Charged ion channel blocker and method of use

Abstract

The present invention provides a compound of the following formula (I) or a pharmaceutically acceptable salt thereof: The compounds, compositions, methods, and kits of the present invention are useful for the treatment of pain, cough, itching, and neurological inflammation.

Description

Technology Field

[0001] Related applications

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 62 / 816,441 filed March 11, 2019 and U.S. Provisional Application Serial No. 62 / 931,599 filed November 6, 2019. The entire contents of the said applications are incorporated herein by reference. Background Technology

[0003] The present invention features compounds, compositions, and methods for treating neuroinflammatory disease by selectively inhibiting sensory neurons (nociceptors, cough receptors, and itch receptors) and targeting nociceptors with small molecule drugs while minimizing effects on non-nociceptor neurons or other types of cells. According to the method of the present invention, small cationic drug molecules access the intracellular compartments of sensory neurons through entry via large pore receptor / ion channels that are present in pain-sensing, cough-sensing, and itch-sensing neurons but are present to lesser extent or not at all in other types of neurons or other types of tissues.

[0004] Local anesthetics, such as lidocaine and articaine, act by inhibiting voltage-dependent sodium channels in neurons. By blocking sodium channels, these anesthetics block the excitability of all neurons, not just pain-sensing neurons (nociceptors). Therefore, while the goal of local or regional anesthesia is to prevent pain by blocking nociceptor signaling, the administration of local anesthetics also causes unwanted or harmful effects, such as generalized numbness due to low threshold pressure and blockade of contact receptors, paralysis due to motor deficits and / or motor axon blockade, and other complications due to autonomic fiber blockade. Local anesthetics are relatively hydrophobic molecules that diffuse through the cell membrane to access blockade sites on sodium channels. Charged derivatives of these compounds, which are not membrane-permeable, do not affect neuronal sodium channels when applied to the outer surface of the nerve membrane; however, they can block sodium channels if they are somehow introduced into the cell, for example, by diffusion from a micropipette used for whole-cell electrophysiological recording from isolated neurons. Pain-sensing, cough-sensing, and itch-sensing neurons differ from other types of neurons (in most cases) in that they express TRPV1 receptors / channels that are activated by painful heat or capsaicin, the spicy component of chili peppers. Other types of channels selectively expressed in various types of pain-sensing, cough-sensing, and itch-sensing (itch receptor) neurons include, but are not limited to, TRPV2-4, TRPA1, TRPM8, ASIC, and P2X(2 / 3) channels. It is well established that some cationic small molecules, such as QX-314, can enter cells by passing through activated large porous channels such as TRPV1.

[0005] Neuropathic, inflammatory, and nociceptive pain differ in their etiology, pathophysiology, diagnosis, and treatment. Nociceptive pain occurs in response to the activation of nociceptors, a specific subset of high-threshold peripheral sensory neurons, by intense or noxious stimuli. It is generally acute, self-limiting, and serves a protective biological function by acting as a warning against potential or ongoing tissue damage. It is typically well-localized. Examples of nociceptive pain include, but are not limited to, traumatic or surgical pain, analgesia, sprains, fractures, burns, bumps, contusions, injections, dental procedures, skin biopsies, and closures.

[0006] Inflammatory pain is pain associated with damage to joints, muscles, and tendons, such as pain occurring in the presence of tissue damage or inflammation, including post-operative pain (i.e., acute pre- and post-operative pain resulting from inflammation caused by tissue trauma (e.g., surgical incision, resection, burn) or direct nerve damage (e.g., nerve transection, stretching, or compression)), post-traumatic pain, arthritis pain (rheumatic; or osteoarthritis (i.e., joint pain and stiffness due to the progressive deterioration of articular cartilage; risk factors including aging, injury, and obesity; generally affected joints are the hands, wrists, neck, knees, hips, and spine)), pain, and axial low back pain (i.e., a pervasive and painful condition affecting the lower part of the back; common causes include muscle strain, vertebral fractures, bulging or ruptured discs, and arthritis), and severe nociceptive pain can transition into inflammatory pain if there is associated tissue damage.

[0007] Neuropathic pain is a common type of chronic, non-malignant pain resulting from damage or dysfunction of the peripheral or central nervous system that does not provide a protective biological function. It is estimated that more than 1.6 million people in the U.S. population are affected. Neuropathic pain has many different etiologies and can occur, for example, due to trauma, surgery, herniated discs, spinal cord injury, diabetes, herpes zoster infection, HIV / AIDS, terminal cancer, amputation (including mastectomy), carpal tunnel syndrome, chronic alcohol use, radiation exposure, and as an unintended side effect of neurotoxic agents such as certain anti-HIV and chemotherapy drugs. Peripheral neuropathy is caused by peripheral nerve damage from injury, trauma, or prolonged pressure, or by inflammation and pain in the affected part of the body that causes numbness.

[0008] Neuropathic pain is often naturally described as "burning," "electric," "tingling," or "stinging." It is often characterized by chronic dynamic allodynia (defined as pain caused by moving stimuli that do not typically trigger a painful response, such as light contact) and hyperalgesia (generally defined as increased sensitivity to painful stimuli), and can persist for months or years even after the apparent healing of any damaged tissue.

[0009] Pain can occur in patients with cancer and can be caused by various factors such as inflammation, compression, invasion, or the spread of metastasis to bone or other tissues.

[0010] There are some conditions in which pain occurs in the absence of harmful stimuli, tissue damage, or lesions to the nervous system referred to as dysfunctional pain, and these include, but are not limited to, fibromyalgia, tension headache, and irritable bowel disorder.

[0011] Migraine is a headache associated with the activation of sensory fibers that innervate the meninges of the brain.

[0012] Pruritus is a dermatological condition that can be localized or systemic and may be associated with skin lesions (rash, atopic eczema, urticaria). Pruritus is associated with many conditions, including but not limited to stress, anxiety, UV radiation from the sun, metabolic and endocrine disorders (e.g., liver or kidney disease, hyperthyroidism), cancer (e.g., lymphoma), reactions to drugs or food, parasitic and fungal infections, allergic reactions, blood disorders (e.g., polycythemia vera), and dermatological conditions. Pruritus is mediated by pruritus receptors, which are a subset of small-diameter primary sensory neurons that share many features of nociceptive neurons, including but not limited to the expression of TRPV1 channels and other large porous channels (e.g., TRPV2-4, TRPA1, TRPM8, ASIC, and P2X(2 / 3)). Certain pruritic mediators, such as eicosanoids, histamine, bradykinin, ATP, and various neurotrophins, possess endovaniloid functions. Topical capsaicin inhibits histamine-induced itching. Therefore, like nociceptors, pruritus receptors are suitable targets for the aforementioned method of delivering ion channel blockers.

[0013] Coughing is a defensive reflex designed to protect the airways from foreign substances and aid in clearing lumen debris. However, in many diseases, this reflex becomes abnormal, which can lead to an unproductive dry cough in which a hyper-tussive or allo-tussive state is present. Hyper-tussive and allo-tussive states are often chronic in nature, lasting for more than three months, and can occur in many airway disease conditions, including asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), and lung cancer. Additionally, an inappropriate cough reflex can appear acutely or chronically following a viral infection. Furthermore, chronic cough can be idiopathic, meaning the etiology is unknown.

[0014] Neuroinflammation is a mode of inflammation mediated by the efferent (motor) function of sensory neurons, where pro-inflammatory mediator molecules released from the periphery by pain sensory neurons (nociceptors) activate various inflammatory pathways in immune cells and also act on the vascular system to alter blood flow and capillary permeability.

[0015] Neuroinflammation is attributed to peripheral inflammation in various tissues caused by tissue damage, autoimmune diseases, infections, allergies, and exposure to irritants, and is thought to play a significant role in the pathogenesis of numerous disorders (e.g., migraines, arthritis, rhinitis, gastritis, colitis, cystitis, and sunburn). One method to reduce neuroinflammation is to block the excitability of nociceptors, thereby preventing the activation of nociceptive terminals and the release of pro-inflammatory chemicals.

[0016] Despite the development of various therapies for pain, itching, and neuroinflammation, additional agents are needed.

[0017] The present invention relates to the following chemical formula that can be used to treat or prevent pain, itching, and neuroinflammatory disease ( I Provides a compound represented by ):

[0018]

[0019] In the above formula,

[0020] Y - is a pharmaceutically acceptable anion;

[0021] R F and R G is N + Together with, to form an optionally substituted heteroaryl ring having one or more heteroatoms or an optionally substituted bicyclic heteroaryl ring having one or more heteroatoms;

[0022] R A , R B and R CEach is independently H, D, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, nitrile, OR I , NR J R K , NR L C(O)R M , S(O)R N , SO2R O , SO2R O R P , SO2NR Q R R , SO3R S , CO2R T , C(O)R U and C(O)NR V R W Selected from; R B and adjacent R C They form substituted or unsubstituted 3 to 7-membered cycloalkyl (C3-C7-cycloalkyl) or substituted or unsubstituted aryl (e.g., phenyl) together with the carbon atoms to which they are attached;

[0023] R I , R J , R K , R L , R M , R N , R O , R P , R Q , R R , R S , R T , R U , R V and R W Each is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, and substituted or unsubstituted heteroalkyl;

[0024] X 1 -CR X R Y -, -NR Z C(O)-, -OC(O)-, -SC(O)-, -NR Z S(O)-, -S(O)NR Z-, -NR X C(O)NR Y -, -C(O)NR 1A Selected from -, -C(O)O-, -C(O)-, -S(O)-, -S(O)2- and -(O)CS-; X 1 It is also -NR Z C(O)CR X R Y -It could be,

[0025] R X , R Y , R Z and R 1A Each is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, and substituted or unsubstituted heteroalkyl;

[0026] R D and R E Each is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl (optionally substituted with halogen, cyclic alkyl, aryl or heteroaryl), and cycloalkyl; R D and R E is formed with the carbon to which they are attached a substituted or unsubstituted 3 to 7-membered cycloalkyl (C3-C7-cycloalkyl) or a substituted or unsubstituted heterocyclic or heteroalkyl ring; R D and R Z They form 5 to 8-membered lactams that are optionally substituted with the carbon to which they are attached and -NC(O)-.

[0027] In a preferred embodiment, X 1 is -NHC(O)- or -C(O)NH-. In a further preferred embodiment, X 1 It is -NHC(O)-.

[0028] In some implementations, R A and R B Each is independently H, D, halogen, substituted or unsubstituted C1-4 (C1-C4) alkyl and NR J R K Selected from; R J and R K Each is independently H and substituted or unsubstituted C 1-4 Selected from alkyl and / or; R C is not H, for example, halogen, C 1-4 Alkyl and NR J R K am.

[0029] In a preferred embodiment, R A and R B Each is -CH3.

[0030] In a specific other implementation example, R D is halogen, oxygen (iodine), C 3-8 (C3-C8) C optionally substituted with a substituent selected from the group consisting of cycloalkyl, aryl, and heteroaryl. 1-4 Alkyl and / or, R E is H, or halogen, oxygen, C 3-8 C optionally substituted with a substituent selected from the group consisting of cycloalkyl, aryl, and heteroaryl 1-4 It is an alkyl.

[0031] In a preferred embodiment, R D and R E Each is independently selected from -H, -CH3, -CH2CH3, and -(CH2)2CH3. In a more preferred embodiment, R E is hydrogen, and R D is -H, -CH3, -CH2CH3, or -(CH2)2CH3.

[0032] In a specific preferred embodiment, R D and R E Both are hydrogen. Also, in an additional preferred embodiment, R D is hydrogen, and R Eis an alkyl, for example, a C1-C6 alkyl or C1-C4 alkyl including, butyl, methyl, ethyl, propyl, and butyl. In certain additional preferred embodiments, R D and R E These are taken together with the carbon to which they are attached to form C3-C6 cycloalkyls including, but not limited to, cyclopropyl or cyclobutyl.

[0033] In some implementations, Y - It includes, but is not limited to, halide ions, substituted or unsubstituted alkylsulfonates, substituted or unsubstituted arylsulfonates, aliphatic carboxylates, substituted aliphatic carboxylates, aryl carboxylates, substituted aryl carboxylates, heterocyclyl carboxylates, or substituted heterocyclyl carboxylates.

[0034] In an additional modality, Y - is a halide ion. In one embodiment, Y - is a halide ion selected from bromide, chloride, and iodide.

[0035] Each embodiment mentioned herein may be taken in combination with one, any, or all other embodiments. Specific details for implementing the invention

[0036] The present invention is based on the chemical formula as described above ( I The present invention provides a compound represented by ) or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, or combination thereof. The present invention also provides the formula ( I A composition comprising a compound having ) or a pharmaceutically acceptable salt thereof, for example, an effective amount of formula ( IThe present invention provides a composition comprising a compound of ) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. The composition of the present invention may further comprise the compound of the present invention and a biological active agent. The composition described herein may be formulated for oral, intravenous, intramuscular, rectal, skin, subcutaneous, topical, transdermal, sublingual, nasal, inhalation, vaginal, intradural, epidural, or ocular administration.

[0037] The present invention also has a chemical formula ( I A method for treating a patient's pain, cough, itching, or neuroinflammatory disorder comprises administering to the patient a composition comprising a compound having a) said compound, wherein the compound inhibits one or more voltage-gated ion channels present in nociceptors and / or cough receptors and / or itch receptors when exposed to or applied to the inner surface of a channel, but does not substantially inhibit the channel when applied to the outer surface of a channel, said compound may enter the nociceptors, cough receptors, or itch receptors through a large porous channel when the channel is activated and may inhibit one or more voltage-gated ion channels present in the nociceptors, cough receptors, or itch receptors.

[0038] In a specific embodiment, the large porous channel is a transient receptor potential ion channel (TRP channel). In another embodiment, the TRP channel is activated by an exogenous or endogenous agonist. In yet another embodiment, the large porous channel is TRPA1, TRPV1-4, TRPM8, ASIC, or P2X. In a specific embodiment, the compound can enter a nociceptor, cough receptor, or itch receptor through the TRPA1, TRPV1-4, TRPM8, ASIC, or P2X receptor / channel when the receptor / channel is activated. In yet another embodiment, the compound inhibits a voltage-gated sodium channel. In another embodiment, the type of pain treated by the method, composition, and kit of the present invention is selected from the group consisting of neuropathic pain, inflammatory pain, nociceptive pain, pain caused by infection, and procedural pain, or the neuroinflammatory disorder is selected from the group consisting of allergic inflammation, asthma, chronic cough, conjunctivitis, rhinitis, psoriasis, inflammatory bowel disease, interstitial cystitis, and atopic dermatitis.

[0039] The inventors have the following chemical formula, which is expressed in nociceptors and / or cough receptors and / or itchiness receptors but is not expressed in motor neurons and can pass through an open, large porous channel ( I We identified compounds having ):

[0040]

[0041] Since the ion channel blocking compounds of the present invention are positively charged, they are not membrane-permeable and therefore cannot enter cells that do not express large porous channels. Since large porous channels are often more active in tissue conditions associated with pain (e.g., inflammation) due to activation by the release of endogenous ligands or thermal stimulation, the ion channel blockers of the present invention may be used alone to selectively target activated nociceptors to effectively treat (e.g., eliminate or alleviate) pain, cough, itching, or neuroinflammatory conditions. The ion channel blockers of the present invention may also be used in combination with one or more exogenous large porous receptor agonists to selectively target nociceptors to effectively treat (e.g., eliminate or alleviate) pain, cough, itching, or neuroinflammatory conditions.

[0042] Voltage-dependent ion channels in pain-sensing neurons are currently attracting significant attention in the development of drugs to treat pain. Blocking voltage-dependent sodium channels in pain-sensing neurons can block pain signals by interfering with the initiation and transmission of action potentials. Additionally, blocking voltage-dependent sodium channels in nociceptors can reduce or eliminate neuroinflammation by preventing the activation of nociceptive terminals and the release of pro-inflammatory chemicals.

[0043] To date, the limitation in treating with molecules that block sodium or calcium channels is that the majority of the aforementioned external application molecules are hydrophobic and can pass through membranes. Because of this, they enter all cells and lack selectivity to affect only nociceptors.

[0044] The inhibitor of the present invention is membrane-impermeable and effective only when present inside nociceptor cells; therefore, to produce an effect, it must pass through the cell membrane via channels or receptors such as large porous channels (e.g., TRPAV1-4, TRPA1, TRPM8, ASIC, and P2X(2 / 3)). Under normal circumstances, most large porous channels of nociceptors are not activated and require harmful thermal, mechanical, or chemical stimuli to activate them. For example, TRP channels of nociceptors can be activated by exogenous TRP ligands (i.e., TRP agonists), such as capsaicin, which open TRPV1 channels. Therefore, one approach to selectively targeting nociceptors is to co-administer a membrane-impermeable ion channel inhibitor with an exogenous TRP ligand that allows the inhibitor to pass into the cell through TRP channels. In addition to capsaicin, the exogenous TRP ligand may also be another capsaicinoid, mustard oil, or lidocaine. In another example, TRP channels can be activated in response to exogenous stimulant activators, such as acrolein inhaled from smoke or chemical warfare agents like tear gas.

[0045] Under certain circumstances, large porous channels can be activated by endogenous inflammatory activators generated by tissue damage, infection, autoimmunity, atopy, ischemia, hypoxia, cellular stress, immune cell activation, immune mediator production, and oxidative stress in the absence of exogenous large porous channel agonists / ligands. Under these conditions, endogenous molecules (e.g., protons, lipids, and reactive oxygen species) can activate large porous channels expressed in nociceptors, allowing membrane-impermeable, voltage-gated ion channel blockers to access the interior of nociceptors through the endogenously activated large porous channels. Endogenous inflammatory activators of large porous channels include, for example, prostaglandins, nitric oxide (NO), peroxide (H2O2), cysteine-reactive inflammatory mediators, for example, 4-hydroxynonenal, protons, ATP, endogenous alkenylaldehyde, endocannabinoids, and immune mediators (e.g., interleukin 1 (IL-1), nerve growth factor (NGF), and bradykinin, whose receptor is coupled to the large porous channel).

[0046] definition

[0047] As used herein, singular terms mean including one or more unless otherwise specified.

[0048] "Biological activity" refers to the exertion of biological, physical, or chemical effects or activity by a molecule, including biological molecules such as nucleic acids, peptides, polypeptides, and proteins, on proteins, enzymes, receptors, ligands, antigens themselves, or other molecules. For example, a "biologically active" molecule may possess, for instance, enzymatic activity, protein-binding activity, or pharmacological activity.

[0049] Biological agents that may be used in the methods and kits described herein include, but are not limited to, TRPA1 receptor agonists, TRPV1-4 receptor agonists, ASIC agonists, TRPM8 agonists, P2X receptor agonists, NSAIDs, glucocorticoids, narcotics, antiproliferative agents and inflammatory modifiers, antibodies or antibody fragments, antibiotics, polynucleotides, polypeptides, proteins, anticancer agents, growth factors, and vaccines.

[0050] "Inflammation" refers to any type of inflammation, such as inflammation caused by the immune system (immune-mediated inflammation) and the nervous system (neuro-inflammation), and any symptoms of inflammation including redness, heat, swelling, pain, and / or loss of function.

[0051] "Neuro-inflammatory" refers to any type of inflammation mediated by or caused by neurons (e.g., nociceptors) or any other component of the central or peripheral nervous system.

[0052] The term "pain" is used in its broadest sense herein and refers to all types of pain, including nociceptive pain, e.g., somatic pain and visceral pain; inflammatory pain, dysfunctional pain, idiopathic pain, neuropathic pain, e.g., centrally originating pain and peripherally originating pain; and acute and chronic pain such as migraines and cancer pain.

[0053] The term "nociceptive pain" is used to encompass all pain caused by noxious stimuli that threaten or actually damage body tissues, including, but not limited to, those resulting from amputations, contusions, fractures, crush injuries, burns, etc. Pain receptors (nociceptors) for tissue damage are primarily located in the skin, musculoskeletal system, or internal organs.

[0054] The term "somatic pain" is used to refer to pain occurring in bones, joints, muscles, skin, or connective tissue. This type of pain is typically well localized.

[0055] The term "visceral pain" is used herein to refer to pain occurring in internal organs such as the respiratory system, gastrointestinal tract and pancreas, urinary tract, and reproductive organs. Visceral pain includes pain resulting from tumor invasion of the tracheal capsule. Another type of visceral pain, typically caused by obstruction of the hollow intestine, is characterized by intermittent cramping pain and non-localized pain. Visceral pain may be associated with inflammation, such as in cystitis or reflux esophagitis.

[0056] The term "inflammatory pain" includes pain associated with active inflammation that can be caused by trauma, surgery, infection, and autoimmune diseases.

[0057] The term "neuropathic pain" is used herein to refer to pain resulting from the abnormal processing of sensory input by the peripheral or central nervous systems caused by lesions in these systems.

[0058] The term "procedure pain" refers to pain that occurs during medical, dental, or surgical procedures that are typically planned or associated with acute trauma.

[0059] The term "pruritus" is used in its broadest sense at this institution and refers to all types of itching and stinging that are localized, generalized, acute, intermittent, or persistent. Pruritus may be idiopathic, allergic, metabolic, infectious, or drug-induced, and may be caused by liver or kidney disease or cancer. "Pruritus" refers to severe itching.

[0060] The term “cough” is used herein to refer to an abnormal cough reflex that causes a chronic, unproductive dry cough and manifests as a hyper- or allo-tussive state. This cough can be found in many disease conditions, including asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), and lung cancer. Additionally, an inappropriate cough reflex can occur acutely and chronically following a viral infection. Furthermore, chronic cough may be idiopathic, meaning the etiology is essentially unknown.

[0061] "Patient" means any animal. In one embodiment, the patient is a human. Other animals that can be treated using the method, composition and kit of the present invention include, but are not limited to, non-human primates (e.g., monkeys, gorillas, chimpanzees), livestock (e.g., horses, pigs, goats, rabbits, sheep, cattle, llamas), and companion animals (e.g., guinea pigs, rats, mice, lizards, snakes, dogs, cats, fish, hamsters, and birds).

[0062] Compounds useful for the present invention include, but are not limited to, any of the compounds described herein in any pharmaceutically acceptable forms, such as their isomers, such as diastereomers and enantiomers, salts, esters, amides, thioesters, solvates, and polymorphs, as well as racemic mixtures and pure isomers of the compounds described herein. As used herein, the term “pharmaceutically acceptable anion” refers to the conjugate base of a pharmaceutically acceptable acid. Such acid is described in the literature [Stahl, PH and Wermuth, CG (eds.), Handbook of Pharmaceutical Salts: Properties, Selection and Use, Wiley VCH (2008)]. Pharmaceutically acceptable acids are acetic acid, dichloroacetic acid, adipic acid, alginic acid, L-ascorbic acid, L-aspartic acid, benzenesulfonic acid, 4-acetamidobenzoic acid, benzoic acid, p-bromophenylsulfonic acid, (+)-camponic acid, (+)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, cyclamic acid, dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, sulfuric acid, boric acid, citric acid, formic acid, fumaric acid, galactar acid, gentisic acid, D-glucoheptonic acid, D-gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphate, glycolic acid, hippuric acid, hydrochloric acid, Hydrobromide, hydroiodide, isobutyric acid, DL-lactic acid, lactobionic acid, lauric acid, maleic acid, (-)-L-malic acid, malonic acid, DL-mandelic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, parmoic acid, phosphoric acid, propionic acid, (-)-L-pyroglutamic acid, salicylic acid, 4-aminosalicylic acid, sebactic acid, stearic acid, succinic acid, (+)-L-tartaric acid, thiocyanate, p-toluenesulfonic acid, and undecylenic acid, but are not limited thereto.Pharmaceutically acceptable anions include the conjugate base of any acid presented above.

[0063] The term "pharmaceuticalally acceptable salt" refers to a salt that is suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, or allergic reactions within the scope of sound medical judgment, and corresponds to a reasonable benefit / risk ratio. The salt may be prepared in situ during the final separation and purification of the compound of the present invention, or separately by reacting a free base functional group with a suitable organic acid. Representative acid salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, isethionate, lactobionate, lactate, laurate, lauryl sulfate, maleate, maleate, malonate, mesylate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, and oxalate. Includes, but is not limited to, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, etc.

[0064] In the general description of the compounds of the present invention, the number of atoms of a specific type in the substituent group is generally in a range, for example, an alkyl group containing 1 to 4 carbon atoms or C 1-4 alkyl or C 1-It is provided as a C4 alkyl. References to the above range are intended to include specific references to groups having each of the integer atoms within the specified range. For example, alkyl groups of 1 to 4 carbon atoms include C1, C2, C3, and C4 alkyls, respectively. Different numbers of atoms and different types of atoms may be indicated in a similar manner.

[0065] "D" is deuterium.

[0066] As used herein, the term "alkyl" and the prefix "alk-" include both straight-chain and branched-chain groups and cyclic groups, namely cycloalkyl. Cyclic groups may be monocyclic or polycyclic and, preferably, may have 3 to 6 cyclic carbon atoms or 3 to 7 carbon atoms. Exemplary cyclic groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups.

[0067] "C 1-4 "alkyl" or "C1-C4alkyl" refers to a branched or unbranched hydrocarbon group having 1 to 4 carbon atoms. Similarly, "C 1-6 "Alkyl" or "C1-C6" is a branched or unbranched hydrocarbon group having 1 to 6 carbon atoms. For example, C 1-4 alkyl or C 1-6Alkyl groups containing alkyl groups may be substituted or unsubstituted. Exemplary substituents include, but are not limited to, alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide, hydroxyl, fluoroalkyl, perfluoroalkyl, amino, alkylamino, disubstituted amino, quaternary amino, alkylcarboxyl, and carboxyl groups. Exemplary substituents are also alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide (F, Cl, Br or I), hydroxyl, fluoroalkyl, perfluoroalkyl, oxo, amino, alkylamino, disubstituted amino, quaternary amino, amido, ester, alkylcarboxyl, alkoxycarbonyl, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxyl, alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, phosphate, phosphonato, phosphinato, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amido, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, It includes cyano, azido, aryl, heterocyclil, alkylaryl, or aromatic or heteroaromatic moiety. C 1-4 Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and cyclobutyl. C 1-6 Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0068] An example of a substituted alkyl is a heteroalkyl. "Heteroalkyl" means a branched or unbranched alkyl, cycloalkyl, alkenyl, or alkynyl group having 1 to 7 or more carbon atoms in addition to 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. "C 1-7 "Heteroalkyl" means a branched or unbranched alkyl, alkenyl, or alkynyl group having 1 to 7 carbon atoms in addition to 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. Heteroalkyl may include, but is not limited to, tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiesters, phosphoramidates, sulfonamides, and disulfides. Heteroalkyl may optionally include monocyclic, bicyclic, or tricyclic rings, each ring preferably having 3 to 6 members. The heteroalkyl group may be substituted or unsubstituted. Examples of substituents are alkyl, alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide (F, Cl, Br, or I), hydroxyl, fluoroalkyl, perfluoroalkyl, oxo, amino, alkylamino, disubstituted amino, quaternary amino, amido, ester, alkylcarboxy, alkoxycarbonyl, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxyl, alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, phosphate, phosphonato, phosphinato, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amido, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, aryl, heterocyclil, alkylaryl, or includes aromatic or heteroaromatic moiety. C 1-7Examples of heteroalkyls include, but are not limited to, methoxymethyl and ethoxyethyl.

[0069] Alkenyls are branched or unbranched hydrocarbon groups containing one or more double bonds. For example, "C 2-6 "Alkenyl" or "C2-C6 alkenyl" refers to a branched or unbranched hydrocarbon group containing one or more double bonds and having 2 to 6 carbon atoms. The alkenyl may optionally comprise a monocyclic or polycyclic ring, wherein each ring preferably has 3 to 6 members. The alkenyl group may be substituted or unsubstituted. Exemplary substituents include those described above for the alkyl group, specifically including alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide, hydroxyl, fluoroalkyl, perfluoroalkyl, amino, alkylamino, disubstituted amino, quaternary amino, alkylcarboxyl, and carboxyl groups. C 2-6 Alkenyls include, but are not limited to, vinyl, allyl, 2-cyclopropyl-1-ethenyl, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl and 2-methyl-2-propenyl.

[0070] Alkynyl is a branched or unbranched hydrocarbon group containing one or more triple bonds. For example, "C 2-6"Alkynyl" or "C2-C6-alkynyl" refers to a branched or unbranched hydrocarbon group containing one or more triple bonds and having 2 to 6 carbon atoms. The alkynyl may optionally comprise a monocyclic, bicyclic, or tricyclic ring, wherein each ring preferably has 5 or 6 members. The alkynyl group may be substituted or unsubstituted. Exemplary substituents include those described above for the alkyl group, specifically including alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide, hydroxy, fluoroalkyl, perfluoroalkyl, amino, alkylamino, disubstituted amino, quaternary amino, alkylcarboxyl, and carboxyl groups. C 2-6 Alkynyls include, but are not limited to, ethinyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl.

[0071] "Heterocyclil," "heterocyclic," or "heterocycloalkyl" means a stable monocyclic or polycyclic (including bicyclic or tricyclic) heterocyclic ring that is saturated, partially unsaturated, or unsaturated (including heteroaryl or aromatic), and comprises two or more carbon atoms and one, two, three, or four or more heteroatoms independently selected from N, O, and S, and any of the heterocyclic rings defined above comprises any bicyclic or polycyclic group fused to a benzene ring, heteroaryl, cycloalkyl, or heterocycloalkyl ring. In certain embodiments, the heterocyclil is a 3 to 15-membered ring system, a 3 to 12-membered ring system, or a 3 to 9-membered ring system. "C 2-6"Heterocyclile" means a stable 5 to 7-membered monocyclic or 7 to 14-membered bicyclic heterocyclic ring that is saturated, partially unsaturated, or unsaturated (including heteroaryl or aromatic) and consists of 2 to 6 carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S, and any of the heterocyclic rings defined above comprises any bicyclic group fused to a benzene ring, heteroaryl, cycloalkyl, or heterocycloalkyl ring. The heterocyclile or heteroaryl group may be substituted or unsubstituted. Exemplary substituents are substituted or unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide, hydroxy, fluoroalkyl, perfluoroalkyl, amino, alkylamino, It includes disubstituted amino, quaternary amino, alkylcarboxyl, and carboxyl groups. Nitrogen and sulfur heteroatoms may be optionally oxidized. The heterocyclic ring may be covalently attached through any heteroatom or carbon atom to form a stable structure; for example, the imidazolinyl ring may be connected at any one of the ring-carbon atom positions or at the nitrogen atom. The nitrogen atom of the heterocycle may be quaternized. Preferably, when the total number of S and O atoms in the heterocycle exceeds 1, these heteroatoms are not adjacent to each other. The heterocycle includes 1H-indazole, 2-pyrrolidonyl, 2H,6H-1,5,2-dithiazinyl, 2H-pyrrolyl, 3H-indolyl, 4-piperidonyl, 4aH-carbazole, 4H-quinolidinyl, 6H-1,2,5-thiadiazinyl, Acridinyl, Azocinyl, Benzimidazolyl, Benzofuranyl, Benzothiofuranyl, Benzothiophenyl, Benzoxazolyl, Benzthiazolyl, Benztriazolyl, Benztetrazollyl, Benzisoxazolyl, Benzisothiazolyl, Benzimidazalonil, Carbazollyl, 4aH-Carbazollyl, b-Carbolinyl, Cromanyl, Chromenyl, Cinnolinyl, Decahydroquinolinyl, 2H,6H-1,5,2-Dithiazinyl, Dihydrofuro[2,3-b]tetrahydrofuran, Furanyl, Furazanyl, Imidazolidinyl, Imidazolininyl, Imidazolyl, 1H-Indazolyl, Indolenyl, Indolinyl, Indoliginyl, Indolyl, Isobenzofuranyl, Isochromanyl, Isoindazolyl, Isoindolinyl, Isoindolyl, Isoquinolinyl, Isothiazolyl, Isoxazolyl, Morfollinyl, Naphthiridinyl, Octahydroisoquinolinyl, Oxadiazolyl, 1,2,3-Oxadiazolyl, 1,2,4-Oxadiazolyl, 1,2,5-Oxadiazolyl, 1,3,4-Oxadiazolyl, Oxazolidinyl, Oxazolyl, Oxazolidinylperimidinyl, Phenanthridinyl, Fenanthrolinyl, Fenarsaginyl, Fenazinyl, Fenothiaginyl, Fenoxatiinyl, Fenoxazinyl, Phthalaginyl, Piperazinyl, Piperidinyl, Pteridinyl, Piperidonil, 4-Piperidonyl, Pteridinyl, Purinyl, Pyranil, Pyrazinyl, Pyrazolidinyl, Pyrazolininyl, Pyrazolil, Pyridazinyl, Pyridoxazole, Pyridomidazole, Pyridothiaazole, Pyridinyl, Pyridyl, Pyrimidinyl, Pyrrolidinyl, Pyrrolininyl, Pyrrolininyl, Pyrrolilil, Quinazolininyl, Quinolinyl, 4H-Quinolidinyl, Quinoxalinyl, Quinuclidinyl, Carbolinyl, Tetrahydrofuranil, Tetrahydroisoquinolinyl, Tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thiantrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, xanthenyl, β-lactam, γ-lactam and δ-lactam are included, but not limited thereto. Preferred 5 to 10-membered heterocycles include pyridinyl, pyrimidinyl, triazinyl, furanyl, thienyl, thiazolyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, tetrazolyl, benzofuranyl, benzothiofuranyl, indolyl, benzimidazolyl, 1H-indazolyl, oxazolidinyl, isoxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolidinyl,It includes, but is not limited to, quinolinyl and isoquinolinyl. Preferred 5 to 6-membered heterocycles include, but are not limited to, pyridinyl, quinolinyl, pyrimidinyl, triazinyl, furanyl, thienyl, thiazolyl, pyrrolyl, piperazinyl, piperidinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, and tetrazolyl. Preferred substituents include phenyl, methyl, ethyl, propyl, butyl, chloro, bromo, fluoro, and iodo.

[0072] "Aryl" refers to an aromatic group (e.g., phenyl) having a ring system containing a carbon atom with conjugated π electrons. "C 6- C 12 "Aryl" or "C6-C 10 "Aryl" is an aryl group having 6 to 12 carbon atoms or 6 to 10 carbon atoms, respectively. The aryl group may optionally comprise a monocyclic, bicyclic, or tricyclic ring, wherein each ring preferably has 5 or 6 members. The aryl group may be substituted or unsubstituted. Exemplary substituents include substituted or unsubstituted alkyl, hydroxy, alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halide, fluoroalkyl, carboxyl, alkylcarboxyl, amino, alkylamino, monosubstituted amino, disubstituted amino, and quaternary amino groups. A preferred aryl group is phenyl.

[0073] "Aralkyl" means a substituted or unsubstituted alkyl group substituted with a substituted or unsubstituted aryl group (e.g., including benzyl, phenethyl, or 3,4-dichlorophenethyl).

[0074] "Heteroaralkyl" means a substituted or unsubstituted alkyl group substituted by a heteroaryl group.

[0075] "C 7-14"Aralkyl" means an alkyl group substituted with an aryl group having 7 to 14 carbon atoms (e.g., benzyl, phenethyl, or 3,4-dichlorophenethyl).

[0076] "C 3-10 "Heterocycloalkyl" means an alkyl-substituted heterocyclic group having 3 to 10 carbon atoms in addition to one or more heteroatoms (e.g., 3-furanylmethyl, 2-furanylmethyl, 3-tetrahydrofuranylmethyl, or 2-tetrahydrofuranylmethyl).

[0077] "Halide" or "halogen" refers to bromine, chlorine, iodine, or fluorine.

[0078] "Fluoroalkyl" refers to an alkyl group substituted with a fluorine atom.

[0079] "Alkyl carboxy" refers to a chemical moiety having the chemical formula -(R)-COOH, where R is C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkinyl, C 2-6 Heterocyclil, C 6-12 Aril, C 7-14 Aralkyl, C 3-10 Heterocycloalkyl or C 1-7 It is selected from heteroalkyls.

[0080] "Alkoxy" refers to a chemical substituent having the chemical formula -OR, where R is a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl, or R is C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkinyl, C 2-6 Heterocyclil, C 6-12 Aril, C 7-14 Aralkyl, C 3-10 Heterocycloalkyl or C 1-7 It can be selected from heteroalkyls.

[0081] "Aryloxy" refers to a chemical substituent of the chemical formula -OR, where R is C 6-12It is Aril Gi.

[0082] "Alkylthio" refers to a chemical substituent of the chemical formula -SR, where R is C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkinyl, C 2-6 Heterocyclil, C 6-12 Aril, C 7-14 Aralkyl, C 3-10 Heterocycloalkyl or C 1-7 It is selected from heteroalkyls.

[0083] "Arylthio" refers to a chemical substituent of the chemical formula -SR, where R is C 6-12 It is Aril Gi.

[0084] "Charged moiety" means a moiety that gains a proton at physiological pH and becomes positively charged (e.g., ammonium, guanidinium, or amidinium) or a moiety containing a net positive charge without protonation (e.g., quaternary ammonium). A charged moiety may be permanently charged or temporarily charged.

[0085] "Therapeutic effective dose" or "effective dose" means an amount sufficient to produce the desired result, e.g., reduction or elimination of pain, cough, itching, or neuroinflammatory inflammation, in a patient (e.g., human) suffering from a condition, disease, or disorder caused wholly or partially by neuroinflammatory inflammation (e.g., asthma, arthritis, colitis, contact dermatitis, diabetes, eczema, cystitis, chronic intractable cough, postviral cough, gastritis, migraine, psoriasis, rhinitis, rosacea, or sunburn).

[0086] "Solvent" refers to a form of solvent addition containing stoichiometric or non-stoichiometric amounts of solvent.

[0087] The compound of the present invention, comprising a salt of the compound, may exist in a solvated form including a hydrated form and a non-hydrated form, as well as in a non-solvent form. Generally, the solvated form is equivalent to the non-solvent form and is included within the scope of the present invention. Non-limiting examples of hydrates include monohydrates, dihydrates, hemihydrates, etc. In certain embodiments, the compound is a hemihydrate. Non-limiting examples of solvates include ethanol solvates, acetone solvates, etc.

[0088] The compounds of the present invention may exist in polycrystalline or amorphous forms. Generally, all physical forms are intended to be equivalent for the uses considered by the present invention and to be within the scope of the present invention.

[0089] The compounds that can be used in the compositions, kits, and methods of the present invention are those with the following chemical formula ( I It includes a compound having ) or a pharmaceutically acceptable salt thereof:

[0090]

[0091] In the above formula,

[0092] Y - is a pharmaceutically acceptable anion;

[0093] R F and R G is N to which they are attached + N together with + In addition to, an optionally substituted heteroaryl ring having zero or one or more heteroatoms or N + In addition, forming an optionally substituted bicyclic heteroaryl ring having zero or one or more heteroatoms;

[0094] R A , R B and R C Each is independently H, D, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, nitrile, OR I , NRJ R K , NR L C(O)R M , S(O)R N , SO2R O , SO2R O R P , SO2NR Q R R , SO3R S , CO2R T , C(O)R U and C(O)NR V R W Selected from; R B and adjacent R C They form substituted or unsubstituted 3 to 7-membered cycloalkyl (C3-C7-cycloalkyl) or substituted or unsubstituted aryl (e.g., phenyl) together with the carbon atoms to which they are attached;

[0095] R I , R J , R K , R L , R M , R N , R O , R P , R Q , R R , R S , R T , R U , R V and R W Each is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkenyl;

[0096] X 1 -CR X R Y -, -NR Z C(O)-, -NR Z C(O)CR X R Y -, -OC(O)-, -SC(O)-, -NR Z S(O)-, -S(O)NR Z -, -NR X C(O)NR Y -, -C(O)NR1A Selected from -, -C(O)O-, -(O)CS-, -S(O)-, -S(O)2- and -C(O)-;

[0097] R X , R Y , R Z and R 1A Each is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkenyl;

[0098] R D and R E Each is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, cycloalkyl (e.g., C3-C6 or C3-C7 cycloalkyl cycloalkyl), aryl, or heteroaryl, or R D and R E , together with the carbon to which they are attached, form a substituted or unsubstituted C3-C7 cycloalkyl or a substituted or unsubstituted heterocyclic ring (e.g., a 5 to 7-membered heterocyclic ring), or R D and R Z They form 5 to 8-membered lactams that are optionally substituted with the carbon to which they are attached and -NC(O)-.

[0099] In some implementations, R F and R G N to which they are attached by + A heteroaryl ring or bicyclic heteroaryl ring formed together with substituted or unsubstituted C 1-6 Alkan, C 1-6 It may be optionally substituted with heteroalkanes, carbocycles, substituted carbocycles, heterocarbocycles, substituted heterocarbocycles, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, carboxamide, hydroxy, ether, amide, ester, sulfonamide, sulfone, amino, aminoalkyl, urea, nitrile, or halogen. In certain embodiments, N+ R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by may be unsubstituted. In a further embodiment, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by is a substituted or unsubstituted C1-C6 alkyl, a substituted or unsubstituted C3-C6 cycloalkyl, a substituted or unsubstituted 3 to 15-membered heterocyclyl, alkoxy, or CO2R 2A It can be substituted as, where R 2A is selected from H, D, substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C1-C6 alkyl) and substituted or unsubstituted alkenyl (e.g., C2-C6 alkenyl). In addition, in a further preferred embodiment, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by is a substituted or unsubstituted C1-C6 alkyl, a substituted or unsubstituted C3-C6 cycloalkyl, a substituted or unsubstituted 3 to 8-membered heterocyclyl, OR 2B or CO2R 2A It can be substituted as, where R 2A is selected from H or substituted or unsubstituted C1-C6 alkyls and R 2B is a substituted or unsubstituted C1-C6 alkyl. In additional embodiments, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by may be substituted or unsubstituted 3 to 8-membered heterocyclils, wherein the 3 to 8-membered heterocyclils contain at least one nitrogen ring atom.

[0100] In a preferred embodiment, N + R together with F and R GThe heteroaryl ring or bicyclic heteroaryl ring formed by C 1-6 The alkane substituted or C1-C6 alkyl group is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl. In a preferred embodiment, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by is substituted with a group selected from -O-methyl, -O-ethyl, -O-propyl, -O-isopropyl, -O-butyl, -O-isobutyl, -O-cyclohexyl, -O-cyclopentyl, and -ethyl-O-methyl. In a preferred embodiment, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by is substituted with a carbocycle, or a C3-C6 cycloalkyl selected from cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In a preferred embodiment, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by is substituted with a heterocarbocycle, or a heterocyclic selected from aziridine, azetidine, furan, pyrrolidine, pyran, piperidine, piperazine, azepine, and diazepine. Also, in a further embodiment, N + R together with F and R G The heteroaryl ring or bicyclic heteroaryl ring formed by is substituted with a heterocarbocyclic or heterocyclic selected from those presented in Tables 2 to 6.

[0101] In a preferred embodiment, N + R together with F and R GThe heteroaryl ring or bicyclic heteroaryl ring formed by can be optionally substituted with methylphenyl, pyridinyl, methylpyridinyl, -C(O)NH2, -C(O)N(CH3)2, -C(O)OCH3, -C(O)OCH2CH3, -S(O)2NH2, -S(O)2N(CH3)2, -S(O)2CH3, -NHS(O)2CH3, -NHC(O)CH3, -C(O)N(CH3)2, -O(CH2)2OCH3, fluorine, chlorine, tetrazolyl, methylpyrrolidinyl, methylamino, pyrrolidine, and dimethylamino.

[0102] In a preferred embodiment, X 1 is -NHC(O)- or -C(O)NH-. In another preferred embodiment, X 1 It is -NHC(O)-.

[0103] In some implementations, R A and R B Each is independently H, D, halogen, substituted or unsubstituted C 1-4 Alkyl and NR J R K Selected from; R J and R K Each is independently H and substituted or unsubstituted C 1-4 Selected from alkyl; and / or R C is not H, for example, halogens, substituted or unsubstituted C 1-4 Alkyl and NR J R K am.

[0104] Also, in an additional desirable embodiment, R A , R B and R C is independently H, D, halogen, OR I , substituted or unsubstituted C1-C4 alkyl and NR J R K Selected from; R I , R J and R KEach is independently selected from H and substituted or unsubstituted C1-C4 alkyls. In a preferred embodiment, R A and R B Each is CH3, and R C is selected from the group consisting of H, CH3, halogen, nitrile (cyano), methoxy, and ethoxy, in a further preferred embodiment, R A and R B Each is CH3, and R C is selected from the group consisting of H, CH3, fluoro, chloro, nitrile, methoxy, and ethoxy. In a further preferred embodiment, R A and R B Each is CH3, and R C is hydrogen.

[0105] In a preferred embodiment, R A and R B Each is -CH3.

[0106] In a specific mode, R B and adjacent R C They form substituted or unsubstituted 3 to 7-membered cycloalkyls (C3-C7-cycloalkyl) or substituted or unsubstituted aryls (e.g., phenyl) together with the carbon atoms to which they are attached.

[0107] In a specific other implementation example, R D is halogen, oxygen, C 3-8 C optionally substituted with a substituent selected from the group consisting of cycloalkyl, aryl, and heteroaryl 1-4 Alkyl and / or R E is H, or halogen, oxygen, C 3-8 C optionally substituted with a substituent selected from the group consisting of cyclic alkyl, aryl, or heteroaryl 1-4 It is an alkyl.

[0108] In a preferred embodiment, R D and R EEach is independently selected from -H, -CH3, -CH2CH3, and -(CH2)2CH3. In a more preferred embodiment, R E is hydrogen, and R D is -H, -CH3, -CH2CH3, or -(CH2)2CH3.

[0109] In a specific preferred embodiment, R D and R E Both are hydrogen. Also, in an additional preferred embodiment, R D is hydrogen, and R E is an alkyl, for example, a C1-C6 alkyl or C1-C4 alkyl including, butyl, methyl, ethyl, propyl, and butyl. In certain additional preferred embodiments, R D and R E These are taken together with the carbon atoms to which they are attached to form C3-C6 cycloalkyls, including cyclopropyl or cyclobutyl, to a non-limiting extent.

[0110] In some implementations, Y - is a halide anion, carboxylate, or sulfonate. Y - It may be, for example, a halide ion, a substituted or unsubstituted alkyl sulfonate, a substituted or unsubstituted aryl sulfonate, a substituted or unsubstituted alkyl or aliphatic carboxylate, a substituted or unsubstituted aryl carboxylate, or a substituted or unsubstituted heterocyclyl carboxylate.

[0111] In a specific implementation example, Y -trifluoroacetate, sulfate, phosphate, acetate, fumarate, formate, carbonate, maleate, citrate, pyruvate, succinate, oxalate, sulfonate (e.g., methanesulfonate, trifluoromethanesulfonate, toluenesulfonate, e.g., p-toluenesulfonate, benzenesulfonate, ethanesulfonate, camphorsulfonate, 2-methylenesulfonate, or naphthalenesulfonate, e.g., 2-naphthalenesulfonate), bisulfate, malonate, cinnapoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, L-lactate, D-lactate, aspartate, maleate, L-tartrate, D-tartrate, stearate, 2-furoate, It is selected from the group consisting of 3-furoate, napadisilate (naphthalene-1,5-disulfonate or naphthalene-1-(sulfonic acid)-5-sulfonate), edicilate (ethane-1,2-disulfonate or ethane-1-(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), D-mandelate, L-mandelate, propionate, tartrate, phthalate, hydrochlorate, hydrobromate, and nitrate. In one embodiment, Y - is a halide anion.

[0112] In one embodiment, Y - is a halide ion selected from bromide, chloride, or iodide.

[0113] Each embodiment mentioned herein may be taken in combination with one, any, or all other embodiments.

[0114] In a specific preferred embodiment, the present invention has the formula ( I It relates to a compound of ) or a pharmaceutically acceptable salt thereof, wherein R F and R G is N + It forms a pyridinium ring that is optionally substituted together with. In a further embodiment, N+ R together with F and R G The pyridinium ring formed by is unsubstituted. Also, in an additional embodiment, N + R together with F and R G The pyridinium ring formed by is a substituted or unsubstituted C1-C6 alkyl, a substituted or unsubstituted C3-C6 cycloalkyl, a substituted or unsubstituted 3 to 15-membered heterocyclyl, alkoxy, and CO2R 2A It can be substituted with a substituent selected from the group consisting of, where R 2A is selected from H, D, substituted or unsubstituted alkyl, and substituted or unsubstituted alkenyl. In addition, in a further preferred embodiment, N + R together with F and R G The pyridinium ring formed by is a substituted or unsubstituted C1-C6 alkyl, a substituted or unsubstituted C3-C6 cycloalkyl, a substituted and unsubstituted 3 to 8-membered heterocyclile, OR 2B or CO2R 2A It can be substituted with a substituent selected from the group consisting of, where R 2A is selected from H or substituted or unsubstituted C1-C6 alkyls, and R 2B is a substituted or unsubstituted C1-C6 alkyl. In additional embodiments, N + R together with F and R G The pyridinium ring formed by may be substituted or unsubstituted with a 3 to 8-membered heterocyclile, wherein the 3 to 8-membered heterocyclile contains a nitrogen ring atom.

[0115] In a preferred embodiment, the present invention has a formula ( I It relates to a compound of ) or a pharmaceutically acceptable salt thereof, wherein R F and R G is N +It forms an optionally substituted heteroaryl ring comprising one or more nitrogen groups selected from one of the following:

[0116]

[0117] Here, each of the monocyclic heteroaryls presented above is optionally substituted where possible.

[0118] In a preferred embodiment, the present invention has a formula ( I It relates to a compound of ) or a pharmaceutically acceptable salt thereof, wherein R F and R G is N + Together with, it forms an optionally substituted bicyclic heteroaryl ring containing one or more nitrogen groups as follows:

[0119]

[0120] Also, in a further preferred embodiment, the present invention has a formula ( I It relates to a compound of ) or a pharmaceutically acceptable salt thereof, wherein R F and R G is N + It forms a heteroaryl ring selected from those presented in Tables 1 to 6 below.

[0121] In a further embodiment, the present invention is based on the following formula ( II Includes a compound of ) or a pharmaceutically acceptable salt thereof:

[0122]

[0123] In the above equation, Y - is the chemical formula ( I As defined for );

[0124] R 1 and R 2 One of them is hydrogen, and R 1 and R 2 The other one is methyl, ethyl, unsubstituted phenyl and C(O)OR 3Selected from a group composed of;

[0125] R 3 It is selected from the group consisting of hydrogen, methyl, and ethyl.

[0126] In certain embodiments, the compound has the chemical formula ( II With ), here Y - is a halide ion, a sulfonate (e.g., including substituted or unsubstituted alkylsulfonates and substituted or unsubstituted arylsulfonates), or a carboxylate (e.g., including substituted or unsubstituted alkyl or aliphatic carboxylates, substituted or unsubstituted aryl carboxylates or substituted or unsubstituted heterocyclyl carboxylates). Also, in further embodiments, the compound is of the formula ( II With ), here Y -trifluoroacetate, sulfate, phosphate, acetate, fumarate, formate, carbonate, maleate, citrate, pyruvate, succinate, oxalate, sulfonate (e.g., methanesulfonate, trifluoromethanesulfonate, toluenesulfonate, e.g., p-toluenesulfonate, benzenesulfonate, ethanesulfonate, camphorsulfonate, 2-methylenesulfonate or naphthalenesulfonate, e.g., 2-naphthalenesulfonate), bisulfate, malonate, cinnapoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, L-lactate, D-lactate, aspartate, maleate, L-tartrate, D-tartrate, stearate, 2-furoate, It is selected from the group consisting of 3-furoate, napadisilate (naphthalene-1,5-disulfonate or 2-naphthalene-1-(sulfonic acid)-5-sulfonate), edicilate (ethane-1,2-disulfonate or ethane-1-(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), D-mandelate, L-mandelate, propionate, tartarate, phthalate, hydrochlorate, hydrobromate, and nitrate. In one embodiment, Y - is a halide anion.

[0127] In an additional embodiment, the compound has the chemical formula ( II With ), here Y - is a halide ion. In one embodiment, Y - is a halide ion selected from bromide, chloride, and iodide.

[0128] In a specific embodiment, the compound has the chemical formula ( II With ), here R 2 is hydrogen, and R 1 It is methyl, ethyl, unsubstituted phenyl and C(O)OR 3 Selected from a group consisting of, where R 3is selected from hydrogen, methyl, and ethyl. In a preferred embodiment, R 1 is an unsubstituted phenyl, and R 2 is hydrogen. Also, in an additional preferred embodiment, R 1 is methyl, and R 2 is hydrogen. In a further preferred embodiment, R 1 is ethyl, and R 2 is hydrogen. In a further preferred embodiment, R 1 is C(O)OR 3 and, here R 3 is methyl or ethyl, and R 2 is hydrogen. In a further preferred embodiment, R 1 is C(O)OR 3 and, here R 3 is ethyl, and R 2 is hydrogen.

[0129] In an additional embodiment, the compound has the chemical formula ( II With ), here R 1 is hydrogen, and R 2 is methyl, ethyl, unsubstituted phenyl and C(O)OR 3 Selected from a group consisting of, where R 3 is hydrogen, methyl, or ethyl. In a preferred embodiment, R 1 is hydrogen, and R 2 is an unsubstituted phenyl. Also, in a further preferred embodiment, R 1 is hydrogen, and R 2 is methyl. In a further preferred embodiment, R 1 is hydrogen, and R 2 is ethyl. In a further preferred embodiment, R 1 is hydrogen, and R 2 is C(O)OR 3 and, here R 3 is methyl or ethyl. In a further preferred embodiment, R 1 is hydrogen, and R 2 is C(O)OR 3 and, here R 3is ethyl.

[0130] In addition, in further embodiments, the present invention has the following chemical formula ( III Includes a compound of ) or a pharmaceutically acceptable salt thereof:

[0131]

[0132] In the above equation, Y - is the chemical formula ( I As defined for );

[0133] R 1 It is selected from the group consisting of methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, iso-butyl, tert-butyl and sec-butyl), cyclohexyl, phenyl, and CH2C(O)NHR3;

[0134] R 2 is hydrogen or methyl;

[0135] R 3 is hydrogen, methyl, or ethyl.

[0136] In certain embodiments, the compound has the chemical formula ( III With ), here Y - is a halide ion, a sulfonate (e.g., including substituted or unsubstituted alkylsulfonates and substituted or unsubstituted arylsulfonates), or a carboxylate (e.g., including substituted or unsubstituted alkyl or aliphatic carboxylates, substituted or unsubstituted aryl carboxylates or substituted or unsubstituted heterocyclyl carboxylates). Also, in further embodiments, the compound is of the formula ( III With ), here Y -trifluoroacetate, sulfate, phosphate, acetate, fumarate, formate, carbonate, maleate, citrate, pyruvate, succinate, oxalate, sulfonate (e.g., methanesulfonate, trifluoromethanesulfonate, toluenesulfonate, e.g., p-toluenesulfonate, benzenesulfonate, ethanesulfonate, camphorsulfonate, 2-methylenesulfonate or naphthalenesulfonate, e.g., 2-naphthalenesulfonate), bisulfate, malonate, cinnapoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, L-lactate, D-lactate, aspartate, maleate, L-tartrate, D-tartrate, stearate, 2-furoate, It is selected from the group consisting of 3-furoate, napadisilate (naphthalene-1,5-disulfonate or naphthalene-1-(sulfonic acid)-5-sulfonate), edicilate (ethane-1,2-disulfonate or ethane-1-(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), D-mandelate, L-mandelate, propionate, tartarate, phthalate, hydrochlorate, hydrobromate, and nitrate. In one embodiment, Y - is a halide anion.

[0137] In an additional embodiment, the compound has the chemical formula ( III With ), here Y - is a halide ion. In one embodiment, Y - is a halide ion selected from bromide, chloride, and iodide.

[0138] In an additional embodiment, the compound has the chemical formula ( III It has ), where R1 is methyl and R2 is hydrogen. Also, in an additional embodiment, the compound has the chemical formula ( III ) having, where both R1 and R2 are methyl.

[0139] Also, in additional embodiments, the compound has the chemical formula ( III It has the formula ), where R1 is ethyl and R2 is hydrogen. In another embodiment, the compound has the chemical formula ( III It has ), where R1 is ethyl and R2 is methyl.

[0140] In an additional embodiment, the compound has the chemical formula ( III It has ), where R1 is n-propyl or iso-propyl and R2 is hydrogen. Also, in additional embodiments, the compound has the chemical formula (III) It has, where R1 is n-propyl or iso-propyl and R2 is methyl.

[0141] In an additional embodiment, the compound has the chemical formula ( III It has ), where R1 is n-butyl, iso-butyl, tert-butyl, or sec-butyl, and R2 is hydrogen. In a further embodiment, the compound has the chemical formula (III) It has the formula, where R1 is n-butyl, iso-butyl, tert-butyl, or sec-butyl, and R2 is methyl. Also, in additional embodiments, the compound has the formula ( III It has the formula ), where R1 is iso-butyl and R2 is hydrogen. In a further embodiment, the compound has the formula ( III It has ), where R1 is iso-butyl and R2 is methyl.

[0142] In an additional embodiment, the compound has the chemical formula ( III It has ), where R1 is cyclohexyl and R2 is hydrogen. Also, in an additional embodiment, the compound has the chemical formula ( III It has ), where R1 is cyclohexyl and R2 is methyl.

[0143] In another embodiment, the compound has the chemical formula ( III It has ), where R1 is phenyl and R2 is hydrogen. Also, in additional embodiments, the compound has the chemical formula ( III It has ), where R1 is phenyl and R2 is methyl.

[0144] In another embodiment, the compound has the chemical formula ( III It has ), where R1 is CH2C(O)NHR3 and R2 is hydrogen. In an additional embodiment, the compound has the chemical formula ( III ) having, where R1 is CH2C(O)NHR3 and R2 is methyl. In further embodiments, R1 is CH2C(O)NHR3, R2 is hydrogen, and R3 is methyl or ethyl. Also in further embodiments, R1 is CH2C(O)NHR3, R2 is methyl, and R3 is methyl or ethyl. In certain further embodiments, R1 is CH2C(O)NHR3, R2 is hydrogen, and R3 is ethyl. In further embodiments, R1 is CH2C(O)NHR3, R2 is methyl, and R3 is ethyl.

[0145] In certain embodiments, the compound is selected from Table A below or pharmaceutically acceptable salts thereof, where Y - is a pharmaceutically acceptable anion:

[0146] Table A

[0147]

[0148]

[0149]

[0150]

[0151] In a further preferred embodiment, the compound is selected from Table B below or pharmaceutically acceptable salts thereof:

[0152] Table B

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161] In certain embodiments, the compound is selected from Table C below or pharmaceutically acceptable salts thereof, where Y - is a pharmaceutically acceptable anion:

[0162] Table C

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173] In a further preferred embodiment, the compound is selected from Table D below or pharmaceutically acceptable salts thereof:

[0174] Table D

[0175]

[0176]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182] In a further preferred embodiment, the compound is selected from Table E below or pharmaceutically acceptable salts thereof:

[0183] Table E

[0184]

[0185]

[0186]

[0187]

[0188] Each preferred embodiment described herein may be taken in combination with one, any, or all other preferred embodiments as presented herein in all permutations.

[0189] The composition of the present invention may comprise a racemic mixture, a pure enantiomer, or an excess of one enantiomer relative to another enantiomer. For example, the alpha atom of compound 3 is chiral. Thus, the compound may have the stereochemistry described in the following structures (3a) and (3b):

[0190]

[0191] The composition of the present invention may comprise a racemic mixture of structure 3a and structure 3b, a pure enantiomer of structure 3a or structure 3b, or an excess of one enantiomer relative to other enantiomers. For example, the composition may comprise a mixture with an excess of at least 5, 10, 20, 30, 40, 50, 60, 70, 80, or 90% of the enantiomer. In one embodiment, the excess of the enantiomer is at least 95%.

[0192] The compounds of the present invention include all enantiomers that can be defined as (R)- or (S)- in terms of absolute stereochemistry, as well as their racemic and optically pure forms, and are not limited to any of the pharmaceutically acceptable forms described herein, including enantiomers, salts, solvates, polymorphs, solvatomorphs, hydrates, anhydrous and other crystalline forms, and combinations thereof. Likewise, all tautomer forms are intended to be included.

[0193] Preferably, the pharmaceutical composition comprises the compound of the present invention as a substantially pure R enantiomer; the pharmaceutical composition comprises the compound of the present invention as a substantially pure S enantiomer; or the pharmaceutical composition comprises the compound of the present invention as an enantiomer mixture containing an excess of the R enantiomer or an excess of the S enantiomer. It is particularly preferable that the pharmaceutical composition contains the compound of the present invention as a substantially pure optical isomer. To avoid any doubt, the compound of the present invention may be used in the form of a solvate if desired.

[0194] synthesis

[0195] Chemical formula ( I Compounds having ) can be prepared using a method similar to the following general synthesis reaction scheme:

[0196]

[0197] Reaction Equation A

[0198] and,

[0199]

[0200] Reaction Equation B

[0201] for example,

[0202]

[0203] and

[0204]

[0205] Chemical formula ( II The compound of ) can be prepared using a method similar to that described in the examples and the general synthesis reaction scheme below:

[0206]

[0207] Reaction equation C

[0208] Chemical formula ( III The compound of ) can be prepared using a method similar to that described in the examples and the general synthesis reaction scheme below:

[0209]

[0210] Reaction Equation D

[0211] Representative heterocycles that can be coupled with halide ions, such as bromide or chloride, include, but are not limited to:

[0212]

[0213] Additional biological active agents and exogenous large porous channel agonists

[0214] As described above, the compounds or compositions of the present invention may be administered together with biological activators. For example, one or more additional biological activators, including those typically used to treat neuroinflammatory diseases, may be used in combination with the compounds or compositions of the present invention described herein. Biological activators include, but are not limited to, TRP1A receptor agonists, TRPV1-4 receptor agonists, TRPM8 agonists, ASIC agonists, P2X receptor agonists, acetaminophen, NSAIDs, glucocorticoids, narcotics, tricyclic antidepressants, amine transporter inhibitors, anticonvulsants, antiproliferative agents and immunomodulators, antibodies or antibody fragments, antibiotics, polynucleotides, polypeptides, proteins, anticancer agents, growth factors, and vaccines.

[0215] TRPV1 agonists that may be used in the methods, kits, and compositions of the present invention include, but are not limited to, any that activate TRPV1 receptors on nociceptors and allow entry of at least one inhibitor (e.g., compounds of the present invention) into voltage-gated ion channels. Suitable TRPV1 agonists are capsaicin or other capsaicinoids that are members of the vanilloid family of molecules. Naturally occurring capsaicinoids are capsaicin itself, dihydrocapsaicin, nordihydrocapsaicin, homodihydrocapsaicin, homocapsaicin, and nonivamid. Other suitable capsaicinoids and capsaicinoid analogs and derivatives for use in the compositions and methods of the present invention are, for example, vanilloids (e.g., N-vanillyl-alkanidienamide, N-vanillyl-alkanidienyl, and N-vanillyl-cis-monounsaturated alkenamide), capsiates, dihydrocapsiates, nordihydrocapsiates and other capsinoids, capsiconiates, dihydrocapsiconiates and other coniferyl esters, capsiconinoids, resiniferatoxin, tinyatoxin, civamide, N-phenylmethylalkenamide capsaicin derivatives, olvanil, N-[(4-(2-aminoethoxy)-3-methoxyphenyl)methyl]-9Z-octa-decaneamide, N-oleyl-homovanilamid, triprenyl It includes naturally occurring and synthetic capsaicin derivatives and analogs, including phenol (e.g., scutigeral), gingerol, piperine, shogaol, guaiacol, eugenol, zingerone, nuvanil, NE-19550, NE-21610, and NE-28345. Additional capsaicinoids, their structures, and methods for their preparation are described in U.S. Patent Nos. 7,446,226 and 7,429,673, which are incorporated herein by reference.

[0216] Additional suitable TRPV1 agonists include eugenol, arvanil (N-arachidonoylvanilamin), anandamide, 2-aminoethoxydiphenyl borate (2APB), AM404, resininiferatoxin, phorbol 12-phenylacetate 13-acetate 20-homovanilate (PPAHV), olvanil (NE 19550), OLDA (N-oleoyldopamine), N-arachidonyldopamine (NADA), 6'-iodoresiniferatoxin (6'-IRTX), C18 N-acylethanolamine, lipoxygenase derivatives, e.g., 12-hydroperoxyeicosatetraenoic acid, inhibitor cysteine ​​knot (ICK) peptide (vanillotoxin), piperine. MSK195 (N-[2-(3,4-dimethylbenzyl)-3-(pivaloyloxy)propyl]-2-[4-(2-aminoethoxy)-3-methoxyphenyl]acetamide), JYL79 (N-[2-(3,4-dimethylbenzyl)-3-(pivaloyloxy)propyl]-N'-(4-hydroxy-3-methoxybenzyl)thiourea), hydroxy-alpha-sanshool, 2-aminoethoxydiphenyl borate, 10-shogaol, oleylgingerol, oleylshogaol and SU200 (N-(4-tert-butylbenzyl)-N'-(4-hydroxy-3-methoxybenzyl)thiourea), but is not limited thereto.Other TRPV1 agonists include amylocaine, articaine, benzocaine, bupivacaine, carbocaine, carticaine, chloroprocaine, cyclomethicaine, dibucaine (cinchocaine), dimethocaine (larocaine), etidocaine, hexylcaine, levobupivacaine, lidocaine, mepivacaine, meprylcaine (oracaine), metabutoxycaine, piperocaine, prilocaine, procaine (novacaine), proparacaine, propoxycaine, and risocaine. Includes ropivacaine, tetracaine (amethocaine) and trimecaine.

[0217] Suitable TRPV2-4 agonists are 2-APB, cannabinol, diphenylboronic anhydride, insulin-like growth factor 1, lysophosphatidylcholine, lysophosphatidylinositol, probenecid, Δ9-tetrahydrocannabinol, vanillin, eugenol, cinnamaldehyde, camphor, carvacrol, thymol, citral, farnesyl diphosphate, tetrahydrocannabivarin, Incenseol acetate, diphenylboronic anhydride, 6-tert-butyl-m-cresol, dihydrocarbeocarbeol, borneol, (-)-menthol, GSK1016790A, 4α-PDH, 5,6-epoxyeicosatrienoic acid, 4α-PDD, bisandrographolide, citric acid, phorbol 12-myristate 13-acetate and Includes, but is not limited to, RN1747.

[0218] Suitable TRPM8 agonists include, but are not limited to, menthol, icilin, eucalyptus, linalool, geraniol, hydroxy-citronellal, WS-3, WS-23, Frescolat MGA, Frescolat ML, PMD 38, CPS125, Coolact P, M8-Ag, AITC, cryosim-3, and Cooling Agent 10.

[0219] Suitable ASIC agonists include, but are not limited to, chlorophenylguanidine hydrochloride, GMQ hydrochloride, tetrahydropapaverolin (THP), reticulin, polyamine agmatine, lysophosphatidylcholine, arachidonic acid, and neuropeptide SF.

[0220] Other biological activators that may be used in the methods, compositions, and kits of the present invention include any that activate TRP1A receptors on nociceptors or itchiness receptors and allow the entry of at least one inhibitor of voltage-gated ion channels. Suitable TRP1A agonists include, but are not limited to, cinnamaldehyde, allyl-isothiocyanate (mustard oil), diallyl disulfide, icyl, cinnamon oil, wintergreen oil, clove oil, acrolein, hydroxy-alpha-sanshool, 2-aminoethoxydiphenyl borate, 4-hydroxynonenal, methyl p-hydroxybenzoate, and 3'-carbamoylbiphenyl-3-yl cyclohexylcarbamate (URB597).

[0221] P2X agonists that may be used in the methods, compositions, and kits of the present invention include any that activate P2X receptors on nociceptors or itchiness receptors and allow entry of at least one inhibitor of voltage-gated ion channels. Suitable P2X agonists include, but are not limited to, ATP, α,β-methylene ATP, 2-methylthio-ATP, 2' and 3'-O-(4-benzoylbenzoyl)-ATP, and ATP5'-O-(3-thiotriphosphate).

[0222] Other biological active agents that can be used in combination with the compounds of the present invention include NSAIDs, glucocorticoids, narcotics, tricyclic antidepressants, amine transporter inhibitors, anticonvulsants, antiproliferative agents and immunomodulators, antibodies or antibody fragments, antibiotics, polynucleotides, polypeptides, proteins, anticancer agents, growth factors, and vaccines.

[0223] Non-steroidal anti-inflammatory drugs (NSAIDs) that may be administered to a patient (e.g., human) suffering from neuroinflammatory disease in combination with the composition of the present invention are acetylsalicylic acid, amoxiprin, benorylate, benorylate, choline magnesium salicylate, diflunisal, ethenzamide, faislamine, methyl salicylate, magnesium salicylate, salicyl salicylate, salicylamide, diclofenac, aceclofenac, acemethacin, aclofenac, bromfenac, etodolac, indomethacin, nabumetone, oxametacin, Proglumetacin, Sulindac, Tolmetin, Ibuprofen, Alminoprofen, Benoxaprofen, Carprofen, Dexibuprofen, Dexketoprofen, Fenbufen, Fenoprofen, Flunoxaprofen, Flurbiprofen, Ibuproxam, Indoprofen, Ketoprofen, Ketorolac, Loxoprofen, Naproxen, Oxaprozin, Pirprofen, Suprofen, Tiaprofenic acid, Mefenamic acid, flufenamic acid, meclofenamic acid, tolfenamic acid,phenylbutazone, ampyrone, azapropazone, clofezone, kebuzone, metamizole, mofebutazone, oxyphenbutazone, phenazone, sulfinpyrazone, piroxicam, droxicam, lornoxicam, meloxicam, tenoxicam, and COX-2 inhibitors celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, valdecoxib, and pharmaceutically acceptable salts thereof, wherein It is not limited.

[0224] Glucocorticoids that may be administered to a patient (e.g., human) suffering from neuroinflammatory disease in combination with the composition of the present invention include, but are not limited to, hydrocortisone, cortisone acetate, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, fludrocortisone acetate, deoxycorticosterone acetate, aldosterone, and pharmaceutically acceptable salts thereof.

[0225] Narcotic agents that may be administered to a patient (e.g., human) suffering from neuroinflammatory disease in combination with the composition of the present invention include, but are not limited to, tramadol, hydrocodone, oxycodone, morphine, and pharmaceutically acceptable salts thereof.

[0226] Antiproliferative agents and immunomodulators that may be administered to a patient (e.g., human) suffering from neuroinflammatory disease in combination with the composition of the present invention include, but are not limited to, alkylating agents, platinum agents, antimetabolites, focal isomerase inhibitors, dihydrofolate reductase inhibitors, antitumor antibiotics, antimitotic agents, aromatase inhibitors, thymidylate synthase inhibitors, DNA antagonists, farnesyltransferase inhibitors, pump inhibitors, histone acetyltransferase inhibitors, metalloproteinase inhibitors, ribonucleoside reductase inhibitors, TNF-alpha agonists, TNF-alpha antagonists or scavengers, interleukin 1 (IL-1) antagonists or scavengers, endothelin A receptor antagonists, retinoic acid receptor agonists, hormones, antihormones, photodynamic agents, and tyrosine kinase inhibitors.

[0227] The biological active agent may be administered before, simultaneously with, or after the administration of the composition of the present invention using any formulation, dosage, or administration known in the art that is therapeutically effective.

[0228] Formulation of the composition

[0229] The administration of the compound of the present invention may be carried out by any suitable means that causes a reduction in perceived pain sensation in the target area. The compound of the present invention may be contained in any suitable amount in any suitable carrier material, which is generally present in an amount of 1-99% by weight of the total weight of the composition. The composition may be provided in a form of administration suitable for oral, parenteral (e.g., intravenous, intramuscular), rectal, skin, subcutaneous, topical, transdermal, sublingual, nasal, vaginal, intrathecal, epidural, or ocular administration, or may be provided by injection, inhalation, or direct contact with the nasal or oral mucosa.

[0230] Accordingly, the composition may be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, plasters, drenches, osmotic delivery devices, suppositories, enemas, injections, implants, sprays, or aerosols. The composition may be formulated according to ordinary pharmaceutical practice (e.g., the literature [Remington: The Science and Practice of Pharmacy, 22nd edition, 2013, ed. LV Allen, Pharmaceutical Press, Philadelphia, and Encyclopedia of Pharmaceutical Technology, 4 th Edition, ed. J. Swarbrick, 2013, CRC Press, New York].

[0231] Each compound may be formulated in various ways known in the art. For example, the compounds of the present invention and biological active agents as defined herein may be formulated together or separately. Preferably, the compounds and biological active agents of the present invention are formulated together for simultaneous or near-simultaneous administration thereof. In another embodiment, two or more biological active agents may be formulated together with or separately from the compounds of the present invention. Other examples include, but are not limited to, two or more compounds of the present invention formulated together, said compounds being formulated with or without one or more biological active agents.

[0232] Preparations formulated individually or separately may be packaged together as a kit. Non-limiting examples include, but are not limited to, a kit containing, for example, two pills, a pill and a powder, a suppository and a liquid in a vial, two topical creams, etc. The kit may include optional components that aid in administering a unit dose to a patient, such as a vial for reconstitution of the powder form, an injectable syringe, a customized IV delivery system, an inhaler, etc. Additionally, the unit dose kit may include instructions for the preparation and administration of the composition.

[0233] The kit may be manufactured as a single-use unit dose for one patient or as a multiple-use for a specific patient (at a constant dose or where the efficacy of individual compounds may vary as the regimen progresses); or the kit may contain multiple doses suitable for administration to a number of patients (“bulk packaging”). Kit components may be assembled in a carton, blister pack, bottle, tube, etc.

[0234] Controlled-release formulation

[0235] Each compound of the present invention, either alone or in combination with one or more of the biological active agents described herein, may be formulated for controlled-release (e.g., sustained or measured) administration as described in U.S. Patent Application Publications No. 2003 / 0152637 and 2005 / 0025765, each incorporated herein by reference. For example, a compound of the present invention, either alone or in combination with one or more of the biological active agents described herein, may be incorporated into a capsule or tablet administered to a patient.

[0236] Any pharmaceutically acceptable vehicle or formulation suitable for topical application and / or injection to a site to be treated (e.g., painful surgical incision, wound, or joint) capable of providing sustained release of the compound of the present invention, either alone or in combination with one or more of the biological active agents described herein, may be used as needed to provide long-term clearance or relief of inflammation. Control-release formulations known in the art comprise specially coated pellets, polymer formulations, or matrices for surgical insertion, or sustained-release microparticles, e.g., microspheres or microcapsules, for implantation, insertion, infusion, or injection, wherein the slow release of the active agent occurs through sustained or controlled diffusion outside the matrix and / or selective degradation of the product coating or selective degradation of the polymer matrix. Other formulations or vehicles for controlled, sustained, or immediate delivery of the agent to a desired local site of a patient include, e.g., suspensions, emulsions, gels, liposomes, and any other suitable delivery vehicle or formulation known in the art that is acceptable for subcutaneous or intramuscular administration.

[0237] A wide variety of biocompatible materials may be used as controlled-release carriers to provide controlled release of the compounds of the present invention, either alone or in combination with one or more biological activators as described herein. Any pharmaceutically acceptable biocompatible polymer known to those skilled in the art may be used. It is desirable that the biocompatible controlled-release material degrades in vivo within about one year, preferably within about three months, and more preferably within about two months. More preferably, the controlled-release material will degrade significantly within one to three months, at least 50% of the material will degrade into non-toxic residues and be eliminated by the body, and 100% of the compounds of the present invention will be released within about two weeks, preferably within about two to seven days. The degradable controlled-release material should preferably degrade by hydrolysis due to surface erosion or bulk erosion to provide not only sustained release but also a desirable release rate. However, the pharmacokinetic release profiles of these formulations may be primary, zero, secondary, or multiple to provide the desired reversible local anti-nourishing effect over the desired period.

[0238] Suitable biocompatible polymers may be used as controlled-release materials. Polymeric materials may include biocompatible, biodegradable polymers, and in certain preferred embodiments, preferably copolymers of lactic acid and glycolic acid. Preferred controlled-release materials useful for the formulations of the present invention include polyanhydrides, polyesters, copolymers of lactic acid and glycolic acid (preferably, the weight ratio of lactic acid to glycolic acid is 4:1 or less, i.e., 80 weight% or less of lactic acid to 20 weight% or more of glycolic acid), and polyorthoesters containing a catalyst or degradation-enhancing compound, and, for example, at least 1 weight% of an anhydride catalyst such as maleic anhydride. Examples of polyesters include polylactic acid, polyglycolic acid, and polylactic acid-polyglycolic acid copolymers. Other useful polymers include protein polymers such as collagen, gelatin, fibrin, and fibrinogen, and polysaccharides such as hyaluronic acid.

[0239] Polymeric materials may be prepared by any method known to those skilled in the art. For example, if the polymeric material comprises a copolymer of lactic acid and glycolic acid, such copolymer may be prepared by the procedure described in U.S. Patent No. 4,293,539, incorporated herein by reference. Alternatively, copolymers of lactic acid and glycolic acid may be prepared by any other procedure known to those skilled in the art. Other useful polymers include polylactide, polyglycolide, polyanhydride, polyorthoester, polycaprolactone, polyphosphazene, polyphosphoester, polysaccharide, proteinaceous polymer, soluble derivative of polysaccharide, soluble derivative of proteinaceous polymer, polypeptide, polyester and polyorthoester, or mixtures or blends of any of these.

[0240] The pharmaceutically acceptable polyanhydrides useful in the present invention have anhydrous bonds that are unstable in water. The drug release rate can be controlled by the specific polyanhydride polymer used and its molecular weight. The polysaccharide may be a poly-1,4-glucan, for example, starch glycogen, amylose, amylopectin, and mixtures thereof. The biodegradable hydrophilic or hydrophobic polymer may be a water-soluble derivative of poly-1,4-glucan, including hydrolyzed amylopectin, derivatives of hydrolyzed amylopectin, for example, hydroxyethyl starch (HES), hydroxyethyl amylose, dialdehyde starch, etc. The polyanhydride polymer may be branched or linear.

[0241] Examples of polymers useful in the present invention include (in addition to homopolymers and copolymers of poly(lactic acid) and / or poly(glycolic acid)) poly[bis(p-carboxyphenoxy)propane anhydride] (PCPP), poly[bis(p-carboxy)methane anhydride] (PCPM), polyanhydrides of oligomerized unsaturated aliphatic acids, polyanhydride polymers prepared from amino acids modified to include additional carboxylic acids, aromatic polyanhydride compositions, and copolymers of polyanhydrides with other materials, e.g., fatty acid-terminated polyanhydrides, e.g., polyanhydrides polymerized from monomers of dimers and / or trimers of unsaturated fatty acids or unsaturated aliphatic acids. Polyanhydrides may be prepared according to the method described in U.S. Patent No. 4,757,128, which is incorporated herein by reference. Polyorthoester polymers may be manufactured, for example, as described in U.S. Patent No. 4,070,347, incorporated herein by reference. Polyphosphoesters may be manufactured and used as described in U.S. Patent Nos. 6,008,318, 6,153,212, 5,952,451, 6,051,576, 6,103,255, 5,176,907, and 5,194,581, each incorporated herein by reference.

[0242] Protein polymers may also be used. Protein polymers and their soluble derivatives include gelling biodegradable synthetic polypeptides, elastin, alkylated collagen, alkylated elastin, etc. Biodegradable synthetic polypeptides include copolymers of poly-(N-hydroxyalkyl)-L-asparagine, poly-(N-hydroxyalkyl)-L-glutamine, N-hydroxyalkyl-L-asparagine, and N-hydroxyalkyl-L-glutamine with other amino acids. The proposed amino acids include L-alanine, L-lysine, L-phenylalanine, L-valine, L-tyrosine, etc.

[0243] In additional embodiments, a release control material that acts as a carrier for a compound of the present invention, either alone or in combination with one or more biological active agents as described herein, may further comprise a bioadhesive polymer, e.g., pectin (polygalacturonic acid), mucopolysaccharide (hyaluronic acid, mucin), or non-toxic lectin, or the polymer itself may be a bioadhesive, e.g., a polysaccharide such as polyanhydride or chitosan.

[0244] In an embodiment in which a biodegradable polymer comprises a gel, one of the useful polymers is a heat-gelling polymer, e.g., a polyethylene oxide, polypropylene oxide (PEO-PPO) block copolymer, e.g., BASF Wyandotte’s Pluronic™ F127. In the above case, the local anesthetic formulation may be injected via a syringe as a free-flowing liquid that gels rapidly at 30°C or higher (e.g., when injected into a patient). Subsequently, the gel system releases a steady dose of the compound of the present invention at the site of administration, either alone or in combination with one or more biological active agents as described herein.

[0245] Dosage form for oral use

[0246] Formulations for oral use comprise tablets containing the active ingredient(s) mixed with non-toxic, pharmaceutically acceptable excipients. These excipients are, for example, inactive diluents or fillers (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulizing agents and disintegrants (e.g., cellulose derivatives including microcrystalline cellulose, starch including potato starch, sodium croscarmellose, alginate, or alginic acid); Binders (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone or polyethylene glycol); and lubricants, lubricants, and anti-adhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients may be colorants, flavorings, plasticizers, wetting agents, buffers, taste masking agents (e.g., hydroxypropyl methylcellulose, hydroxypropyl cellulose), etc.

[0247] One or more compounds of the present invention and one or more biological active agents as defined herein may be mixed with or divided into tablets, capsules, or other vehicles. In one example, the compound of the present invention is contained inside a tablet and the biological active agent is contained outside the tablet, so that a substantial portion of the biological active agent is released before the release of the compound of the present invention.

[0248] Formulations for oral use may also be provided as chewable tablets, or as hard gelatin capsules in which the active ingredient is mixed with an inactive solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with a water or oil medium, e.g., peanut oil, liquid paraffin, or olive oil. Powders, granules, and pellets may be prepared using the aforementioned ingredients in tablets and capsules in a conventional manner, for example, using a mixer, a fluid bed device, or spray drying equipment.

[0249] Formulations for oral administration into the mouth may also be provided as mouthwash, oral spray, oral rinse solution, oral ointment, or oral gel.

[0250] Dissolution or diffusion-controlled release may be achieved by a suitable coating of a tablet, capsule, pellet, or granule formulation of the compound, or by incorporating the compound into a suitable matrix. The controlled release coating may comprise one or more of the coating materials mentioned above and / or, for example, shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resin, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-hydroxymethacrylate, methacrylate hydrogel, 1,3-butylene glycol, ethylene glycol methacrylate, and / or polyethylene glycol. In a controlled-release matrix formulation, the matrix material may also include, for example, hydrated methylcellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbons.

[0251] Liquid forms in which compounds and compositions of the present invention may be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions having edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0252] Generally, when administered to humans, the oral dosage of any compound of the combination of the present invention will depend on the properties of the compound and can be easily determined by a person skilled in the art. Typically, the dosage is generally about 0.001 mg to 2000 mg per day, preferably about 1 mg to 1000 mg per day, more preferably about 5 mg to 500 mg per day. A maximum dosage of 200 mg per day may be required.

[0253] As described herein, in combination therapy, the administration of each drug may be independent, from one to four times daily for one day to one year, and even for the patient's lifetime. In many cases, chronic long-term administration will be indicated.

[0254] parenteral formulation

[0255] Formulations suitable for parenteral administration (e.g., by injection) comprise aqueous or non-aqueous, isotonic, pyrogen-free, sterile liquids (e.g., solutions, suspensions), wherein the compound is dissolved, suspended, or otherwise provided (e.g., in liposomes or other microparticles). The liquid may further contain other pharmaceutically acceptable components, e.g., antioxidants, buffers, preservatives, stabilizers, bacteriostatic agents, suspending agents, thickeners, and solutes that make the formulation isotonic with the blood (or other related body fluids) of the intended recipient. Examples of excipients include, e.g., water, alcohol, polyol, glycerol, vegetable oil, etc. Examples of suitable isotonic carriers for use in the formulation include sodium chloride injection, Ringer's solution, or lactated Ringer's injection. Typically, the concentration of the compound in the liquid is about 1 ng / ml to about 10 μg / ml, for example, about 10 ng / ml to about 1 μg / ml. The formulation may be provided in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored under freeze-drying conditions requiring only the addition of a sterile liquid carrier, for example, water for injection, immediately before use. Immediate injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0256] Topical formulations

[0257] Compositions of the present invention, alone or in combination with one or more of the biological active agents described herein, may also be adapted for topical use with a topical vehicle containing 0.0001% to 25% (w / w) or more of active ingredient(s).

[0258] In a preferred combination, the active ingredient is preferably 0.0001% to 10% (w / w) each, more preferably 0.0005% to 4% (w / w) of active ingredient. Topical formulations, including but not limited to creams, gels, or ointments, may be applied 1 to 4 times a day or as needed. By carrying out the method described herein, a topical vehicle containing a composition of the present invention or a combination therapy containing a composition of the present invention is preferably applied to the inflamed area of ​​a patient. For example, a cream may be applied to the hand of a patient suffering from an arthritic finger.

[0259] The composition may be formulated using any dermatologically acceptable carrier. Exemplary carriers include solid carriers, e.g., alumina, clay, microcrystalline cellulose, silica, or talc; and / or liquid carriers, e.g., alcohol, glycol, or aqueous alcohol / glycol blends. The therapeutic agent may also be administered in a liposomal formulation that allows the therapeutic agent to enter the skin. The liposomal formulations are U.S. Patent Nos. 5,169,637; 5,000,958; 5,049,388; 4,975,282; 5,194,266; 5,023,087; 5,688,525; 5,874,104; 5,409,704; 5,552,155; 5,356,633; 5,032,582; It is described in Patent No. 4,994,213; Patent No. 8,822,537, and PCT Publication No. WO 96 / 40061. Examples of other suitable vehicles are described in U.S. Patent No. 4,877,805, Patent No. 8,822,537, and EP Publication No. 0586106A1. Suitable vehicles of the present invention may also include inorganic oil, petroleum jelly, polydecene, stearic acid, isopropyl myristate, polyoxyl 40 stearate, stearyl alcohol, or vegetable oil.

[0260] The composition may further comprise a skin penetration enhancer such as that described in the literature ["Percutaneous Penetration enhancers", (eds. Smith EW and Maibach H I. CRC Press 1995)]. Exemplary skin penetration enhancers are alkyl (N,N-disubstituted aminoalkanoate) esters described in U.S. Patent Nos. 6,083,996 and 6,118,020, both of which are incorporated herein by reference, e.g., dodecyl 2-(N,N dimethylamino)propionate (DDAIP); Water-dispersible acid polymers, e.g., polyacrylic acid polymers, carbomers (e.g., Carbopol™ or Carbopol 940P™, available from BF Goodrich Company (Akron, Ohio)), copolymers of polyacrylic acid (e.g., Pemulen™ from BF Goodrich Company or Polycarbophil™ from AH Robbins, Richmond, Va.); polysaccharide gums, e.g., agar gum, alginate, carrageenan gum, ghatti gum, karaya gum, kadaya gum, ramicane gum, xanthan gum, and galactomannan gum (e.g., guar gum, carob gum, and locust bean gum), as well as other gums known in the art (e.g., Industrial Gums: Polysaccharides & Their Derivatives, Whistler RL, BeMiller JN (eds.), 3rd Ed. Academic Press (1992) and Davidson, RL, Handbook of Water-Soluble [See Gums & Resins, McGraw-Hill, Inc., NY (1980)]); or a combination thereof.

[0261] Other suitable polymeric skin penetration enhancers are cellulose derivatives, e.g., ethyl cellulose, methyl cellulose, and hydroxypropyl cellulose. Additionally, known transdermal penetration enhancers may also be added if desired. Examples include dimethyl sulfoxide (DMSO) and dimethyl acetamide (DMA), 2-pyrrolidone, N,N-diethyl-m-toluamide (DEET), 1-dodecyl azacycloheptan-2-one (Azone™, a registered trademark of Nelson Research), N,N-dimethylformamide, N-methyl-2-pyrrolidone, calcium thioglycolate, and other enhancers, e.g., dioxolane, cyclic ketones, and derivatives thereof.

[0262] Also examples are a group of biodegradable absorption enhancers that are alkyl N,N-2-(disubstituted amino)alkanoates, such as those described in U.S. Patent No. 4,980,378 and U.S. Patent No. 5,082,866, both of which are incorporated herein by reference, including tetradecyl (N,N-dimethylamino) acetate, dodecyl (N,N-dimethylamino) acetate, decyl (N,N-dimethylamino) acetate, octyl (N,N-dimethylamino) acetate, and dodecyl (N,N-diethylamino) acetate.

[0263] Particularly preferred skin penetration enhancers are isopropyl myristate; isopropyl palmitate; dimethyl sulfoxide; decyl methyl sulfoxide; dimethylalanine amide of a medium-chain fatty acid; dodecyl 2-(N,N-dimethylamino)propionate as described in U.S. Patent No. 6,118,020 or a salt thereof, e.g., its organic salts (e.g., hydrochloric acid, hydrobromide, sulfuric acid, phosphoric acid, and nitric acid addition salts) and inorganic salts (e.g., acetic acid, benzoic acid, salicylic acid, glycolic acid, succinic acid, nicotinic acid, tartaric acid, maleic acid, malic acid, parmolic acid, methanesulfonic acid, cyclohexanesulfamic acid, picric acid, and lactic acid addition salts); and includes alkyl 2-(N,N-disubstituted amino)-alkanoates as described in U.S. Patent No. 4,980,378 and U.S. Patent No. 5,082,866.

[0264] The skin penetration enhancer of this composition will be in the range of 0.5% to 10% (w / w) by weight. The most preferred range will be 1.0% to 5% (w / w). In another embodiment, the skin penetration enhancer constitutes 0.5%-1%, 1%-2%, 2%-3%, 3%-4%, or 4%-5% (w / w) of the composition.

[0265] The composition may be provided in any useful form. For example, the composition of the present invention may be formulated as a solution, emulsion (including microemulsion), suspension, cream, ointment, foam, lotion, gel, powder, or other typical solid, semi-solid, or liquid composition (e.g., topical spray) used for application to skin or other tissues where the composition may be used. The above composition comprises other ingredients typically used in the product, e.g., colorants, fragrances, thickeners (e.g., xanthan gum, fatty acids, fatty acid salts or esters, fatty alcohols, modified cellulose, modified inorganic materials, Krisgel 100™, or synthetic polymers), antimicrobial agents, solvents, surfactants, detergents, gelling agents, antioxidants, fillers, dyes, viscosity modifiers, preservatives, wetting agents, emollients (e.g., natural or synthetic oils, hydrocarbon oils, waxes or silicones), wettable agents, chelating agents, emollients, solubilizing excipients, adjuvants, dispersants, skin penetration enhancers, plasticizers, preservatives, stabilizers, anti-emulsifiers, humidifiers, UV blockers, emulsifiers, moisturizers, astringents, deodorizers, and optionally anesthetics, anti-itch agents, plant extracts, conditioning agents, darkening or lightening agents, glitter, wetting agents, mica, inorganic materials, polyphenols, It may include silicone or derivatives thereof, sunscreens, vitamins, and phytomedicines.

[0266] The composition may also include other similar ingredients to provide additional benefits and to improve the feel and / or appearance of the topical formulation. Specific classes of additives commonly used in these formulations include isopropyl myristate, sorbic acid NF powder, polyethylene glycol, phosphatidylcholine (a mixture of phosphatidylcholine, e.g., Phospholipone G), Krisgel 100™ distilled water, sodium hydroxide, decyl methyl sulfoxide (as a skin penetration enhancer), menthol crystals, lavender oil, butylated hydroxytoluene, ethyl diglycol reagent, and 95% (190 proof) ethanol.

[0267] Ophthalmic formulation

[0268] The compounds of the present invention may also be formulated with an ophthalmologically acceptable carrier at a concentration sufficient to deliver an effective amount of the active compound or compounds to the optic nerve area of ​​the eye. Preferably, the ophthalmic therapeutic solution contains one or more active compounds in a concentration range of approximately 0.0001% to approximately 5% (weight / volume), more preferably approximately 0.0005% to approximately 0.1% (weight / volume).

[0269] Ophthalmologically acceptable carriers do not cause significant irritation to the eye and do not negate the pharmacological activity and properties of charged sodium channel blockers.

[0270] Ophthalmologically acceptable carriers are generally sterile and essentially free of foreign particles, and generally have a pH in the range of 5 to 8. Preferably, the pH is as close as possible to the pH of tears (7.4). Ophthalmologically acceptable carriers are, for example, sterile isotonic solutions, for example, isotonic sodium chloride or boric acid solutions. The carrier is typically an aqueous solution containing sodium chloride or boric acid. Additionally, phosphate-buffered saline (PBS) solutions are useful.

[0271] Various preservatives may be used in ophthalmic formulations. Preferred preservatives include, but are not limited to, potassium benzalkonium, chlorobutanol, thimerosal, phenylmercury acetate, and phenylmercury nitrate. Likewise, various preferred vehicles may be used in the ophthalmic formulations. These vehicles include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropyl methylcellulose, poloxamer, carboxymethylcellulose, and hydroxyethylcellulose.

[0272] Tense modifiers may be added as needed or for convenience. These include, but are not limited to, salts, particularly sodium chloride, potassium chloride, etc., mannitol and glycerin, or any other suitable ophthalmologically acceptable tone modifier.

[0273] Various buffers and means for adjusting pH may be used as long as the resulting product is sufficiently acceptable. Thus, buffers include, but are not limited to, acetate buffers, citrate buffers, phosphate buffers, and borate buffers. Acids or bases may be used to adjust the pH of these formulations as needed. Ophthalmologically acceptable antioxidants may also be included. Antioxidants include, but are not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole, and butylated hydroxytoluene.

[0274] Formulations for nasal and inhalation administration

[0275] The pharmaceutical composition of the present invention may be formulated for nasal or intranasal administration. When the carrier is a solid, a formulation suitable for nasal administration comprises, for example, a coarse powder having a particle size in the range of approximately 20 to 500 microns, which is administered by rapid inhalation through the nasal cavity. When the carrier is a liquid, for example, a nasal spray or nasal drop, one or more formulations may be mixed in an aqueous or oily solution and may be inhaled or sprayed into the nasal cavity.

[0276] In the case of administration by inhalation, the active ingredient may be conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve for delivering a metered amount. Capsules and cartridges, for example, of gelatin, containing a powder mixture of the compound and a suitable powder base, e.g., lactose or starch, may be formulated for use in an inhaler or blower.

[0277] A dry powder composition for local delivery to the lungs by inhalation may be provided in capsules and cartridges, for example, of laminated aluminum foil, for example, gelatin, or blister, for use in an inhaler or blower. Powder blend formulations generally contain a powder mixture for inhalation of the compounds of the present invention and a suitable powder base (carrier / diluent / excipient material), for example, a monosaccharide, disaccharide, or polysaccharide (e.g., lactose or starch). The use of lactose is preferred. In one embodiment, each capsule or cartridge contains about 2 µg to about 100 mg of the formula (optionally combined with another therapeutic active ingredient) IIt may contain a compound of ). In a preferred embodiment, each capsule or cartridge contains about 10 µg to about 50 mg of the formula (optionally combined with another therapeutic active ingredient) I It may contain a compound of ). In another embodiment, each capsule or cartridge contains about 20 µg to about 10 mg of the formula (optionally combined with another therapeutic active ingredient) I It may contain a compound of ). Alternatively, the compound of the present invention may be delivered without excipients.

[0278] Suitablely, the packaging / drug dispenser is a type selected from the group consisting of reservoir dry powder inhalers (RDPI), unit dose dry powder inhalers (e.g., capsule or blister pack inhalers), multi-dose dry powder inhalers (MDPI), and metered-dose inhalers (MDI).

[0279] A solution or suspension for use in a pressurized vessel, pump, spray, atomizer, or nebulizer may be formulated to contain an aqueous medium, ethanol, aqueous ethanol, or suitable alternative formulation for dispersing, solubilizing, or extending the release of the active ingredient(s); a propellant as a solvent; and / or a surfactant, e.g., sorbitan trioleate, oleic acid, or oligolactic acid.

[0280] Compositions formulated for nasal or inhalation administration may comprise one or more taste masking agents, e.g., flavoring agents, sweeteners, and other agents, e.g., sucrose, dextrose, and lactose, carboxylic acids, menthol, amino acids or amino acid derivatives, e.g., arginine, lysine, and monosodium glutamate, and / or synthetic flavoring oils and flavoring agents and / or natural oils, extracts from plants, leaves, flowers, fruits, etc., and combinations thereof. These may comprise cinnamon oil, wintergreen oil, peppermint oil, clover oil, bay oil, anise oil, eucalyptus, vanilla, citrus oils, e.g., lemon oil, orange oil, grape and grapefruit oil, and fruit essences including apple, peach, pear, strawberry, raspberry, cherry, plum, pineapple, apricot, etc. Additional sweeteners include sucrose, dextrose, aspartame, acesulfame-K, sucralose and saccharin, and organic acids (non-limiting examples include citric acid and aspartic acid). The flavoring agent may be present in an amount of about 0.05 to about 4 weight percent and may be present in a smaller or larger amount due to one or more factors of efficacy of the effect on the flavoring agent, the solubility of the flavoring agent, the solubility of other formulation components or the effect of the flavoring agent on other physicochemical or pharmacokinetic properties, or other factors.

[0281] Indications

[0282] The compounds, compositions, methods, and kits of the present invention are for trigeminal neurotrophic syndrome, erythematous limb pain, back and neck pain, low back pain, cancer pain, gynecological pain and analgesia, abdominal wall pain, chronic abdominal wall pain, fibromyalgia, allergic rhinitis, arthritis, rheumatoid arthritis, osteoarthritis, rheumatic pain, orthopedic pain, acute and post-herpetic neuralgia and other neuropathic pain (including peripheral neuropathy), sickle cell crisis, myalgia, vulvodynia, rectal pain, levator ani syndrome, transient rectal pain, perianal pain, hemorrhoidal pain, stomach pain, ulcer, inflammatory bowel disease, irritable bowel disease, irritable bowel syndrome, oral mucositis, esophagitis, interstitial cystitis, urethritis and other urological pain, toothache, burn pain, headache, ophthalmic irritation, conjunctivitis (e.g., allergic conjunctivitis), eye hyperemia, dry eye, dry eye syndrome (chronic eye pain), complex It can be used to treat pain, cough, or itching associated with any of the following conditions, including regional pain syndrome, acute postoperative pain, postoperative pain, postoperative ocular pain, and procedure pain (i.e., injection, abscess drainage, surgery, dental procedure, ophthalmic procedure, ophthalmic irritation, conjunctivitis (e.g., allergic conjunctivitis), eye redness, dry eye, pain associated with the use of arthroscopy and other medical devices, cosmetic surgery procedures, dermatological procedures, bone setting, biopsy, etc.).

[0283] Since a subclass of nociceptors mediates the sensation of itching, the compounds, compositions, methods, and kits of the present invention may also be used to treat itching in patients with conditions such as pruritus (including but not limited to brachioradialis muscle, chronic idiopathic, genital / anal, paresthetic back pain, and scalp), allergic dermatitis, atopic dermatitis, contact dermatitis, poison ivy, infection, parasite, insect bite, pregnancy, metabolic disorder, hepatic or renal failure, drug reaction, allergic reaction, eczema, hand eczema, genital and anal itching, hemorrhoidal itching, and cancer.

[0284] Since subclasses of nociceptors can initiate an abnormal cough reflex, the compounds, compositions, methods, and kits of the present invention can also be used to treat cough in patients with diseases such as asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), idiopathic pulmonary fibrosis, postviral cough, post-infectious cough, chronic idiopathic cough, and lung cancer.

[0285] The compounds, compositions, methods, and kits of the present invention may also be used to treat neuroinflammatory and neuroinflammatory disorders. Inflammation is a complex set of responses to noxious stimuli that cause localized redness, swelling, and pain. Inflammation can be innate or adaptive; the latter is induced by antigens and mediated by immune cells (immune-mediated inflammation). Neuroinflammatory inflammation arises from the efferent function of pain-sensing neurons (nociceptors), and neuropeptides and other chemicals that are pro-inflammatory mediators are released from them when the peripheral terminals of nociceptors are activated. This release process is mediated by calcium influx and the exocytosis of peptide-containing vesicles, and pro-inflammatory neuropeptides include substance P, neurokinins A and B (collectively known as tachykinins), calcitonin gene-associated peptide (CGRP), and vasoactive intestinal polypeptide (VIP).

[0286] The release of peripheral endogenous chemicals stimulates various inflammatory responses. First, the release of substance P increases capillary permeability, causing plasma proteins to leak from intravascular compartments into the extracellular space (plasma leakage), which can induce edema. This can be detected as wheals (firm, raised swelling of the skin), which are one of the three components of the inflammatory response known as the Lewis triple reaction: wheals, red spots, and redness. Second, the release of CGRP causes vasodilation, resulting in increased blood flow. This can be detected as redness, another component of the Lewis triple reaction.

[0287] Substance P also exerts pro-inflammatory effects on immune cells (e.g., macrophages, T-cells, mast cells, and dendritic cells) through neurokinin-1 (NK1) receptors. These effects have been recorded in allergic rhinitis, gastritis, and colitis, representing the interface between the neurogenic and immune-mediated components of inflammation. Substance P released from one nociceptor can also act on NK1 receptors on neighboring nociceptors to sensitize or activate them, thereby inducing activation and the diffusion of afferent / efferent functions. These efferent functions of nociceptors include 1) direct activation of the nociceptor terminal by appropriate peripheral stimulation (e.g., stinging pain) applied to the terminal; 2) indirect reverse activation of an unstimulated nociceptor terminal by an axonal reflex, wherein an action potential input from one nociceptor terminal reaches an axonal branch converging at the periphery, thereby generating an action potential moving from the branch to the unstimulated terminal's periphery; and 3) activation as a result of activation at the nociceptor central terminal of the CNS moving to the periphery (e.g., primary afferent depolarization of the central terminal generated by GABA may be sufficient to initiate an action potential moving in the "wrong direction").

[0288] Genomic analysis of lung-resident ILC2 cells revealed the expression of receptors for various neuropeptides released by sensory neurons, including SP, CGRP, and VIP, providing an opportunity for nociceptors to communicate directly with these cells. In particular, VIP was found to be expressed in NaV1.8+ nodular ganglion neurons, including lung afferents, in OVA-exposed mice. Cultured nodular ganglion neurons stimulated with capsaicin or IL5 also released VIP, whereas BALF from OVA-exposed mice contained elevated VIP compared to vehicle-attacked mice (Talbot et al., Neuron. 2015 July 15; 87(2): 341-354). These data show that VIP is released in inflamed lungs and can be blocked by silencing neurons using the charged sodium channel blocker of the present invention. Also, T H 2. When CD4+ T cells cultured under biased conditions were exposed to recombinant mouse VIP, transcript levels of IL-13 and IL-5 increased, indicating that VIP is responsible for the transcription of these type II regulatory cytokines in T cells. H 2. It suggests that it contributes to the ability of cells.

[0289] The release of immune mediators from immune cells can also activate nociceptors. Mast cells are found near primary nociceptor neurons and contribute to nociceptor sensitization in many situations. Injection of the secretagogue compound 48 / 80 promotes the degranulation of mast cells in the dura mater and excites meningeal nociceptors. Mast cell degranulation also contributes to the rapid onset of nerve growth factor-induced heat hyperalgesia. Macrophages contribute to nociceptor sensitization by releasing various soluble mediators. The expression of chemokine macrophage inflammatory protein-1α (MIP-1α) and its receptors CCR1 and CCR5 increases in macrophages and Schwann cells following partial ligation of the sciatic nerve, which contributes to the development of neuropathic pain. Lymphocytes contribute to the sensitization of peripheral nociceptors. T cells infiltrate the sciatic nerve and dorsal root ganglion (DRG) after nerve injury. Neuroinjury-induced hyperalgesia and allodynia are significantly attenuated or abolished in T-cell-deficient rodents, and the immunosuppressant rapamycin attenuates neuropathic pain in rats, partly due to its effect on T cells. Among the subsets of T cells, type 1 and type 2 helper T cells (T H 1 and T H 2 cells) were shown to have different roles in neuropathic pain. HWhile 1 cells promote neuropathic pain behavior by releasing pro-inflammatory cytokines (IL-2 and interferon-γ (IFNγ)), T H 2. Cells inhibit this by releasing anti-inflammatory cytokines (IL-4, IL-10, and IL-13). The complement system also plays a role in inflammatory hyperalgesia and neuropathic pain. The anaphylactic toxin C5a is an important effector of the complement cascade, and upon binding to the C5aR1 receptor on neutrophils, it becomes a potent neutrophil attractor (Ren & Dubner, Nat. Med. 16:1267-1276 (2010)).

[0290] Bacterial infection directly activates nociceptors, and immune responses mediated through TLR2, MyD88, T cells, B cells, neutrophils, and monocytes in mice Staphylococcus aureus ( Staphylococcus aureus It was found that it is not necessary for induced pain (Chiu et al., Nature Mechanical and thermal hyperalgesia in mice correlates with live bacterial load rather than tissue swelling or immune activation. Bacteria induce calcium flux and action potentials in nociceptor neurons through bacterial N-formylated peptides and porosiogenic toxins α-hemolysins, partly via distinct mechanisms. Specific depletion of Nav1.8-lineage neurons containing nociceptors abolishes pain during bacterial infection but simultaneously increased local immune infiltration and lymphadenopathy of draining lymph nodes. Thus, bacterial pathogens generate pain by directly activating sensory neurons that regulate inflammation, which represents an unexpected role of the nervous system in host-pathogen interactions. Literature [Talbot et al., ( Neuron Data from . 2015 July 15; 87(2): 341-354.)] also suggested that nociceptors are activated during exposure to allergens in sensitized animals.

[0291] In certain disorders, neuroinflammatory disorders are attributed to peripheral inflammation caused by tissue damage, autoimmune diseases, infections, and exposure to irritants in soft tissues, skin, respiratory system, joints, genitourinary and gastrointestinal tracts, liver, and brain. Neuroinflammatory disorders include, but are not limited to, allergic inflammation as described herein, inflammatory bowel disease, interstitial cystitis, atopic dermatitis, asthma, conjunctivitis, arthritis, colitis, contact dermatitis, diabetes mellitus, eczema, cystitis, gastritis, migraine, psoriasis, rhinitis, rosacea, sunburn, pancreatitis, chronic cough, chronic rhinosinusitis, traumatic brain injury, polybacterial sepsis, tendinopathy, chronic urticaria, rheumatic disease, acute lung injury, exposure to irritants, and inhalation of irritants, contaminants, or chemical warfare agents.

[0292] Evaluation of pain, cough, itching, and neuroinflammation

[0293] Measurement indices may be used to assess the efficacy of any compound, composition, method, and kit of the present invention in the treatment of pain associated with musculoskeletal, immuno-inflammatory, and neuropathic disorders. Useful indices include visual analog scales (VAS), Likert scales, categorical pain scales, descriptors, Lequesne indices, WOMAC indices, and AUSCAN indices, each of which is widely known in the art. These indices may be used to measure pain, itching, function, stiffness, or other variables.

[0294] Visual Analog Scales (VAS) provide one-dimensional quantitative measurements. VAS typically utilizes distance representations, such as line drawings with hash marks drawn at regular distance intervals, for example, at 10 intervals of 1-cm. For example, a patient may be asked to rank the sensation of pain or itching by selecting the point on the line that best corresponds to the sensation of pain or itching, where one end of the line corresponds to "no pain" (score of 0 cm) or "no itching," and the other end corresponds to "unbearable pain" or "unbearable itching" (score of 10 cm). This procedure provides a simple and rapid approach to obtaining quantitative information about how a patient experiences pain or itching. VAS scales and their uses are described, for example, in U.S. Patents No. 6,709,406 and 6,432,937.

[0295] The Likert scale similarly provides one-dimensional positive measurements. Generally, the Likert scale has discrete integer values ​​ranging from a low value (e.g., 0, meaning no pain) to a high value (e.g., 7, meaning severe pain). Patients experiencing pain are asked to select a number between the low and high values ​​to indicate the degree of pain experienced. The Likert scale and its uses are described, for example, in U.S. Patent Nos. 6,623,040 and 6,766,319.

[0296] The Requisnais Index and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index use self-management questionnaires to assess pain, function, and stiffness in the knees and hips of OA patients. While both the knee and hip are included by WOMAC, there is a single Requisnais questionnaire for the knee and a separate questionnaire for the hip. These questionnaires are useful because they contain more informative content compared to the VAS or Likert scales. Both the WOMAC Index and the Requisnais Index questionnaires have been extensively validated in OA, including in surgical settings (e.g., knee and hip arthroplasty). Their metric characteristics do not differ significantly.

[0297] The AUSCAN (Australia-Canada Hand Arthritis) index uses validated, reliable, and responsive patient self-report questionnaires. In one example, these questionnaires include 15 questions in three dimensions (pain, 5 questions; stiffness, 1 question; and physical function, 9 questions). The AUSCAN index may use, for example, Likert or VAS scales.

[0298] Indices useful in the method, composition, and kit of the present invention for measuring pain include the Pain Descriptor Scale (PDS), Visual Analog Scale (VAS), Verbal Descriptor Scale (VDS), Numeric Pain Intensity Scale (NPIS), Neuropathic Pain Scale (NPS), Neuropathic Pain Symptom Inventory (NPSI), Present Pain Inventory (PPI), Geriatric Pain Measure (GPM), McGill Pain Questionnaire (MPQ), Mean Pain Intensity (Descriptor Differential Scale), Numeric Pain Scale (NPS), Global Evaluation Score (GES), Short-Form McGill Pain Questionnaire, Minnesota Multiphasic Personality Inventory, Pain Profile, and Multidimensional Pain Inventory. It includes the Multidimensional Pain Inventory, the Child Health Questionnaire, and the Child Assessment Questionnaire.

[0299] Itching can be measured using subjective scales (VAS, Likert, Technician). Another approach is to measure scratching, which is an objective correlation of itching, using vibration transducers or motion sensors.

[0300] Coughing can be measured using standard questionnaires such as the Leicester Cough Questionnaire, as well as validated objective tools that measure cough frequency (e.g., VitaloJAK).

[0301] Examples

[0302] The following examples are intended to illustrate the invention and are not intended to limit it.

[0303] Example 1 - Compound Synthesis

[0304] Definition of common abbreviations

[0305] ACN Acetonitrile

[0306] aq. Mercury

[0307] ℃ Celsius temperature

[0308] δ Chemical shift (ppm)

[0309] DCM dichloromethane

[0310] DMSO dimethyl sulfoxide

[0311] ESI Electro-spray ionization

[0312] Et 2 O diethyl ether

[0313] EtOAc ethyl acetate

[0314] h city

[0315] MeOH methanol

[0316] mHz megahertz

[0317] min minute

[0318] ml milliliter

[0319] MS Mass Spectrometry

[0320] m / z Mass to charge ratio

[0321] NMR nuclear magnetic resonance

[0322] Pet ether petroleum ether

[0323] RT room temperature

[0324] TLC Thin-layer chromatography

[0325] UV ultraviolet rays

[0326] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide:

[0327]

[0328] Compound 1A

[0329] ● Synthesis of the intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide.

[0330] Bromoacetyl bromide (23.8 mL, 272.31 mmol, 1.1 eq) was added to a suspension of 2,6-dimethylaniline (30.5 mL, 247.56 mmol, 1.0 eq) in water (300 mL) at 10°C. The reaction mixture was maintained at pH 9-10 for 1 hour in a 15% Na2CO3 (aq.) solution while monitoring the progress of the reaction by TLC (mobile phase: 30% EtOAc in hexane, visualization by UV). The reaction mixture was extracted with EtOAc (2 x 600 mL), the combined organic extract was washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude mixture was ground with Et2O (2 x 200 mL) to obtain 2-bromo-N-(2,6-dimethylphenyl)acetamide (21 g) as a white solid. MS (ESI): m / z 244.02 [M + 2] + . 1 H NMR (400 MHz, CDCl3) δ 7.75 (br s, 1H), 7.20 - 7.02 (m, 3H), 4.07 (s, 2H), 2.24 (s, 6H).

[0331] ● Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide.

[0332] Pyridine (0.166 mL, 2.06 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.25 g, 1.03 mmol, 1.0 eq) in EtOAc (10 mL), and the resulting mixture was stirred at 80°C in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). After 16 hours, the crude solution was cooled to room temperature, the resulting precipitate was collected by filtration, washed with EtOAc (2 x 10 mL), and dried under vacuum to obtain N-(2,6-dimethylphenyl)-2-(pyridinium-1-yl)acetamide bromide (0.1 g) as a white solid. MS (ESI): m / z 241.1 [M]+ . 1 H NMR (400 MHz, DMSO-d6) δ 10.01 (s, 1H), 9.08 (d, J = 5.5 Hz, 2H), 8.69 (t, J = 7.8 Hz, 1H), 8.22 (dd, J = 6.8, 7.5 Hz, 2H), 7.16 - 6.99 (m, 3H), 5.75 (s, 2H), 2.20 (s, 6H).

[0333] Synthesis of 3-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-methyl-1H-imidazole-3-ium bromide:

[0334]

[0335] Compound 2A

[0336] ● Synthesis of 3-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-methyl-1H-imidazole-3-ium bromide.

[0337] The synthesis of the intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide was as described above for Compound 1. 1-methyl-1H-imidazole (0.072 mL, 0.88 mmol, 1.1 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.2 g, 0.8 mmol, 1.0 eq) in EtOAc (3 mL), and the resulting mixture was stirred at 80°C in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). After 16 hours, the solution was concentrated under reduced pressure, and the resulting crude product was ground with EtOAc (10 mL) to obtain 3-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-methyl-1H-imidazole-3-ium bromide (0.15 g). MS (ESI): m / z 244.1 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 9.15 (s, 1H), 7.77 (s, 1H), 7.72 (s, 1H), 7.14 - 7.03 (m, 3H), 5.28 (s, 2H), 3.91 (s, 3H), 2.17 (s, 6H).

[0338] Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutane-2-yl)pyridine-1-um bromide:

[0339]

[0340] Compound 3A

[0341] ● Synthesis of the intermediate 2-bromobutanoyl chloride.

[0342] Thionyl chloride (150 mL) was added to 2-bromobutanoic acid (25 g, 149.7 mmol, 1.0 eq) at 0°C, and the reaction mixture was subsequently stirred at 80°C for 2 h. The reaction mixture was cooled to RT and concentrated under reduced pressure to obtain unpurified 2-bromobutanoyl chloride (27.7 g, 99.5%) as a brown residue, which was used immediately in the next step without further purification. 1 H NMR (400 MHz, CDCl3) δ 4.52 - 4.44 (m, 1H), 2.29 - 2.16 (m, 1H), 2.15 - 2.02 (m, 1H), 1.10 (t, J = 7.3 Hz, 3H).

[0343] ● Synthesis of the intermediate 2-bromo-N-(2,6-dimethylphenyl)butanamide.

[0344] A solution of 2,6-dimethylaniline (15 g, 121.18 mmol, 1.0 eq) and pyridine (15 mL, 189.6 mmol, 1.5 eq) in DCM (400 mL) was cooled to 0°C in a freezing bath. A solution of 2-bromobutanoyl chloride (27.5 g, 148.6 mmol, 1.2 eq) in DCM (50 mL) was slowly added to this solution, and the resulting mixture was heated to room temperature while stirring for 2 hours. The reaction mixture was adjusted to a pH of about 5 to 6 using 2 N HCl and extracted with DCM (2 x 200 mL). The combined organic extract was washed with water (250 mL) and brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude product was ground with n-pentane (150 mL) to obtain 2-bromo-N-(2,6-dimethylphenyl)butanamide (30 g). MS (ESI): m / z 272.13 [M + 2] + . 1 H NMR (400 MHz, DMSO-d6) δ 9.69 (s, 1H), 7.15 - 7.02 (m, 3H), 4.51 (t, J = 7.3 Hz, 1H), 2.22 - 2.04 (m, 7H), 2.03 - 1.91 (m, 1H), 0.98 (t, J = 7.3 Hz, 3H).

[0345] ● Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutane-2-yl)pyridine-1-um bromide.

[0346] Pyridine (0.12 mg, 1.4 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)butanamide (0.2 g, 0.7 mmol, 1.0 eq) in acetonitrile (3 mL), and the resulting mixture was heated in a sealed tube at 90°C for 36 hours. After cooling to rt, the solution was concentrated under reduced pressure, and the resulting crude mixture was ground with EtOAc (5 mL) to obtain 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutane-2-yl)pyridine-1-um bromide (0.17 g). MS (ESI): m / z 269.2 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 0.92 (t, J=7.23 Hz, 3H), 2.10 (br s, 6H), 2.28-2.45 (m, 1H), 2.59 (dt, J=13.98, 6.93 Hz, 1H), 5.75 (dd, J=9.43, 5.92 Hz, 1H), 7.00-7.20 (m, 3H), 8.25 (t, J=7.13 Hz, 2H), 8.73 (t, J=7.78 Hz, 1H), 9.23 (d, J=5.70 Hz, 2H), 10.12 (s, 1H).

[0347] Synthesis of 3-(1-((2,6-dimethylphenyl)amino)-1-oxobutane-2-yl)-1-methyl-1H-imidazole-3-ium bromide:

[0348]

[0349] Compound 4A

[0350] ● Synthesis of 3-(1-((2,6-dimethylphenyl)amino)-1-oxobutane-2-yl)-1-methyl-1H-imidazole-3-um bromide.

[0351] The synthesis of the intermediates 2-bromobutanoyl chloride and 2-bromo-N-(2,6-dimethylphenyl)butanamide was as described above for Compound 3. 1-methyl-1H-imidazole (0.12 ml, 1.4 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)butanamide (0.2 g, 0.7 mmol, 1.0 eq) in acetonitrile (3 mL), and the resulting reaction mixture was heated at 90°C for 36 hours in a sealed tube. After cooling to rt, the solution was concentrated under reduced pressure, and the resulting crude product was ground with EtOAc (10 mL) to obtain 3-(1-((2,6-dimethylphenyl)amino)-1-oxobutane-2-yl)-1-methyl-1H-imidazole-3-ium bromide (0.09 g). MS (ESI): m / z 272.2 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 0.95 (t, J=7.27 Hz, 3H), 2.04-2.26 (m, 7H), 2.29-2.43 (m, 1H), 3.91 (s, 3H), 5.34 (dd, J=9.54, 5.72 Hz, 1H), 7.04-7.18 (m, 3H), 7.79 (t, J=1.79 Hz, 1H), 7.93 (t, J=1.79 Hz, 1H), 9.40 (s, 1H), 9.97 (s, 1H).

[0352] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-2-methylpyridine-1-um bromide:

[0353]

[0354] Compound 5A

[0355] ● Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-2-methylpyridine-1-um bromide.

[0356] The synthesis of the intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide was as described above for compound 1. EtOAc2-methylpyridine (0.149 g, 1.6 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.2 g, 0.8 mmol, 1.0 eq) in 5 mL, and the resulting mixture was stirred at 80°C in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). After 16 hours, the solution was concentrated under reduced pressure, and the resulting crude product was ground with EtOAc (2 x 15 mL) to obtain 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-2-methylpyridine-1-um bromide (0.13 g) as a grayish-white solid. MS (ESI): m / z 255.2 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 2.20 (s, 6H), 2.79 (s, 3H), 5.73 (s, 2H), 6.99-7.17 (m, 3H), 7.99-8.10 (m, 1H), 8.12 (d, J=7.89 Hz, 1H), 8.58 (td, J=7.78, 1.10 Hz, 1H), 9.05 (d, J=5.48 Hz, 1H), 10.11 (s, 1H).

[0357] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-2,6-dimethylpyridine-1-um bromide:

[0358]

[0359] Compound 6A

[0360] ● Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-2,6-dimethylpyridine-1-um bromide.

[0361] The synthesis of the intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide was as described above for Compound 1. 2,6-dimethylpyridine (0.17 g, 1.6 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.2 g, 0.8 mmol, 1.0 eq) in EtOAc (5 mL), and the resulting mixture was stirred at 80°C in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). After 32 hours, the solution was concentrated under reduced pressure, and the resulting crude product was ground with EtOAc (2 x 10 mL) to obtain 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-2,6-dimethylpyridine-1-um bromide (0.157 g) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1 H), 8.45-8.41 (m, 1 H), 7.98-7.96 (d, 2 H), 7.11-7.10 (d, 3 H), 5.65 (s, 2 H), 2.83 (s, 6H), 2.20 (s, 6H).

[0362] Synthesis of 1-(2-((4-fluoro-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide:

[0363]

[0364] Compound 7A

[0365] ● Synthesis of the intermediate 2-bromo-N-(4-fluoro-2,6-dimethylphenyl)acetamide.

[0366] 2-bromoacetyl bromide (1.453 g, 7.2 mmol, 2.0 eq) was added to a stirred solution of 4-fluoro-2,6-dimethylaniline (0.5 g, 3.6 mmol, 1.0 eq) in water (10 mL) at 0 °C, and the resulting mixture was stirred at RT for 16 h while monitoring the progress of the reaction by TLC (mobile phase: 50% EtOAc in petroleum ether, visualization by UV). The pH of the reaction mixture was adjusted with a 15% Na2CO3 (aq.) solution and extracted with EtOAc (2 x 25 mL). The combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 2-bromo-N-(4-fluoro-2,6-dimethylphenyl)acetamide (0.45 g). MS (ESI): m / z 260.11 [M + H] + . 1 H NMR (400 MHz, CDCl3) δ 7.65 (br s, 1H), 6.81 (d, J=8.94 Hz, 2H), 4.07 (s, 2H), 2.23 (s, 6H).

[0367] ● Synthesis of 1-(2-((4-fluoro-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide.

[0368] Pyridine (0.126 g, 1.6 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(4-fluoro-2,6-dimethylphenyl)acetamide (0.2 g, 0.8 mmol, 1.0 eq) in EtOAc (5 mL), and the resulting mixture was stirred at 80°C for 16 h in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). The solution was concentrated under reduced pressure, and the crude product was ground with EtOAc (15 mL) to obtain 1-(2-((4-fluoro-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide (0.15 g). MS (ESI): m / z 259.2 [M] + . 1H NMR (400 MHz, DMSO-d6) δ 9.98 (s, 1H), 9.07 (d, J=5.48 Hz, 2H), 8.68 (t, J=7.89 Hz, 1H), 8.22 (dd, J=7.67, 6.80 Hz, 2H), 6.95 (d, J=9.43) Hz, 2H), 5.75 (s, 2H), 2.20 (s, 6H).

[0369] Synthesis of 1-(2-(methylamino)-2-oxoethyl)pyridine-1-um bromide:

[0370]

[0371] Compound 8A

[0372] ● Synthesis of the intermediate 2-bromo-N-methylacetamide.

[0373] 2-bromoacetyl bromide (1.497 g, 7.4 mmol, 2.0 eq) was added to a stirred solution of 2,4,6-trimethylaniline (0.5 g, 3.7 mmol, 1.0 eq) in water (10 mL) at 0 °C, and the resulting mixture was stirred at RT for 16 h while monitoring the progress of the reaction by TLC (mobile phase: 50% EtOAc in petroleum ether, visualization by UV). The pH of the reaction mixture was adjusted with a 15% Na2CO3 (aq.) solution and extracted with EtOAc (2 x 25 mL). The combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 2-bromo-N-methylacetamide (0.65 g). MS (ESI): m / z 256.18 [M+H]. 1 H NMR (400 MHz, CDCl3) δ 7.67 (br s, 1H), 6.91 (s, 2H), 4.07 (s, 2H), 2.24-2.29 (m, 3H), 2.20 (s, 6H).

[0374] ● Synthesis of 1-(2-(methylamino)-2-oxoethyl)pyridine-1-um bromide.

[0375] Pyridine (0.126 g, 1.6 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-methylacetamide (0.2 g, 0.8 mmol, 1.0 eq) in ethyl acetate (5 mL), and the reaction mixture was stirred at 80°C for 16 h in a sealed tube while monitoring the progress of the reaction by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). The reaction mixture was concentrated under reduced pressure, and the crude product was ground with EtOAc (15 mL) to obtain 1-(2-(methylamino)-2-oxoethyl)pyridine-1-um bromide (0.13 g). MS (ESI): m / z 255.2 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 9.90 (s, 1H), 9.07 (d, J=5.48 Hz, 2H), 8.68 (t, J=7.78 Hz, 1H), 8.22 (t, J=7.13 Hz, 2H), 6.89 (s, 2H), 5.73 (s, 2H), 2.22 (s, 3H), 2.15 (s, 6H).

[0376] Synthesis of 1-(2-((4-chloro-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide:

[0377]

[0378] Compound 9A

[0379] ● Synthesis of the intermediate 2-bromo-N-(4-chloro-2,6-dimethylphenyl)acetamide.

[0380] 2-bromoacetyl bromide (0.797 g, 3.952 mmol, 1.1 eq) was added to a stirred solution of 4-chloro-2,6-dimethylaniline (0.5 g, 3.592 mmol, 1 eq) in water (5 mL) at 0 °C, and the resulting mixture was stirred for 1 hour while monitoring the reaction progress by TLC (mobile phase: 50% EtOAc in petroleum ether, visualization by UV). The pH of the reaction mixture was adjusted with a 15% Na2CO3 (aq.) solution, and extracted with EtOAc (2 x 25 mL). The combined organic extract Na 2 SO 4 2-bromo-N-(4-chloro-2,6-dimethylphenyl)acetamide (0.45 g) was obtained by drying on a surface, filtering, and concentrating under reduced pressure. MS (ESI): m / z 276.05 [M+H]. 1 H NMR (400 MHz, CDCl3) δ 7.66 (br s, 0.5 H), 7.09 (s, 1H), 6.89-6.96 (m, 2H), 4.07 (s, 1H), 2.22 (s, 3H), 2.12-2.19 (m, 6H).

[0381] ● Synthesis of 1-(2-((4-chloro-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide.

[0382] Pyridine (0.085 g, 1.1 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(4-chloro-2,6-dimethylphenyl)acetamide (0.15 g, 0.5 mmol, 1.0 eq) in EtOAc (5 mL), and the reaction mixture was stirred at 80°C for 16 h in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). The solution was concentrated under reduced pressure, and the crude product was ground with EtOAc (15 mL) to obtain 1-(2-((4-chloro-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide (0.1 g). MS (ESI): m / z 275.1 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.06 (s, 1H), 9.07 (d, J=5.48 Hz, 2H), 8.68 (t, J=7.89 Hz, 1H), 8.22 (t, J=7.23 Hz, 2H), 7.19 (s, 2H), 5.75 (s, 2H), 2.20 (s, 6H).

[0383] Synthesis of 1-(2-((4-methoxy-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide:

[0384]

[0385] Compound 10A

[0386] ● Synthesis of the intermediate 2-bromo-N-(4-methoxy-2,6-dimethylphenyl)acetamide.

[0387] 2-bromoacetyl bromide (0.734 g, 3.6 mmol, 1.1 eq) was added to a stirred solution of 4-methoxy-2,6-dimethylaniline (0.5 g, 3.3 mmol, 1 eq) in water (5 mL) at 0 °C, and the resulting mixture was stirred for 16 hours while being heated to RT. The progress of the reaction was monitored by TLC (mobile phase: 40% EtOAc in petroleum ether, visualization by UV). The pH of the reaction mixture was adjusted with a 15% Na2CO3 (aq.) solution and extracted with EtOAc (3 x 30 mL). The combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The generated crude product was purified by column chromatography (100-200 mesh silica gel; 20% EtOAc in petroleum ether) to obtain 2-bromo-N-(4-methoxy-2,6-dimethylphenyl)acetamide (0.09 g). MS (ESI): m / z 272.08 [M+H]. 1 1H NMR (400 MHz, CDCl 3 ) δ 7.62 (br s, 1H), 6.64 (s, 2H), 4.07 (s, 2H), 3.77 (s, 3H), 2.22 (s, 6H), 1.3 (m, 1H).

[0388] ● Synthesis of 1-(2-((4-methoxy-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide.

[0389] Pyridine (34 mg, 0.4 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(4-methoxy-2,6-dimethylphenyl)acetamide (60 mg, 0.2 mmol, 1.0 eq) in EtOAc (1 mL), and the reaction mixture was stirred at 80°C for 16 h in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). The solution was concentrated under reduced pressure, and the crude product was ground with EtOAc (2 x 5 mL) to obtain 1-(2-((4-methoxy-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide (0.07 g). MS (ESI): m / z 271.1 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1H), 9.07 (d, J=5.70 Hz, 2H), 8.68 (t, J=7.89 Hz, 1H), 8.22 (t, J=7.13 Hz, 2H), 6.66 (s, 2H), 5.72 (s, 2H), 3.71 (s, 3H), 2.16 (s, 6H).

[0390] Synthesis of 1-(2-((4-cyano-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide:

[0391]

[0392] Compound 11A

[0393] ● Synthesis of the intermediate 2-bromo-N-(4-cyano-2,6-dimethylphenyl)acetamide.

[0394] K2CO3 (0.563 g, 4.08 mmol, 1.2 eq) was added to a stirred solution of 4-amino-3,5-dimethylbenzonitrile (0.5 g, 3.4 mmol, 1.0 eq) in DCM (20 mL), and the resulting mixture was cooled to 0°C. Subsequently, 2-bromoacetyl bromide (0.823 g, 4.08 mmol, 1.2 eq) was added dropwise, and the reaction mixture was heated to RT and stirred for 16 h. The progress of the reaction was monitored by TLC (mobile phase: 50% ethyl acetate in PET ether, Rf: 0.41, UV visualization). The reaction mixture was basicized with a 15% Na2CO3 solution at 0°C to obtain a precipitate, which was filtered and dried under vacuum to obtain 2-bromo-N-(4-cyano-2,6-dimethylphenyl)acetamide (0.35 g). MS (ESI): m / z 267.36 [M+H]. 1 H NMR (400 MHz, CDCl3) δ 7.80 (br s, 1H), 7.41 (s, 2H), 4.09 (s, 2H), 2.29 (s, 6H).

[0395] ● Synthesis of 1-(2-((4-cyano-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide.

[0396] Pyridine (0.1106 g, 1.6 mmol, 2.0 eq) was added to a stirred solution of 2-bromo-N-(4-cyano-2,6-dimethylphenyl)acetamide (0.2 g, 0.7 mmol, 1.0 eq) in EtOAc (5 mL), and the reaction mixture was stirred at 80°C for 16 h in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). The solution was concentrated under reduced pressure, and the crude product was ground with EtOAc (15 mL) to obtain 1-(2-((4-cyano-2,6-dimethylphenyl)amino)-2-oxoethyl)pyridine-1-um bromide (0.14 g). MS (ESI): m / z 266.1 [M] + . 1H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 9.08 (d, J=5.70 Hz, 2H), 8.69 (t, J=7.78 Hz, 1H), 8.23 ​​(t, J=7.23 Hz, 2H), 7.60 (s, 2H), 5.79 (s, 2H), 2.25 (s, 6H).

[0397] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-phenylpyridine-1-um bromide:

[0398]

[0399] Compound 12A

[0400] ● Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-phenylpyridine-1-um bromide.

[0401] The synthesis of the intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide was as described above for Compound 1. 3-phenylpyridine (0.144 g, 0.9 mmol, 1.5 eq) was added to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.150 g, 0.6 mmol, 1.0 eq) in ACN (2.0 ml), and the reaction mixture was stirred at 80°C in a sealed tube while monitoring the consumption of the starting material by TLC (mobile phase: 10% MeOH in DCM, visualization by UV). After stirring for 54 hours, the solution was cooled to RT, concentrated under reduced pressure, and the crude product was collected by filtration and ground with EtOAc (2 x 5 mL) to obtain 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-phenylpyridine-1-um bromide (0.15 g). MS (ESI): m / z 317.2 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.02 (s, 1H), 9.56 (s, 1H), 9.09 - 8.95 (m, 2H), 8.30 (dd, J = 6.1, 8.1 Hz, 1H), 7.90 (d, J = 7.0 Hz, 2H), 7.70 - 7.54 (m, 3H), 7.15 - 7.01 (m, 3H), 5.81 (s, 2H), 2.22.

[0402] Synthesis of Compounds 13-28: Table F.

[0403] Table F provides additional representative examples of the present invention synthesized according to the method described for the synthesis of compound 12 from 2-bromo-N-(2,6-dimethylphenyl)acetamide and a suitable heterocycle as presented above.

[0404] Table F

[0405]

[0406]

[0407] Synthesis of 2-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-phenyl-1H-pyrazol-2-um formate:

[0408]

[0409] Compound 29A

[0410] 1-phenyl-1H-pyrazole (595 mg, 4.12 mmol) was added from a microwave vial to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (500 mg, 2.06 mmol) in ACN (10 mL). The resulting reaction mixture was stirred at 100°C for 2 hours in a microwave reactor (CEM Corporation, Matthews, NC). The reaction mixture was concentrated under reduced pressure to obtain an unpurified product, which was subjected to flash column chromatography (silica gel 100-200 mesh; 10%-25% MeOH / DCM gradient elution It was purified by ). The collected pure fraction was concentrated under reduced pressure to obtain (120 mg, LCMS 48%), and this was analyzed by flash column chromatography (silica gel 100-200 mesh; 10%-25% MeOH / DCM gradient elutionIt was purified by ) and then further purified by the reverse-phase Prep-HPLC method (Column: X-Select CSH C18 (250*19) mm, 5 µ, Temperature: Ambient, Mobile phase (A): 0.1% formic acid in water; Mobile phase (B): ACN; (T / %B) 0 / 10, 2 / 10, 10 / 35, 14 / 35, 14.2 / 98, 17 / 98, 17.2 / 10, 20 / 10, Flow rate: 13 mL / min). The pure fractions were combined and concentrated by freeze-drying to deliver 2-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-phenyl-1H-pyrazole-2-um (20.3 mg). MS (ESI): m / z 306.2 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 10.06 (s, 1 H), 8.92 (dd, J=9.54, 2.69 Hz, 2 H), 8.51 (s, 1 H), 7.64 - 7.85 (m, 5 H), 7.21 (t, J=2.69 Hz, 1 H), 6.93 - 7.10 (m, 3 H), 5.58 (s, 2 H), 1.87 (s, 6 H).

[0411] Synthesis of 2-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(2-(ethylamino)-2-oxoethyl)-1H-pyrazol-2-um bromide:

[0412]

[0413] Compound 30A

[0414] N-ethyl-2-(1H-pyrazole-1-yl)acetamide (94.9 mg, 0.619 mmol) was added to a solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (100 mg, 0.413 mmol) in ACN (2 mL) at RT, and the resulting reaction mixture was stirred in a sealed tube at 90°C for 72 h while monitoring the progress of the reaction by TLC (visualization by UV in 10% MeOH in DCM). The reaction mixture was cooled to RT and concentrated under reduced pressure. The crude product was ground with EtOAc (2 x 8 mL) to obtain the product 2-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(2-(ethylamino)-2-oxoethyl)-1H-pyrazole-2-um bromide (90 mg). Mass (ESI): m / z 315.1 [M] + . UPLC: 97.79%. 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.92 (s, 1 H), 8.68 (dd, J=2.86, 0.95 Hz, 1 H), 8.53 - 8.64 (m, 2 H), 7.05 - 7.16 (m, 3 H), 7.01 (t, J=2.98 Hz, 1 H), 5.58 (s, 2 H), 5.28 (s, 2 H), 3.09 - 3.25 (m, 2 H), 2.18 (s, 6 H), 1.09 (t, J=7.27 Hz, 3 H).

[0415] Synthesis of Compounds 31-105: Table G.

[0416] Table G below provides further representative examples of the present invention synthesized according to the method described from 2-bromo-N-(2,6-dimethylphenyl)acetamide or 2-chloro-N-(2,6-dimethylphenyl)acetamide and a suitable heterocycle as presented above. The compounds were purified by grinding or reverse-phase preparative HPLC.

[0417] Table G

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425] Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxo-3-phenylpropane-2-yl)pyridine-1-um chloride

[0426]

[0427] Compound 106

[0428] ● Synthesis of 2-Chloro-N-(2,6-Dimethylphenyl)-3-Phenylpropanamide

[0429] 2,6-dimethylaniline (0.116 g, 0.960 mmol) was added to a stirred solution of 2-bromo-3-phenylpropanoic acid (0.2 g, 0.873 mmol), EDC.HCl (0.836 g, 4.365 mmol), and DMAP (0.085 g, 0.698 mmol) in DCM (5 ml) at room temperature, and the resulting reaction mixture was stirred in a sealed tube at room temperature for 60 h while monitoring the progress of the reaction by TLC (20% EtOAc-hexane, visualization: UV). The reaction mixture was concentrated under reduced pressure to obtain an unpurified product, which was diluted with EtOAc (100 ml), washed with water (20 ml x 3), dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain 0.26 g of unpurified product. The crude product was purified by normal-phase flash chromatography (eluted with 10%–50% EtOAc / Pet ether) to obtain 2-chloro-N-(2,6-dimethylphenyl)-3-phenylpropanamide (0.1 g) as a grayish-white solid. MS (ESI): m / z 287.82 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ ppm 9.61 (s, 1 H), 7.35 - 7.23 (m, 5 H), 7.08-6.99 (m, 3 H), 4.83 (t, 1 H), 3.39-3.35 (m, 1 H), 3.21-3.14 (m, 1 H), 1.92 (s, 6 H).

[0430] ● Synthesis of 1-1-(1-((2,6-dimethylphenyl)amino)-1-oxo-3-phenylpropane-2-yl)pyridine-1-um chloride

[0431] Pyridine (0.059 g, 0.745 mmol) was added at room temperature to a stirred solution of 2-chloro-N-(2,6-dimethylphenyl)-3-phenylpropanamide (0.05 g, 0.174 mmol) in 5 ml of EtOAc, and the resulting mixture was stirred in a sealed tube at 120°C for 16 hours while monitoring the reaction progress by TLC (10% MeOH in DCM, visualization: UV). The reaction mixture was concentrated under reduced pressure to obtain an unpurified product, which was ground in ethyl acetate (20 ml x 3) to obtain 1-1-(1-((2,6-dimethylphenyl)amino)-1-oxo-3-phenylpropane-2-yl)pyridine-1-ium chloride (42.3 mg) as a grayish-white solid. MS (ESI): m / z 331.25 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 10.50 (s, 1 H), 9.28 (d, 2 H), 8.64-8.60 (m, 1 H), 8.18-8.14 (m, 2 H), 7.34-7.19 (m, 5 H), 7.13-7.05 (m, 3) H), 6.33-6.29 (m, 1 H), 4.03-3.98 (m, 1 H), 3.73-3.67 (m, 1H), 2.03 (s, 6 H).

[0432] Synthesis of Compounds 107-108: Table H.

[0433] Table H below provides additional representative examples of the present invention synthesized from appropriately substituted 2-bromo-propanoic acid, 2,6-dimethylaniline, and pyridine.

[0434] Table H

[0435]

[0436] Synthesis of 1,2-bis(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1H-pyrazol-2-um bromide

[0437]

[0438] Compound 109

[0439] A solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (200 mg, 0.8260 mmol) and 1-(tert-butyl)-1H-pyrazole (0.5 mL) was stirred in a sealed tube at 120°C for 16 h while monitoring the reaction progress by TLC (10% MeOH in DCM, detection: UV). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain an unpurified product, which was purified by column chromatography (eluted with 20% MeOH in DCM) to obtain 1,2-bis(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1H-pyrazole-2-um bromide (111.9 mg). MS (ESI): m / z 391.0 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 10.17 (s, 2 H), 8.74 (d, 2 H), 7.04 - 7.12 (m, 7 H), 5.65 (s, 4 H), 2.19 (s, 12 H).

[0440] Synthesis of 1,3-Bis(2-((2,6-Dimethylphenyl)amino)-2-oxoethyl)-1H-imidazole-3-yum bromide

[0441]

[0442] Compound 110

[0443] 1H-imidazole (0.084 g, 1.238 mmol) was added at room temperature to a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.15 g, 0.619 mmol) in acetonitrile (2.0 ml), and the resulting reaction mixture was stirred at 90°C for 16 h while monitoring the progress of the reaction mixture by TLC (10% MeOH in DCM, visualization: UV). The reaction mixture was concentrated under reduced pressure to obtain an unpurified product, which was ground with 5% MeOH in DCM (50 ml) to obtain 1,3-bis(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1H-imidazole-3-um bromide (56 mg) as a grayish-white solid. Mass (ESI): m / z 391.38 [M] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.82 (s, 2 H), 9.24 (s, 1 H), 7.80 (d, 2 H), 7.12-7.06 (s, 6 H), 5.34 (s, 4 H), 2.17 (s, 12 H).

[0444] Synthesis of 1-cyclohexyl-2-(2-(2,6-dimethylphenoxy)-2-oxoethyl)-1H-pyrazol-2-um bromide

[0445]

[0446] Compound 111

[0447] ● Synthesis of the intermediate 2,6-dimethylphenyl 2-bromoacetate.

[0448] Pyridine (0.647 g, 8.184 mmol) and 2-bromoacetyl bromide (1.23 g, 6.138 mmol) were added to a stirred solution of 2,6-dimethylphenol (0.5 g, 4.092 mmol) in acetonitrile (5 ml) at 0°C, and the resulting reaction mixture was stirred for 15 minutes at 0°C while monitoring the progress of the reaction by TLC (10% EtOAc in PET ether, visualization: UV). The reaction mixture was diluted with water (20 ml), extracted with ethyl acetate (2 x 25 ml), washed with brine (30 ml), dried over Na2SO4, and concentrated under reduced pressure to obtain 2,6-dimethylphenyl 2-bromoacetate (500 mg) as a pale yellow liquid. 1 H NMR (400 MHz, CDCl3) δ ppm 7.07 (s, 3 H), 4.07 (s, 2 H), 2.18 (s, 6 H).

[0449] ● Synthesis of 1-cyclohexyl-2-(2-(2,6-dimethylphenoxy)-2-oxoethyl)-1H-pyrazol-2-um bromide

[0450] 1-cyclohexyl-1H-pyrazole (0.617 g, 4.112 mmol) was added to a stirred solution of 2,6-dimethylphenyl 2-bromoacetate (0.5 g, 2.056 mmol) in ACN (5 ml) at room temperature, and the resulting reaction mixture was heated at 90°C for 16 h while monitoring the progress of the reaction by TLC (10% MeOH in DCM, visualization: UV). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain an unpurified product, which was purified by normal-phase flash chromatography (10%-50% MeOH in DCM) to obtain 1-cyclohexyl-2-(2-(2,6-dimethylphenoxy)-2-oxoethyl)-1H-pyrazole-2-um bromide (20 mg) as a grayish-white solid. Mass (ESI): m / z 313.2 [M] + . 1H NMR (400 MHz, DMSO-d6) δ ppm 8.95 (d, 1 H), 8.68 (d, 1 H), 7.17-7.11 (m, 4 H), 6.32 (s, 2 H), 4.66-4.61 (m, 1 H), 2.16 (s, 6 H), 2.05-2.02 (m, 2 H), 1.87-1.78 (m, 4 H), 1.71-1.67 (m, 1 H), 1.44-1.41 (m, 2 H), 1.25-1.22 (m, 1 H).

[0451] Synthesis of Compounds 112-130: Table I.

[0452] Table I below provides additional representative examples of the present invention synthesized according to the method described from 2-bromoacetyl bromide and a suitably substituted aniline and heterocycle as presented above. The compounds were purified by grinding or reverse-phase preparative HPLC.

[0453] Table I

[0454]

[0455]

[0456]

[0457] Example 2 - Representative Compounds

[0458] Representative compounds according to the present invention, their enantiomers, and pharmaceutically acceptable salts thereof are described below, wherein Y - is a pharmaceutically acceptable anion as defined above, and Z is a heteroaryl structure selected from one of the structures in the following table. The compound can be prepared according to the generally described method above.

[0459]

[0460]

[0461]

[0462] Table 1 - Representative Z structures

[0463]

[0464]

[0465]

[0466] Table 2 - Representative Z structures

[0467]

[0468]

[0469]

[0470]

[0471]

[0472]

[0473]

[0474]

[0475]

[0476]

[0477]

[0478]

[0479] Table 3 - Representative Z structures

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486] Table 4 - Representative Z structures

[0487]

[0488]

[0489]

[0490] Table 5 - Representative Z structures

[0491]

[0492]

[0493]

[0494] Table 6 - Representative Z structures

[0495]

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502]

[0503] The preferred compounds according to the present invention, their enantiomers, and pharmaceutically acceptable salts have the following chemical formula ( IV It is displayed as ),

[0504]

[0505] In the above formula, the preferred compound is a substituent combination R as defined in Table 7. C , R D Composed of and Z, and Y - is a pharmaceutically acceptable anion as defined above. The compound can be prepared according to the generally described method above.

[0506] Table 7 - Chemical Formula ( IV R according to ) C , R D A desirable combination of and Z substituents.

[0507]

[0508]

[0509]

[0510]

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521]

[0522]

[0523]

[0524]

[0525]

[0526]

[0527]

[0528]

[0529]

[0530]

[0531]

[0532]

[0533]

[0534]

[0535] Example 3 - Nav1.7 Current Suppression

[0536] Representative compounds of the present invention were synthesized according to the described method and tested for their ability to inhibit voltage-gated sodium channels.

[0537] cell culture

[0538] NaV1.7 was expressed upon tetracycline induction. Cells were induced with 10% dialyzed fetal bovine serum (VWR, Radnor, PA), 1% glutamex (VWR, Radnor, PA), 1% penicillin-streptomycin (VWR, Radnor, PA), 100 mg / L hygromycin (Thermo Fisher Scientific, Waltham, MA), and 5 mg / L blasticidin (Alfa Aesar, Haverhill, MACells were cultured in DMEM containing ). Cells were grown and maintained at 37°C in a humidified environment containing 10% CO2 in the air. Cells were detached from culture flasks for subculturing and harvested using 0.05% trypsin-EDTA (Thermo Fisher Scientific, Waltham, MA). To induce NaV1.7, cells were induced with tetracycline (0.1–1 μg / mL, IBI Scientific, Peosta, IA) the day before recording and plated in 24-well plates. Cells were washed with DPBS (VWR, Radnor, PA), and after trypsin treatment, cell aggregates were separated by pulverizing five times in 10 mL of growth medium. For one 24-well plate, 2 mL of cell suspension was mixed with 23 mL of fresh growth medium, and 0.1–1 μg / mL of tetracycline was added. Subsequently, 1 ml of medium mixed with cells was added to each well of a 24-well plate in which a 12 mm coverslip was already placed at the bottom of the wells. Then, the cells were incubated overnight at 37°C and 10% CO2.

[0539] Patch Clamp Solution & Drug

[0540] The intracellular solution contained the following (in mM units): CsCl 135, NaCl 10, EGTA 10, HEPES 10, and MgCl22, and was adjusted to pH 7.2 with CsOH. The ex vivo solution was normal Ringer's solution containing (in mM units): NaCl 155, HEPES 10, glucose 10, KCl 3.5, CaCl21.5, and MgCl21, adjusted to pH 7.4 using NaOH. CsCl was Alfa Aesar, Haverhill, MA It is from. All other chemicals are from Sigma-Aldrich, St. Louis, MOIt is from. To test the degree of internal blockade by the test compound, the compound was dissolved in an internal solution at the indicated test concentration. In the control experiment, the internal solution did not contain any compound. To test the degree of external blockade by the test compound, the compound was dissolved in an external solution at the indicated test concentration.

[0541] Whole Cell Patch Clamp Protocol

[0542] 18–24 hours after inducing cells with tetracycline, the coverslip was placed in a chamber filled with normal Ringer's solution at room temperature, and the chamber was placed under the microscope. The pipette was removed from the borosilicate glass of a P97 puller (Sutter Instrument, Novato, CA) and polished with an MF-830 Microforge (Narishige International USA, Inc, Amityville, NY) to have a resistance of 1.5–2.5 MΩ when filled with an internal CsCl solution at room temperature. Healthy cells (spherical, translucent cells without visible defects) were selected for seal formation. Once a seal was formed between the pipette and the cells, they were "broken in" using a short suction pulse, and the total cell composition was established. The membrane potential was maintained at -100 mV before the voltage protocol began. Only cells with a series resistance of 1.5 to 5 MΩ were retained for analysis. The voltage protocol was as follows: the cells were maintained at -100 mV for 12 ms, followed by a hyperpolarization step at -105 mV for 12 ms to monitor for leakage. Subsequently, the cells were reverted to -100 mV for 40 ms. Afterward, the cells were depolarized to -20 mV for 10 ms, followed by a return to -100 mV for 26 ms.

[0543] Internal blockage by test compound

[0544] When recording began, the voltage protocol was executed at 30-second intervals for 5 minutes to acquire a stable baseline. This was followed by four 30-second cycles of 5 Hz stimulation of the same voltage protocol, separated by a 1-minute rest, and then a 0.33 Hz stimulation after the last training. Current was recorded using PatchMaster software with the Heka EPC10 (HEKA Electronics, Lambrecht, Germany). Only cells with an internal current amplitude of -20 mV between 400 pA and 4 nA were allowed. Additionally, cells with a leakage current exceeding 10% of the current amplitude were discarded.

[0545] Data Analysis: Internal Blocking

[0546] The data were plotted using Patchmaster software (HEKA Electronics, Lambrecht, Germany), and the minimum current during voltage steps up to -20 mV (peak internal current) was plotted and analyzed as a function of time. To determine the degree of rundown throughout the experiment, the average peak internal current amplitude (2-3 points) prior to the 5 Hz stimulation was set as the baseline (I baseline It was designated as ). The average peak internal current during the last 2 seconds of the last 5 Hz train was measured (I test ). I test ul I baseline The remaining control fraction current was calculated by dividing by... Three cells were tested with the control internal solution on each recording day, and the average fraction of the remaining current was calculated (control fraction current).

[0547] To determine the % blocking generated by the internally applied test compound, the following was performed. The average peak internal current amplitude (2-3 points) before 5 Hz stimulation was 0% blocking (I 0%block It was designated as ). To correct for current changes under control group conditions, I 0%blockThe modified 0% cutoff current was obtained by multiplying it by the remaining average control fraction current. The average peak internal current during the last 2 seconds of the last 5 Hz train was the uncutoff current (I unblocked It was designated as ). The blocking % was calculated using the following equation: (1 - I unblocked / ( I 0%block * Remaining control fraction current) x 100).

[0548] Representative examples of the present invention were tested for intracellular inhibition of NaV 1.7. The range of activity is % inhibition at 32 μM: "++++" (>95%), "+++" 95-70%, "++" (70-40%), or "+" (< 40%). The results are presented below.

[0549] Table J

[0550]

[0551] Representative examples of the present invention were tested for intracellular inhibition of NaV 1.7. The range of activity is % inhibition at 10 μM: "++++" (>95%), "+++" 95-70%, "++" (70-40%), or "+" (< 40%). The results are presented below.

[0552] Table K

[0553]

[0554] External blockage by the test compound

[0555] Once recording began, a voltage protocol was executed at 30-second intervals for 5 minutes to establish a stable baseline. A 5 Hz stimulus of the same voltage protocol was followed until the end of the experiment. The test compound was added during the 5 Hz stimulus train to allow cells to wait until they exhibited a stable current rundown rate before compound addition. After adding the test compound for 5 minutes, the cells were washed with standard Ringer's solution. Current was recorded using PatchMaster software with a Heka EPC10 (HEKA Electronics, Lambrecht, Germany). Only cells with an internal current amplitude of -20 mV between 400 pA and 4 nA were allowed. Additionally, cells with a leakage current exceeding 10% of the current amplitude were discarded.

[0556] Data Analysis: External Blocking

[0557] The data were plotted using Patchmaster software (HEKA Electronics, Lambrecht, Germany), and the minimum current during voltage steps up to -20 mV (peak internal current) was plotted and analyzed as a function of time. To determine the % cutoff generated by the externally applied test compound, the following was performed: After a stable current rundown rate was established during a 5 Hz excitation train, the rate was calculated by dividing the change in peak current amplitude by time. rundown was calculated. Using the average peak internal current amplitude (2-3 sec) before compound addition, 0% cutoff (I 0%block Decided on ). To modify the rundown, I 0%block (speed rundown The modified 0% cutoff current was obtained by subtracting * 5 min). The average peak internal current during the last 2-3 seconds of the 5-minute compound application time before washing was the uncutoff current (I unblocked ...is. Subsequently, the blocking % was calculated using the following equation: Fractional current blocking = 1 - I unblocked / ( I0%block - Speed rundown * 5 minutes).

[0558] Representative examples of the present invention were tested for extracellular inhibition of NaV 1.7. The range of activity is inhibition %: "++++" (>95%), "+++" 95-70%, "++" (70-40%), or "+" (< 40%). The results are presented below.

[0559] Table I

[0560]

[0561] Automated Patch Clamp:

[0562] cell culture

[0563] NaV1.7 was expressed in HEK293 cells upon tetracycline induction. Cells were cultured in DMEM containing 10% dialyzed fetal bovine serum (VWR, Radnor, PA), 1% glutamax (VWR, Radnor, PA), 1% penicillin-streptamicin (VWR, Radnor, PA), 100 mg / L hygromycin (Thermo Fisher Scientific, Waltham, MA), and 5 mg / L blasticidin (Alfa Aesar, Haverhill, MA). Cells were grown and maintained at 37°C in a humidified environment containing 10% CO2 in the air. Cells were detached from culture flasks for passage and harvested using 0.05% trypsin-EDTA (Thermo Fisher Scientific, Waltham, MA). To induce NaV1.7, cells were induced with tetracycline (0.1 - 1 μg / mL, IBI Scientific, Peosta, IA) the day before recording.

[0564] Before the experiment, cells were washed with DPBS (VWR, Radnor, PA), digested with Detachin, and then resuspended by grinding 10 times in CHO serum-free medium (VWR, Radnor, PA) to separate cell aggregates. Cells were counted, and the final concentration was set to 2 million to 5 million cells per mL.

[0565] Patch Clamp Solution & Drug

[0566] The intracellular solution contained the following: 140 mM CsF, 1 mM / 5 mM EGTA / CsOH, 10 mM HEPES, 10 mM NaCl, adjusted to pH 7.3 using CsOH and an osmolal concentration of 320 using sucrose. The extracellular solution contained the following: 145 mM NaCl, 4 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM HEPES, 10 mM glucose, adjusted to pH 7.4 using CsOH and an osmolal concentration of 305 using sucrose. All chemicals were from Sigma-Aldrich (St. Louis, MO). To test the degree of internal blockade by the test compound, the compound was dissolved in the internal solution at the indicated test concentration. In the control experiment, the internal solution contained no compound. To test the degree of external blockade by the test compound, the compound was dissolved in the external solution at the indicated test concentration.

[0567] Automated Patch Clamp Protocol

[0568] Automated patch clamping was performed on a Qube 384 (Sophion Bioscience, Woburn MA) equipped with a multi-hole Qchip at a temperature setting of 22°C. The entire cell configuration was formed using default Qube sealing and break-in parameters. The membrane potential was maintained at -100 mV before the voltage protocol was initiated. Two voltage protocols were followed.

[0569] Step 1: Cells were maintained at -100 mV with a depolarization pulse of -20 mV for 10 ms, and the interval was set to 5 s. The current was modified by subtracting the default leakage from all pulses. The duration was set to 5 minutes.

[0570] Step 2: The cells were maintained at -100 mV with a depolarization pulse of -20 mV for 10 ms. The frequency was 5 Hz. The current was modified by the leakage deduction calculated prior to Step 2. The duration was set to 4 minutes.

[0571] Internal blockage by test compound

[0572] After Step 2, the Q-chip was removed from the recording chamber. The internal solution was changed to a solution containing the test compound. After replacing the Q-chip in the recording chamber, it was maintained at -100mV without pulses. The total solution switching time was 8 minutes. After the internal solution exchange, the cells were recorded, and Step 2 was repeated for 10 minutes.

[0573] Data analysis was performed using a Sophion Analyzer. Cells were filtered with a minimum sealing resistance of 50 MOhm and a minimum starting current of 5 nA. The current rundown was adjusted for control cells (non-drug). The remainder was calculated by averaging the last three points at the end of the experiment. The baseline was calculated as the average of the last three points of Step 2. IC 50 The curve was plotted using the DR-plot / Hill function (dose-response plot with Hill fit). Data analysis was performed using Sophion Analyzer.

[0574] A representative example of the present invention was tested for the intracellular inhibition of NaV 1.7 in an automated patch clamp assay. The active range is IC50. 50 It is reported as: "++++" (< 1 μM), "+++" (1-3 μM), "++" (3-10 μM) or "+" (10-30 μM). The results are presented in Table M.

[0575] Table M

[0576]

[0577] External blockage by the test compound

[0578] After Step 2, the external solution was changed to a solution containing the test compound. The Qchip was maintained at -100mV without pulses. The total solution switching time was 8 minutes. After the external solution exchange, cells were recorded for 10 minutes using the same procedure as in Step 2.

[0579] Data analysis was performed using Sophion Analyzer. The data was modified to control cells (non-drug). IC 50 The data was plotted using the DR-plot / Hill function (dose-response plot with Hill fit).

[0580] A representative example of the present invention was tested for the extracellular inhibition of NaV 1.7 in an automated patch clamp assay. The active range is IC50. 50 It is reported as: "++++" (<10 μM), "+++" (10-30 μM), "++" (30-100 μM) or "+" (>100 μM). The results are presented in Table N.

[0581] Table N

[0582]

[0583] Example 4 - Membrane Permeability

[0584] The ability of the compound of the present invention to cross an artificial lipid membrane by passive diffusion was determined using the PAMPA assay (pION, Inc., Woburn MA). The test compound was dissolved in DMSO (10 mM) and diluted 200-fold in buffer (pION Inc., pH 7.4) to provide a 50 μM stock solution. Buffer (150 μL) was added to a UV blank plate, and the stock solution (150 μL) was transferred to a UV reference plate. The blank and reference spectra were read using a spectrophotometer. Stock solution (200 μL) was added to the donor plate of a PAMPA sandwich plate, and a receptor plate painted with GIT lipid (pION Inc., 5 μL) was placed on top. Buffer (200 μL) was added to the receptor plate, and the PAMPA sandwich plate was incubated for 4 hours. An aliquot (150 μL) from the receptor plate was added to a UV plate and read using the receptor spectrum. An aliquot (150 μL) of the donor solution was added to a UV analysis plate and read as a donor spectrum. Based on the AUC of the reference plate, donor plate, and acceptor plate, the penetration coefficient of the test compound was calculated using PAMPA Explorer™ software (version 3.5.0.4).

[0585] PAMPA penetration results of representative compounds (10 -6 cm / s) is "+" (< 0.1 10 -6 cm / s), "++"0.1-2.0 10 -6 cm / s), "+++" (2.0-10.0 10 -6 cm / s) or "++++" (>10.0 10 -6 It is reported as cm / s (Table O).

[0586] Table O

[0587]

[0588] The patents and scientific literature cited herein establish the knowledge available to those skilled in the art. All U.S. patents and published or unpublished U.S. patent applications cited herein are incorporated by reference. All published foreign patents and patent applications cited herein are incorporated by reference. All other published references, literature, manuscripts, and scientific literature cited herein are incorporated by reference.

[0589] Although the present invention has been specifically presented and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various variations in form and detail may be made within the scope of the invention as set forth in the appended claims without departing from the scope of the invention. Furthermore, it will be understood that none of the embodiments described herein are mutually exclusive and may be combined in various ways without departing from the scope of the invention as set forth in the appended claims.

Claims

Claim 1 The following chemical formula ( I Compounds represented by ): In the above equation, Y - is a pharmaceutically acceptable anion; R F and R G is N to which they are attached + Together with, it forms a pyridinium ring substituted with one or more substituents, wherein the one or more substituents are C3-C6 cycloalkyl, phenyl, C 1-6 Alkoxy, or CO2R 2A Selected from a group composed of, and R 2A is C 1-6 Selected from alkyl; R A is a halogen, C 1-6 Alkyl, CN, and -OC 1-6 Selected from alkyls; R B is C 1-6 It is alkyl and;R C is H, halogen, C 1-6 Selected from alkyl, or CN; X 1 is -NHC(O)- and;R D and R E Each independently H, and C 1-6 Selected from alkyls. Claim 2 In paragraph 1, Y - A compound that is iodide, bromide, or chloride. Claim 3 In paragraph 1, R A and R B Each independently C 1-4 Alkyl compound. Claim 4 In paragraph 1, R A and R B Each is CH3, and R C A compound selected from the group consisting of H, CH3, halogen, and CN. Claim 5 In paragraph 1, R D Ga C 1-4 Alkyl compound. Claim 6 In paragraph 1, R E a H, or C 1-4 Alkyl compound. Claim 7 In paragraph 1, R D and R E A compound in which both are hydrogen. Claim 8 In paragraph 1, R D is hydrogen, and R E A compound in which α is a C1-C6 alkyl. Claim 9 In paragraph 1, R F and R G The ring formed together by is a C3-C6 cycloalkyl, or CO2R 2A It is replaced with, where R 2A Ga C 1-6 A compound selected from alkyls. Claim 10 Compounds selected from the following: . Claim 11 A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, for treating pain, cough, itching, or neuroinflammatory disorders in a patient. Claim 12 A composition according to claim 11, wherein the composition is formulated for oral, intravenous, intramuscular, rectal, skin, subcutaneous, topical, transdermal, sublingual, nasal, inhalation, vaginal, intradural, epidural, or ocular administration. Claim 13 A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, for treating pain, cough, itching, or neuroinflammatory disorders in a patient. Claim 14 In Paragraph 13, the above pain includes lower back pain and neck pain, lumbago, cancer pain, gynecological pain and analgesia, fibromyalgia, arthritis, rheumatoid arthritis, osteoarthritis, rheumatic pain, orthopedic pain, acute and post-herpetic neuralgia and other neuropathic pain, sickle cell crisis, vulvodynia, perianal pain, irritable bowel disease, irritable bowel syndrome, inflammatory bowel disease, oral mucositis, esophagitis, interstitial cystitis, urethritis and other urological pain, toothache, headache, trigeminal neurodystrophic syndrome, erythematous limb pain, abdominal wall pain, chronic abdominal wall pain, allergic rhinitis, myalgia, rectal pain, levator ani syndrome, transient rectalgia, hemorrhoidal pain, stomach pain, skin ulcer, gastric ulcer, burn pain, ophthalmic irritation, conjunctivitis, eye hyperemia, dry eye, dry eye syndrome, complex regional pain syndrome, postoperative ocular pain, postoperative pain, acute surgery A pharmaceutical composition selected from the group consisting of post-procedure pain and pain caused by procedure pain. Claim 15 A pharmaceutical composition according to claim 13, wherein the cough is selected from the group consisting of asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), idiopathic pulmonary fibrosis, postviral cough, post-infectious cough, chronic idiopathic cough, and cough in patients with lung cancer. Claim 16 A pharmaceutical composition according to claim 13, wherein the above-mentioned itching is selected from the group consisting of pruritus, brachioradialis pruritus, chronic idiopathic pruritus, genital / anal pruritus, paresthetic back pain, scalp pruritus, allergic dermatitis, contact dermatitis, atopic dermatitis, hand eczema, poison ivy, infection, parasite, insect bite, pregnancy, metabolic disorder, hepatic or renal failure, drug reaction, allergic reaction, eczema, genital and anal itching, hemorrhoidal itching, and itching caused by cancer. Claim 17 A pharmaceutical composition according to claim 13, wherein the neuroinflammatory disorder is selected from the group consisting of allergic inflammation, asthma, chronic cough, conjunctivitis, rhinitis, psoriasis, inflammatory bowel disease, interstitial cystitis, arthritis, colitis, contact dermatitis, diabetes mellitus, eczema, cystitis, gastritis, migraine, rosacea, sunburn, pancreatitis, chronic rhinosinusitis, traumatic brain injury, polybacterial sepsis, tendinopathy, chronic urticaria, rheumatic disease, acute lung injury, exposure to irritants, irritants, contaminants, inhalation of chemical warfare agents, and atopic dermatitis. Claim 18 In Clause 13, chemical formula ( I A pharmaceutical composition in which a compound represented by ) is used in combination with one or more exogenous large porous receptor agonists. Claim 19 delete Claim 20 delete Claim 21 delete Claim 22 delete Claim 23 delete

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