0.5 h. Then, BnBr (3.48 g, 2.42 mL, 3.00 equivalent) and NaI (10.1 mg, 0.010 equivalent) were added at 0 °C. The reaction mixture was stirred at 20 °C for 12 h. TLC (petroleum ether:ethyl acetate = 5:1) indicated approximately 0% of compound 17 remaining, and a major new spot with low polarity was detected. The reaction mixture was quenched at 0 °C by adding saturated aqueous NH4Cl solution (100 mL) and extracted with EtOAc (50 mL × 3). The organic extract was washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0 to 0:1) to produce compound 27 (3.60 g), which was a yellow oil. 1H NMR:400MHz DMSO-d6δ(ppm)7.37-7.25(m,20H),4.85(d,J=6.8Hz,1H),4.63(t,J=1.2Hz,1H),4.54-4.50(m,2H),4.30(t,J=5.4Hz, 1H), 3.16 (d, J = 10Hz, 1H), 2.81 (d, J = 9.6Hz, 1H), 1.62-1.60 (m, 1H), 1.41 (s, 3H), 1.21-1.19 (m, 4H), 0.51-0.48 (m, 1H).

[0478] Step 2

[0479]

[0480] At 30 °C, a mixture of compound 27 (3.60 g, 1.00 equivalent) and AcOH (113.4 g, 108.0 mL, 80.0% aqueous solution) in acetonitrile (33 mL) was stirred for 12 h. TLC (dichloromethane:methanol = 10:1) indicated approximately 0% of compound 27 remaining and detected a major new spot with high polarity. The reaction mixture was alkalized to pH approximately 8 with ammonia and concentrated under reduced pressure to produce a residue. The residue was stirred with EtOAc (20 mL) and filtered. The filtrate was concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 1:0 to 0:1) to produce compound 28 (2.00 g, crude), which was a yellow oil. 1 H NMR:400MHz DMSO-d6δ(ppm)7.38-7.33(m,4H),7.30-7.26(m,1H),4.57(d,J=12Hz,1H),4.49-4.40(m,3H),4.20-4.18(m,1H),4.06-3.98(m, 2H),3.88-3.85(m,1H),3.78(d,J=12Hz,1H),3.01(d,J=11.6Hz,1H),1.38-1.36(m,1H),1.30(t,J=4Hz,1H),0.29-0.26(m,1H).

[0481] Step 3

[0482]

[0483] At 20°C, 2,2-dimethoxypropane (4.99 g, 5.87 mL, 6.00 equivalents) and TsOH·H₂O (1.38 g, 1.00 equivalents) were added to a solution of compound 28 (2.00 g, crude) in acetone (200 mL). The resulting mixture was stirred at 20°C for 2 h. TLC analysis (petroleum ether:ethyl acetate = 0:1) indicated approximately 0% of compound 28 remaining, and a major new spot with low polarity was detected. The reaction mixture was alkalized to pH approximately 8 with ammonia, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO₂, petroleum ether:ethyl acetate = 1:0 to 0:1) to produce compound 29 (1.50 g) as a pale yellow solid. 1 H NMR:400MHz DMSO-d6δ(ppm)7.35(d,J=4.4Hz,4H),7.31-7.27(m,1H),4.81(d,J=6.4Hz,1H),4.61-4.56(m,2H),4.56-4.48(m,2H),4.21(d,J=8H z,1H),3.64-3.60(m,1H),3.11-3.07(m,1H),1.62-1.58(m,1H),1.42(s,3H),1.21(s,3H),1.15(t,J=4.4Hz,1H),0.53-0.50(m,1H).

[0484] Step 4

[0485]

[0486] CBr4 (1.14 g, 2.00 equivalent), PPh3 (903.3 mg, 2.00 equivalent), and TEA (522.8 mg, 719.1 μL, 3.00 equivalent) were added to a solution of compound 29 (500.0 mg, 1.00 equivalent) in DCM (10 mL). The mixture was stirred at 25 °C for 16 h. TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction was complete. The mixture was quenched by adding H2O (20 mL) and extracted with DCM (10 mL × 3). The organic extract was washed with saturated aqueous NaCl solution (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by silica gel chromatography (SiO2, petroleum ether: ethyl acetate = 100:1 to 50:1) to obtain compound 30 (500.0 mg) as a white solid. 1H NMR:400MHz DMSO-d6δ(ppm)7.36-7.29(m,5H),4.82(d,J=6.0Hz,1H),4.62(t,J=6.4Hz,1H),4.56-4.48(m,2H),4.24-4.21 (m,1H),3.74-3.59(m,2H),1.84-1.80(m,1H),1.49-1.47(m,1H),1.43(s,3H),1.22(s,3H),0.87-0.84(m,1H).

[0487] Step 5

[0488]

[0489] At 0 °C, LiBHEt3 (1 M, 8.49 mL, 4.00 equivalent) was added to a solution of compound 30 (750.0 mg, 1.00 equivalent) in THF (10 mL). The mixture was stirred at 25 °C for 1 h. TLC analysis (petroleum ether: ethyl acetate = 8:1) showed that the reaction was complete. The mixture was quenched by adding H2O (20 mL) and extracted with ethyl acetate (10 mL × 3). The organic extract was washed with saturated aqueous NaCl solution (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by preparative TLC (petroleum ether: ethyl acetate = 5:1) to obtain compound 31 (600.0 mg, 90.0% purity) as a white solid. 1 H NMR:400MHz DMSO-d6δ(ppm)7.35-7.27(m,5H),4.55-4.67(m,4H),44.24(s,1H),1.51-1.48(m,1H),1.42(s,3H),1.17-1.14(m,7H),0.39-0.36(m,1H).

[0490] Step 6

[0491]

[0492] Under a nitrogen atmosphere, 10% Pd / C (500.0 mg) was added to a solution of compound 31 (600.0 mg, 1.00 equivalent) in MeOH (15 mL). The suspension was degassed and purged three times with H2. The mixture was stirred for 6 h at 25 °C under H2 (15 Psi). TLC analysis (petroleum ether:ethyl acetate = 5:1) showed approximately 1 / 3 of compound 31 remaining. The solid was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 3:1) to obtain compound 32 (150.0 mg) as a white solid.1 H NMR: 400MHz DMSO-d6δ(ppm)4.49-4.48(m,1H),4.37-4.33(m,3H),1.42-1.39(m,4H),1.78(s,3H),1.13-1.10(m,4H),0.30-0.26(m,1H).

[0493] Step 7

[0494]

[0495] At 0 °C and under N2, PPh3 (484.1 mg, 2.00 equivalent) and DIAD (373.1 mg, 358.8 μL, 2.00 equivalent) were added to a solution of compound 32 (170.0 mg, 1.00 equivalent) in THF (4 mL). The mixture was stirred at 20 °C for 15 min, and compound 4 (209.3 mg, 1.20 equivalent) was added to the mixture in one go. The resulting mixture was stirred at 20 °C for 1 h. TLC (petroleum ether:ethyl acetate = 1:1) showed that the reaction was complete. The mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 1:1) to obtain compound 33 (130.0 mg, 55.0% purity) as a yellow oil. MS: 355.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.75(s,1H),5.03-4.99(m,2H),4.78-4.74(m,1H),1.56-1.52(m,1H) ),1.45(s,3H),1.39(s,3H),1.15(s,3H),0.90(t,J=4.8Hz,1H),0.74-0.70(m,1H).

[0496] Step 8

[0497]

[0498] Compound 6 (67.8 mg, 3.00 equivalent) and TEA (84.6 mg, 116.4 μL, 3.00 equivalent) were added to a solution of compound 33 (180.0 mg, 55.0% purity, 1.00 equivalent) in t-BuOH (5 mL). The mixture was stirred at 65 °C for 1 h. TLC (petroleum ether:ethyl acetate = 0:1) showed that the reaction was complete. The mixture was concentrated under reduced pressure. The residue was diluted with ethyl acetate (10 mL × 3) and extracted with water (15 mL). The organic layer was washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to produce a residue to give compound 34 (150.0 mg) as a yellow oil. MS: 400.0 (M + H) + . 1 HNMR:400MHz DMSO-d6δ(ppm)8.60(s,1H),8.25(s,1H),6.33-6.04(m,1H),5.01(d,J=6.8Hz,1H),4.86(s,1H),4.63(d, J=7.2Hz,1H),3.85(s,2H),1.44(s,3H),1.37(s,3H),1.27-1.19(m,2H),1.16(s,3H),0.70-0.67(m,1H).

[0499] Example 11. Preparation of (1R,2R,3S,4R,5S)-4-(2-chloro-6-((2,2-difluoroethyl)amino)-9H-purin-9-yl)bicyclo[3.1.0]hexane-2,3-diol (35)

[0500]

[0501] Compound 7.1 (110.0 mg, 1.00 equivalent) was stirred in 10 mL of 20% TFA aqueous solution at 20 °C for 1 h. LC-MS analysis showed that the starting material was completely consumed. The reaction mixture was concentrated under reduced pressure to produce a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.05% NH3H2O ​​+ 10 mM NH4HCO3)-ACN]; B%: 5%-35%, 8 min) to produce compound 35 (87.2 mg) as a white solid. MS: 346.1 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.69-8.40(m,1H),8.28(s,1H),6.37-6.01(m,1H),5.15(d,J=4.4Hz,1H),4.70-4.49( m,3H),3.91-3.72(m,3H),1.87-1.78(m,1H),1.55-1.43(m,1H),1.12-1.05(m,1H),0.67-0.59(m,1H).

[0502] The following compounds were prepared similarly by substituting compound 7.1 in the method of Example 11:

[0503] Compound 7.2 produced (1R,2R,3S,4R,5S)-4-(2-chloro-6-((2-fluoroethyl)amino)-9H-purin-9-yl)bicyclo[3.1.0]hexane-2,3-diol (36). MS: 328.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.46-8.40(m,1H),8.23(s,1H),5.13(d,J=4.4Hz,1H),4.70-4.48(m,5H),3. 84-3.66(m,3H),1.85-1.78(m,1H),1.54-1.46(m,1H),1.13-1.06(m,1H),0.68-0.58(m,1H);

[0504] Compound 7.3 produced (1R,2R,3S,4R,5S)-4-(2-chloro-6-((2,2,2-trifluoroethyl)amino)-9H-purin-9-yl)bicyclo[3.1.0]hexane-2,3-diol (37). MS: 364.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.88(s,1H),8.32(s,1H),5.14(d,J=4.4Hz,1H),4.66(d,J=1.2Hz,1H),4.63-4.58(m,1H),4.55-4.53( m,1H),4.26(s,2H),3.80(t,J=5.2Hz,1H),1.84-1.81(m,1H),1.53-1.51(m,1H),1.12-1.09(m,1H),0.64-0.62(m,1H);

[0505] Compound 11.1 yielded (1R,2R,3S,4R,5S)-4-(2-(cyclopropylethynyl)-6-((2,2-difluoroethyl)amino)-9H-purin-9-yl)bicyclo[3.1.0]hexane-2,3-diol (38). MS: 376.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.27(s,1H),8.13(s,1H),6.34-5.99(m,1H),5.13(d,J=4.4Hz,1H),4.69(s,1H),4.62-4.50(m,2H),3.96-3.72(m,3H) ,1.84-1.80(m,1H),1.63-1.53(m,1H),1.52-1.45(m,1H),1.13-1.10(m,1H),0.96-0.88(m,2H),0.84-0.75(m,2H),0.64-0.59(m,1H);

[0506] Compound 11.1 produced (1R,2R,3S,4R,5S)-4-(2-(cyclopropylethynyl)-6-(ethylamino)-9H-purin-9-yl)bicyclo[3.1.0]hexane-2,3-diol (39). MS: 340.2 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.18(s,1H),7.79(s,1H),5.11(d,J=4.4Hz,1H),4.68(s,1H),4.62-4.51(m,2H),3.75(t,J=4.8Hz,1H),3.4 7(s,2H),1.84-1.83(m,1H),1.57-1.49(m,2H),1.18-1.12(m,4H),0.93-0.88(m,2H),0.79-0.78(m,2H),0.62-0.60(m,1H);

[0507] Compound 9.1 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2,2-difluoroethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (40). MS: 345.0 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.20(s,1H),7.40(t,J=6.4Hz,1H),6.55(s,1H),6.31-6.16(m,1H) ,5.09(d,J=2.8Hz,1H),4.71(s,1H),4.62-4.60(m,1H),4.52-4.51(m,1H),3.98(br s,2H),3.75(d,J=5.6Hz,1H),1.84-1.81(m,1H),1.50-1.47(m,1H),1.13-1.10(s,1H),0.62-0.60(m,1H);

[0508] Compound 9.2 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2-fluoroethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (41). MS: 327.1 (M+H) + . 1 H NMR: 400MHzDMSO-d6δ(ppm)8.17(s,1H),7.35-7.32(m,1H),6.46(s,1H),5.11(s,1H),4.71-4.53(m, 5H),3.81-3.75(m,3H),1.86-1.81(m,1H),1.50-1.48(m,1H),1.14-1.11(m,1H),0.63-0.61(m,1H);

[0509] Compound 9.3 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2,2,2-trifluoroethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (42). MS: 363.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.23(s,1H),7.67(t,J=6.8Hz,1H),6.64(s,1H),5.10(d,J=4.4Hz,1H),4.71(s,1H),4.61-4.59(m,1H) ,4.54-4.47(m,3H),3.76(t,J=5.2Hz,1H),1.84-1.81(m,1H),1.52-1.50(m,1H),1.13-1.10(m,1H),0.63-0.60(m,1H);

[0510] Compound 9.4 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (43). MS: 309.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.12(s,1H),7.13(t,J=5.6Hz,1H),6.33(s,1H),5.08(t,J=4.4Hz,1H),4.70(s,1H),4.62-4.59(m,1H),4.51(d, J=7.2Hz,1H),3.75(t,J=4.8Hz,1H),3.40(brs,2H),1.84-1.82(m,1H),1.49-1.48(m,1H),1.19-1.11(m,4H),0.62-0.60(m,1H);

[0511] Compound 9.5 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-(cyclobutylamino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (44). MS: 335.0 (M+H) + . 1 H NMR: 400MHz DMSO-d6δ(ppm)8.14(s,1H),7.41(d,J=7.2Hz,1H),6.28(s,1H),5.09(d,J=4. 4Hz,1H),4.69(s,1H),4.63-4.57(m,1H),4.51(d,J=7.2Hz,1H),4.46-4.16(m, 1H),3.76-3.71(m,1H),2.35-2.28(m,2H),2.09-1.97(m,2H),1.85-1.78(m,1H ),1.74-1.64(m,2H),1.51-1.43(m,1H),1.13-1.08(m,1H),0.65-0.58(m,1H);

[0512] Compound 9.6 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2,5-difluorobenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (45). MS: 407.0 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.19(s,1H),7.77-7.74(m,1H),7.28-7.26(m,1H),7.17-7.13(m,2H),6.37(s,1H),5.08(d,J=4.4Hz,1H),4.70-4.69(m,3H ),4.62-4.59(m,1H),4.52-4.50(m,1H),3.76(t,J=4.8Hz,1H),1.84-1.80(m,1H),1.50-1.48(m,1H),1.13-1.10(m,1H),0.62-0.60(m,1H);

[0513] Compound 14.1 yielded (1R,2R,3S,4R,5S)-4-(5-(cyclopropylethynyl)-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (46). MS: 339.1 (M+H) + . 1 H NMR: 400MHz DMSO-d6δ(ppm)8.12(s,1H),6.75(t,J=5.8Hz,1H),6.39(s,1H),5.07(d,J=4.4H z,1H),4.76(s,1H),4.62-4.57(m,1H),4.48(d,J=7.2Hz,1H),3.72(t,J=5.4Hz, 1H),3.37(t,J=6.6Hz,2H),1.82-1.81(m,1H),1.57-1.53(m,1H),1.48-1.44(m, 1H),1.19-1.12(m,4H),0.91-0.87(m,2H),0.78-0.74(m,2H),0.63-0.59(m,1H);

[0514] Compound 16.1 yielded (1R,2R,3S,4R,5S)-4-(7-((2,2-difluoroethyl)amino)-5-methyl-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (47). MS: 325.0 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.06(s,1H),6.93-6.82(m,1H),6.37(s,1H),6.32-6.01(m,1H),5.04(d,J=4.4Hz,1H),4.81(s,1H),4.67-4.61(m,1H),4.50(d ,J=7.3Hz,1H),3.99-3.86(m,2H),3.74-3.71(m,1H),2.39(s,3H),1.83 -1.79(m,1H),1.49-1.40(m,1H),1.13-1.10(m,1H),0.63-0.55(m,1H);

[0515] Compound 16.2 yielded (1R,2R,3S,4R,5S)-4-(7-((2-fluoroethyl)amino)-5-methyl-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (48). MS: 307.1 (M+H) + . 1 H NMR: 400MHzDMSO-d6δ(ppm)8.03(s,1H),6.75(t,J=6.0Hz,1H),6.30(s,1H),5.06(d,J=4.4Hz,1H),4.81(s,1H),4.67-4.64(m,2H ),4.55-4.51(m,2H),3.78-3.70(m,3H),2.40(s,3H),1.84-1.80(m,1H),1.46-1.44(m,1H),1.14-1.10(m,1H),0.61-0.59(m,1H);

[0516] Compound 16.3 yielded (1R,2R,3S,4R,5S)-4-(7-(ethylamino)-5-methyl-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (49). MS: 289.0 (M+H) + . 1H NMR: 400MHzDMSO-d6δ(ppm)7.99(s,1H),6.52(t,J=5.6Hz,1H),6.20(s,1H), 5.04(d,J=4.4Hz,1H),4.80(s,1H),4.67-4.60(m,1H),4.50(d,J=7.2Hz,1H), 3.73(t,J=5.2Hz,1H),3.41-3.34(m,2H),2.39(s,3H),1.83-1.79(m,1H),1. 47-1.40(m,1H),1.18(t,J=7.2Hz,3H),1.12-1.09(m,1H),0.63-0.55(m,1H);

[0517] Compound 16.4 yielded (1R,2R,3S,4R,5S)-4-(7-((2,5-difluorobenzyl)amino)-5-methyl-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (50). MS: 387.1 (M+H) + . 1 HNMR:400MHz DMSO-d6δ(ppm)8.05(s,1H),7.28-7.23(m,2H),7.14-7.11(m,2H),6.21(s,1H),5.03(d,J=4.4Hz,1H),4.80(s,1H),4.68-4.62(m,3H), 4.50(d,J=7.6Hz,1H),3.74(t,J=4.8Hz,1H),2.35(s,3H),1.83-1.80(m,1H),1.46-1.44(m,1H),1.13-1.10(m,1H),0.60-0.58(m,1H);

[0518] Compound 18.1 yielded (1R,2R,3S,4R,5S)-4-(7-((2,5-difluorobenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (51). MS: 373.0 (M+H) + . 1H NMR: 400MHzDMSO-d6δ(ppm)8.15(s,1H),7.89(d,J=5.6Hz,1H),7.37(t,J=6.4Hz,1H),7. 29-7.23(m,1H),7.17-7.08(m,2H),6.31(d,J=5.6Hz,1H),5.07(d,J=4.4Hz,1H),4.79(d, J=0.8Hz,1H),4.69(d,J=6.0Hz,2H),4.63-4.57(m,1H),4.48(d,J=7.3Hz,1H),3.78(t,J =5.4Hz,1H),1.84-1.80(m,1H),1.55-1.46(m,1H),1.14-1.10(m,1H),0.65-0.57(m,1H);

[0519] Compound 23 produced (1S,2R,3S,4R,5S)-4-(2-chloro-6-((2,2-difluoroethyl)amino)-9H-purin-9-yl)-1-(fluoromethyl)bicyclo[3.1.0]hexane-2,3-diol (52). MS: 378.0 (M+H) + . 1 H NMR: 400MHzDMSO-d6δ(ppm)8.62(s,1H),8.13(s,1H),6.19(t,J=55.8Hz,1H),5.36(d,J=7.2Hz,1H),4.91-4.76(m,2H),4.69 (s,1H),4.58(t,J=6Hz,1H),4.49-4.34(m,1H),3.89-3.85(m,3H),1.71-1.68(m,1H),1.51-1.47(m,1H),0.83-0.79(m,1H);

[0520] Compound 26.1 produced (1S,2R,3S,4R,5S)-4-(2-chloro-6-(ethylamino)-9H-purin-9-yl)-1-(difluoromethyl)bicyclo[3.1.0]hexane-2,3-diol (53). MS: 360.0 (M+H) + . 1H NMR: 400MHzDMSO-d6δ(ppm)8.31(s,1H),8.05(s,1H),6.38-6.09(m,1H),5.46(s,1H),4.86(d,J=7.2Hz,1H),4.73(t,J=6.4Hz, 1H), 4.62 (s, 1H), 3.97 (s, 1H), 3.45 (t, J = 6.4Hz, 2H), 1.81-1.78 (m, 1H), 1.49 (s, 1H), 1.16 (t, J = 7.0Hz, 3H), 1.07-1.05 (m, 1H). 1 H NMR:400MHz MeODδ(ppm)8.00(s,1H),6.36-6.07(m,1H),4.88(d,J=6.8Hz,1H),4.78(s,1H),4.02(d,t=6.8H z,1H),3.59(s,2H),1.92-1.89(m,1H),1.68(s,1H),1.28(t,J=14.8Hz,3H),1.17-1.15(m,1H);

[0521] Compound 26.2 yielded (1S,2R,3S,4R,5S)-4-(5-chloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-1-(difluoromethyl)bicyclo[3.1.0]hexane-2,3-diol (54). MS: 359.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.01(s,1H),7.18(t,J=6.0Hz,1H),6.39-6.10(m,2H),5.43(d,J=5.2Hz,1H),4.85-4.83(m,1H),4.77-4.73(m,1H) ,4.68(s,1H),3.94(t,J=5.2Hz,1H),3.40(s,2H),1.78-1.75(m,1H),1.52-1.51(m,1H),1.18(t,J=7.2Hz,3H),1.07-1.04(m,1H);

[0522] Compound 11.3 yielded (1S,2R,3S,4R,5S)-4-(5-(cyclopropylethynyl)-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-1-(difluoromethyl)bicyclo[3.1.0]hexane-2,3-diol (55). MS: 389.0 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.00(s,1H),6.81(s,1H),6.44-6.11(m,2H),5.42(d,J=4.8Hz,1H),4.84-4.80(m,1H),4.77-4.71(m,2H),3.89(s,1H),3 .42-3.36(m,2H),1.78-1.70(m,1H),1.59-1.50(m,2H),1.16(t,J=7.2Hz,3H),1.07-1.01(m,1H),0.93-0.87(m,2H),0.79-0.72(m,2H);

[0523] Compound 34 produced (1R,2R,3S,4R,5S)-4-(2-chloro-6-((2,2-difluoroethyl)amino)-9H-purin-9-yl)-1-methylbicyclo[3.1.0]hexane-2,3-diol (56). MS: 360.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.59(s,1H),8.12(s,1H),6.32-6.04(m,1H),5.11(d,J=4.0Hz,1H),4.64(s,1H),4.50(d,J=8.0Hz,1 H),4.40(t,J=7.2Hz,1H),3.87-3.75(m,3H),1.34(s,3H),1.29-1.26(m,1H),1.23-1.21(m,1H),0.46-0.43(m,1H);

[0524] Compound 7.4 yielded (1R,2R,3S,4R,5S)-4-(2-chloro-6-(methyl-d3-amino)-9H-purin-9-yl)bicyclo[3.1.0]hexane-2,3-diol (57). MS 299 (M+H) + . 1 H NMR: 400MHz DMSO d6δ8.18(s,1H),7.99(b,1H),4.63–4.58(m,3H). 5.25(d,J=5.2Hz,1H),5.1(t,J=5.2Hz,1H),1.84–1.80(m,1H),1.51.1.49(m,1H),1.12–1.09(m,1H),0.625–0.61(m,1H).

[0525] Compound 9.7 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((5-chloro-2-((3-methylisoxazol-5-yl)methoxy)benzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (BIO-0461) (75). LCMS: MS: 516.1 (M+H) + . 1 H NMR: 400MHz DMSO-d6δ(ppm)8.17(s,1H),7.72-7.66(m,1H),7.32-7.29(m,1H),7.23-7.18 (m,2H),6.51(s,1H),6.26(s,1H),5.35(s,2H),5.08(d,J=4.4Hz,1H),4.70(s ,1H),4.63-4.59(m,3H),4.52(d,J=6.8Hz,1H),3.78-3.75(m,1H),2.24(s,3H ),1.84-1.80(m,1H),1.50-1.48(m,1H),1.13-1.10(m,1H),0.62-0.60(m,1H)

[0526] Compound 9.8 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-(((6-methoxypyridin-2-yl)methyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (88). LCMS: MS: 402.1 (M+H) + . 1 H NMR: 400MHz DMSO-d6.δ(ppm)8.17(s,1H),7.73(t,J=6.4Hz,1H),7.64(t,J=7.6Hz,1H),6.90 (d,J=7.2Hz,1H),6.68(d,J=8.4Hz,1H),6.41(s,1H),5.09(d,J=4.4Hz,1H),4.7 0(s,1H),4.63-4.59(m,3H),4.52(d,J=6.4Hz,1H),3.85(s,3H),3.77-3.74(m,1 H),1.84-1.80(m,1H),1.50-1.48(m,1H),1.13-1.10(m,1H),0.63-0.60(m,1H).

[0527] Compound 110.1 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2,2-difluoroethyl)amino)-6-fluoro-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (61). It was a white solid in 71.5% yield. LCMS: 363.1 (M+H)+. 1 H NMR:400MHz DMSO-d6δ(ppm)8.31(s,1H),7.48-7.40(m,1H),6.39-6.06(m,1H),5.11(s,1H),4.70(s,1H),4.56(d,J=17.6Hz,2H),4.34(tt,J=5.6, 14.8Hz,2H),3.76(d,J=5.6Hz,1H),1.84-1.80(m,1H),1.49(td,J=4.4,8.8Hz,1H),1.12(q,J=4.4Hz,1H),0.62(dt,J=4.8,8.4Hz,1H).

[0528] Compound 103.1 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2,2-difluoroethyl)amino)-6-methyl-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (60). 43.6% yield, as a yellow solid. LCMS: 359.1 (M+H)+. 1 H NMR:400MHz DMSO-d6δ(ppm)8.21(s,1H),6.72(t,J=6.4Hz,1H),6.40-6.07(m,1H),5.10(d,J=4.4Hz,1H),4.70(s,1H),4.64-4.57(m,1H),4.56-4.5 1(m,1H),4.40-4.38(m,2H),3.76-3.71(m,1H),2.22(s,3H),1.82-1.79(m,1H),1.54-1.40(m,1H),1.13-1.10(m,1H),0.62-0.59(m,1H)

[0529] Compound 150.1 yielded (1R,2R,3S,4R,5S)-4-(5,6-dichloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (71) (57% yield). LCMS 343.0 (M+H)+. 1HNMR:400MHz DMSO-d6δ(ppm)8.23(s,1H),6.96(t,J=6.4Hz,1H),5.10(d,J=4.4Hz,1H),4.69(s,1H),4.63-4.57(m,1H),4.55-4.51(m,1H),4.11-4 .00(m,2H),3.76(t,J=5.2Hz,1H),1.85-1.78(m,1H),1.51-1.46(m,1H),1.22(t,J=7.2Hz,3H),1.12-1.10(m,1H),0.63-0.59(m,1H).

[0530] Compound 9.9 produced ((1R,2R,3S,4R,5S)-4-(5-chloro-7-(propylamino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (66). LCMS: 323.1 (M+H) + . 1 H NMR: DMSO+D2O_400MHzδ(ppm)8.13(s,1H),7.18-7.16(m,1H),6.35(s,1H),4.69(s,1H),4.60(t,J=5.6Hz,1H),3.74(d,J=6.4Hz,1H) ,3.41(s,2H),1.83-1.82(m,1H),1.62-1.56(m,2H),1.51-1.49(m,1H),1.12-1.11(m,1H),0.92(t,J=7.6Hz,3H),0.63-0.61(m,1H).

[0531] Compound 9.10 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((cyclobutylmethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (67). LCMS: 349.2 (M+H) + . 1H NMR: DMSO400MHz.δ(ppm)8.12(s,1H),7.15(t,J=6.00Hz,1H),6.35(s,1H),5.08 (d,J=4.8Hz,1H),4.69(s,1H),4.63-4.58(m,1H),4.51(d,J=7.2Hz,1H),3.74(t, J=5.2Hz,1H),3.43(s,2H),2.52-2.51(m,1H),2.00-1.98(m,2H),1.85-1.82(m, 3H),1.80-1.72(m,2H),1.51-1.48(m,1H),1.13-1.11(m,1H),0.62-0.60(m,1H).

[0532] Compound 9.11 yielded (1R,2R,3S,4R,5S)-4-(5-chloro-7-((2,5-dichlorobenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (68). LCMS: 439.1 (M+H) + . 1 HNMR: DMSOδ(ppm)8.20(s,1H),7.80(t,J=5.6Hz,1H),7.53(d,J=8.8Hz,1H ),7.40-7.35(m,2H),6.33(s,1H),5.09(d,J=4.4Hz,1H),4.75(s,2H),4.71 (s,1H),4.62-4.59(m,1H),4.51(d,J=7.2Hz,1H),3.77(t,J=4.8Hz,1H),1 .83-1.81(m,1H),1.51-1.49(m,1H),1.13-1.10(m,1H),0.63-0.61(m,1H).

[0533] Example 12. Preparation of 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine (100)

[0534]

[0535] At -70 °C and under N2, LiHMDS (1 M, 13.23 mL, 2.5 equivalents) was added dropwise to a solution of compound 5.3 (1.8 g, 5.29 mmol, 1.0 equivalent), TIPSCl (1.70 mL, 7.94 mmol, 1.5 equivalent), and HMPA (9.30 mL, 52.91 mmol, 10 equivalent) in 20 mL of THF. The mixture was stirred at -70 °C for 3 h. TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction was complete. The reaction was quenched by adding saturated NH4Cl solution (40 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (40 mL × 1), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography (SiO2, petroleum ether) to give compound 100 (2 g, 4.03 mmol, 76.1% yield) as a white solid. LCMS:MS:496.1(M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)7.61(s,1H),5.45-5.40(m,1H),4.89(d,J=7.2Hz,1H),4.72(s,1H),1.98- 1.94(m,1H),1.64-1.50(m,4H),1.20-1.11(m,24H),0.89-0.81(m,1H),0.71-0.69(m,1H).

[0536] Example 13. Preparation of 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine-6-carboxynitrile (101)

[0537]

[0538] At -70 °C and under N2, LDA (2 M, 1.01 mL, 2 equivalents) was added dropwise to a solution of compound 100 (0.5 g, 1.01 mmol, 1 equivalent) in THF (10 mL). After the addition, the mixture was stirred at -70 °C for 0.5 h. TosCN (364.9 mg, 2.01 mmol, 2 equivalents) was added dropwise in THF (10 mL) at -70 °C. The resulting mixture was stirred at -70 °C for 2 h. LC-MS showed approximately 7.0% of compound 100 remaining. Several new peaks were observed on LC-MS, and approximately 17.2% of the desired compound was detected. The reaction mixture was quenched by adding saturated aqueous NH4Cl solution (40 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (40 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1, 8 / 1 to 0 / 1) to obtain compound 101 as a white solid (0.23 g, 440.99 μmol, 43.79% yield). LCMS: 521.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)5.37(t,J=6.4Hz,1H),4.91(d,J=7.2Hz,1H),4.72(s,1H),2.01-1.99(m,1H),1.69-1.68(m, 1H),1.59-1.55(m,3H),1.43(s,3H),1.18-1.14(m,18H),0.98(s,3H),0.91-0.89(m,1H),0.72-0.71(m,1H).

[0539] The following compounds were prepared similarly according to the method of Example 13:

[0540] Replacing TosCN with NFSI yielded 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropeno[3,4]cyclopentadieno[1,2-d][1,3]dioxacyclopenten-5-

[0541] 6-Fluoro-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine (101.1)

[0542] ·use The substitution of TosCN produced 5,7-dichloro

[0543] -3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenyl[3,4]cyclopentadiene

[0544] [1,2-d][1,3]dioxacyclopenten-5-yl)-6-methyl-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine (102.1). LCMS: 510.2 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)5.44-5.43(m,1H),4.86(d,J=6.8Hz,1H),4.71(s,1H),1.98-1.93(m,1H), 1.59-1.56(m,4H),1.43(s,3H),1.19-1.10(m,24H),0.86-0.84(m,1H),0.70-0.73(m,1H).

[0545] Replacing TosCN with Cl2Cl6 yielded 5,6,7-trichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine (104.1 LCMS: 532.1 (M+H) + . 1 H NMR:400MHz CDCl3δ(ppm)5.63-5.58(m,1H),4.82(t,J=3.2Hz,2H),2.17-2.10(m,1H),1.67-1.59 (m,3H),1.56-1.53(m,3H),1.53-1.45(m,1H),1.23-1.17(m,21H),0.95-0.83(m,2H).

[0546]

[0547] Example 14. Preparation of 5-chloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-7-(ethylamino)-3H-imidazo[4,5-b]pyridine-6-carboxynitrile (102)

[0548]

[0549] Under N2, DIEA (1.92 mmol, 0.35 mL, 5 equivalents) and EtNH2·HCl (156.3 mg, 1.92 mmol, 5 equivalents) were added dropwise to a solution of compound 101 (0.2 g, 383.47 μmol, 1 equivalent) in NMP (5 mL). The resulting mixture was stirred at 130 °C for 16 h. LC-MS showed approximately 0% of compound 101 remaining. Several new peaks were observed on LC-MS, and approximately 14.9% of the desired compound was detected. The reaction mixture was quenched by adding water (15 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (40 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to yield compound 102 (0.2 g, crude) as a white solid. LC-MS: MS: 374.1 (M+H) + .

[0550] Example 15. Preparation of 5-chloro-3-((1S,2R,3S,4R,5R)-3,4-dihydroxybicyclo[3.1.0]hexane-2-yl)-7-(ethylamino)-3H-imidazo[4,5-b]pyridine-6-carboxynitrile (93)

[0551]

[0552] TFA (60.78 mmol, 15.0 mL, 30% purity, 113.6 equivalents) was added dropwise to a solution of compound 102 (0.20 g, 534.99 μmol, 1 equivalent) at 20 °C. After addition, the mixture was stirred at 40 °C for 2 h. LC-MS showed approximately 0% of compound 102 remaining. Several new peaks were observed on LC-MS, and approximately 97.8% of the desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove the solvent. The crude product was purified by preparative HPLC (column: Phenomenex C1875 30 mm 3 μm; mobile phase: [water (NH3H2O ​​+ NH4HCO3)-ACN]; B%: 10%-40%, 8 min) to obtain BIO-0458 (52.0 mg, 155.79 μmol, 29.1% yield) as a white solid. LCMS:MS:334.0(M+H) + . 1H NMR: DMSO 400MHz δ (ppm) 8.32 (s, 1H), 7.72 (t, J = 6.0Hz, 1H), 5.13 (d, J = 4.4Hz 1H),4.70(s,1H),4.60-4.54(m,2H),4.03-3.98(m,2H),3.78-3.76(m,1H),1.83-1.8 1(m,1H),1.51-1.48(m,1H),1.24-1.20(m,3H),1.12-1.11(m,1H),0.64-0.61(m,1H).

[0553] Example 16. Preparation of 5-chloro-N-(2,2-difluoroethyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-6-fluoro-3H-imidazo[4,5-b]pyridine-7-amine (110.1)

[0554]

[0555] A mixture of compound 101.1 (300 mg, 583.1 μmol, 1.0 equivalent) and compound 8 (472.6 mg, 5.83 mmol, 10 equivalent) in NMP (2 mL) was stirred for 40 h at 120 °C. LC-MS showed the reaction was complete. The reaction mixture was partitioned between ethyl acetate (15 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over Na₂SO₄, filtered, concentrated, and then purified by preparative TLC (petroleum ether:ethyl acetate = 2:1) to give compound 110.1 (130 mg, 322.7 μmol, 55.3% yield) as a yellow solid. LC-MS: 403.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.36(s,1H),7.48-7.45(m,1H),6.38-6.06(m,1H),5.27-5.22(m,1H),4.92(s,1H),4.58(d,J=7. 2Hz,1H),4.34-4.30(m,2H),2.00-1.94(m,1H),1.75-1.69(m,1H),1.44(s,3H),1.16(s,3H),0.88-0.76(m,2H).

[0556] The following compounds were prepared similarly according to the method of Example 16:

[0557] Replacing 101.1 with 102.1 yielded 5-chloro-N-(2,2-difluoroethyl)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-6-methyl-3H-imidazo[4,5-b]pyridine-7-amine (103.1). Yield: 42.6%, as a yellow solid. LCMS: 399.1 (M+H) + . 1 HNMR:400MHz DMSO-d6δ(ppm)8.25(s,1H),6.74(t,J=6.5Hz,1H),6.38-6.07(m,1H),5.29-5.23(m,1H),4 .90(s,1H),4.56(d,J=6.8Hz,1H),4.49-4.32(m,2H),2.22(s,3H),1.98-1.90(m,1H),1.70 -1.68(m,1H),1.44(s,3H),1.16(s,3H),0.88-0.76(m,2H).

[0558] Replacing 101.1 with 104.1 and adding HCl yielded 5,6-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-N-ethyl-3H-imidazo[4,5-b]pyridine-7-amine (150.1). LCMS: 383.1 (M+H) + . 1 H NMR: 400MHz DMSO-d6δ (ppm) 7.81 (s, 1H), 5.41-5.30 (m, 1H), 5.25 (t, J = 5.6Hz, 1H), 5.01 (s, 1H),

[0559] 4.65(d,J=7.2Hz,1H),4.25-4.07(m,2H),2.09-2.07(m,1H),1.72-

[0560] 1.64(m,1H),1.55(s,3H),1.37(t,J=7.2Hz,3H),1.24(s,3H),1.00-

[0561] 0.86(m,2H).

[0562]

[0563] Example 17. Preparation of 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine-6-carboxaldehyde (110)

[0564]

[0565] At -78°C and under N2, LDA (2M, 1.01mL, 2.0 equivalent) was added dropwise to a solution of compound 100 (500mg, 1.01mmol, 1.0 equivalent) in THF (8mL), and the mixture was stirred for 30 min. Compound 7 (373.0mg, 5.03mmol, 5.0 equivalent) was added dropwise to THF (1mL), and the mixture was stirred at -78°C for 1 h. TLC (petroleum ether:ethyl acetate = 20:1) showed that the reaction was complete. The reaction mixture was quenched at 20°C by adding saturated NH4Cl solution (20mL), followed by extraction with ethyl acetate (15mL × 3). The combined organic layers were washed with brine (20mL), dried over Na2SO4, filtered, concentrated, and then purified by preparative TLC (petroleum ether:ethyl acetate = 20:1) to give 110 (470mg, 896.00μmol, 88.9% yield) as a white solid. LCMS: 524.1 (M+H) + . 1 H NMR:400MHz CDCl3.δ(ppm)10.62(s,1H),5.66-5.60(m,1H),4.87-4.80(m,2H),2.18-2.17(m,1H),1.71 -1.59(m,3H),1.56-1.54(m,3H),1.53-1.48(m,1H),1.24-1.20(m,21H),0.97-0.84(m,2H).

[0566] Example 18. Preparation of 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine-6-ol (111)

[0567]

[0568] At 25 °C, m-CPBA (77.41 mg, 381.28 μmol, 85% purity, 1.0 equivalent) was added to a solution of compound 110 (200 mg, 381.28 μmol, 1.0 equivalent) in DCM (5 mL), and the reaction mixture was stirred at 40 °C for 20 h. LCMS showed that compound 110 was consumed. The mixture was cooled to room temperature, and NH3-MeOH (2 M, about 2.5 mL, until a clear solution appeared) was added and stirred at the same temperature. LCMS showed the desired MS. The reaction mixture was quenched by adding saturated Na2SO3 (20 mL) and then extracted with DCM (10 mL × 3). The combined organic layers were washed with brine (15 mL), dried over Na₂SO₄, filtered, concentrated, and then purified by preparative TLC (petroleum ether:ethyl acetate = 5:1) to yield compound 111 as a white solid (120 mg, 234.13 μmol, 61.4% yield). LCMS: 512.2 (M+H) + . 1 H NMR: 400MHz CDCl3δ(ppm)5.64-5.58(m,1H),4.85-4.79(m,2H),2.14 -2.11(m,1H),1.67-1.60(m,4H),1.54(s,3H),1.53-1.47(m,1H),1.23-1.17(m,21H),0.94-0.82(m,2H).

[0569] Example 19. Preparation of 5,7-dichloro-6-((3-chlorobenzyl)oxy)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-2-(triisopropylsilyl)-3H-imidazo[4,5-b]pyridine (112).

[0570]

[0571] Compound 10 (75.0 mg, 364.85 μmol, 1.1 equivalent) and Cs₂CO₃ (216.1 mg, 663.4 μmol, 2.0 equivalent) were added to a solution of compound 111 (170 mg, 331.68 μmol, 1.0 equivalent) in DMF (3 mL). The mixture was stirred at 25 °C for 2 h. TLC (petroleum ether:ethyl acetate = 20:1) showed that the reaction was complete. The reaction mixture was partitioned between ethyl acetate (10 mL × 3) and water (20 mL). The combined organic layers were washed with brine (15 mL), dried over sodium sulfate, filtered, concentrated, and then purified by preparative TLC (petroleum ether:ethyl acetate = 20:1) to give compound 112 (170 mg, 266.83 μmol, 80.4% yield) as a white solid. LCMS: 638.1 (M+H) + . 1 H NMR:400MHz CDCl3δ(ppm)7.63(s,1H),7.54-7.48(m,1H),7.40-7.35(m,2H),5.68-5.59(m,1H),5.04(s,2H),4.90-4.79(m,2 H),2.17-2.14(m,1H),1.68-1.61(m,3H),1.55(s,3H),1.53-1.49(m,1H),1.25-1.88(m,21H),0.96-0.82(m,2H).

[0572] Example 20. Preparation of 5-chloro-6-((3-chlorobenzyl)oxy)-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethylhexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine-7-amine (113)

[0573]

[0574] A solution of compound 112 (170 mg, 266.83 μmol, 1.0 equivalent) in NMP (1 mL) was added with NH3·H2O (342.58 μL, 2.67 mmol, 30% purity, 10 equivalents). The mixture was stirred at 140 °C for 50 h. LCMS showed that the reaction was complete. The reaction mixture was partitioned between ethyl acetate (10 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over Na2SO4, filtered, concentrated, and then purified by preparative TLC (petroleum ether:ethyl acetate = 1:1) to give compound 113 (30 mg, 65.03 μmol, 24.3% yield) as a yellow oil. LCMS: 461.1 (M+H)+ .

[0575] Example 21. Preparation of (1R,2R,3S,4R,5S)-4-(7-amino-5-chloro-6-((3-chlorobenzyl)oxy)-3H-imidazo[4,5-b]pyridin-3-yl)bicyclo[3.1.0]hexane-2,3-diol (92)

[0576]

[0577] Compound 113 (30 mg, 65.03 μmol, 1.0 equivalent) was dissolved in TFA (1 mL, 30% purity) and stirred at 25 °C for 3 h. The mixture was concentrated under reduced pressure and purified by preparative HPLC (column: Phenomenex C18 75 × 30 mm × 3 μm; mobile phase: [water (NH3H2O ​​+ NH4HCO3)-ACN]; B%: 10%-65%, 8 min) to yield 92 (4 mg, 9.49 μmol, 14.6% yield) as a white solid. LCMS: 421.1 (M+H)+. 1 H NMR:400MHz DMSO-d6δ(ppm)8.17(s,1H),7.72(s,1H),7.58-7.51(m,1H),7.49-7.38(m,2H),6.68(s,2H),5.09(d,J=4.4Hz,1H),4.88(s,2H),4.68(s,1H) ,4.66-4.57(m,1H),4.51(d,J=7.2Hz,1H),3.76(t,J=5.2Hz,1H),1.84 -1.81(m,1H),1.53-1.45(m,1H),1.13-1.10(m,1H),0.63-0.60(m,1H).

[0578] Example 22. Preparation of 5,7-dichloro-3-((3aR,3bR,4aS,5R,5aS)-2,2-dimethyl-3b-((triphenylmethyloxy)methyl)hexahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-5-yl)-3H-imidazo[4,5-b]pyridine (3)

[0579]

[0580] Under N2, PPh3 (34.6 g, 131.91 mmol, 2.0 equivalent) and DIAD (26.67 g, 131.91 mmol, 25.65 mL, 2.0 equivalent) were added to a solution of compound 1 (35.0 g, 79.09 mmol, 1.2 equivalent) in THF (440 mL). The mixture was stirred at 20 °C for 15 min. Then, compound 2 (12.4 g, 65.96 mmol, 1.0 equivalent) was added to the mixture, and the mixture was stirred at 20 °C for 16 h. TLC (petroleum ether / ethyl acetate = 3 / 1) showed that the reaction was complete, and LCMS showed the same result. The reaction mixture was concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 0 / 1) to obtain compound 3 (61.0 g, crude) as a yellow solid. LCMS: 612.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.68(s,1H),7.62(s,1H),7.38-7.24(m,15H),5.24(d,J=7.2Hz,1H),5.05(s,1H),4.79(d,J=7.2H z,1H),3.38-3.28(m,2H),1.75-1.72(m,1H),1.46(s,3H),1.19(s,3H),1.03(t,J=4.8Hz,1H),0.95-0.93(m,1H).

[0581] Example 23. Preparation of ((3aR,3bR,4aS,5R,5aS)-5-(5,7-dichloro-3H-imidazo[4,5-b]pyridin-3-yl)-2,2-dimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-yl)methanol (4)

[0582]

[0583] At 30 °C, AcOH (8.53 mol, 610 mL, 80% purity, 85.7 equivalents) was added to a solution of compound 3 (61.0 g, crude) in ACN (160 mL). The mixture was stirred at 30 °C for 16 h. TLC (petroleum ether:ethyl acetate = 1:1) showed that the reaction was complete. The reaction mixture was alkalized with NH3·H2O to adjust the pH to 8, followed by extraction with ethyl acetate (400 mL × 3). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by silica gel chromatography (SiO2, petroleum ether:ethyl acetate = 5:1 to 1:3) to obtain compound 4 (13.3 g, 35.9 mmol, 36.0% yield) as a yellow solid. LCMS: 370.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.75(s,1H),7.67(s,1H),5.20(d,J=7.2Hz,1H),5.01(s,1H),4.95(t,J=5.2Hz,1H),4 .66-4.65(m,2H),3.47-3.43(m,1H),1.72-1.69(m,1H),1.45(s,3H),1.16(s,3H),1.00-0.92(m,2H).

[0584] Example 24. Preparation of (3aR,3bS,4aS,5R,5aS)-5-(5,7-dichloro-3H-imidazo[4,5-b]pyridin-3-yl)-2,2-dimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopentene-3b(3aH)-formaldehyde (5)

[0585]

[0586] At 20 °C, IBX (11.1 g, 39.68 mmol, 1.3 equivalents) was added to a solution of compound 4 (11.3 g, 30.52 mmol, 1.0 equivalent) in 115 mL of ACN. The mixture was stirred at 80 °C for 1 h. TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction was complete. The solid was removed by filtration, and the filtrate was used directly for the next step.

[0587] Example 25. Preparation of (3aR,3bS,4aS,5R,5aS)-5-(5,7-dichloro-3H-imidazo[4,5-b]pyridin-3-yl)-2,2-dimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-carboxylic acid (6).

[0588]

[0589] At 20°C, a solution of compound 5 (11.24 mg, 20.53 mmol, 1.0 equivalent) in MeCN (as described above) was added to a mixture of H₂O (23 mL) and H₂O₂ (3.46 g, 30.53 mmol, 30% purity, 1.0 equivalent) in NaH₂PO₄ (7.32 g, 61.05 mmol, 2.0 equivalent). Then, a solution of NaClO₂ (3.31 g, 36.63 mmol, 80% purity, 1.2 equivalent) in H₂O (30 mL) was added at 0°C. The mixture was stirred at 20°C for 1 h. TLC (petroleum ether:ethyl acetate = 1:1) showed the reaction was complete. The reaction mixture was quenched at 0°C by adding saturated aqueous Na₂S₂O₃ solution (500 mL) and extracted with ethyl acetate (150 mL × 3). The combined organic layers were washed with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain compound 6 (11.3 g, crude product) as a yellow solid. LCMS: 384.0 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.58(s,1H),7.63(s,1H),5.68(d,J=7.2Hz,1H),5.09(s,1H),4.83(d,J=6.8 Hz,1H),2.24-2.22(m,1H),1.56-1.54(m,1H),1.46(s,3H),1.35-1.32(m,1H),1.19(s,3H).

[0590] Example 26. Preparation of (3aR,3bS,4aS,5R,5aS)-5-(5,7-dichloro-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopentene-3b(3aH)-formamide (7).

[0591]

[0592] At 0 °C, DIEA (20.13 g, 155.77 mmol, 4.5 equivalents) and T3P (33.04 g, 51.92 mmol, 50% purity, 1.5 equivalents) were added to a solution of compound 6 (13.3 g, 34.62 mmol, 1.0 equivalents) in THF (240 mL). The mixture was stirred at 25 °C for 1 h. TLC (dichloromethane:methanol = 10:1) showed that the reaction was complete and the desired substance was detected. The reaction mixture was partitioned between ethyl acetate (80 mL × 3) and water (300 mL). The organic phase was separated, washed with saturated NaCl (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to produce a residue, which was purified by silica gel chromatography (SiO₂, petroleum ether:ethyl acetate = 50:1 to 0:2) to obtain compound 7 as a yellow solid (11.7 g, 29.45 mmol, 85.08% yield). LCMS: 397.0 (M+H) + . 1 H NMR: 400MHzDMSO-d6δ(ppm)8.57(s,1H),7.78(d,J=4.4Hz,1H),7.64(s,1H),6.64(s,1H),5.64(d,J=6.8Hz,1H), 5.08(s,1H),4.83(d,J=6.0Hz,1H),3.16(d,J=5.2Hz,1H),2.64(d,J=4.4Hz,3H),1.47(s,3H),1.25-1.20(m,4H).

[0593] Example 27. Preparation of (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopentene-3b(3aH)-formamide (209.1)

[0594]

[0595] At 20 °C, DIEA (21.47 g, 166.14 mmol, 6.0 equivalent) and compound 8 (11.29 g, 138.45 mmol, 5.0 equivalent) were added to a solution of compound 7 (11 g, 27.69 mmol, 1.0 equivalent) in NMP (110 mL). The mixture was stirred at 120 °C for 16 h. The reaction was shown to be almost complete. The mixture was diluted with H2O (400 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (150 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 50:1 to 0:1) to obtain compound 209.1 (4.5 g, 11.09 mmol, 40.04% yield) as a yellow solid. LCMS: 406.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.06(s,1H),7.73-7.70(m,1H),7.21(t,J=6.0Hz,1H),6.35(s,1H),5.62(d,J=7.2Hz,1H),4.94(s,1H ), 4.71 (d, J = 6.4Hz, 1H), 3.40 (s, 2H), 2.64 (d, J = 4.4Hz, 3H), 2.07-2.03 (m, 1H), 1.47-1.44 (m, 4H), 1.23-1.15 (m, 7H).

[0596] By the method of Example 27, (5-chloro-2-((3-methylisoxazol-5-yl)methoxy)phenyl)methylamine was used to replace 8 to similarly prepare (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-((5-chloro-2-((3-methylisoxazol-5-yl)methoxy)benzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (201.1). LCMS: 613.2 (M+H) + .

[0597] By means of Example 27, 8 was prepared similarly by replacing 8 with diethylamino-HCl to form (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-ethyl-2,2-dimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (202.1). LCMS: 420.2 (M+H)+.

[0598] By analogy with the method of Example 27, using diethylamino-HCl instead of 8, (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-((2-hydroxyethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (203.1). LCMS: 422.1 (M+H) + .

[0599] The method of Example 27 was used to similarly prepare (3aR,3bS,4aS,5R,5aS)-5-(7-amino-5-chloro-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (204.1), using NH3-H2O instead of 8. LCMS: 378.1 (M+H) + .

[0600] The method of Example 27 was similarly used, replacing 8 with cyclobutylmethylamine, to form (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-((cyclobutylmethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (205.1). LCMS: 446.1 (M+H) + .

[0601] By means of Example 27, 2-methylpropyl-1-amine was substituted for 8 to similarly prepare (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-(isobutylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (206.1). LCMS 434.2 (M+H) + .

[0602] By means of Example 27, propan-1-amine was substituted for 8 to similarly prepare (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-(propylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (207.1). LCMS: 420.2 (M+H) +

[0603] The method of Example 27 was similarly used, replacing 8 with cyclopropylmethylamine, to form (3aR,3bS,4aS,5R,5aS)-5-(5-chloro-7-((cyclopropylmethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopenten-3b(3aH)-formamide (208.1). LCMS: 432.2 (M+H) + .

[0604]

[0605] Example 28. Preparation of (3aR,3bS,4aS,5R,5aS)-5-(5-cyano-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-N,2,2-trimethyltetrahydrocyclopropenzo[3,4]cyclopentadienozo[1,2-d][1,3]dioxacyclopentene-3b(3aH)-formamide (11).

[0606]

[0607] A mixture of compound 209.1 (0.1 g, 246.38 μmol, 1 equivalent), Zn(CN)2 (20.3 mg, 172.47 μmol, 0.7 equivalent), and PdCl2(dppf) (18.0 mg, 24.64 μmol, 0.1 equivalent) in DMF (2 mL) was degassed and purged three times with N2, followed by stirring at 130 °C under an N2 atmosphere for 16 h. Several new peaks were observed on LC-MS, and approximately 19% of the desired compound was detected. The reaction mixture was quenched by adding water (10 mL) and extracted with EtOAc (5 mL × 3). The combined organic layers were washed with brine (5 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to produce the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1, 1 / 1 to 0 / 1) to obtain compound 11 (0.162 g) as a yellow solid. LCMS: 397.2 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.27(s,1H),7.77-7.76(m,1H),7.42(t,J=6.0Hz,1H),6.91(s,1H),5.62(d,J=6.8Hz,1H),5.00(s,1H) ,4.72(d,J=6.4Hz,1H),3.45-3.44(m,2H),2.65-2.64(m,3H),2.08-2.06(m,1H),1.48-1.46(m,4H),1.24-1.18(m,7H).

[0608] Example 29. Preparation of (1S,2R,3S,4R,5S)-4-(5-cyano-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (11)

[0609]

[0610] Compound 11 (0.162 g, 408.63 μmol, 1 equivalent) was stirred in TFA (81.04 mmol, 20 mL, 30% purity, 198.3 equivalent) for 16 h at 25 °C. LC-MS showed approximately 0% of compound 11 remaining. Several new peaks were observed on LC-MS, and approximately 58% of the desired compound was detected. The mixture was concentrated under reduced pressure to produce the residue. The residue was purified by preparative HPLC (column: Phenomenex C18 75*30 mm*3 μm; mobile phase: [water (NH3H2O+NH4HCO3)-CAN]; B%: 5%-35%, 8 min) to obtain 70 (63.77 mg, 178.94 μmol, 43.7% yield, 100% purity) as a white solid. LCMS: 357.0 (M+H) + . 1 H NMR:400MHz CDCl3δ(ppm)8.03(s,1H),6.75-6.74(m,1H),6.71(s,1H),5.63-5.61(m,1H),4.95-4.91(m,2H),4.58-4.56(m,1H),4.14-4.12 (m,1H),3.44-3.34(m,2H),3.12-3.10(m,1H),2.95(d,J=4.8Hz,3H),2.33-2.29(m,1H),1.82-1.79(m,1H),1.40-1.36(m,4H).

[0611] By means of Example 29, 11 was prepared similarly using 201.1 instead of 11 to form (1S,2R,3S,4R,5S)-4-(5-chloro-7-((5-chloro-2-((3-methylisoxazol-5-yl)methoxy)benzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (62). LCMS: 573.1 (M+H) + . 1HNMR:400MHz DMSO-d6δ(ppm)8.05(s,1H),7.69-7.72(m,1H),7.60-7.57(m,1H),7.32-7.29(m,1H),7 .22-7.18(m,2H),6.51(s,1H),6.26(s,1H),5.40(d,J=4.4Hz,1H),5.35(s,2H),4.92(t, J=6.4Hz,1H),4.80(d,J=8.0Hz,1H),4.71(s,1H),4.61(s,2H),3.87-3.85(m,1H),2.67 (d,J=4.4Hz,3H),2.34(s,3H),1.81-1.79(m,1H),1.63-1.60(m,1H),1.31-1.29(m,1H).

[0612] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(5-chloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-ethyl-2,3-dihydroxybicyclo[3.1.0]hexane-1-carboxamide (69), replacing 11 with 202.1. LCMS: 380.2 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)7.98(s,1H),7.57(t,J=5.6Hz,1H),7.14(t,J=6.0Hz,1H), 6.34(s,1H),4.93(d,J=6.0Hz,1H),4.69(s,1H),3.84(d,J=6.0Hz,1H),3.5 0-3.48(m,2H),3.38(s,2H),3.18-3.16(m,2H),1.82-1.79(m,1H),1.62-1. 61(m,1H),1.31-1.29(m,1H),1.16(t,J=7.2Hz,3H),1.05(t,J=7.2Hz,3H).

[0613] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(5-chloro-7-((2-hydroxyethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (91), replacing 11 with 203.1. LCMS: 382.1 (M+H) + . 1H NMR:400MHz DMSO-d6δ(ppm)8.02(s,1H),7.60-7.59(m,1H),7.07(s,1H),6.42(s,1H),5.43(t,J=4.8Hz,1H),4.93(d,J=6.8Hz,1H),4.84-4.80(m,2H),4.72 (s,1H),3.85(d,J=4.8Hz,1H),3.60-3.56(m,2H),3.42(s,2H),2.67(d, J=4.4Hz,3H),1.80-1.78(m,1H),1.64-1.61(m,1H),1.32-1.30(m,1H).

[0614] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(7-amino-5-chloro-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (90), replacing 11 with 204.1. LCMS: 338.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.00(s,1H),7.58-7.57(m,1H),6.78(s,2H),6.37(s,1H),5.41(d,J=4.8Hz,1H),4.94(t,J=7.2Hz,1H),4.80(d,J =8.0Hz,1H),4.70(s,1H),3.86-3.83(m,1H),2.66(d,J=4.8Hz,3H),1.79-1.77(m,1H),1.61(t,J=4.4Hz,1H),1.30-1.29(m,1H).

[0615] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(5-chloro-7-((cyclobutylmethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (65), replacing 11 with 205.1. LCMS: 406.1 (M+H) + . 1H NMR: 400MHz DMSO-d6δ(ppm)7.99(s,1H),7.58-7.57(m,1H),7.21(t,J=6.0Hz,1H),6.36(s, 1H),5.41(d,J=4.8Hz,1H),4.92(t,J=7.2Hz,1H),4.80(d,J=8.4Hz,1H),4.70(s ,1H),3.86-3.83(m,1H),3.45(s,2H),2.66(d,J=4.4Hz,3H),2.52-2.50(m,1H), 2.05-1.97(m,2H),1.85-1.71(m,5H),1.04(t,J=4.8Hz,1H),1.30-1.29(m,1H).

[0616] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(5-chloro-7-(isobutylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (83), replacing 11 with 206.1. LCMS: 394.1 (M+H) + . 1 H NMR: 400MHz DMSO-d6δ(ppm)8.01(s,1H),7.59-7.57(m,1H),7.28(t,J=6.4Hz,1H),6.37(s,1 H),5.42(t,J=4.8Hz,1H),4.95(t,J=6.8Hz,1H),4.82(d,J=8.4Hz,1H),4.71(s,1 H),3.86(t,J=4.8Hz,1H),3.21(s,2H),2.68(d,J=4.8Hz,3H),1.91-1.93(m,1H) ,1.80-1.78(m,1H),1.64-1.61(m,1H),1.30-1.29(m,1H),0.91(d,J=6.4Hz,6H).

[0617] By analogy with the method of Example 29, replacing 11 with 208.1, (1S,2R,3S,4R,5S)-4-(5-chloro-7-((cyclopropylmethyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (63) was prepared. LCMS: 392.1 (M+H)+. 1H NMR: 400MHz CDCl3δ(ppm)7.85(s,1H),6.98-6.96(m,1H),6.31(s,1H),5.71(t,J=6.0Hz,1H),5 .17(s,1H),4.94(d,J=6.8Hz,1H),4.89(s,1H),4.13(d,J=6.4Hz,1H),3.59(s,1H) ,3.15(t,J=5.2Hz,2H),2.92(d,J=4.8Hz,3H),2.28-2.24(m,1H),1.77(t,J=4.8Hz ,1H),1.38-1.36(m,1H),1.18-1.15(m,1H),0.64-0.62(m,2H),0.33-0.31(m,2H).

[0618] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(5-chloro-7-(propylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (64), replacing 11 with 207.1. LCMS: 380.1 (M+H) + . 1 H NMR:400MHz CDC13δ(ppm)7.83(s,1H),6.97-6.94(m,1H),6.32(s,1H),5.60(t,J=5.2Hz,1H),5.17(s,1H),4.93(d,J=6.8Hz,1H),4.88(s,1H),4.13(d,J=6 .4Hz,1H),3.58(s,1H),3.30-3.25(m,2H),2.92(d,J=4.8Hz,3H),2.27 -2.25(m,1H),1.78-1.71(m,3H),1.36-1.35(m,1H),1.06-1.02(m,3H).

[0619] The method of Example 29 was used to similarly prepare (1S,2R,3S,4R,5S)-4-(5-chloro-7-(ethylamino)-3H-imidazo[4,5-b]pyridin-3-yl)-2,3-dihydroxy-N-methylbicyclo[3.1.0]hexane-1-carboxamide (59) by replacing 11 with 209.1. (39.6% yield), as a white solid. LCMS: 366.1 (M+H) + . 1 H NMR:400MHz DMSO-d6δ(ppm)8.00(s,1H),7.55(d,J=4.8Hz,1H),7.17(t,J=5.6Hz,1H),6. 34(s,1H),5.39(d,J=4.4Hz,1H),4.94-4.92(m,1H),4.79(d,J=8.4Hz,1H),4 .71(s,1H),3.85(t,J=5.2Hz,1H),3.40(s,2H),2.67(d,J=4.4Hz,3H),1.80- 1.77(m,1H),1.61(t,J=4.8Hz,1H),1.31-1.29(m,1H),1.17(t,J=6.8Hz,3H).

[0620] Table 10. Compounds prepared by the above examples and measurement data

[0621]

[0622]

[0623]

[0624]

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633] Example 30: Adenosine Receptor Binding Assay

[0634] Materials and reference compounds: [ 125 I]-AB-MECA and [ 3 H]CCPA was obtained from Amersham Biosciences (Little Chalfont, UK). IB-MECA was obtained from Tocris (Bristol, UK), and N6-cyclopentyladenosine (CPA) was obtained from Sigma-Alrich (Poole, UK). Cell culture medium, G418, and hygromycin were obtained from Invitrogen (Paisley, UK). Fetal bovine serum was obtained from Perbio Science (Cheshire, UK). All other reagents were obtained from Fisher Chemicals (Loughborough, UK).

[0635] Cell Culture and Membrane Preparation: This example demonstrates the binding affinity of the compound according to one embodiment of the invention for human A3 or A1 receptors stably expressed in Chinese hamster ovary (CHO-K1) cells (for A1 receptors) or HEK-293 cells (for A3 receptors). The method followed was that of Yates L, Clark JH, Martin TJ, James S, Broadley KJ, Kidd EJ. Radioligand binding and functional responses of ligands for human recombinant adenosine A3 receptors. Auton Autacoid Pharmacol. 2006; 26(2):191-200. Cells stably transfected with human adenosine A3 or A1 receptors were grown in Dulbecco's modified Eagles medium containing a nutrient mixture F12 of NaHCO3 and Glutamax. The culture medium was supplemented with 10% fetal bovine serum, genimycin (G418; 0.67 mg / mL), hygromycin (500 μg / mL), and adenosine deaminase (1 U / mL). Cells were maintained at 37°C in air containing 5% CO2. Cells were collected and homogenized using a Polytron homogenizer in ice-cold Tris buffer [50 mM Tris, 150 mM NaCl, 1 mM EDTA, pH 7.4] supplemented with the peptidase inhibitor 4-(2-aminoethyl)benzenesulfonyl fluoride (1 mM) and bacitracin (0.1 mg / mL). The homogenate was rotated at 500 g for 10 min (4°C). The supernatant was retained, and the precipitate was resuspended in buffer and homogenized and rotated as described above. The combined supernatants were centrifuged at 48,000 g for 15 min (4°C). The resulting precipitate was resuspended in buffer and centrifuged again at 48,000 g. The membrane precipitate was then resuspended in Tris buffer without peptidase inhibitors or adenosine deaminase and stored at -80°C at a protein concentration of 1 mg / mL.

[0636] Binding assay: using adenosine A3 receptor agonists [ 125 I]-AB-MECA and A1 receptor agonists [ 3 Radioligand binding assays were performed using H]CCPA. The assay was performed using a buffer solution containing 50 mM Tris pH 7.4 (with or without 10 mM MgCl2), 20 μg of membrane, and 0.15 nM [ 125I]-AB-MECA (for A3 receptors) or 1 nM [ 3 Binding assays were performed using a total volume of 100 μL of H]CCPA (for the A1 receptor). Assays were conducted at room temperature for 60 min (A1 receptor) or 120 min (A3 receptor), and terminated by adding 2 ml of ice-cold wash buffer (50 mM Tris, pH 7.4, with or without 10 mM MgCl2) and rapidly filtering through a Whatman GF / C filter treated with 0.03% polyethyleneimine using a Brandel cell collector (Semat International Ltd., St Albans, UK). The filter was then washed three times with 2 ml of ice-cold buffer. The radioactivity bound to the filter was counted using a Compugamma counter (LKB Wallac, Turku, Finland). Competitive experiments were performed to investigate the inhibition of […] by the reference compound and embodiments of the present invention. 125 The ability of I]-AB-MECA to bind. Non-specific binding was determined using the corresponding compound at its highest concentration.

[0637] Data Analysis: Based on Binding and Free [ 125 I]-AB-MECA molar quantity calculation combined with data K d and B 最大 The values ​​are then used to fit unit points and curves to these data using Prism (GraphPad), and this is used to derive K. d and B 最大 Value. The IC50 value for the inhibition of radioligand binding by the compounds of this invention was calculated from the ligand competition curve using an Excel spreadsheet. 50 and K i The value and Hill slope, and calculate [ 125 I]-AB-MECA or [ 3 H]CCPA's K d The results are expressed as the percentage of control-specific binding and the percentage of inhibition of control-specific binding obtained in the presence of the compounds of the present invention. IC was determined by nonlinear regression analysis of the competition curves generated using the average repeatability values ​​fitted with the following Hill equation. 50 Value (concentration that results in half-maximal inhibition of specific binding to the control) and Hill coefficient (nH):

[0638]

[0639] Where Y = specific binding, A = left asymptote of the curve, D = right asymptote of the curve, C = compound concentration, and C50 = IC50. 50And nH = slope factor. This analysis was performed using software developed by Cerep (Hillsoftware), and compared with commercial software. 4.0 (for) The data generated was compared for verification. The suppression constant (K) was calculated using the following Cheng-Prusoff equation. i value):

[0640]

[0641] Where L = the concentration of the radioactive ligand in the determination, and K D = Affinity of the radioligand to the receptor. K is determined using a Scatchard plot. D .for[ 125 I]-AB-MECA, L=0.15nM, and K D =0.22nM. Table 10 includes the K values ​​of the compounds described herein. i value.

[0642] Example 31: Intracellular cAMP Levels

[0643] This embodiment demonstrates the effect of a compound according to an embodiment of the invention on intracellular cAMP levels in Chinese hamster ovarian (CHO-K1) cells. The method followed is disclosed in Yates L, Clark JH, Martin TJ, James S, Broadley KJ, Kidd EJ. Radioligand binding and functional responses of ligands for human recombinant adenosine A3 receptors. Auton Autacoid Pharmacol. 2006 Apr; 26(2):191-200.

[0644] Materials and Cell Culture: Adenosine deaminase was obtained from Roche (Lewes, UK). Forskolin was obtained from Sigma-Aldrich (Poole, UK). The sources of other materials are provided above in Example 1. CHO-K1 cells stably transfected with human adenosine A3 receptor were grown in Dubecalch modified Igner medium F12 containing a nutrient mixture of NaHCO3 and Glutamax. The medium was supplemented with 10% fetal bovine serum, genistein (G418; 0.67 mg / mL), hygromycin (500 μg / mL), and adenosine deaminase (1 U / mL). Cells were maintained at 37°C in air containing 5% CO2.

[0645] cAMP measurement: Intracellular cAMP production was directly measured using a non-acetylated cAMP enzyme immunoassay (Amersham Biosciences, LittleChalfont, UK). Cultured CHO cells expressing human adenosine A3 receptor were seeded overnight in 96-well plates containing adenosine deaminase. After 24 hours, they were incubated at 37°C for 5 min with an adenosine analog or the compound of this invention (0.1 nM–1 μM), followed by incubation with trichodin (10 μM) for 10 min. Cells were then lysed for 10 min with 200 μL of the lysing reagent 1 provided with the kit to release intracellular cAMP. Next, aliquots of the supernatant were transferred to 96-well microtiter plates coated with donkey anti-rabbit IgG antibody; 100 μL of cAMP standard (12.5–3200 fmol / well) was also added to the microtiter plates; 100 μL of rabbit anti-cAMP antibody was added to all wells, and the plates were incubated at 2–5 °C for 2 h. Antiserum was prepared in dissolving reagent 2, which chelates the key components in dissolving reagent 1 and ensures that cAMP is free for subsequent analysis. Then, 50 μL of cAMP peroxidase conjugate was added to all wells and incubated at 2–5 °C for 60 min to allow competition for the antibody between unlabeled cAMP and horseradish peroxidase (HRP)-labeled cAMP. The wells were then washed with buffer, and 150 μL of enzyme substrate containing 3,3',5,5'-tetramethylbenzidine / hydrogen peroxide in 20% (v / v) dimethylformamide was added to each well. The wells were then incubated at room temperature for 60 min on a Titramax 100 microtiter plate shaker (Heidolph Instruments, Schwabach, Germany) for color development. The reaction was terminated by adding 100 μL of 1M sulfuric acid, and the optical density (OD) was read at 450 nm using a spectrophotometer MRX microplate reader (Dynex Technologies, Chantilly, VA, USA).

[0646] Analysis: Analyze each sample / standard in duplicate and calculate the average of the resulting OD values. Calculate the binding percentage (B / B0) between each standard and sample using the following relationship:

[0647]

[0648] Where B is the specific OD of the standard / sample, and B0 is the specific OD of the zero standard. A standard curve was generated by plotting B / B0% as a function of log fmol cAMP standard per well. The fmol cAMP value per well for each sample was then read from the graph. The result is expressed as the percentage of the control agonist response in the presence of the compounds of the present invention: (measured response / control response) * 100. Concentration-response curves for inhibition of cAMP production were constructed for all compounds of the present invention and reference compounds. EC was determined by nonlinear regression analysis of the concentration-response curves generated by curve fitting with the average repeatability using the Hill equation according to the following equation. 50 Value (concentration that produces the half-maximal response):

[0649]

[0650] Where Y = response, A = left asymptote of the curve, D = right asymptote of the curve, C = compound concentration, and C 50 =EC 50 And nH = slope factor. This analysis was performed using software developed by Cerep (Hill Software), and compared with commercial software. 4.0 (for) The data generated were compared to verify this. Table 10 includes the EC50 of the compounds described herein. 50 Value. The maximum response reported is the percentage of responses generated by the 100nM IB-MECA.

[0651] Example 32: Intracellular calcium release assay

[0652] This embodiment demonstrates the effect on intracellular calcium levels in CHO-K1 or HEK293 cells that stably express four subtypes of adenosine receptors induced by a test compound according to an embodiment of the invention.

[0653] Cell Culture: The table below lists the four cell lines that stably express the adenosine receptor and the growth medium used for each cell line:

[0654]

[0655] For experiments using ADORA1- and ADORA3-expressing cells, the culture medium was aspirated, and the cells were washed twice with 10 mL of DPBS. 2 mL of trypsin was added, and the cells were incubated at 37°C for 1 minute. Then, 10 mL of plating medium was added to terminate the digestion, and 1 mL of the solution was collected for cell counting. ADORA2A- and ADORA2B-expressing cells were rapidly thawed in a 37°C water bath. The cell suspension was transferred to 50 mL conical tubes. Plating medium was added to a 10 mL mark, and 1 mL was collected for cell counting. The 50 mL tubes containing each cell type were centrifuged at 1000 g. The supernatant was then aspirated, taking care not to aspirate the cells. The cell pellet was resuspended in 3–5 mL of plating medium, and 0.5 mL was collected for cell counting. The cell suspension was then diluted to 1 × 10⁻⁶ in plating medium. 6 20,000 cells per 20 μL per well, and seeded into 384-well cell culture plates (Greiner-781946). The 384-well plates were then incubated at 37°C / 5% CO2 for 16–20 hours in a Thermo-371, Thermo Scientific.

[0656] Intracellular calcium imaging agent (Fluo-4 Direct) TM Preparation of the calcium assay kit (Invitrogen-F10471, Thermo Scientific): Add 1 mL of FLIPR assay buffer to 77 mg of probenecid to prepare a 250 mM solution, which should be used fresh daily. Add one vial of Fluo-4 Direct... TM Thaw the crystals and add 10 mL of FLIPR assay buffer to the vial. Add 0.2 mL of probenecid to Fluo-Direct. TM Divide the solution into 10 mL vials to achieve a final determination concentration of 2.5 mM. Vortex the vial, allow it to stand for >5 min, and protect it from light. Prepare the dye fresh daily.

[0657] Compound preparation: The compound plates were prepared as follows. First, the test compound was diluted to prepare a 2 mM solution. Then, 10 serial dilutions (3-fold) were prepared using ECHO (ECHO 555, LabCyte), and 900 nmL was transferred to the compound plate. For all targets, the final maximum concentration of the test compound was 10 μM. For the agonist reference compound, 10 serial dilutions (3-fold) were prepared, and 900 nmL was transferred to the assay plate. Next, 30 μL of assay buffer was added to the compound plate, and the plate was centrifuged at 1000 rpm for 1 min.

[0658] FLIPR assay: Remove the cell plate from the incubator and add 20 μL of 2×Fluo-4Direct to it. TM The solution was then incubated in a 5% CO2, 37°C incubator for 50 min, followed by incubation at room temperature for 10 min. To run the protocol on a Fluorescence Imaging Plate Reader (FLIPR) TETRA (MD-FT0249, Molecular Devices, Inc.), 10 μL of assay buffer was transferred from a 384-well plate (Greiner-781280) to a cell plate, and the fluorescence signal was read. Then, 10 μL of the compound solution was transferred from the compound plate to the cell plate, and the fluorescence signal was read. The "Max-Min" value was calculated, starting from a reading of 91 to the maximum permissible signal. For agonists, activation % = (RLU - LC) / (HC - LC) * 100, based on the following: RLU = relative light units, 91 to the maximum permissible signal; HC = average signal of high concentrations of agonist; LC = average signal of DMSO wells. Using GraphPad Prism 5, the model "log(agonist) vs. response - variable slope" was fitted to the data to determine EC. 50 Values. The receptor efficacy of the test compound relative to the agonist NECA was calculated as a percentage (%) of response to 1 μM NECA. Table 10 includes the EC50 values ​​of the compounds described herein. 50 value.

[0659] Example 33: Animal Model of Neuropathic Pain

[0660] This embodiment evaluates the performance of compounds embodying the invention in an in vivo model of neuropathic pain. As described in Little JW, Chen Z, Doyle T, Porreca F, Ghaffari M, Bryant L, Neumann WL, Salvemini D. Supraspinal peroxynitrite modulates pain signaling by suppressing the endogenous opioid pathway. J Neurosci. 2012; 32(32):10797-808. Animals and neuropathic injury: Male Sprague Dawley rats (225–270 g) (Harlan Laboratories) were used for all studies. Rats were housed in climate-controlled rooms with a 12-hour light / dark cycle and unlimited food and water. All experiments were conducted in accordance with the guidelines for laboratory animal welfare of the International Association for the Study of Pain (Seattle, MD) and the National Institutes of Health (NIH; Bethesda, MD). All observers were unaware of any treatments they received. Rats were anesthetized and subjected to chronic sciatic nerve compression injury (CCI) achieved using a modification of the commonly used method (Bennett and Xie, 1988).

[0661] Shave the hair on the outer back of the thigh, using The rats were scrubbed, and a 2 cm incision was made in the skin. The left common sciatic nerve was exposed in the mid-thigh by blunt dissection. Approximately 7 mm of the nerve was detached from its adhesions near the three bifurcations of the sciatic nerve, followed by loose tethering of the nerve with three 4-0 silk sutures, resulting in a slight restraint spaced approximately 1 mm apart. The incision was then closed with 5-0 silk sutures. Mechanical anomalous pain was measured using a von Frey filament to determine the mechanical claw withdrawal threshold (PWT) in grams. The rats were acclimatized to a Plexiglas chamber with a wire mesh bottom for 15 min. PWT was assessed three times at each time point, and the mean mechanical PWT (in grams) for both the ipsilateral and contralateral claws was reported. PWT was assessed on day 0 (D0) prior to surgery and subsequently on the required postoperative days. Mechanical anomalous pain was defined as a significant (p<0.05) decrease in mean PWT compared to D0. Seven days (D7) provided the maximum reduction in PWT on the same side and was used as the target time point for compound evaluation.

[0662] Compound administration: The compound was administered orally to rats by dissolving it in distilled water containing 0.5% methylcellulose and 0.1% DMSO. The solution was sonicated until all substances were dissolved. The administered dose volume was 2 mL / kg. Oral dose levels from 0.3 to 30 mg / kg were evaluated in CCI rats. Five rats were administered dose levels from 0.3 to 3 mg / kg, and four rats were administered dose levels of 10 mg / kg.

[0663] Data analysis: The compounds of this invention reduced the mean mechanical abnormal pain in limbs with CCI, as evidenced by a statistically significant increase in claw withdrawal threshold at 1 and 2 hours after oral administration. The compounds of this disclosure do not affect the claw withdrawal threshold in limbs that have not undergone CCI surgery. In this animal model, treatment with the mediator alone had no effect on mechanical abnormal pain.

[0664] Example 34: GTPγ[ 35 Scintillation Proximity Detection (SPA)

[0665] This assay was used to monitor the activation of the A3 adenosine receptor (A3AR) by the test compound using a protocol similar to that disclosed for A1AR (Langemeijer et al., Purinergic Signalling 9:91 (2013)). Membrane homogenates of membrane extracts from CHO-K1 cells expressing recombinant human A3AR were equilibrated in assay buffer (20 mM HEPES pH 7.4; 100–200 mM NaCl, 10 μg / ml saponin, MgCl2, 0–0.1% BSA). The membrane was mixed with GDP. In parallel, GTPg[ 35 [S] was mixed with SPA imaging beads (PerkinElmer, RPNQ001) containing polyvinyltoluene and wheat germ lectin (PVT-WGA), and then diluted in assay buffer at 50 mg / ml (0.5 mg / 10 μl). The following reagents were subsequently added sequentially to the wells of the Optiplate (Perkin Elmer): 50 μl of test or reference ligand, 10 μl of assay buffer, 20 μl of membrane:GDP mixture, and 20 μl of GTPg[ 35 S]: Bead mixture. Cover the plate with a top sealant, mix on an orbital shaker for 2 min, then incubate at room temperature for 1 hour. Next, centrifuge the plate at 2000 rpm for 10 min, incubate at room temperature for 1 hour, and count using a PerkinElmer TopCount reader for 1 min / well.

[0666] Measurements were performed in duplicate, and the test compounds were evaluated at 10 concentrations ranging from 10 mM to 0.3 nM through semi-log dilution. The percentage of activation was fitted relative to a ligand binding reaction model with four free parameters using Prism software (Graphpad).

[0667] Example 35: Inhibition of the release of pro-inflammatory cytokines from human peripheral blood mononuclear cells (PBMCs).

[0668] The anti-inflammatory activity of the test compounds was assessed by quantifying the secretion of TNFα and IL-1β from human PBMCs.

[0669] Cell preparation

[0670] Human PBMCs (StemCell Technologies 70025.1) cryopreserved are typically purchased from a recognized supplier. Thaw the vials in a 37°C water bath until only small ice crystals remain. Gently transfer the cells to 50 ml Falcon tubes and wash the vials with 1 ml of pre-warmed assay medium (DMEM [Sigma 6456] + 10% FCS [Sigma F9665] + 1×Glutamax [Invitrogen 350500380]), then gently mix with the cells. Dilute the cells dropwise in 20 ml of warm medium by gentle mixing, followed by centrifugation (300×g for 10 min). After discarding the supernatant, resuspend the pellet in the remaining volume by gentle tapping. Add 20 ml of fresh medium as described above and gently mix with the pellet. Add DNase (Sigma). Mix the cells (10104159001) to a final concentration of 0.1 mg / ml (from a 1 mg / ml stock prepared and filtered sterilized in PBS), and incubate at room temperature for 10 minutes. Then centrifuge the cells (300 × g for 10 minutes) and discard the supernatant, resuspending the pellet in the remaining liquid by gentle tapping. Next, dilute the cells in 1 ml of culture medium and count them using a Luna II cell counter at a 1:2 dilution in trypan blue. Then dilute the cells to the target density in culture medium; typically seed 40 μl / well, 15,000 or 25,000 cells / well in black clear-bottomed 384-well plates (Greiner 781091). Allow the cells to recover overnight in an incubator (37°C, 5% CO2) before performing experiments.

[0671] Cytokine assay

[0672] All cell treatment agents were typically added in 5 μl of an 11-fold concentrated stock prepared in assay medium and incubated at 37°C in a 5% CO2 incubator. Cells were usually pretreated with the test compound for 1 h in the presence of adenosine deaminase (Sigma A5043-250UN), followed by sensitization with LPS (1 ng / ml Sigma L3129) for an additional 4 h. NLRP3 inflammasomes were then activated by adding 6.7 μM nigericin (Invivogen tiri-nig) and incubating for 3 h. The supernatant was sampled for cytokine quantification, and 4 μl was added to a white low-volume 384-well assay plate (Greiner 784075); for IL-1β, this was after nigericin stimulation, and for TNFα, this was before nigericin stimulation. The supernatant was diluted in culture medium to fall within the detection range of the AlphaLISA (typically 1:50 for IL-1β and 1:10 for TNFα). A mixture of AlphaLISA receptor beads and biotinylated antibody was prepared in AlphaLISA immunoassay buffer, and 4 μl was added to the supernatant and incubated overnight at 4°C. Then, 2 μl of streptavidin donor beads were added and incubated at room temperature with shaking for 30–60 minutes. The plates were then read using Pherastar, and the AlphaLISA signal was interpolated according to a standard curve for each cytokine at known concentrations (prepared individually for each assay plate). The AlphaLISA kit was from Perkin Elmer (IL-1β; AL220C, TNFα; AL208C).

[0673] Data analysis was performed using Prism software (GraphPad). The concentration-response curves were fitted against a four-parameter nonlinear function describing inhibition.

Claims

1. A compound represented by Formula (II’): or a pharmaceutically acceptable salt thereof, wherein: R 4’ selected from the group consisting of: (a) hydrogen, (b) unsubstituted C1-C6 alkyl, (c) C1-C6 alkyl substituted with one or more fluorines, (d) C1-C6 alkyl substituted with one or more hydroxyls, (e) C1-C6 alkyl substituted with an unsubstituted C3-C6 carbocycle, and (f) R 5’ selected from -CN and halogen; R 6’ is -C(O)(NR 50 2), wherein each R 50 is independently selected from hydrogen and C1-C3alkyl; and R 12’ selected from hydrogen, halogen and unsubstituted C 1-6 alkyl.

2. The compound or salt of claim 1, wherein the compound of Formula (II’) is represented by Formula (IIA’):

3. The compound or salt of claim 1 or 2, wherein: R 4’ is selected from CH2CH3, CH2CFH2, CH2CF2H and CH2CF3.

4. The compound or salt of claim 1 or 2, wherein: R 5’ is halogen.

5. The compound or salt of claim 1 or 2, wherein: R 5’ is Cl.

6. The compound or salt of claim 1, wherein: R 6’ is -C(O)(NHMe).

7. The compound or salt of claim 1 or 2, wherein: R 12’ is hydrogen.

8. The compound or salt of claim 1, selected from the following compounds:

9. The compound or salt of claim 1, selected from the following compounds:

10. The compound or salt of claim 1, selected from the following compounds:

11. The compound or salt of claim 1, selected from the following compounds:

12. The compound or salt of claim 1, selected from the following compounds:

13. The compound or salt of claim 1, selected from the following compounds:

14. The compound or salt of claim 1, selected from the following compounds:

15. A pharmaceutical composition comprising a compound or salt of any one of claims 1 to 14 and a pharmaceutically acceptable excipient.

16. Use of a compound or salt of any one of claims 1 to 14 or a pharmaceutical composition of claim 15 in the manufacture of a medicament for treating a disease in a subject in need thereof.

17. The use of claim 16, wherein the medicament is for agonizing A3 adenosine receptors in the subject.

18. Use of a compound or salt of any one of claims 1 to 14 or a pharmaceutical composition of claim 15 in the manufacture of a medicament for treating a condition in a subject in need thereof, wherein: the condition is selected from vascular inflammation, arthritis, allergy, asthma, wound healing, stroke, heart failure, acute spinal cord injury, acute head injury or trauma, seizure, neonatal hypoxia, cerebral palsy, chronic hypoxia due to arteriovenous malformations and occlusive cerebral arterial disease, ischemia and reperfusion injury in skeletal muscle, severe neurological conditions associated with excitotoxicity, Parkinson’s disease, Huntington’s chorea, CNS diseases, heart disease, kidney disease, glaucoma, cancer, neuropathic pain, transient ischemic attack, bone marrow protection, dry eye syndrome, osteoarthritis, rheumatoid arthritis, loss of cutaneous pigmentation, inflammatory bowel disease, pulmonary inflammation, uveitis, and septic shock; or the condition is selected from vascular inflammation, arthritis, allergy, asthma, wound healing, stroke, heart failure, acute spinal cord injury, acute head injury or trauma, seizure, neonatal hypoxia, cerebral palsy, chronic hypoxia due to arteriovenous malformations and occlusive cerebral arterial disease, ischemia and reperfusion injury in skeletal muscle, severe neurological conditions associated with excitotoxicity, Parkinson’s disease, Huntington’s chorea, CNS diseases, heart disease, kidney disease, glaucoma, cancer, neuropathic pain, transient ischemic attack, bone marrow protection, dry eye syndrome, osteoarthritis, rheumatoid arthritis, loss of cutaneous pigmentation, inflammatory bowel disease, pulmonary inflammation, uveitis, and septic shock. The condition is selected from chemotherapy-induced peripheral neuropathy, diabetic peripheral neuropathy, neurodegeneration, drug-induced ototoxicity, spinocerebellar degeneration, symptoms associated with traumatic brain injury, chemotherapy-induced cognitive impairment, pain and discomfort of irritable bowel syndrome, and neuropathic pain.

19. The use of claim 18, wherein: The condition is neuropathic pain.

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