Charged ion channel blockers and methods of use thereof

Abstract

The invention provides a charged ion channel blocker and a use method thereof. The present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof: # imgabs0. The compounds, compositions, methods, or kits of the invention can be effectively used for the treatment of pain, cough, pruritus, and neurogenic inflammation.

Description

[0001] This application is a divisional application of application number 202080033901.4 filed on March 11, 2020, and the invention name is “Charged ion channel blockers and methods of use thereof”.

[0002] Related Applications

[0003] This application claims priority to U.S. Provisional Patent Application No. 62 / 816,434 filed on March 11, 2019 and U.S. Provisional Patent Application No. 62 / 931,590 filed on November 6, 2019. The entire contents of the above applications are hereby incorporated by reference in their entirety. Technical Field

[0004] The present invention generally relates to quaternary ammonium compounds, pharmaceutical compositions, and methods for use as selective inhibitors of pain, cough and itch sensing neurons (nociceptive neurons, cough neurons and itch receptor neurons) and for treating neurogenic inflammation. Background Art

[0005] The invention features compounds, compositions and methods for selectively inhibiting sensory neurons (nociceptive neurons, cough receptor neurons and itch receptor neurons), and therapies for treating neurogenic inflammation by targeting nociceptors with small molecule drugs while minimizing effects on non-nociceptive neurons or other types of cells. According to the methods of the invention, small cationic drug molecules enter the intracellular compartment of sensory neurons through macroporous receptors / ion channels that are present in pain-neurons, cough receptor neurons and itch receptor neurons, but to a lesser extent or not at all in other types of neurons or other types of tissues.

[0006] Local anesthetics, such as lidocaine and articaine, work by inhibiting voltage-dependent sodium channels on neurons. These anesthetics can inhibit sodium ion channels, thereby affecting the excitability of all neurons, not just pain neurons (nociceptive neurons). Therefore, although the purpose of local or regional anesthesia is to prevent pain by blocking the signals transmitted by nociceptive neurons, the use of local anesthetics can also produce unwanted or harmful effects, such as general numbness caused by blocking low threshold pressure and touch sensory neurons, movement disorders and / or paralysis caused by blocking motor axes, and other complications caused by blocking autonomic nerve fibers. Local anesthetics are relatively hydrophobic molecules that diffuse through the cell membrane into the blocking points on the sodium channels. Charged derivatives of these compounds are cell membrane impermeable and have no effect on the sodium channels of nerve cells when applied to the outer surface of the nerve membrane, but if introduced into the cell in some way, such as by diffusion from a micropipette used for whole-cell electrophysiological recording of isolated neurons, they can affect the blocking of sodium channels. Pain-, cough-, and itch-receptor neurons differ from other types of neurons (in most cases) in expressing TRPV1 receptors / channels, which are activated by painful heat or capsaicin (the spicy ingredient in chili peppers). Other types of channels are selectively expressed in various types of pain-, cough-, and itch-receptor neurons, including but not limited to TRPV2-4, TRPA1, TRPM8, ASIC, and P2X(2 / 3) channels. It is well known that some cationic small molecules, such as QX-314, are able to enter cells through activated large pore channels, such as TRPV1.

[0007] Neuropathic, inflammatory, and nociceptive pain differ in etiology, pathophysiology, diagnosis, and treatment. Nociceptive pain is produced by the activation of a specific high-threshold subset of peripheral sensory neurons, the nociceptive neurons, by intense or noxious stimuli. It is typically acute, self-limited, and serves to protect biological function by serving as a warning of potential or ongoing tissue damage. It is usually well localized. Examples of nociceptive pain include, but are not limited to, traumatic or surgical pain, labor pain, sprains, fractures, burns, masses, bruises, injections, dental procedures, skin biopsies, and obstructions.

[0008] Inflammatory pain is pain that occurs in the presence of tissue damage or inflammation, including postoperative pain (i.e., pain associated with acute perioperative pain caused by inflammation resulting from tissue trauma (e.g., surgical incision, dissection, burns) or direct nerve injury (e.g., nerve severance, stretching, or compression)), posttraumatic pain, arthritis (rheumatoid; or osteoarthritis (i.e., joint pain and stiffness caused by the gradual deterioration of joint cartilage; risk factors include aging, injury, and obesity; commonly affected joints include the hands, wrists, neck, knees, hips, and spine), pain associated with damage to joints, muscles, and tendons, such as axial low back pain (i.e., a generalized painful condition affecting the lower back; common causes include muscle strain, spinal fracture, bulging or ruptured disc, and arthritis), and severe nociceptive pain that may transform into inflammatory pain if there is associated tissue damage.

[0009] Neuropathic pain is a common, chronic, non-malignant pain condition that is the result of damage or malfunction of the peripheral or central nervous system without protective biological functions. It is estimated to affect more than 1.6 million people in the U.S. population. Neuropathic pain has many different etiologies, for example, due to trauma, surgery, herniated disks, spinal cord injuries, diabetes, herpes zoster infection (shingles), HIV / AIDS, advanced disease / cancer, amputation (including mastectomy), carpal tunnel syndrome, chronic alcohol use, exposure to radiation, and unintended side effects of neurotoxic therapeutic drugs, such as certain anti-HIV and chemotherapy drugs. Peripheral neuropathic pain is damage to the peripheral nerves due to injury, trauma, prolonged pressure, or inflammation that causes numbness and pain in the corresponding area of ​​the body.

[0010] Neuropathic pain is often described as a "burning," "electrical," "tingling," or "throbbing" sensation. It is often characterized by chronic dynamic allodynia (defined as pain evoked by movement stimuli that do not normally evoke a painful response, such as light touch) and hyperalgesia (defined as increased sensitivity to normally painful stimuli) and may persist for months or years until any damaged tissue has significantly healed.

[0011] Patients with cancer also experience pain, which is caused by a variety of reasons: inflammation, compression, invasion, and metastasis to bones or other tissues.

[0012] In some cases, pain occurs in the absence of noxious stimulation, tissue damage, or nervous system injury, known as functional pain, including but not limited to fibromyalgia, tension-type headaches, and irritable bowel disorder.

[0013] Migraine is a type of headache associated with activation of sensory fibers innervating the meninges.

[0014] Pruritus (itch) is a skin disorder that may be localized or systemic and may be associated with skin lesions (rash, atopic eczema, urticaria). Itch accompanies many conditions, including but not limited to stress, anxiety, ultraviolet radiation from the sun, metabolic and endocrine diseases (e.g., liver or kidney disease, hyperthyroidism), cancer (e.g., lymphoma), reactions to medications or foods, parasitic and fungal infections, allergic reactions, blood disorders (e.g., polycythemia vera), and skin diseases. Itch is mediated by a subset of small-diameter primary sensory neurons, the pruritoreceptors, which share many features of nociceptor neurons, including but not limited to the expression of TRPV1 channels, as well as other large-pore channels such as TRPV2-4, TRPA1, TRPM8, ASICs, and P2X(2 / 3). Certain itch mediators—such as eicosanoids, histamine, bradykinin, ATP, and various neurotrophins—function as endogenous anthocyanins. Topical capsaicin can inhibit histamine-induced itch. Therefore, itch receptor neurons, similar to nociceptive receptor neurons, are suitable targets for this approach to deliver ion channel blockers.

[0015] Cough is a defensive reflex designed to protect the airways from foreign bodies and help clear luminal debris. However, this reflex can become abnormal in some diseases, resulting in an ineffective, dry cough with the presence of an excessive cough or ectopic cough state. Excessive cough and ectopic cough states are often chronic, lasting for more than 3 months, and can manifest in many airway disease states, including asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), and lung cancer. In addition, an inappropriate cough reflex can manifest acutely and chronically following viral infections. Furthermore, chronic cough can be idiopathic with unknown etiology.

[0016] Neurogenic inflammation is a mode of inflammation mediated by the efferent (motor) function of sensory neurons. Proinflammatory mediator molecules released peripherally by pain neurons (nociceptive neurons) not only activate multiple inflammatory pathways in immune cells, but also act on the vascular system to change blood flow and capillary permeability.

[0017] Neurogenic inflammation is peripheral inflammation caused by tissue injury, autoimmune diseases, infection, allergy, exposure to irritants in various tissues, and is thought to play an important role in the pathogenesis of many diseases such as migraine, arthritis, rhinitis, gastritis, colitis, cystitis, and sunburn. One way to reduce neurogenic inflammation is to block the excitability of nociceptive neurons, thereby preventing the activation of the peripheral terminals of nociceptive neurons and the release of pro-inflammatory chemicals.

[0018] Despite the diversity of approaches to treat pain, itch, and neurogenic inflammation, additional agents are needed. Summary of the invention

[0019] The present invention provides a compound of formula (I) which can be used to treat or prevent pain, itching and neurogenic inflammation:

[0020]

[0021] in,

[0022] Y - is a pharmaceutically acceptable anion;

[0023] R A , R B , and R C are independently selected from H, D, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, OR I , CN, NR J R K , NR L C(O)R M ,S(O)R N , S(O)2R N , SO2R O R P , SO2NR Q R R , SO3R S ,CO2R T ;C(O)R U , and C(O)NR V R W ; (preferably H, F, Cl, or CN and more preferably H);

[0024] 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 WEach of R is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl; R J and R K or R V and R W or R Q and R R They can also form a substituted or unsubstituted 5-, 6-, 7- or 8-membered ring together with the nitrogen atom to which they are attached.

[0025] R A , R B and / or R C They can form fused bicyclic or tricyclic ring systems together with the benzene ring to which they are attached, such as naphthyl, indanyl, tetrahydronaphthyl, quinolyl, indolyl, etc.;

[0026] X 1 Selected from—CR X R Y —,—NR Z C(O)—, —OC(O)—, —SC(O)—, —C(O)NR 1A —, —C(O)O—, -C(O)-, —(O)CS—, —NR 1A S(O)—,—S(O)NR 1A —,—NR 1A C(O)NR 1A —, —S(O)— and —S(O)2—; X 1 Can be -NR Z C(O)CR X R Y —;

[0027] R X , R Y , R Z , and R 1A Each of the above is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl and substituted or unsubstituted heteroalkyl;

[0028] R D and R E each independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl; or R D and RE Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted 3-6-membered cycloalkyl (C3-C6 cycloalkyl) or a substituted or unsubstituted heterocyclyl or a substituted or unsubstituted heteroalkyl ring;

[0029] or R D and R Z Together with the carbon atom to which they are attached or with -NC(O)- they form an optionally substituted 5- to 8-membered lactam.

[0030] R F and R G With N + together form an optionally substituted heterocyclyl ring having one or more nitrogen atoms, or, R F and R G are independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted C3-C6 cycloalkyl; and

[0031] R H is a substituted or unsubstituted aromatic ring, or a substituted or unsubstituted heteroaromatic ring.

[0032] In other embodiments, R H It may be a substituted alkyl. The substituent preferably has an ester group, such as -OC(O)R 1B , where R 1B is substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. 1B Preferred is substituted or unsubstituted phenyl. H Preferred is -CH2OC(O)-phenyl. DETAILED DESCRIPTION

[0033] The present invention provides a compound represented by formula (I) as described above, or a pharmaceutically acceptable salt, stereoisomer, solvent, hydrate or a combination thereof. The present invention also provides a composition comprising a compound having formula (I) or a pharmaceutically acceptable salt thereof, for example, a composition comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. The composition of the present invention may also include a compound of the present invention and a bioactive agent. The composition described herein may be formulated for oral, intravenous, intramuscular, rectal, dermal, subcutaneous, topical, transdermal, sublingual, nasal, inhalation, vaginal, intrathecal, epidural or intraocular administration.

[0034] The present invention also provides a method for treating pain, cough, pruritus or neurogenic inflammatory disorders in a patient, comprising administering to the patient a composition comprising a compound of formula (I), wherein the compound inhibits one or more voltage-gated ion channels present in nociceptive neurons and / or cough receptor neurons and / or itch receptor neurons when exposed or applied to the inner surface of the channel, but does not substantially inhibit the channel when applied to the outer surface of the channel, and wherein, when the channel is activated, the compound is able to enter the nociceptive neurons, cough receptor neurons or itch receptor neurons through the macroporous channel, and the compound is able to inhibit one or more voltage-gated ion channels on the nociceptive neurons, cough receptor neurons and / or itch receptor neurons.

[0035] In certain embodiments, macroporous channel is transient receptor potential ion channel (Trp channel). In other embodiments, Trp channel is activated by exogenous or endogenous agonist. Still in other embodiments, macroporous channel is TRPA1, TRPV1-4, TRPM8, ASIC or P2X. In specific embodiments, when receptor / channel is activated, the compound can enter nociceptive neurons, cough sensory neurons or itch sensory neurons through TRPA1, TRPV1-4, TRPM8, ASIC or P2X receptor / channel. Still in another embodiment, the compound inhibits voltage-gated sodium channels. Still in another embodiment, the pain type treated by the method, composition and kit of the present invention is selected from neuropathic pain, inflammatory pain, nociceptive pain, pain caused by infection and procedural pain, or wherein neurogenic inflammatory disorder is selected from allergic inflammation, asthma, chronic cough, conjunctivitis, rhinitis, psoriasis, inflammatory bowel disease, interstitial cystitis and atopic dermatitis.

[0036] Here we identify the compound represented by formula (I):

[0037]

[0038]

[0039] The compound can pass through the open macropore channel expressed on pain sensory neurons and / or cough sensory neurons and / or itch sensory neurons but not expressed on motor neurons. Because the ion channel blocking compounds of the present invention are positively charged, they are not membrane permeable, so they cannot enter cells that do not express macropore channels. Due to the release of endogenous ligands or the activation of thermal stimulation, macropore channels are usually more active in the tissue conditions related to pain (such as inflammation), so the ion channel blockers of the present invention can be used alone for selectively targeting the nociceptive neurons activated, so as to effectively treat (for example, eliminate or alleviate) pain, cough, pruritus or neurogenic inflammation. The ion channel blockers of the present invention can also be used in combination with one or more exogenous macroporous receptor agonists, to selectively target nociceptive neurons, so as to effectively treat (for example, eliminate or alleviate) pain, cough, pruritus or neurogenic inflammation.

[0040] Voltage-dependent ion channels in pain neurons are currently of interest in the development of drugs to treat pain. Blocking voltage-dependent sodium channels in pain neurons can block pain signals by blocking the initiation and transmission of action potentials. In addition, blocking voltage-dependent sodium channels in nociceptive neurons can reduce or eliminate neurogenic inflammation by preventing the activation of nociceptor peripheral terminals and the release of their pro-inflammatory chemicals.

[0041] One limitation of treatments using molecules that block sodium or calcium channels to date is that the vast majority of these externally applied molecules are hydrophobic and can pass through cell membranes. They will therefore enter all cells and thus have no selectivity for affecting only nociceptive neurons.

[0042] The inhibitors of the present invention cannot penetrate the cell membrane and are only effective when present in the nociceptive neuron cells. Therefore, they must pass through the cell membrane through channels or receptors, such as large pore channels (such as TRPAV1-4, TRPA1, TRPM8, ASIC and P2X (2 / 3)) to produce an effect. Under normal circumstances, most large pore channels in nociceptive neurons are not active, but require harmful thermal, mechanical or chemical stimulation to activate them. For example, TRP channels in nociceptors can be activated by exogenous TRP ligands (i.e., TRP agonists) such as capsaicin to open TRPV1 channels. Therefore, one method of selectively targeting nociceptive neurons is to co-administer a membrane-impermeable ion channel inhibitor with an exogenous TRP ligand that allows the inhibitor to enter the cell through the TRP channel. In addition to capsaicin, the exogenous TRP ligand can also be another capsaicin, mustard oil or lidocaine. In another embodiment, the TRP channel may be activated in response to an exogenous stimulus activator (such as acrolein inhaled from smoke) or a chemical agent (such as tear gas).

[0043] In some cases, when there is no exogenous macroporous channel agonist / ligand, endogenous inflammatory activators due to tissue damage, infection, autoimmunity, atopy, ischemia, hypoxia, cell stress, immune cell activation, immune mediator production, and oxidative stress can also activate macroporous channels. Under such conditions, endogenous molecules (such as protons, lipids, and reactive oxygen) can activate macroporous channels expressed on nociceptive neurons, allowing membrane-impermeable voltage-gated ion channel blockers to enter the interior of nociceptive neurons through endogenously activated macroporous channels. Endogenous inflammatory activators of macroporous channels include prostaglandins, nitric oxide (NO), hydrogen peroxide (H2O2), cysteine-reactive inflammatory mediators, such as 4-hydroxynonenol, protons, ATP, endogenous alkenyl aldehydes, and immune mediators (such as interleukin 1 (IL-1), nerve growth factor (NGF), and bradykinin, whose receptors are coupled to macroporous channels).

[0044] definition

[0045] As used herein, the terms "a" and "an" are intended to include one or more than one unless expressly stated otherwise.

[0046] "Biological activity" refers to a molecule, including a biomolecule, such as a nucleic acid, peptide, polypeptide, and protein, that exerts a biological, physical, or chemical effect or activity on a protein, enzyme, receptor, ligand, antigen, itself, or other molecule. For example, a "biologically active" molecule may have, for example, enzymatic activity, protein binding activity, or pharmacological activity.

[0047] Bioactive agents that can 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, anesthetics, antiproliferative and immunomodulatory agents, antibodies or antibody fragments, antibiotics, polynucleotides, polypeptides, proteins, anticancer agents, growth factors, and vaccines.

[0048] "Inflammation" refers to any type of inflammation, such as that caused by the immune system (immune-mediated inflammation) and the nervous system (neurogenic inflammation) and any symptoms of inflammation, including redness, heat, swelling, pain, and / or loss of function.

[0049] "Neurogenic inflammation" refers to any type of inflammation mediated or contributed to by neurons (eg, nociceptive neurons) or any other component of the central or peripheral nervous system.

[0050] The term "pain" is used here in the broadest sense to refer to all types of pain, including acute and chronic pain, such as nociceptive pain, such as somatic pain and visceral pain; inflammatory pain, functional pain, idiopathic pain, neuropathic pain, such as central pain and peripheral pain, migraine, and cancer pain.

[0051] The term "nociceptive pain" is used to include all pain caused by noxious stimuli that threaten or actually injure body tissue, including but not limited to wounds, abrasions, fractures, crush injuries, burns, etc. Pain receptors for tissue damage (nociceptors) are primarily located in the skin, musculoskeletal system, or internal organs.

[0052] The term "somatic pain" refers to pain arising from the bones, joints, muscles, skin, or connective tissue. This type of pain is usually well localized.

[0053] The term "visceral pain" is used herein to refer to pain arising from visceral organs, such as the respiratory tract, gastrointestinal tract and pancreas, urinary tract and reproductive organs. Visceral pain includes pain caused by tumors invading the capsule of an organ. Another type of visceral pain, usually caused by obstruction of a hollow viscus, is characterized by intermittent cramping and severe localized pain. Visceral pain may be associated with inflammation, such as cystitis or reflux esophagitis.

[0054] The term "inflammatory pain" includes pain associated with activated inflammation, which may be caused by trauma, surgery, infection, and autoimmune disease.

[0055] The term "neuropathic pain" is used herein to refer to pain resulting from abnormal processing of sensory input by the peripheral or central nervous system due to damage to these systems.

[0056] The term "procedural pain" refers to pain experienced during a medical, dental, or surgical procedure, where the procedure is usually planned or associated with an acute trauma.

[0057] The term "pruritus" is used here in the broadest sense to refer to all types of itching and tingling sensations, localized and generalized, acute intermittent and persistent. Pruritus may be idiopathic, allergic, metabolic, infectious, drug-induced, due to liver or kidney disease or cancer. "Pruritus" refers to severe itching.

[0058] "Patient" refers to any animal. In one embodiment, the patient is a human. Other animals that can be treated using the methods, compositions, and kits of the invention include, but are not limited to, non-human primates (e.g., monkeys, gorillas, chimpanzees), domesticated animals (e.g., horses, pigs, goats, rabbits, sheep, cattle, camels), and companion animals (e.g., guinea pigs, mice, rats, lizards, snakes, dogs, cats, fish, hamsters, and birds).

[0059] Compounds useful in the present invention include, but are not limited to, compounds described herein in any pharmaceutically acceptable form thereof, including isomers, such as diastereomers and enantiomers, salts, esters, amides, thioesters, solvates, and polymorphs thereof, 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 acids are described in Stahl, PH and Wermuth, CG (eds.), Handbook of Pharmaceutical Salts: Properties, Selection and Use, Wiley VCH (2008). Pharmaceutically acceptable acids include, but are not limited to, acetic acid, dichloroacetic acid, adipic acid, alginic acid, L-ascorbic acid, L-aspartic acid, benzenesulfonic acid, 4-acetamidobenzoic acid, benzoic acid, p-bromobenzenesulfonic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, cyclohexyl acid, dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, sulfuric acid, boric acid, citric acid, formic acid, fumaric acid, galactic acid, gentisic acid, D-glucoheptanoic acid, D-glucuronic acid, glutamic acid, glutaric acid, 2-oxopentanoic acid, Diacid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, 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, palmitic acid, phosphoric acid, propionic acid, (-)-L-pyroglutamic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, (+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, and undecylenic acid. Pharmaceutically acceptable anions include conjugate bases of any of the above acids.

[0060] The term "pharmaceutically acceptable salts" means salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit / risk ratio. The salts may be prepared in situ during the final isolation and purification of the compounds of the invention, or separately prepared by reacting a free base function with a suitable organic acid. Representative acid salts include, but are not limited to, acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphor ester, camphor sulfonate, citrate, cyclopentylpropionate, digluconate, dodecyl sulfate, ethane sulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxyethane sulfonate, isothiocyanate, lactobionate, lactic acid, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, and the like.

[0061] In general descriptions of compounds of the invention, the number of atoms of a particular type in a substituent is often given as a range, e.g., an alkyl group containing 1 to 4 carbon atoms or a C 1-4 Alkyl or C1-C4 alkyl. Reference to such ranges is intended to include specific reference to groups having each of the integer number of atoms within the specified range. For example, an alkyl group from 1 carbon atom to 4 carbon atoms includes each of C1, C2, C3, and C4 alkyl groups. Other numbers of atoms and other types of atoms may be represented in a similar manner.

[0062] "D" is for deuterium.

[0063] As used herein, the term "alkyl" and the prefix "alkyl-" include straight chain groups, branched groups and cyclic groups, for example, cycloalkyl. The cyclic radical group can be monocyclic or polycyclic, and preferably has 3 to 6 ring carbon atoms or 3 to 7 carbon atoms. Exemplary cyclic radical groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0064] “C 1-4 "C1-C4 alkyl" or "C1-C4 alkyl" refers to a branched or unbranched hydrocarbon group having 1 to 4 carbon atoms. Similarly, "C 1-6 "Alkyl" or "C1-C6" refers to a branched or unbranched hydrocarbon group having 1 to 6 carbon atoms. Alkyl includes, for example, C 1-4 Alkyl or C 1-6Alkyl groups, which 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 amine, alkylcarboxyl and carboxyl. Exemplary substituents also include alkoxy, aryloxy, thiol, alkylthio, arylthio, halide (F, Cl, Br or I), hydroxy, fluoroalkyl, perfluoroalkyl, oxo, amino, alkylamino, disubstituted amino, quaternary amino, amino, ester, alkylcarboxyl, alkoxycarbonyl, alkoxycarbonyl, aryloxycarbonyl, carboxyl, alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, phosphate, phosphonate, phosphonate, amide (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amino, imino, thiol, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfoxide, sulfonate, sulfamoyl, sulfonamide, nitro, trifluoromethyl, cyano, azido, aryl, heterocyclyl, alkylaryl, or aromatic or heteroaromatic moiety. 1-4 Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl, n-butyl, isobutyl, sec-butyl, and cyclobutyl. 1-6 Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0065] An example of a substituted alkyl group is a heteroalkyl group. "Heteroalkyl" refers to branched and unbranched alkyl, cycloalkyl, alkenyl, or alkynyl groups having 1 to 7 or more carbon atoms and additional 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O, S and P. "C 1-7"Heteroalkyl" refers to branched and unbranched alkyl, alkenyl, or alkynyl groups having 1 to 7 carbon atoms and additionally 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O, S and P. Heteroalkyl groups include, but are not limited to, tertiary amines, secondary amines, ethers, thioethers, amides, sulfamides, carbamates, thiocarbamates, hydrazones, imides, phosphodiesters, phosphoamides, sulfonamides, and disulfides. Heteroalkyl groups may optionally include monocyclic, bicyclic or tricyclic rings, wherein each ring preferably has three to six ring atoms. The heteroalkyl group may be substituted or unsubstituted. Exemplary substituents include alkyl, alkoxy, aryloxy, mercapto, alkyl sulfur, arylsulfur, halide (F, Cl, Br or I), hydroxy, fluoroalkyl, perfluoroalkyl, oxo, amino, alkylamino, disubstituted amino, quaternary amino, amino, ester, alkylcarboxyl, alkoxycarbonyl, alkoxycarbonyl, aryloxycarbonyl, carboxyl, alkylcarbonyl, arylcarbonyl, alkylthiocarbonyl, phosphate, phosphonate, phosphonate, amide (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amino, imino, sulfhydryl, alkylsulfide, arylsulfide, sulfocarboxylate, sulfate, alkylsulfoxide, sulfonate, sulfamoyl, sulfonamide, nitro, trifluoromethyl, cyano, azide, aryl, heterocyclyl, alkylaryl, or aromatic or heteroaromatic group. C 1-7 Examples of heteroalkyl groups include, but are not limited to, methoxymethyl and ethoxyethyl.

[0066] Alkenyl refers to a branched or unbranched hydrocarbon group containing one or more double bonds. 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 group may optionally include a single ring or multiple rings, wherein each ring ideally has three to six ring atoms. The alkenyl group may be substituted or unsubstituted. Exemplary substituents include those described above for alkyl, specifically, including but not limited to alkoxy, aryloxy, mercapto, alkylthio, arylthio, halide, hydroxyl, fluoroalkyl, perfluoroalkyl, amino, alkylamino, disubstituted amino, quaternary amine, alkylcarboxyl and carboxyl. C 2-6 Alkenyl groups include, but are not limited to, vinyl, allyl, 2-cyclopropyl-1-ethylene, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl and 2-methyl-2-propenyl.

[0067] Alkynyl refers to a branched or unbranched hydrocarbon group containing one or more triple bonds. 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 group may optionally include a monocyclic, bicyclic or tricyclic ring, wherein each ring ideally has five to six ring atoms. The alkynyl group may be substituted or unsubstituted. Exemplary substituents include those described above for alkyl, specifically, including but not limited to alkyl, as well as alkoxy, aryloxy, thiol, alkylthio, arylthio, halide, hydroxyl, fluoroalkyl, perfluoroalkyl, amino, alkylamino, disubstituted amino, quaternary amine, alkylcarboxyl and carboxyl. C 2-6 Alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl and 3-butynyl.

[0068] "Heterocyclyl", "heterocycle" or "heterocycloalkyl" refers to a stable monocyclic or polycyclic (including bicyclic or tricyclic) heterocyclic ring, which may be saturated, partially unsaturated or unsaturated (including heteroaryl or aromatic), and is composed of 2 or more carbon atoms and 1, 2, 3, 4 or more heteroatoms independently selected from N, O and S, including any bicyclic or polycyclic group, wherein any of the above-defined heterocycles is fused to a benzene ring, heteroaryl, cycloalkyl or heterocycloalkyl. In certain aspects, the heterocyclyl 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"Heterocyclyl" refers to a stable 5-7 membered monocyclic or 7-14 membered bicyclic heterocyclic ring, which may be saturated, partially unsaturated or unsaturated (including heteroaryl or aromatic), consisting of 2-6 carbon atoms and 1, 2, 3 or 4 heteroatoms independently selected from N, O and S, including any bicyclic group, wherein any of the above heterocyclic rings is fused to a benzene ring, heteroaryl, cycloalkyl or heterocycloalkyl. The heterocyclyl or heteroaryl group may be substituted or unsubstituted. Exemplary substituents include substituted or unsubstituted alkyl, aryl, cycloalkyl, heterocycloalkyl, heterocycloalkyl, The term "heterocyclic ring" refers to a group consisting of aryl, alkylthio, arylthio, halogenide, hydroxyl, fluoroalkyl, perfluoroalkyl, amino, alkylamino, disubstituted amino, quaternary amine, alkylcarboxyl and carboxyl. Nitrogen and sulfur heteroatoms can be optionally oxidized. Heterocycle can be covalently linked by any heteroatom or carbon atom that causes stable structure, for example, imidazoline ring can be connected to the carbon atom position of the ring or the nitrogen-atom. The nitrogen-atoms in the heterocycle can be quaternized. Preferably, when the sum of S and O atoms in the heterocycle exceeds 1, these heteroatoms are not adjacent to each other.Heterocyclic rings include, but are not limited to, 1H indole, 2-pyrrolidinyl, 2H, 6H-1,5,2-dithiazinyl, 2H-pyrrolyl, 3H-indolyl, 4-piperidinoyl, 4aH carbazole, 4H-quinolizinyl, 6H-1,2,5-thiadiazinyl, acridinyl, oxazolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothiazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, carbazolyl, 4aH carbazolyl, b-carbonyl, chromyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1,5,2-dithiazolyl, dihydrofuran [2,3-b] tetrahydrofuran, furanyl, furazolidinyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H indolazolyl, indollinyl, indolazolyl, indolizinyl, indolazolyl, indolizinyl, isobenzylfuranyl, isochromanyl, isoindolazolyl, isoindolyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl oxazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl piperidinyl, phenanthridinyl, phenanthrolinyl, phenanthrosazinyl, phenanthrazinyl, phenothiyl, phenothiazinyl, benzoxazinyl, o-phthalazinyl, piperazinyl, piperidinyl, pteridinyl, piperidyl, 4-piperidyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridinoxazole, pyridinimidazole, pyridinthiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, pyrrole, quinazolinyl, quinolinyl, 4H-quinazolinyl, quinoxaline 1,2,3-thiazolyl, 1,2,4-thiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thienyl, thiazolyl, thiophene, thienthiazolyl, thienoxyzolyl, thienyimidazolyl, phenylthio, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, flavonyl, β-lactam, γ-lactam and δ-lactam. Preferred 5 to 10 membered heterocycles include, but are not limited to, pyridyl, pyrimidyl, triazine, furanyl, thiazolyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, tetrazolyl, benzofuranyl, benzothiophenyl, indolyl, benzimidazolyl, 1H indolazolyl, oxazolidinyl, benzotriazolyl, benzisoxazolyl, octanol, benzoxazolinyl, quinolyl and isoquinolyl. Preferred 5 to 6 membered heterocycles include, but are not limited to, pyridyl, quinolyl, pyrimidyl, triazine, furanyl, thiazolyl, pyrrolyl, piperazinyl, piperidinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl and tetrazolyl. Preferred substituents include phenyl, methyl, ethyl, propyl, butyl, chlorine, bromine, fluorine and iodine.

[0069] "Aryl" refers to an aromatic group having a ring system consisting of carbon atoms with conjugated π electrons (e.g., phenyl). 12 Aryl" or "C6-C 10 "Aryl" is an aromatic radical having 6 to 12 carbon atoms or 6 to 10 carbon atoms, respectively. The aryl radical may optionally include a monocyclic, bicyclic or tricyclic ring, wherein each ring desirably has five or six members. The aryl radical may be substituted or unsubstituted. Exemplary substituents include substituted or unsubstituted alkyl, hydroxy, alkoxy, aryloxy, thiol, alkylthio, arylthio, halide, fluoroalkyl, carboxyl, alkylcarboxyl, amino, alkylamino, monosubstituted amino, disubstituted amino and quaternary amino. Preferred aryl is phenyl.

[0070] "Aralkyl" refers to a substituted or unsubstituted alkyl group substituted with a substituted or unsubstituted aryl group (including, for example, benzyl, phenethyl or 3,4-dichlorophenethyl).

[0071] "Heteroaralkyl" refers to an unsubstituted or substituted alkyl group substituted with a heteroaryl group.

[0072] “C 7-14 The "aralkyl group" means an alkyl group substituted by an aryl group having 7 to 14 carbon atoms (for example, benzyl, phenethyl or 3,4-dichlorophenethyl).

[0073] “C 3-10 "Heterocycloalkyl" refers to an alkyl substituted heterocyclic group having from 3 to 10 carbon atoms in addition to one or more heteroatoms (e.g., 3-furanylmethyl, 2-furanylmethyl, 3-tetrahydrofuranylmethyl or 2-tetrahydrofuranylmethyl).

[0074] "Halide" or "halogen" refers to bromine, chlorine, iodine or fluorine.

[0075] "Fluoroalkyl" refers to an alkyl group substituted with a fluorine atom.

[0076] "Alkylcarboxyl" refers to a chemical moiety having the formula -(R)-COOH, wherein R is selected from C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkynyl, C 2-6 Heterocyclic group, C 6-12 Aryl, C 7-14 Aralkyl, C 3-10 Heterocycloalkyl or C 1-7 Heterocycloalkyl.

[0077] "Alkoxy" refers to a chemical substituent of the formula --OR, where R is a substituted or unsubstituted alkyl, a substituted or unsubstituted alkenyl, or a substituted or unsubstituted alkynyl, or R can be selected from C 1-7 Alkyl, C 2-7Alkenyl, C 2-7 Alkynyl, C 2-6 Heterocyclic group, C 6-12 Aryl, C 7-14 Aralkyl, C 3-10 Heterocycloalkyl, or C 1-7 Heteroalkyl.

[0078] "Aryloxy" refers to a chemical substituent of the formula -OR, where R is C 6-12 Aryl.

[0079] "Alkylthio" refers to a chemical substituent of the formula -SR, wherein R is selected from C 1-7 Alkyl, C 2-7 Alkenyl, C 2-7 Alkynyl, C 2-6 Heterocyclic group, C 6-12 Aryl, C 7-14 Aralkyl, C 3-10 Heterocycloalkyl or C 1-7 Heterocycloalkyl.

[0080] "Arylthiol" refers to a chemical substituent of the formula -SR, where R is C 6-12 Aryl.

[0081] "Charged group" refers to a moiety that acquires a proton and thereby becomes positively charged at physiological pH (eg, ammonium, guanidine, or amide) or a moiety that contains a net formal positive charge without protonation (eg, quaternary ammonium). Charged groups may be permanently or transiently charged.

[0082] "Therapeutically effective amount" or "effective amount" refers to an amount sufficient to produce the desired result, for example, to reduce or eliminate pain, cough, itching or neurogenic inflammation in a patient (e.g., a human) suffering from a certain condition, disease, or a disease caused in whole or in part by neurogenic inflammation (such as asthma, arthritis, colitis, contact dermatitis, diabetes, eczema, cystitis, chronic refractory cough, post-viral cough, gastritis, migraine, psoriasis, rhinitis, rosacea or sunburn).

[0083] "Solvate" refers to a solvent addition form containing either stoichiometric or non-stoichiometric amounts of solvent.

[0084] Compounds of the present invention, including salts of compounds, can exist in unactivated forms and solvated forms, including hydrated forms and unhydrated forms. Generally speaking, solvated forms are equivalent to unsolvated forms and are included within the scope of the present invention. Non-limiting examples of hydrates include monohydrates, dihydrates, hemihydrates, etc. In some aspects, the compound is a hemihydrate. Non-limiting examples of solvates include ethanol solvates, acetone solvates, etc.

[0085] The compounds of the present invention may exist in a variety of crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0086] Compounds useful in the compositions, kits, and methods of the present invention include compounds of formula (I) or pharmaceutically acceptable salts thereof:

[0087]

[0088] in,

[0089] Y - is a pharmaceutically acceptable anion;

[0090] R A , R B , and R C are independently selected from H, D, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, OR I , NR J R K , NR L C(O)R M ,S(O)R N , S(O)2R N , SO2R O R P , SO2NR Q R R , SO3R S ,CO2R T , C(O)R U and C(O)NR V R W ;

[0091] 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 of which is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl;

[0092] X 1 Selected from—CRX R Y —,—NR Z C(O)—,—NR Z C(O)CR X R Y —, —OC(O)—, —SC(O)—, —C(O)NR 1A —, —C(O)O—, -C(O)-, —(O)CS—, —NR 1A S(O)—,—S(O)NR 1A —,—NR 1A C(O)NR 1A —, —S(O)— and —S(O)2—;

[0093] R X , R Y , R Z , and R 1A Each of which is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl;

[0094] 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 cycloalkyl; or R D and R E Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted C3-C6 cycloalkyl or a substituted or unsubstituted heterocyclyl (eg, a 5- to 7-membered heterocyclyl ring);

[0095] R F and R G N + together form an optionally substituted heterocyclyl ring having zero, one or more nitrogen atoms other than N+, or, R F and R G are independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted C3-C6 cycloalkyl; and

[0096] R H is a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group.

[0097] In a preferred embodiment, R H Independently selected from substituted or unsubstituted C 5-10Aryl or substituted or unsubstituted C 5-10 Heteroaryl.

[0098] In a further preferred embodiment, R H Independently selected from substituted or unsubstituted C 6-10 Aryl or substituted or unsubstituted 5-membered to 10-membered heteroaryl.

[0099] In certain embodiments, R H is replaced by C 5-10 Aryl or substituted C 5-10 Heteroaryl, which is optionally substituted by C 1-6 Alkanes, C 1-6 In a preferred embodiment, C 1-6 The alkane is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl and n-hexyl. 1-6 The heteroalkanes are 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, the carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. In a preferred embodiment, the heterocyclic compound is selected from aziridine, aziridine, furan, pyrrolidine, pyran, piperidine, piperazine, aziridine and diaziridine.

[0100] In a still further preferred embodiment, R H Independently selected from substituted or unsubstituted C 6-10 aryl or substituted or unsubstituted 5-membered to 10-membered heteroaryl. In certain cases, R H is the unsubstituted C 6-10 aryl or unsubstituted 5-membered to 10-membered heteroaryl. In other aspects, R H It is C 6-10 aryl or 5-membered to 10-membered heteroaryl, which is optionally substituted by: substituted or unsubstituted C 1-6 alkyl, halogen, nitrile, hydrogen and alkoxy. In still other aspects, R H It is C 6-10 aryl or 5-membered to 10-membered heteroaryl, which is optionally substituted by: substituted or unsubstituted C 1-6 Alkyl, halogen, nitrile and OR 2B , where R 2Bis hydrogen or substituted or unsubstituted C1-C6 alkyl. In a further preferred embodiment, R H is unsubstituted phenyl. In other embodiments, R H is selected from substituted or unsubstituted C 1-6 alkyl, halogen, nitrile, hydrogen and alkoxy groups. H is selected from substituted or unsubstituted C 1-6 Alkyl, halogen, nitrile and OR 2B The substituents in the group consisting of optionally substituted phenyl, wherein R 2B is hydrogen or substituted or unsubstituted C1-C6 alkyl. In still another embodiment, R H is selected from the unsubstituted C 1-6 phenyl substituted with a substituent selected from the group consisting of alkyl, halogen, nitrile, hydrogen and alkoxy. H is selected from the unsubstituted C 1-6 Alkyl, halogen, nitrile and OR 2B The group consisting of phenyl substituted with a substituent, wherein R 2B It is hydrogen or substituted or unsubstituted C1-C6 alkyl.

[0101] In still further aspects, R H is selected from the Z groups described in Tables 1 to 3.

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

[0103] In a preferred embodiment, R A , R B , and R C are independently selected from H, D, halogen, substituted or unsubstituted C 1-4 Alkyl, and NR J R K ; and, R J and R K Each of which is independently selected from H and substituted or unsubstituted C 1-4 alkyl.

[0104] In a preferred embodiment, R A and R B is CH3, and R C Yes H, CH 3， Halogen, nitrile, methoxy or ethoxy.

[0105] In still other preferred aspects, R A , R B and R C Independently selected from H, D, halogen, OR I , substituted or unsubstituted C 1- C4 alkyl, or NR J R K ; Among them, R I , R J and R K Each of which is independently selected from H and substituted or unsubstituted C 1- In a preferred embodiment, R A and R B Each of 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 of is CH3, and R C is selected from the group consisting of: H, CH3, fluorine, chlorine, nitrile, methoxy and ethoxy. A and R B Each of is CH3, and R C It's hydrogen.

[0106] In certain other embodiments, R D It is C 1-4 Alkyl, which is optionally substituted by a substituent selected from the group consisting of halogen, oxygen, C 3-8 Cycloalkyl, aryl or heteroaryl, and / or R E Is H or C 1-4 Alkyl, which is optionally substituted by a substituent selected from the group consisting of halogen, oxygen, C 3-8 Cycloalkyl, aryl or heteroaryl.

[0107] In a preferred embodiment, R D and R E Each of R is independently selected from H, D, CH3, CH2CH3, (CH2)2CH3 and (CH2)3CH3. In a more preferred embodiment, R E is hydrogen, and R D is CH3, CH2CH3, (CH2)2CH3 or (CH2)3CH3. In certain or other preferred embodiments, R D and R E Together they form a substituted or unsubstituted C3-C6 cycloalkyl group.

[0108] In certain preferred embodiments, R D and R E In still another preferred embodiment, R D is hydrogen and R E is an alkyl group, for example, a C1-C6 alkyl group or a C1-C4 alkyl group, including but not limited to methyl, ethyl, propyl and butyl. In certain other preferred embodiments, R D and R E Together with the carbon atom to which it is attached, it forms a C3-C6 cycloalkyl group, including, but not limited to, cyclopropyl or cyclobutyl.

[0109] In a preferred embodiment, R F and R G With their attached N + Together they form a five-, six-, seven- or eight-membered heterocyclyl ring, which may be optionally substituted, to give compounds of formula Ia:

[0110]

[0111] wherein each variable is as defined above, including preferred or alternative embodiments, n is 1, 2, 3, 4 or 5; and R 1B is H or a substituent, for example, H, D, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, OR 1I , CN, NR 1J R 1K , NR 1L C(O)R 1M ,S(O)R 1N , S(O)2R 1N , SO2R 1O R 1P , SO2NR 1Q R 1R , SO3R 1S ,CO2R 1T , C(O)R 1U and C(O)NR 1V R 1W ; and, where R 1I , R 1J , R 1K , R 1L , R 1M , R 1N , R 1O , R 1P , R 1Q, R 1R , R 1S , R 1T , R 1U , R 1V and R 1W Each of R is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, R 1J and R 1K or R 1V and R 1W or R 1Q and R 1R Together with the nitrogen atom to which they are attached they may form a substituted or unsubstituted 5-, 6-, 7- or 8-membered ring.

[0112] In a preferred embodiment, R F and R G N + Together they form a five-membered, six-membered, seven-membered or eight-membered nitrogen-containing heterocyclic ring, including, but not limited to:

[0113] or are optionally substituted. Preferred substituents include phenyl, CO2R 1T and C(O)NR 1V R 1W .

[0114] In some respects, R F and R G are independently C1-C4 alkyl. F and R G are independently selected from CH3 and CH2CH3. In certain other aspects, R F and R G are the same, and are substituted or unsubstituted C1-C4 alkyl. In other aspects, R F and R G are the same, all being methyl, ethyl, propyl or butyl. In still other embodiments, R F and R G are the same, both are CH3 or CH2CH3.

[0115] In some embodiments, Y - is a halogen anion, a carboxylate, or a sulfonate. -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 heterocyclic carboxylate.

[0116] In certain embodiments, Y- is selected from the group consisting of trifluoroacetate, sulfate, phosphate, acetate, fumarate, formate, carbonate, maleate, citrate, pyruvate, succinate, oxalate, sulfonate (e.g., methanesulfonate, trifluoromethanesulfonate, toluenesulfonate, such as p-toluenesulfonate, benzenesulfonate, ethanesulfonate, camphorsulfonate, 2-mesitylenesulfonate, or naphthalenesulfonate, such as 2-naphthalenesulfonate), sodium bisulfate, sodium malonate, sodium octanoate, sodium ascorbate, sodium oleate, sodium nicotinate, sucrose In one embodiment, Y- is a halogen anion.

[0117] In one embodiment, Y - is a halogen anion selected from bromide, chloride and iodide.

[0118] Each embodiment described herein may be combined with one, more than one, or all of the other embodiments.

[0119] In a preferred embodiment, the present invention relates to compounds of formula (I), or pharmaceutically acceptable salts thereof, wherein R H is an optionally substituted aryl or an optionally substituted heteroaryl selected from the following groups:

[0120]

[0121]

[0122] In certain preferred aspects, R H is a substituted or unsubstituted phenyl group.

[0123] In certain other aspects, the present invention has a compound of formula (II):

[0124]

[0125] in,

[0126] Y - A pharmaceutically acceptable anion;

[0127] q is 0, 1, 2, or 3;

[0128] R D is hydrogen, methyl or ethyl, and

[0129] R 1B As defined above.

[0130] In certain embodiments, the compound has a structure as shown in formula (II), wherein Y - is a halide ion, a carboxylate ion or a sulfate ion. - It may be, for example, a halogen ion, a substituted or unsubstituted alkyl sulfate ion, a substituted or unsubstituted aryl sulfate ion, a substituted or unsubstituted alkyl or aliphatic carboxylate ion, a substituted or unsubstituted aryl carboxylate ion, or a substituted or unsubstituted heterocyclic carboxylate ion.

[0131] In certain embodiments, the compound is as shown in formula (II), wherein Y- is selected from the group consisting of trifluoroacetate, sulfate, phosphate, acetate, fumarate, formate, carbonate, maleate, citrate, pyruvate, succinate, oxalate, sulfonate (e.g., methanesulfonate, trifluoromethanesulfonate, toluenesulfonate, such as p-toluenesulfonate, benzenesulfonate, ethanesulfonate, camphorsulfonate, 2-mesitylenesulfonate, or naphthalenesulfonate, such as 2-naphthalenesulfonate), sodium bisulfate, sodium malonate, sodium octanoate, sodium ascorbate, sodium oleate, Sodium nicotinate, sodium sucrose, sodium adipate, sodium formate, sodium glycolate, sodium L-lactate, sodium D-lactate, sodium aspartate, sodium malate, sodium L-tartrate, sodium D-tartrate, sodium stearate, sodium 2-furoate, sodium 3-furoate, sodium naphthalene dicarboxylate sulfonate or naphthalene-1-(sulfonic acid)-5-sulfonate), oxalyl (ethane-1,2-disulfonate or ethane-1-(sulfonic acid)-2-sulfonate), isothiocyanate (2-hydroxyethyl sulfonate), D-mandelate, L-mandelate, propionate, tartrate, phthalate, hydrochloride, hydrobromide, and nitrate.

[0132] In one embodiment, the compound is as shown in formula (II), and Y - is a halogen anion. In a preferred embodiment, Y - It is a halogen ion selected from bromide, chloride and iodide.

[0133] It is understood that when q is 0, 1, 2 or 3, N+ The formula (II) are:

[0134] or In certain preferred embodiments, q is 0. In other preferred aspects, q is 1. In other preferred aspects, q is 2. In still further preferred aspects, q is 3.

[0135] In still other preferred aspects, the compound is as shown in formula (II) and R D It's hydrogen.

[0136] In a further aspect, the compound is as shown in formula (II) and R D It's methyl.

[0137] In still other aspects, the compound is as shown in formula (II) and R D It is ethyl.

[0138] In a further aspect, the compound is as shown in formula (II), q is 0 and R D It's hydrogen.

[0139] In other preferred aspects, the compound is as shown in formula (II), q is 1 and R D It's hydrogen.

[0140] In a further preferred aspect, the compound is as shown in formula (II), q is 2 and R D It's hydrogen.

[0141] In still other aspects, the compound is as shown in formula (II), q is 3 and R D It's hydrogen.

[0142] In still another preferred embodiment, the compound is as shown in formula (II), q is 0 and R D It's methyl.

[0143] In another preferred aspect, the compound is as shown in formula (II), q is 1 and R D It's methyl.

[0144] In a further preferred aspect, the compound is as shown in formula (II), q is 2 and R D It's methyl.

[0145] In other aspects, the compound is as shown in formula (II), q is 3 and R D It's methyl.

[0146] In a further preferred embodiment, the compound is as shown in formula (II), q is 0 and R D It is ethyl.

[0147] In another preferred aspect, the compound is as shown in formula (II), q is 1 and R D It is ethyl.

[0148] In a further preferred aspect, the compound is as shown in formula (II), q is 2 and R D It is ethyl.

[0149] In other aspects, the compound is as shown in formula (II), q is 3 and R D It is ethyl.

[0150] In still other aspects, the compound is selected from the following Table A, or a pharmaceutically acceptable salt thereof, wherein Y - It is a pharmaceutically acceptable anion.

[0151] Table A

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158] In other preferred aspects, the compound of the present invention is selected from the compounds shown in the following table, or a pharmaceutically acceptable salt thereof, wherein Y- is as shown or a pharmaceutically acceptable anion thereof:

[0159] Table B

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178] The representative compounds of the present invention and their isomers and pharmaceutically acceptable salts are selected from the following Table C, wherein Y - is a pharmaceutically acceptable anion, as defined above, and Z is either an aryl or heteroaryl structure selected from one of the structures shown in Table 1, or a substituted aryl or substituted heteroaryl structure selected from one of the structures shown in Tables 2-3.

[0179] Table C - Representative compounds of the present invention

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195] Table 1 - Representative Z structures

[0196]

[0197]

[0198] Table 2 - Representative Z structures

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207] Table 3 - Representative Z structures

[0208]

[0209]

[0210]

[0211]

[0212] The preferred compounds of the present invention and their isomers and pharmaceutically acceptable salts are represented by formula (III):

[0213]

[0214] Among them, the preferred substituent combination R C , R D , N + / R F / R Gand Z are as defined in Table 4, and Y - is a pharmaceutically acceptable anion as defined above. The compounds may also be prepared according to the general methods described below.

[0215] Table 4 - Preferred R of compounds of formula (III) C , R D , N + / R F / R G and Z substitution combination.

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252] Each preferred embodiment described herein may be used in combination with one, any or all other preferred embodiments, as if presented in each arrangement herein.

[0253] The compositions of the invention may comprise a racemic mixture, a pure enantiomer, or an excess of one enantiomer over another. For example, the composition may comprise a compound having an enantiomeric excess of at least 5, 10, 20, 30, 40, 50, 60, 70, 80, or 90%. In one embodiment, the enantiomeric excess is at least 95%.

[0254] The compounds of the present invention include all enantiomers defined as (R)- or (S)- according to absolute stereochemistry and their racemic and optically pure forms, as well as those described herein in any pharmaceutically acceptable form thereof, including enantiomers, salts, solvates, polymorphs, solvates, hydrates, anhydrous and other crystalline forms and combinations thereof. Likewise, all tautomeric forms are also intended to be included.

[0255] Preferably, the pharmaceutical composition comprises a substantially pure R isomeric form of the compound of the invention; alternatively, a pharmaceutical composition comprises a substantially pure S isomeric form of the compound of the invention; alternatively, a pharmaceutical composition comprises the compound of the invention in the form of an isomeric mixture, which may include an excess of the R enantiomer or an excess of the S enantiomer. It is particularly preferred that the pharmaceutical composition contains a substantially pure optical isomeric form of the compound of the invention. For the avoidance of doubt, the compound of the invention may be used in the form of a solvate if desired.

[0256] synthesis

[0257] The compounds of formula (I) can be prepared using methods similar to the following general synthetic schemes:

[0258]

[0259] as well as

[0260]

[0261] For example,

[0262]

[0263] The compounds of formula (II) can be prepared using methods similar to those described in the examples and the following general synthetic scheme:

[0264]

[0265] Other bioactive agents and exogenous macropore channel agonists

[0266] As described above, the compounds of the invention or the compositions of the invention may be administered with a biologically active agent, for example, one or more other biologically active agents, including agents commonly used to treat neurogenic inflammation, which may be administered in combination with the compounds of the invention or the compositions described herein. Biologically active agents include, but are not limited to, TRP1A receptor agonists, TRPV1-4 receptor agonists, TRPM8 agonists, ASIC agonists, P2X receptor agonists, acetaminophen, NSAIDs, glucocorticoids, anesthetics, tricyclic antidepressants, amine transporter inhibitors, anticonvulsants, antiproliferative agents and immunomodulators, antibodies or antibody fragments, antibiotics, polynucleotides, polypeptides, proteins, anticancer agents, growth factors and vaccines.

[0267] TRPV1 agonists that can be used in the methods, kits and compositions of the present invention include, but are not limited to, any drug (e.g., a compound of the present invention) that can activate TRPV1 receptors on nociceptive neurons and allow entry of at least one voltage-gated ion channel inhibitor. Suitable TRPV1 agonists are capsaicin or another capsaicinoid, which are members of the vanillin family of molecules. Naturally occurring capsaicinoids are capsaicinoids themselves, dihydrocapsaicinoids, dedihydrocapsaicinoids, homodihydrocapsaicinoids, homocapsaicinoids, and nonanamide. Other suitable capsaicin and capsaicin analogs and derivatives suitable for use in the compositions and methods of the present invention include naturally occurring and synthetic capsaicin derivatives and analogs, including, for example, vanillins (e.g., N-vanillinyl-alkadienamide, N-vanillinyl-alkadienyl, and N-vanillinyl-cis-monounsaturated alkanamide), capsic acid, dihydrocapsic acid, nordihydrocapsic acid and other capsic acids, capsic esters, dihydrocapsic acid and other arosinic acid esters, capsic acid, resin ester toxins, tintoxins, zucassin, N-phenylmethylalkanamide capsaicin derivatives, avanafil, N-[(4-(2-aminoethoxy)-3-methoxyphenyl)methyl]-9Z octadecanoamide, N-oleyl homofragrance, trienols (e.g., cod liver oil), gingerol, pepper, citronella oil, guaiacol, eugenol, zinc ketone, newanil, NE-19550, NE-21610, and NE-28345. Other capsaicinoids, their structures, and methods for making them are described in U.S. Pat. Nos. 7,446,226 and 7,429,673, which are incorporated herein by reference.

[0268] Other suitable TRPV1 agonists include, but are not limited to, eugenol, Avanil (N-arachidonoyl amphetamine), Anandamide, 2-aminoethoxydiphenyl borate (2APB), AM404, resiniferatoxin, phorbol 12-phenylacetic acid 13-acetic acid 20-pervanadate (PPAHV), Avanil (NE 19550), Oda (N-oleoyl dopamine), N-arachidonoyl dopamine (NADA), 6′-iodothiocytoxan (6′-IRTX), C18 N-acylethanolamines, lipoxygenase derivatives such as 12-hydroperoxyeicosatetraenoic acid, inhibitor cysteine ​​knot (ICK) peptides (vanillinotoxins), piperine, MSK195 (N-[2-(3,4-dimethylbenzyl)-3-(pivaloyloxy)propyl]-2-[4-(2-aminoethoxy)-3-methoxyphenyl]acetamide), JYL79 (4-dimethylbenzyl)-3-(pivaloyloxy)propyl]-N′-(4-hydroxy-3-methoxybenzyl)thiourea), hydroxy-α-trihydrate, 2-aminoethoxydiphenylborate, 10-sesame oil, oleyl gingerol, oleyl sesame oil and SU200 (N-(4-tert-butylbenzyl)-N′-(4-hydroxy-3-methoxybenzyl)thiourea). Other TRPV1 agonists include: isopentocaine, articaine, benzocaine, bupivacaine, carbocaine, carticaine, chloroprocaine, cyclomethoxycaine, dibuvacaine (sinchocaine), dimethocaine (larocaine), eticaine, hexylcaine, levobupivacaine, lidocaine, mepivacaine, meprevacaine (orracaine), metabutocaine, pivacaine, procaine (novacaine), procaine, propoxycaine, lidocaine, ropivacaine, tetracaine (aminocaine), and tamoxifen.

[0269] Suitable TRPV2-4 agonists include, but are not limited to, 2-APB, diphenylboronic anhydride, insulin-like growth factor 1, lysophosphatidylcholine, lysophosphatidylinositol, probenecid, vanillin, eugenol, cinnamaldehyde, camphor, carvacrol, thymol, citral, farnesyl diphosphate, sodium acetate, diphenylboronic anhydride, 6-tert-butyl-m-cresol, dihydrocavitin, borneol, (-)-menthol, GSK1016790A, 4α-PDH, 5,6-epoxyeicosatrienoic acid, 4α-PDD, dipyridographolide, citric acid, phorbol 12 tetradecanoate 13 acetate and RN1747.

[0270] Suitable TRPM8 agonists include, but are not limited to, menthol, eucalyptus, linalool, geraniol, hydroxycitronellal, WS-3, WS-23, Frescolat MGA, Frescolat ML, PMD 38, CPS125, Coolact P, M8 Ag, AITC, cryosim-3, and Coolant 10.

[0271] Suitable ASIC agonists include, but are not limited to, clofenac hydrochloride, GMQ hydrochloride, reticulin, the polyamine agmatine, lysophosphatidylcholine, arachidonic acid, and neuropeptide SF.

[0272] Other bioactive agents suitable for use in the methods, compositions and kits of the present invention include bioactive agents that can activate TRP1A receptors on nociceptive neurons or allergic neurons and allow entry of at least one voltage-gated ion channel inhibitor. Suitable TRP1A agonists include, but are not limited to, cinnamaldehyde, allyl isothiocyanate (mustard oil), diallyl disulfide, cinnamon oil, wintergreen oil, clove oil, acrolein, hydroxy α-trimethoxybenzaldehyde, 2-aminoethoxydiphenyl borate, 4-hydroxynonanal, methyl paraben, and 3′-carbamoylbiphenyl-3-cyclohexylcarbamate (URB597).

[0273] P2X agonists used in the methods, compositions, and kits of the present invention include any agonist that can activate P2X receptors on nociceptors or allergic neurons and allow entry of at least one voltage-gated ion channel inhibitor. 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).

[0274] Other biologically active agents that can be used in combination with the compounds of the invention include NSAIDs, glucocorticoids, anesthetics, tricyclic antidepressants, amine transporter inhibitors, anticonvulsants, antiproliferative and immunomodulatory agents, antibodies or antibody fragments, antibiotics, polynucleotides, polypeptides, a protein, an anticancer agent, a growth factor and vaccines.

[0275] Nonsteroidal anti-inflammatory drugs (NSAIDs) that can be administered to patients (e.g., humans) suffering from neurogenic inflammation in combination with the compositions of the present invention include, but are not limited to, acetylsalicylic acid, amoxicillin, benolide, benolide, choline magnesium salicylate, diflunisal, ethobenzamide, farisin, methyl salicylate, magnesium salicylate, salicylate salicylate, salicylamide, diclofenac, aceclofenac, acetylmethacin, alclofenac, bromfenac, etodolac, indomethacin, nabumetone, oxametacin, promethacin, sulindac, tolmetin, ibuprofen, amorolfen, benoxaprofen, carprofen, dexibuprofen, fenbufen, non- Nurofen, fluniprofen, flurbiprofen, ibuprofen, indolepropylamine, ketoprofen, ketorolac, loxoprofen, naproxen, oxaprozin, pirprofen, supprofen, tiaprofenic acid, mefenamic acid, flufenamic acid, meclofenamic acid, toluene acid, phenbutazone, amprolone, azaprotone, clofetonone, kebuzone, analgin, imidazole, mofebutone, oxyphenbutazone, benazoline, thiopyrazolone, piroxicam, deroxicam, lornoxicam, meloxicam, tenoxicam, and COX-2 inhibitors celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, valdecoxib, and pharmaceutically acceptable salts thereof.

[0276] Glucocorticoids that can be administered to patients (e.g., humans) suffering from neurogenic inflammation in combination with the compositions of the present invention include, but are not limited to, hydrocortisone, cortisone acetate, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone acetonide, beclomethasone, fludrocortisone acetate, deoxycortisone acetate, aldosterone, and pharmaceutically acceptable salts thereof.

[0277] Anesthetic agents that can be administered to a patient (eg, a human) suffering from neurogenic inflammation in combination with the compositions of the present invention include, but are not limited to, tramadol, hydrocodone, oxycodone, and pharmaceutically acceptable salts thereof.

[0278] Antiproliferative and immunomodulatory agents that can be administered to patients (e.g., humans) with neurogenic inflammation in combination with the compositions of the present invention include, but are not limited to, alkyl antineoplastic drugs, platinum drugs, antimetabolites, topoisomerase inhibitors, dihydrofolate reductase inhibitors, antitumor antibiotics, antimitotic drugs, aromatase inhibitors, thymidylate synthase inhibitors, DNA antagonists, farnesyl transferase inhibitors, pump inhibitors, histone acetyltransferase inhibitors, metalloproteinase inhibitors, ribonucleoside reductase inhibitors, TNF-α agonists, TNF-α antagonists or scavengers, interleukin 1 (IL-1) antagonists or scavengers, endothelin A receptor antagonists, retinoic acid receptor agonists, hormones, antihormones, photodynamic drugs, and tyrosine kinase inhibitors.

[0279] The above-mentioned biologically active agents may be administered before, simultaneously with, or after administration of the compositions of the present invention, using any formulation, dosage, or administration known in the art to have a therapeutic effect.

[0280] Preparation of the composition

[0281] The administration of the compounds of the present invention can be carried out by any suitable method that can lead to a reduction in the receptive pain sensation of the target area. The compounds of the present invention can be contained in any suitable carrier material in any appropriate amount, and are generally present in an amount of 1-99% by total weight of the total weight of the composition. The composition can be provided in a dosage form suitable for oral, parenteral administration (e.g., intravenous administration, intramuscular administration), rectal administration, dermal administration, subcutaneous administration, topical administration, transdermal administration, sublingual administration, nasal administration, vaginal administration, intrathecal administration, epidural administration or ocular administration, or administered by injection, inhalation or direct contact with the nasal or oral mucosa.

[0282] Therefore, the composition can be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels, including hydrogels, pastes, ointments, creams, plasters, wet agents, osmotic delivery devices, suppositories, enemas, injections, implants, sprays or aerosols. The composition can be prepared according to conventional pharmaceutical methods (e.g., see Remington: Pharmaceutical Science and Practice, 2013, 22nd edition, LVAllen, Pharmaceutical Publishing House, Philadelphia; and Encyclopedia of Pharmaceutical Technology, 4th edition, J edition. Swarbrick, 2013, China Resources Publishing House, New York).

[0283] Each compound can be formulated in a variety of ways known in the art. For example, the compounds of the invention and the bioactive agents defined herein can be formulated together or separately. Ideally, the compounds of the invention and the bioactive agents are formulated together for simultaneous or near-simultaneous administration. In another embodiment, two or more bioactive agents can be formulated together with the compounds of the invention or separately. Other embodiments include, but are not limited to, formulating two or more compounds of the invention together, wherein the compounds are formulated with or without one or more bioactive agents.

[0284] The reagents, which are formulated separately or separately, can be packaged together as a kit. Non-limiting examples include, but are not limited to, kits containing, for example, two pills, one pill and powder, a suppository and a liquid in a medicine bottle, two topical creams, and the like. The kit may include optional components that facilitate administration of a unit dose to a patient, such as a medicine bottle for reconstitution of the powder form, a syringe for injection, a customized IV delivery system, an inhaler, and the like. In addition, the unit dose kit may contain instructions for preparation and administration of the composition.

[0285] The kit may be formulated as a unit dosage form for single use by a single patient, or may be formulated for multiple use by a specific patient (at a constant dose or the dose of a single compound may vary as the treatment progresses to vary potency); or, the kit may contain multiple doses suitable for multiple patients ("bulk packaging"). The components of the kit may be assembled in cartons, blister packs, bottles, tubes, and the like.

[0286] Controlled Release Formulations

[0287] Each compound of the present invention can be prepared separately or can be formulated as controlled release (for example, sustained release or quantitative release) preparation together with one or more bioactive agents described herein, as described in U.S. Patent Application Publication No. 2003 / 0152637 and No. 2005 / 0025765, which are all incorporated herein by reference. For example, the compounds of the present invention can be prepared separately or can be formulated as capsules or tablets together with one or more bioactive agents described herein, and administered to patients.

[0288] Any pharmaceutically acceptable carrier or formulation suitable for topical application and / or for injection into the site to be treated (e.g., painful surgical incisions, wounds or joints) may be used as desired, which is capable of sustained release of the compounds of the invention, which may be used alone or in combination with one or more bioactive agents described herein, to eliminate or alleviate inflammatory responses over a long period of time. Controlled release formulations known in the art include special coated pellets, polymer formulations or matrices for surgical implantation, or as sustained release microparticles for implantation, insertion, infusion or injection, such as microspheres or microcapsules, wherein sustained release of the active drug is achieved by sustained or controlled diffusion from the matrix and / or selective degradation of the formulation coating or selective degradation of the polymer matrix. Other formulations or carriers for controlled, sustained or immediate delivery of the agent to a preferred local site of the patient include, for example, suspensions, emulsions, gels, liposomes and any other suitable known technology delivery carriers or formulations acceptable for subcutaneous or intramuscular administration.

[0289] A variety of biocompatible materials can be used as controlled release carriers to control the release of the compounds of the present invention (alone or in combination with one or more bioactive agents described herein). Any pharmaceutically acceptable biocompatible polymer known to those skilled in the art can be used. Preferably, the biocompatible controlled release material degrades in vivo within about one year, preferably within about 3 months, and more preferably within about two months. More preferably, the controlled release material will degrade significantly within one to three months, wherein at least 50% of the material degrades into non-toxic residues that are cleared by the body, and 100% of the compounds of the present invention are released within about two weeks, preferably within a period of about 2 days to about 7 days. Degradable controlled release materials are preferably degraded by hydrolysis, whether by surface erosion or volume erosion, so that not only sustained release can be achieved, but also a desired release rate can be provided. However, the pharmacokinetic release profiles of these preparations can be first order, zero order, biphasic or multiphasic to provide the desired reversible local anti-injury effect within the desired time period.

[0290] Suitable biocompatible polymers can be used as controlled release materials. The polymer material may include biocompatible, biodegradable polymers, and, in certain preferred embodiments, copolymers of lactic acid and glycolic acid are preferred. Preferred controlled release materials useful in the formulations of the present invention include polyanhydrides, polyesters, copolymers of lactic acid and glycolic acid (preferably wherein the weight ratio of lactic acid to glycolic acid does not exceed 4:1, i.e., 80% or less lactic acid to 20% or more glycolic acid (by weight) and polyorthogonal esters containing catalysts or degradation enhancing compounds, for example, containing at least 1% (by weight) of anhydride catalysts, such as maleic anhydride. Examples of polyesters include polylactic acid, polyglycolic acid, and polylactic-polyglycolic acid copolymers. Other useful polymers include protein polymers such as collagen, gelatin, fibrin and fibrinogen, and polysaccharides such as hyaluronic acid.

[0291] The polymer material can be prepared by any method known to those skilled in the art. For example, if the polymer material is composed of a copolymer of lactic acid and glycolic acid, the copolymer can be prepared by the method specified in U.S. Patent No. 4,293,539, which is incorporated herein by reference. Alternatively, the copolymer of lactic acid and glycolic acid can be prepared by any other method known to those skilled in the art. Other useful polymers include polylactic acid, polyglycerol ethers, polyanhydrides, polyorthogonal esters, polycaprolactones, polyphosphazenes, polyphosphates, polysaccharides, protein polymers, soluble derivatives of polysaccharides, soluble derivatives of protein polymers, polypeptides, polyesters, and polyorthogonal esters or mixtures or combinations thereof.

[0292] Pharmaceutically acceptable polyanhydrides useful in the present invention have anhydride bonds that are not resistant to water. The drug release rate can be controlled by the specific polyanhydride polymer used and its molecular weight. Polysaccharides can be poly-1,4-glucans, such as starch glycogen, straight-chain starch, amylopectin, and mixtures thereof. Biodegradable hydrophilic or hydrophobic polymers can be water-soluble derivatives of poly-1,4-glucans, including hydrolyzed amylopectin, derivatives of hydrolyzed amylopectin, such as hydroxyethyl starch (HES), hydroxyethyl straight-chain starch, dialdehyde starch, etc. The polyanhydride polymer can be branched or linear.

[0293] Examples of polymers useful in the present invention include (in addition to homopolymers and copolymers of polylactic acid and / or polyglycolic acid), poly[bis(p-carboxyphenoxy)propane anhydride] (PCPP), poly[bis(p-carboxy)methane anhydride] (PCPM), polyanhydrides of oligounsaturated fatty acids, polyanhydride polymers prepared from amino acids modified to include additional carboxylic acids, aromatic polyanhydride compositions, and copolymers of polyanhydrides with other substances, such as polyanhydrides of terminal fatty acids, such as polyanhydrides polymerized from monomers of unsaturated fatty acids or dimers and / or trimers of unsaturated fatty acids. Polyanhydrides can be prepared according to the methods set forth in U.S. Pat. No. 4,757,128, which is incorporated herein by reference. Polyorthogonal ester polymers can be prepared, for example, as described in U.S. Pat. No. 4,070,347, which is incorporated herein by reference. Polyphosphates can be prepared and used as described in U.S. Patents 6,008,318, 6,153,212, 5,952,451, 6,051,576, 6,103,255, 5,176,907, and 5,194,581, which are incorporated herein by reference in their entireties.

[0294] Protein polymers may also be used. The protein polymers and their soluble derivatives include gelled biodegradable synthetic polypeptides, elastin, alkylated collagen, alkylated elastin, and the like. Biodegradable synthetic polypeptides include poly(N-hydroxyalkyl)-L-asparagine, poly-(N-hydroxyalkyl)-L-glutamine, copolymers of N-hydroxyalkyl-L-asparagine and N-hydroxyalkyl-L-glutamine and other amino acids. Suggested amino acids include L-alanine, L-lysine, L-phenylalanine, L-valine, L-tyrosine, and the like.

[0295] In other embodiments, the controlled release material, which actually serves as a carrier for the compounds of the invention (used alone or in combination with one or more biologically active agents described herein), may further include a bioadhesive polymer such as pectin (polygalacturonic acid), mucopolysaccharides (hyaluronic acid, mucin) or non-toxic lectins, or the polymer itself may be a bioadhesive, such as a polyanhydride or a polysaccharide, such as chitosan.

[0296] In some embodiments, the biodegradable polymer comprises a gel. One such useful polymer is a thermogelling polymer, such as a polyethylene oxide, polypropylene oxide (PEO-PPO) block copolymer, such as Pluronic® available from BASF Windott Corporation. TM F127. In this case, the local anesthetic can be injected via a syringe as a free-flowing liquid that quickly solidifies above 30°C (e.g., when injected into a patient). The gel system then releases a steady dose of the compound of the invention, alone or in combination with one or more of the bioactive agents described herein, at the site of administration.

[0297] Oral dosage form

[0298] Oral formulations include tablets containing the active ingredient mixed with non-toxic pharmaceutically acceptable excipients. These excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch, including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate); granulating agents and disintegrants (e.g., cellulose derivatives, including microcrystalline cellulose, starch, including potato starch, cross-linked sodium carboxymethyl cellulose, sodium alginate or alginic acid); binders (e.g., sucrose, glucose, sorbitol, gum arabic, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinyl pyrrolidone or polyethylene glycol); and lubricating factors, lubricants, and anti-adhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silicon dioxide, hydrogenated vegetable oil or talc). Other pharmaceutically acceptable excipients may be colorants, flavoring agents, plasticizers, humectants, buffers, taste masking agents (eg, hydroxypropyl methylcellulose, hydroxypropyl cellulose), and the like.

[0299] As defined herein, one or more compounds of the present invention and one or more bioactive agents can be mixed together in tablets, capsules or other carriers, or can be separated into individual parts. In one embodiment, the compounds of the present invention are contained inside the tablet, and the bioactive agent is located outside the tablet, so that the bioactive agent part is continuously released before the compounds of the present invention are released.

[0300] Oral preparations can also be made into chewable tablets, hard gelatin capsules (where the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin)), or soft gelatin capsules (where the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil) in the form of provided. Powders, granules, and pills can be prepared in a conventional manner using the ingredients under the above tablets and capsules, for example, using a mixer, a fluidized bed apparatus or a spray drying device.

[0301] Formulations for oral administration in the buccal cavity may also be provided in the form of a mouthwash, mouth spray, mouth rinse, oral ointment or oral gel.

[0302] Dissolving or diffusion control release can be realized by the granular preparation of suitable coating coating tablet, capsule, granule or compound, or by compound being added to suitable matrix.Controlled release coating can comprise above-mentioned one or more coating materials, and / or, for example shellac, beeswax, ethanol wax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, palmitostearin, ethylcellulose, acrylic resin, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, pyrrolidone, polyethylene, polymethyl methacrylate, methyl methacrylate, 2-hydroxymethylacrylic acid, methacrylic acid hydrogel, 1,3-butylene glycol, ethylene glycol methacrylate and / or polyethylene glycol.In controlled release matrix formula, matrix material can also comprise for example hydrated methylcellulose, carnauba wax and stearyl alcohol, Carbopol 934, silicone resin, tristearin, methyl acrylate, methyl methacrylate, polyvinyl chloride, polyethylene and / or halogenated fluorocarbon.

[0303] Liquid forms in which the compounds and compositions of the invention may be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions and flavored emulsions containing edible oils such as cottonseed, sesame, coconut or peanut oil, as well as tinctures and similar pharmaceutical carriers.

[0304] When generally 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 those skilled in the art. Generally, the dosage is generally about 0.001 mg to 2000 mg per day, ideally about 1 mg to 1000 mg per day and more ideally about 5 mg to 500 mg per day. If necessary, the daily intake dosage is up to 200 mg.

[0305] As described herein, in combination therapy, each drug may be administered independently one to four times daily for one day to one year and possibly even continuously throughout the patient's life. In many cases, slow, long-term administration is required.

[0306] Parenteral preparations

[0307] Preparations suitable for parenteral administration (e.g., by injection) include aqueous or non-aqueous, isotonic, pyrogen-free, sterile liquids (e.g., solutions, suspensions) in which the compound is dissolved, suspended or otherwise provided (e.g., in liposomes or other microparticles). Such liquids may contain other pharmaceutically acceptable ingredients, such as antioxidants, buffers, preservatives, stabilizers, antibacterial agents, suspending agents, thickeners, and solutes that make the preparation isotonic with the blood (or other relevant body fluids) of the intended recipient. Examples of excipients include, for example, water, alcohol, polyols, glycerol, vegetable oils, and the like. Examples of suitable isotonic carriers for such preparations include sodium chloride injection, Ringer's solution, or lactated Ringer's injection. Typically, the concentration of the compound in the liquid is from about 1 ng / ml to about 10 μg / ml, such as from about 10 ng / ml to about 1 μg / ml. The preparation can be present in unit dose or multi-dose sealed containers, for example, in ampoules and vials, and can be stored under freeze-dried (lyophilized) conditions, requiring only the addition of a sterile liquid carrier, such as water for injection, before use. Sterile powders, granules, and tablets can be used to prepare injection solutions or suspensions for immediate use.

[0308] Topical preparations

[0309] The compositions of the present invention (alone or in combination with one or more biologically active agents described herein) may also be adapted for topical use, with a topical excipient containing from 0.0001% to 25% (w / w) or more of the active ingredient.

[0310] In preferred combinations, the active ingredients are preferably present at between 0.0001% and 10% (w / w) each, more preferably between 0.0005% and 4% (w / w) of the active agent. The topical formulations, including but not limited to creams, gels or ointments, can be applied one to four times a day, or as needed. To perform the methods described herein, a topical excipient containing a composition of the invention or a combination therapy containing a composition of the invention is preferably applied to the site of inflammation in a patient. For example, a cream can be applied to the fingers of a patient suffering from arthritis in the fingers.

[0311] The composition can be formulated using any dermatologically acceptable carrier. Exemplary carriers include solid carriers such as alumina, clay, microcrystalline cellulose, silica, or talc; and / or liquid carriers such as alcohol, glycol, or water-alcohol / glycol mixtures. The therapeutic agent can also be administered in a liposomal formulation to allow the therapeutic agent to enter the skin. Such liposome formulations are described in U.S. Pat. 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; 4,994,213; 8,822,537, and PCT Application WO 96 / 40061. Examples of other suitable carriers are described in U.S. Pat. Nos. 4,877,805, 8,822,537, and EP Patent Publication No. 0586106A1. Suitable carriers of the present invention may also include mineral oil, petrolatum, polydecene, stearic acid, isopropyl myristate, polyoxyethylene stearate 40, stearyl alcohol, or vegetable oils.

[0312] The composition may also include a skin penetration enhancer, such as those described in "Transdermal Permeation Enhancers" (Editors: Smith EW and Maibach H I. CRC Press 1995). Exemplary skin penetration enhancers include alkyl (N, N-disubstituted aminoalkanoate) esters, such as dodecyl 2-(N, N-dimethylamino) propionate (DDAIP), which is described in U.S. Pat. Nos. 6,083,996 and 6,118,020, which are incorporated herein by reference; a water-dispersible acid polymer, such as a polyacrylic acid polymer, a carbomer (e.g., carbomer TM or Carbomer 940P TM , available from BF Goodrich Co., Akron, Ohio, polyacrylic acid copolymers (such as Pemulen TM or Polycarbophil from AHRobbins, Richmond, Virginia TMa polysaccharide gum, such as agar gum, alginate, carrageenan, gati gum, karaya gum, kadaya gum, rhamnose gum, xanthan gum, and galactomannan gums (e.g., guar gum, carob gum, and locust bean gum), as well as other gums known in the art (e.g., see Industrial Gums: Polysaccharides and Their Derivatives, Whistler RL, BeMiller JN (Eds.), 3rd ed. Academic Press (1992) and Davidson RL, Handbook of Water-Soluble Gums and Resins, McGraw Hill, Inc., NY (1980)); or a combination thereof.

[0313] Other suitable polymer skin penetration enhancers are cellulose derivatives, such as ethylcellulose, methylcellulose, hydroxypropylcellulose. In addition, if necessary, known transdermal penetration enhancers can also be added. Exemplary transdermal penetration enhancers are dimethyl sulfoxide (DMSO) and dimethylacetamide (DMA), 2-pyrrolidone, N, N-diethyl-meta-toluidine (DEET), 1-dodecylazacycloheptane-2-one (Azone TM , a registered trademark of Nelson Research Company), N,N-dimethylformamide, N-methyl-2-pyrrolidone, calcium thioglycolate and other enhancers such as dioxanalytes, cyclic ketones and their derivatives, etc.

[0314] Also illustrative is a group of biodegradable absorption enhancers which are alkyl N,N-2-(disubstituted amino)alkanoates, as described in U.S. Pat. No. 4,980,378 and U.S. Pat. No. 5,082,866, both of which are incorporated herein by reference, including: tetradecyl (N,N-dimethylamino)ethyl ester, dodecyl (N,N-dimethylamino)ethyl ester, octadecyl (N,N-dimethylamino)ethyl ester, octyl (N,N-dimethylamino)ethyl ester and dodecyl (N,N-diethylamino)ethyl ester.

[0315] Particularly preferred skin penetration enhancers include isopropyl myristate; isopropyl palmitate; dimethyl sulfoxide; decyl methyl sulfoxide; dimethylalanine amides of medium chain fatty acids; dodecyl 2-(N,N-dimethylamino)propionate or its salts, for example, its organic salts (e.g., hydrochloride, hydrobromide, sulfate, phosphate and nitrate addition salts) and inorganic salts (e.g., acetate, benzoate, salicylate, glycolate, succinate, nicotinate, tartrate, maleate, malate, pamoate, methanesulfonate, cyclohexanesulfonate, picrate, and lactic acid addition salts), as described in U.S. Pat. No. 6,118,020; and alkyl 2-(N,N-disubstituted amino)-alkanoic acid esters, as described in U.S. Pat. No. 4,980,378 and U.S. Pat. No. 5,082,866.

[0316] By weight, the content range of the skin penetration enhancer in the composition is 0.5% to 10% (w / w). The optimal range is 1.0% to 5% (w / w). In another embodiment, the skin penetration enhancer accounts for 0.5%-1%, 1%-2%, 2%-3%, 3%-4% or 4%-5% (w / w) of the composition.

[0317] The composition can be provided in any useful form. For example, the composition of the present invention can be formulated into a solution, emulsion (including microemulsion), suspension, cream, ointment, foam, lotion, gel, powder or other typical solid, semisolid or liquid composition (e.g., topical spray) for application to the skin or other tissues to which the composition of the present invention can be used. Such compositions may include other ingredients commonly used in such products, such as colorants, fragrances, thickeners (e.g., xanthan gum, fatty acids, fatty acid salts or esters, fatty alcohols, modified cellulose, modified mineral materials, Krisgel 100) TM , or synthetic polymers), antimicrobial agents, solvents, surfactants, detergents, gelling agents, antioxidants, fillers, dyes, viscosity control agents, preservatives, humectants, emollients (such as natural or synthetic oils, hydrocarbon oils, waxes or silicones), hydrating agents, chelating agents, retarders, solubilizing excipients, adjuvants, dispersants, skin penetration enhancers, plasticizers, preservatives, stabilizers, demulsifiers, wetting agents, sunscreens, emulsifiers, moisturizers, astringents, deodorants, and optionally include anesthetics, antipruritic actives, plant extracts, conditioning agents, darkening or whitening agents, shimmering agents, moisturizers, mica, minerals, polyphenols, silicones or their derivatives, sunscreens, vitamins, and botanicals.

[0318] The composition may also include other similar ingredients to provide additional benefits and improve the feel and / or appearance of the topical formulation. Specific categories of additives commonly used in these formulations include: isopropyl myristate, sorbic acid NF powder, polyethylene glycol, phosphatidylcholine (including mixtures of phosphatidylcholine such as phospholipid G), Krisgel 100 TM Distilled water, sodium hydroxide, decyl methyl sulfoxide (as a skin penetration enhancer), menthol crystals, lavender oil, butylated hydroxytoluene, ethyl diglycol reagent, and 95% ethanol (190 degrees).

[0319] Ophthalmic preparations

[0320] The compounds of the present invention can also be formulated with an ophthalmologically acceptable carrier at a sufficient concentration to deliver an effective amount of the active substance compound to the optic nerve of the eye. Preferably, the ophthalmic treatment solution contains one or more active compounds in a concentration range of about 0.0001% to about 5% (weight per unit volume), more preferably about 0.0005% to about 0.1% (weight per unit volume).

[0321] The ophthalmologically acceptable carrier does not cause significant irritation to the eye and does not abrogate the pharmacological activity and properties of the charged sodium channel blocker.

[0322] Ophthalmologically acceptable carriers are generally sterile, substantially free of foreign particles, and generally have a pH in the range of 5-8. Preferably, the pH is as close as possible to the pH of tear fluid (7.4). Ophthalmologically acceptable carriers include, for example, sterile isotonic solutions, such as isotonic sodium chloride or boric acid solutions. Such carriers are generally aqueous solutions containing sodium chloride or boric acid. Phosphate buffered saline (PBS) solutions are also useful.

[0323] Various preservatives can be used in ophthalmic preparations. Preferred preservatives include, but are not limited to, benzalkonium chloride potassium, chlorobutanol, thimerosal, phenylmercuric acetate, and phenylmercuric nitrate. Similarly, various preferred carriers can also be used in such ophthalmic preparations. These carriers include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropyl methylcellulose, poloxamer, carboxymethyl cellulose, and hydroxyethyl cellulose.

[0324] Tonicity adjusters may be added as needed or convenient and include, but are not limited to, salts, particularly sodium chloride, potassium chloride, and the like, mannitol and glycerin, or any other suitable ophthalmologically acceptable tonicity adjuster.

[0325] Various buffers and methods for adjusting the pH value can be used, as long as the resulting preparation is ophthalmologically acceptable. Therefore, buffers include, but are not limited to, acetate buffers, citrate buffers, phosphate buffers, and borate buffers. Acids or bases can be used to adjust the pH value of these preparations as needed. Ocularly acceptable antioxidants may also be included. Antioxidants include, but are not limited to, sodium pyrosulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole, and butylated hydroxytoluene.

[0326] Preparations for nasal administration or inhalation

[0327] The pharmaceutical composition of the present invention can be formulated for nasal or intranasal administration. When the carrier is solid, preparations suitable for nasal administration include coarse powders having a particle diameter, for example, in the range of about 20 to 500 microns, for rapid inhalation through the nasal passages. When the carrier is liquid, for example, as a nasal spray or nasal drops, one or more preparations can be mixed in an aqueous or oily solution and inhaled or sprayed into the nasal cavity through the nasal cavity.

[0328] For administration by inhalation, the active ingredient may conveniently be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using 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 to deliver a metered amount of capsules and cartridges, e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0329] The dry powder composition for local delivery can be delivered to the lungs by inhalation, using an inhaler or an inflator, for example, in the form of capsules and cartridges, for example, gelatin or blisters, for example, laminated aluminum foil. The powder mixture preparation generally comprises a powder mixture of the compound of the present invention for inhalation and a suitable powdered matrix (carrier / diluent / excipient material), such as a monosaccharide, disaccharide or polysaccharide (such as lactose or starch). Lactose is preferably used. In one embodiment, each capsule or cartridge may contain about 2ug to about 100mg of the compound of formula (I), which is optionally combined with another therapeutically active ingredient. In a preferred embodiment, each capsule or cartridge may contain about 10 micrograms to about 50 milligrams of the compound of formula (I), which is optionally combined with another therapeutically active ingredient. In another embodiment, each capsule or cartridge may contain about 20ug to about 10mg of the compound of formula (I), which is optionally combined with another therapeutically active ingredient. Alternatively, the compound of the present invention may be delivered without excipients.

[0330] Suitably, the package / medicament dispenser is selected from the group consisting of a reservoir dry powder inhaler (RDPI), a unit dose dry powder inhaler (eg a capsule or blister pack inhaler), a multi-dose dry powder inhaler (MDPI) and a metered dose inhaler (MDI).

[0331] Solutions or suspensions for use in pressurized containers, pumps, sprays, atomizers or nebulizers can be formulated to contain an aqueous medium, ethanol, liquid ethanol or other agents suitable for dispersing, dissolving or prolonging the release of the active ingredient; a propellant as a solvent; and / or a surfactant, such as sorbitan trioleate, oleic acid or oligolactic acid.

[0332] Compositions formulated as nasal or inhaled formulations may include one or more taste masking agents, such as flavoring agents, sweeteners and other solutions, such as sucrose, glucose, and lactose, carboxylic acids, menthol, amino acids or amino acid derivatives, such as arginine, lysine, and sodium glutamate, and / or synthetic flavor oils and flavors and / or natural oils, extracts of plants, leaves, flowers, fruits, etc. and combinations thereof. These may include cinnamon oil, wintergreen oil, peppermint oil, clover oil, bay oil, anise oil, eucalyptus oil, vanilla oil, citrus oils (such as lemon oil, orange oil, grape oil and grapefruit oil), fruit flavors (including apples, peaches, pears, strawberries, raspberries, cherries, plums, pineapples, apricots, etc.). Other sweeteners include sucrose, glucose, aspartame, acesulfame-K, sucralose and saccharin, organic acids (non-limiting examples include citric acid and aspartic acid). Such flavorants are present in an amount of about 0.05% to about 4% by weight, and may be present in lower or higher amounts depending upon factors affecting flavor effect, the solubility of the flavoring, the effect of the flavoring on the solubility or other physicochemical or pharmacokinetic properties of other formulation ingredients, or other factors.

[0333] instruct

[0334] The compounds, compositions, methods and kits of the invention can be used to treat pain, cough or pruritus associated with any of a variety of diseases, including trigeminal neurotrophic syndrome, erythromelalgia, back and neck pain, lower back pain, cancer pain, gynecological and labor pain, abdominal wall pain, chronic abdominal wall pain, fibromyalgia, allergic rhinitis, arthritis, rheumatoid arthritis, osteoarthritis, rheumatic pain, orthopedic pain, acute and postherpetic neuralgia and other neuropathic pain (including peripheral neuropathy), sickle cell crisis, myalgia, vulvodynia, rectal pain, levator anus syndrome, rectal pain, perianal pain, hemorrhoidal pain, stomach pain, ulcers, inflammatory bowel disease, intestinal Irritable syndrome, irritable bowel syndrome, oral mucositis, esophagitis, interstitial cystitis, urethritis and other urinary tract pain, toothache, burning pain, headache, eye irritation, conjunctivitis (such as allergic conjunctivitis), eye redness, dry eyes, dry eyes (chronic eye pain), complex regional pain syndrome, acute postoperative pain, postoperative pain, postoperative eye pain, and procedural pain (i.e. pain associated with injections, abscess drainage, surgery, dental surgery, ophthalmic surgery, eye irritation, conjunctivitis (such as allergic conjunctivitis)), eye redness, dry eyes, arthroscopy and other medical device use, cosmetic surgery, dermatologic surgery, fixation of fractures, biopsies, etc.).

[0335] Because a subset of nociceptive neurons mediates itch sensations, the compounds, compositions, methods and kits of the invention can also be used to treat itch in patients with conditions such as pruritus (including, but not limited to, brachioradial, chronic idiopathic, genital / anal, paresthesia and scalp itch), allergic dermatitis, atopic dermatitis, contact dermatitis, poison ivy, infections, parasites, insect bites, pregnancy, metabolic disorders, liver or kidney failure, drug reactions, allergic reactions, eczema, hand eczema, genital and anal itch, hemorrhoidal itch, and cancer.

[0336] Since a subset of nociceptive neurons can trigger an abnormal cough reflex, the compounds, compositions, methods and kits of the present invention can also be used to treat asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), idiopathic pulmonary fibrosis, post-viral cough, post-infectious cough, chronic idiopathic cough and cough in patients with lung cancer.

[0337] The compounds, compositions, methods and kits of the present invention can also be used to treat neurogenic inflammation and neurogenic inflammatory diseases. Inflammation is a series of complex responses to noxious stimuli, resulting in local redness, swelling and pain. Inflammation can be innate or adaptive, the latter driven by antigens and mediated by immune cells (immune-mediated inflammation). Neurogenic inflammation originates from the efferent function of pain neurons (nociceptive neurons), in which neuropeptides and other chemicals that serve as proinflammatory mediators are released from the peripheral terminals of nociceptors when activated. This release process is mediated by calcium influx and exocytosis of peptide-containing vesicles, and proinflammatory neuropeptides include substance P, neurokinin A and B (collectively referred to as tachykinins), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP).

[0338] The release of peripheral chemicals stimulates multiple inflammatory responses. First, the release of substance P can lead to increased capillary permeability, which allows plasma proteins to leak from the intravascular compartment into the extracellular space (plasma extravasation), resulting in edema. This can be detected as a wheal (a firm, raised swelling of the skin), which is one of three inflammatory responses - wheal, erythema, and flare, known as the Lewis triple reaction. Second, the release of calcitonin gene-related peptide causes vasodilation, resulting in increased blood flow. This can be detected as a flare, another component of the Lewis triple reaction.

[0339] Substance P has pro-immune effects on immune cells such as macrophages, T cells, mast cells, and dendritic cells via their neurokinin-1 (NK1) receptors. This effect has been documented in allergic rhinitis, gastritis, and colitis and represents the dividing line between neurogenic and immune-mediated inflammatory components. Substance P released by one nociceptive neuron may also act on NK1 receptors on adjacent nociceptive neurons, sensitizing or activating them, leading to spread of activation and afferent / efferent function. These efferent functions of nociceptive neurons can be triggered by: 1) direct activation of nociceptive neuron terminals by applying sufficient peripheral stimulation (e.g., pressure) to the nociceptive neuron terminals; 2) indirect antidromic activation of non-stimulated nociceptive neuron terminals through axonal reflexes, in which action potential input from one terminal of a nociceptive neuron, upon reaching a converging axon branch point in the periphery, causes the action potential to move downward from the branch point to the peripheral terminal of the non-stimulated terminal; and 3) activation caused by activity of the central terminals of nociceptive neurons in the central nervous system as they move toward the periphery (e.g., GABA-generated primary afferent depolarization of central terminals is sufficient to initiate an action potential that moves the "wrong way").

[0340] Genomic analysis of ILC2 cells in the lung revealed expression of receptors for several neuropeptides released by sensory neurons, including SP, CGRP, and VIP, providing an opportunity for nociceptive neurons to communicate directly with these cells. In particular, VIP was found to be expressed in NaV1.8+ nodose ganglion neurons, including pulmonary afferents in OVA-exposed mice. Stimulation of cultured nodose ganglion neurons with capsaicin or IL5 also released VIP, and VIP levels were elevated in the BALF of mice exposed to OVA compared with vehicle-challenged mice (Talbot et al., Neuron. 2015 July 15;87(2):341–354). These data suggest that VIP is released in the inflamed lung and can be blocked by silencing the neurons using charged sodium channel blockers of the invention. Furthermore, when CD4+ T cells cultured under TH2-biased conditions were exposed to recombinant mouse VIP, transcriptional levels of IL-13 and IL-5 were increased, suggesting that VIP facilitates transcription of these type II regulatory cytokines by TH2 cells.

[0341] Immune mediators released by immune cells can also activate nociceptive neurons. Mast cells are in close proximity to primary nociceptive neurons and contribute to the sensitization of nociceptive neurons in many cases. Injection of the secretagogue compound 48 / 80 can promote mast cell degranulation in the dura mater, leading to the excitation of meningeal nociceptors. Mast cell degranulation also contributes to the rapid onset of heat hyperalgesia induced by nerve growth factor. Macrophages promote the sensitization of nociceptive neurons by releasing several soluble mediators. After partial ligation of the sciatic nerve, the expression of the chemokine macrophage inflammatory protein-1α (MIP-1α) and its receptors CCR1 and CCR5 in macrophages and Schwann cells increases and contributes to the occurrence of neuropathic pain. Lymphocytes contribute to the sensitization of peripheral nociceptive neurons. After nerve injury, T cells infiltrate the sciatic nerve and dorsal root ganglia (DRG). In rodents lacking T cells, hyperalgesia and allodynia caused by nerve injury are significantly reduced or eliminated, and the immunosuppressant rapamycin alleviates neuropathic pain in rats, partly due to its effect on T cells. Among T cell subsets, type 1 and type 2 helper T cells (TH1 and TH2 cells) have different roles in neuropathic pain. TH1 cells promote neuropathic pain behavior by releasing proinflammatory cytokines (IL-2 and interferon-γ (IFN-γ)), while TH2 cells inhibit neuropathic pain behavior 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. C5a is an anaphylatoxin and an important effector of the complement cascade. After binding to the C5aR1 receptor on neutrophils, it becomes an effective neutrophil attractant (Ren & Dubner, Nat. Med. 16: 1267-1276 (2010)).

[0342] Bacterial infection can directly activate nociceptive neurons, and pain caused by Staphylococcus aureus in mice does not require immune responses mediated by TLR2, MyD88, T cells, B cells, and neutrophils and monocytes (Chiu et al., Nature 501: 52-57 (2013)). Mechanical and thermal hyperalgesia in mice is associated with live bacterial load rather than tissue swelling or immune activation. Bacteria induce calcium flux and action potentials in nociceptor neurons through different mechanisms, in part through bacterial N-formyl peptides and the pore-forming toxin α-hemolysin. Specific ablation of Nav1.8 lineage neurons, including nociceptive neurons, abolished pain during bacterial infection but simultaneously increased local immune infiltration and lymphadenopathy in draining lymph nodes. Therefore, bacterial pathogens produce pain by directly activating sensory neurons that regulate inflammation, an unexpected role for the nervous system in host-pathogen interactions. Data from Talbot et al. (Neuron. 2015 July 15; 87(2): 341–354) also demonstrated that nociceptive neurons are activated during exposure to allergens in sensitized animals.

[0343] In certain diseases, neurogenic inflammation results in peripheral inflammation caused by tissue injury, autoimmune disease, infection, and exposure to soft tissue, skin, respiratory system, joints, genitourinary and gastrointestinal tracts, liver, and brain. Neurogenic inflammatory diseases include, but are not limited to, allergic inflammation, inflammatory bowel disease, interstitial cystitis, atopic dermatitis, asthma, conjunctivitis, arthritis, colitis, contact dermatitis, diabetes, eczema, cystitis, gastritis, migraine, psoriasis, rhinitis, rosacea, sunburn, pancreatitis, chronic cough, chronic sinusitis, traumatic brain injury, polymicrobial sepsis, tendinopathy, chronic urticaria, rheumatic disease, acute lung injury, exposure to irritants, inhaled irritants, pollutants, or chemical weapons, as described herein.

[0344] Assessment of pain, cough, pruritus, and neurogenic severity

[0345] In order to measure the efficacy of the compounds of the invention, compositions, methods and kits of the invention in treating pain associated with musculoskeletal, immune inflammation and neuropathy, a measurement index table can be used. Useful indexes include visual analog scales (VAS), Likert scales, categorical pain scales, descriptors, Lequesne index, WOMAC index, and AUSCAN index, each of which is well known in the art. These indices can be used to measure pain, pruritus, function, stiffness or other variables.

[0346] Visual analog scale (VAS) provides a one-dimensional quantitative measurement method. VAS is usually expressed using distance, such as a picture of a line with hash marks drawn at regular distance intervals (e.g., ten 1-cm intervals). For example, the patient can be asked to rank the pain or itch sensation by selecting the point on the line that best matches the pain or itch sensation, where one end of the line corresponds to "no pain" (0 cm) or "no itch" and the other end of the line corresponds to "unbearable pain" or "unbearable itch" (10 cm). This method provides a simple and quick way to obtain quantitative information about how the patient experiences pain or itch. VAS scales and their uses are described in U.S. Patents 6,709,406 and 6,432,937.

[0347] The Likert scale also provides a one-dimensional quantitative measurement method. Typically, the Likert scale has discrete integer values ​​ranging from a low value (e.g., 0, indicating no pain) to a high value (e.g., 7, indicating extreme pain). The pain patient is asked to select a number between the low and high values ​​to indicate the degree of pain. The Likert scale and its use are described in U.S. Patents 6,623,040 and 6,766,319.

[0348] The Lequesne Index and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index assess pain, function, and stiffness of the knee and hip joints using self-administered questionnaires in patients with OA. The knee and hip are both included in the WOMAC, while there is a separate Lequesne questionnaire for the knee and hip. These questionnaires are very useful as they contain more information content compared to the VAS or Likert. The WOMAC Index and the Lequesne Index questionnaires have been extensively validated in osteoarthritis, including in the surgical setting (e.g., knee and hip replacements). There are no significant differences in their measurement characteristics.

[0349] The AUSCAN (Australian-Canadian Hand Arthritis) index uses a valid, reliable and responsive patient self-report questionnaire. In one instance, the questionnaire contains 15 questions in three dimensions (pain, 5 questions; stiffness, 1 question; physical function, 9 questions). The AUSCAN index can utilize Likert or VAS scales, etc.

[0350] Scales for pain measurement used in the methods, compositions and kits of the present invention include the Pain Description Scale (PDS), the Visual Analog Scale (VAS), the Verbal Description Scale (VDS), the Numeric Pain Intensity Scale (NPI), the Neuropathic Pain Scale (NPS), the Neuropathic Pain Symptom Inventory (NPSI), the Current Pain Inventory (PPI), the Geriatric Pain Scale (GPM), the McGill Pain Questionnaire (MPQ), the Average Pain Intensity (Descriptor Difference Scale), the Numeric Pain Scale (NPS) Global Assessment Score (GES), the Short Form McGill Pain Questionnaire, the Minnesota Multiphasic Personality Questionnaire, the Pain Profile and the Multidimensional Pain Questionnaire, the Children's Health Questionnaire, and the Children's Assessment Questionnaire.

[0351] Itch can be measured by subjective measures (VAS, Likert, descriptive scales). Another method is to measure traces, which are objective correlates of itch, using vibration sensors or motion-sensitive meters.

[0352] Cough can be measured using standard questionnaires such as the Leicester Cough Questionnaire as well as validated objective instruments such as the VitaloJAK.

[0353] Example

[0354] The following examples are intended to illustrate the present invention and should not be construed as limiting the present invention.

[0355] Definitions of Common Abbreviations

[0356] ACN Acetonitrile

[0357] AcOH Acetic acid

[0358] aq. Aqueous solution

[0359] Bn Benzyl

[0360] Boc tert-Butyloxycarbonyl

[0361] brine A saturated solution of sodium chloride in water

[0362] ℃ Celsius

[0363] δ Chemical shift (ppm)

[0364] d Deuterium

[0365] DCM Dichloromethane

[0366] DIPEA Diisopropylethylamine

[0367] DMAP 4-dimethylaminopyridine

[0368] DMSO Dimethyl sulfoxide

[0369] ESI Electrospray ionization

[0370] Et2O Diethyl ether

[0371] EtOAc Ethyl acetate

[0372] g

[0373] h hour

[0374] HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxygen

[0375] MeOH Methanol

[0376] mHz Megahertz

[0377] min

[0378] ml milliliters

[0379] MS

[0380] m / z mass to charge ratio

[0381] NMR Nuclear Magnetic Resonance

[0382] Pet ether

[0383] RT Room temperature

[0384] TEA Triethylamine

[0385] TLC Thin Layer Chromatography

[0386] UV lamp

[0387] 1. Synthesis of N-benzyl-2-((2,6-dimethylphenyl)amino)-N,N-diethyl-2-oxyethyl-1-ammonium bromide:

[0388]

[0389] To a stirred solution of 2-(diethylamino)-N-(2,6-dimethylphenyl)acetamide (0.15 g, 0.640 mmol, 1 eq; available from Comb-Blocks, San Diego, CA, Cat. No. QA-3221) in ACN (3 mL) was added benzyl bromide (0.164 g, 0.960 mmol, 1.5 eq) and the reaction mixture was stirred at 75°C for 12 h. The progress of the reaction mixture was monitored by TLC (mobile phase 10% methanol in DCM; UV observation). The reaction mixture was evaporated under reduced pressure to give a crude product which was triturated with ethyl acetate (2 x 5 mL) to give the product N-benzyl-2-((2,6-dimethylphenyl)amino)-N,N-diethyl-2-oxoethyl-1-ammonium bromide (0.08 g) as a white solid. MS (ESI): m / z 325.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δ10.03 (br s, 1H), 7.66-7.39 (m, 5H), 7.13 (s, 3H), 4.82 (s, 2H), 4.17 (s, 2H), 3.52 (q, J=6.7Hz, 4H), 2.20 (s, 6H), 1.42 (br t, J=6.9Hz, 6H).

[0390] 2. Synthesis of N-benzyl-2-((2,6-dimethylphenyl)amino)-N,N-dimethyl-2-oxyethyl-1-ammonium bromide:

[0391]

[0392] ·Synthesize the intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide.

[0393] To a suspension of 2,6-dimethylaniline (30.5 mL, 247.56 mmol, 1.0 eq) in (300 mL) water was added bromoacetyl bromide (23.8 mL, 272.31 mmol, 1.1 eq) at 10°C. The pH of the reaction mixture was maintained at pH 9-10 using 15% Na2CO3 (aq.) aqueous solution for 1 hour and the progress of the reaction mixture was monitored by TLC (mobile phase: 30% ethyl acetate in hexane, UV observation). The reaction mixture was extracted with ethyl acetate (2×600 mL), the combined organic extracts were washed with brine (200 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The resulting crude reaction product was triturated with Et2O (2×200 mL) to produce 2-bromo-N-(2,6-dimethylphenyl)acetamide (21 g) as a white solid. MS (ESI): m / z 244.02 [M+2] + . 1H NMR (400MHz, CDCl3) δ7.75 (br s, 1H), 7.20-7.02 (m, 3H), 4.07 (s, 2H), 2.24 (s, 6H).

[0394] ·Synthesize the intermediate 2-(dimethylamino)-N-(2,6-dimethylphenyl)acetamide.

[0395] To a stirred solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (5 g, 20.7 mmol, 1.0 eq) in EtOAc (75 mL) was added a 2.0 M solution of dimethylamine in THF (51.8 mL, 103.5 mmol, 5 eq) and the reaction mixture was stirred in a stainless steel bowl at 75° C. for 18 hours. The reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (2×200 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to yield 2-(dimethylamino)-N-(2,6-dimethylphenyl)acetamide (4.1 g). 1 H NMR (400MHz, CDCl3-d) δppm8.63 (1H, br s), 7.02-7.15 (3H, m), 3.15 (2H, s), 2.45 (6H, s), 2.24 (6H, s).

[0396] ●Synthesis of N-benzyl-2-((2,6-dimethylphenyl)amino)-N,N-dimethyl-2-oxoethyl-1-ammonium bromide.

[0397] To a stirred solution of 2-(dimethylamino)-N-(2,6-dimethylphenyl)acetamide (0.2 g, 0.969 mmol, 1.0 eq) in ACN (3 mL) was added benzyl bromide (0.23 mL, 1.936 mmol, 2.0 eq), and the reaction mixture was stirred at 90° C. for 16 hours, and the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The resulting reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product, which was triturated with ethyl acetate (5 mL) to give the product N-benzyl-2-((2,6-dimethylphenyl)amino)-N,N-dimethyl-2-oxoethyl-1-ammonium bromide (0.14 g). MS (ESI): m / z 297.2 [M] + . 1H NMR (400MHz, DMSO-d6) δppm 9.95 (1H, s), 7.49-7.66 (5H, m), 7.06-7.19 (3H, m), 4.81 (2H, s), 4.30 (2H, s), 3.26 (6H, s), 2.19 (6H, s).

[0398] 3. Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)nitrogen-1-mustard bromide

[0399]

[0400] ●Synthetic intermediate 2-(aza-1-yl)-N-(2,6-dimethylphenyl)acetamide.

[0401] According to the method for synthesizing compound 2 described above, intermediate 2-bromo-N-(2,6-dimethylphenyl) acetamide was synthesized. K was added to the solution of 2-bromo-N-(2,6-dimethylphenyl) acetamide (0.5g, 1.9mmol, 1.0eq) after stirring in ACN (5.0mL) CO (0.639g, 4.6mmol, 2.5eq) and azo (0.4mL, 4.0mmol, 2.0eq), the reaction mixture was stirred at 90°C for 20 hours and the reaction progress of the reaction mixture was monitored by TLC (mobile phase: 5% methanol solution in DCM; ultraviolet observation). The reaction crude product was concentrated under reduced pressure, and the resulting residue was layered in water and ethyl acetate (2x50mL). The combined organic extracts were washed with brine solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was triturated with ethyl acetate (20 mL) to give 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)acetamide (0.47 g). MS (ESI): m / z 261.57 [M+H] + . 1 H NMR (400MHz, CHLOROFORM-d) δ8.83 (br s, 1H), 7.09 (s, 3H), 3.32 (s, 2H), 2.93-2.73 (m, 4H), 2.24 (s, 6H), 1.87-1.52 (m, 9H).

[0402] ●Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)azepine-1-bromide.

[0403] To a stirred solution of 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)acetamide (0.2 g, 0.8 mmol, 1.0 eq) in ACN (1.0 mL) was added benzyl bromide (0.1 mL, 0.6 mmol, 2.0 eq) and the reaction was heated to reflux for 16 hours. The progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The crude reaction mixture was concentrated under reduced pressure and the reaction product was triturated with EtOAc (15 mL) to give the product 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)azepine-1-mustard bromide (0.12 g). MS (ESI): m / z 351.2 [M] +.1 H NMR (400MHz, DMSO-d6) δ10.04 (s, 1H), 7.71-7.62 (m, 2H), 7.60-7.48 (m, 3H), 7.20-7.03 (m, 3H), 4.93(s, 2H), 4.15(s, 2H), 3.86-3.70(m, 2H), 3.66-3.42(m, 2H), 2.23(s, 6H), 1.98(br d, J=4.8Hz, 4H), 1.67 (br s, 4H).

[0404] 4. Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-1-pyridine bromide:

[0405]

[0406] ●Synthetic intermediate N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)acetamide.

[0407] According to the method for synthesizing compound 2 described above, intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide was synthesized. Piperidine (0.851g, 10.0mmol, 2.0eq) was added to the solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (1.2g, 5.0mmol, 1.0eq) stirred in EtOAc (40mL), and the reaction mixture was stirred at 75°C for 16 hours, while the reaction progress of the reaction mixture was monitored by TLC (mobile phase: 50% ethyl acetate solution in petroleum ether; ultraviolet observation). The reaction mixture was diluted with ethyl acetate (20mL) and washed with water (2x50ml). Combined with organic extracts, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to produce a crude product, which was ground with n-pentane (25mL) and dried under vacuum to produce product N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)acetamide (1.05g). MS (ESI): m / z 247.36 [M+H] + .1 H NMR (400MHz, CDCl3-d) δppm 8.81 (1H, br s), 7.04-7.11 (3H, m), 3.14 (2H, s), 2.62 (4H, br s), 2.24 (6H, s), 1.65 (4H, quin, J=5.61Hz), 1.44-1.54 (2H, m).

[0408] ●Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-1-bromide.

[0409] To a stirred solution of N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)acetamide (0.15 g, 0.6 mmol, 1.0 eq) in ACN (3 mL) was added benzyl bromide (0.205 g, 1.2 mmol, 2.0 eq) and the reaction mixture was stirred at 80° C. for 16 hours in a sealed test tube. The progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product, which was triturated with ethyl acetate (15 mL) and dried under vacuum to give the product 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-1-bromide (0.15 g). MS (ESI): m / z 337.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.07 (1H, s), 7.49-7.66 (5H, m), 7.07-7.21 (3H, m), 4.96 (2H, s), 4.21 (2H, s), 3.65 (2H, br d, J=12.50Hz), 3.50 (2H, dt, J=12.93, 6.69Hz), 2.21 (6H, s), 1.91-2.05 (4H, m), 1.64-1.76 (1H, m), 1.55 (1H, dt, J=13.76, 7.04Hz).

[0410] 5. Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)pyrrolidine-1-bromide:

[0411]

[0412] ●Synthetic intermediate N-(2,6-dimethylphenyl)-2-(pyrrolidin-1-yl)acetamide.

[0413] The intermediate 2-bromo-N-(2,6-dimethylphenyl)acetamide was synthesized according to the method described above for synthesizing compound 2. Pyrrolidine (0.533 g, 7.5 mmol, 1.5 eq) was added to a solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (1.2 g, 5.0 mmol, 1.0 eq) stirred in EtOAc (40 mL), and the reaction mixture was stirred at 75° C. for 3 hours while monitoring the progress of the reaction mixture using TLC (mobile phase: 50% ethyl acetate in petroleum ether; ultraviolet observation), the reaction mixture was diluted with ethyl acetate (25 mL), washed with water (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to produce N-(2,6-dimethylphenyl)-2-(pyrrolidin-1-yl)acetamide (1.0 g). MS (ESI): m / z 233.44 [M+H] + . 1 H NMR (400MHz, CDCl3-d) δppm 8.63 (1H, br s), 7.00-7.15 (3H, m), 3.35 (2H, s), 2.69-2.84 (4H, m), 2.24 (6H, s), 1.80-1.93 (4H, m).

[0414] ●Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)pyrrolidine-1-bromide.

[0415] To a stirred solution of N-(2,6-dimethylphenyl)-2-(pyrrolidin-1-yl)acetamide (0.15 g, 0.6 mmol, 1.0 eq) in ACN (5 mL) was added benzyl bromide (0.205 g, 1.2 mmol, 2.0 eq) and the reaction mixture was stirred in a sealed tube at 80° C. for 16 hours while the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The reaction was cooled to room temperature and concentrated under reduced pressure to give a crude product which was triturated with ethyl acetate (15 mL) and dried under vacuum to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)pyrrolidine-1-bromide (0.14 g). MS (ESI): m / z 323.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 2.11-2.29(10H,m), 3.60-3.74(2H,m), 3.76-3.88(2H,m), 4.20(2H,s), 4.83(2H,s), 7.08-7.18(3H,m), 7.50-7.66(5H,m), 9.97(1H,s).

[0416] 6. Synthesis of 1-benzyl-1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidin-1-bromide Object:

[0417]

[0418]

[0419] ●Synthetic intermediate 2-bromobutyryl chloride.

[0420] Thionyl chloride (150 mL) was added to 2-bromobutyric acid (25 g, 149.7 mmol, 1.0 eq) at 0° C., and the reaction solution was subsequently stirred for 2 hours at 80° C. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give crude 2-bromobutyryl chloride (27.7 g, 99.5%) as a brown residue, which was directly carried to the next step without further purification. 1 H NMR (400MHz, CDCl3) δ4.52-4.44 (m, 1H), 2.29-2.16 (m, 1H), 2.15-2.02 (m, 1H), 1.10 (t, J=7.3Hz, 3H).

[0421] ·Synthesize the intermediate 2-bromo-N-(2,6-dimethylphenyl)butyramide.

[0422] A stirred 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 an ice bath. To this mixture was slowly added a solution of 2-bromobutyryl chloride (27.5 g, 148.6 mmol, 1.2 eq) in DCM (50 mL) and the resulting mixture was warmed to room temperature while stirring for 2 hours. The pH of the reaction solution was adjusted to pH 5 to 6 with 2N HCl and extracted with DCM (2X200 mL). The combined organic extracts were washed with water (250 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was triturated with n-pentane (150 mL) to give 2-bromo-N-(2,6-dimethylphenyl)butyramide (30 g). MS (ESI): m / z 272.13 [M+2] + . 1 H NMR (400MHz, DMSO-d6) δ9.69 (s, 1H), 7.15-7.02 (m, 3H), 4.51 (t, J=7.3Hz, 1H), 2.22-2.04 (m, 7H), 2.03-1.91 (m, 1H), 0.98 (t, J=7.3Hz, 3H).

[0423] ·Synthesize intermediate N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide.

[0424] To a solution of 2-bromo-N-(2,6-dimethylphenyl)butanamide (50 g, 259.1 mmol, 1.0 eq) stirred in toluene (1 L) was added piperidine (54 mL, 544.1 mmol, 2.1 eq) at room temperature. The resulting reaction mixture was stirred at 110° C. for 16 hours while monitoring the progress of the reaction mixture using TLC (mobile phase: 30% ethyl acetate in hexane; ultraviolet observation). The reaction mixture was concentrated under reduced pressure to give a crude product, which was triturated with n-pentane (500 mL) to give N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide (55 g). MS (ESI): m / z275.27[M+H]+.1H NMR (400MHz, DMSO-d6) δ9.16 (s, 1H), 7.06 (s, 3H), 3.06 (dd, J=5.7, 8.6Hz, 1H), 2.60 (br t, J=4.8Hz, 4H), 2.16 (s, 6H), 1.79-1.59 (m, 2H), 1.51 (br s, 4H), 1.44-1.35 (m, 2H), 0.93 (t, J=7.5Hz, 3H).

[0425] ●Synthesis of 1-benzyl-1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidin-1-bromide.

[0426] To a stirred solution of N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide (1.5 g, 5.5 mmol, 1.0 eq) in ACN (25.0 mL) was added benzyl bromide (1.872 g, 10.9 mmol, 2.0 eq) and the reaction mixture was heated to 80° C. for 16 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The reaction mixture was concentrated under reduced pressure and the crude product was triturated with EtOAc (3×10 mL) to give 1-benzyl-1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidin-1-bromide (1.2 g). MS(ESI): m / z 365.4[M].+.1H NMR (400MHz, DMSO-d6) δ10.36 (s, 1H), 7.57-7.54 (m, 5H), 7.17-7.11 (m, 3H), 5.41 (d, 1H), 4.62 (d, 1H), 4.39 (bs, 1H), 3.72(bs, 2H), 3.22(bs, 2H), 2.43-2.39(m, 1H), 2.15-2.07(m, 9H), 1.96-1.95(m, 2H), 1.61-1.45(m, 2H), 1.18(t, 3H).

[0427] 7. Synthesis of Compounds 7A-27A: Table 5

[0428] Table 5 provides other representative examples of compounds of the present invention, which were synthesized from the intermediate N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide and a suitable alkyl halide according to the method for synthesizing compound 6 described above.

[0429] Table 5

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436]

[0437] 28. Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-(2-hydroxybenzyl)piperidin Pyridin-1-yl 2,2,2-trifluoroacetate:

[0438]

[0439] To a solution of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-(2-methoxybenzyl)piperidin-1-bromide (compound 24, 0.1 g, 0.21 mmol, 1.0 eq) in DMC (2 mL) was added boron tribromide (1 M in DCM) (4 mL, 4.0 mmol, 20 eq) at 0°C. The reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; ultraviolet observation). The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by preparative HPLC (mobile phase A: 0.1% TFA (aq.); mobile phase B: ACN; column: Synergy Polar 250x21 mm, 4.7u (purchased from Phenomenex); flow rate: 18 mL / min, method: 0 / 15, 2 / 15, 10 / 50, 15 / 80, 15.2 / 98, 18 / 98, 18.2 / 15, 22 / 15; temperature: room temperature) and the pure fractions were freeze-dried to give the product 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-(2-hydroxybenzyl)piperidin-1-yl 2,2,2-trifluoroacetate (0.056 g). MS(ESI): m / z 381.4[M]+.1H NMR(500MHz, DMSO-d6)δppm1.17(3H, br t, J=7.02Hz), 1.32-1.47 (1H, m), 1.55-1.67 (1H, m), 1.76-2.01 (3H, m), 2. 05-2.19(2H,m), 2.25-2.3(6H,m), 2.30-2.41(1H,m), 2.99-3.26(1H,m), 3. 64-3.85(2H,m), 4.35-4.61(2H,m), 5.41-5.50(1H,m), 6.88-6.95(1H,m), 6.99 (1H, d, J = 8.24Hz), 7.10-7.19 (3H, m), 7.31-7.45 (2H, m), 10.34 (2H, br d, J=10.07Hz).

[0440] 29. Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-(3-hydroxybenzyl)piperidin Pyridine-1-carbonate:

[0441]

[0442] To a stirred solution of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-(3-methoxybenzyl)piperidin-1-bromide (Compound 23, 0.06 g, 0.1 mmol, 1 eq) in DCM (2 mL) at 0°C was added boron tribromide (1 M in DCM) (2 mL), and the reaction mixture was stirred at room temperature for 16 h while the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was further purified by reverse phase preparative-HPLC (mobile phase (A): 10 mM ammonium bicarbonate (aq.); mobile phase (B): 100% CAN; column: X-selective C18 (19*250) 5u, method: 0 / 35, 2 / 40, 20 / 40; flow rate: 18 ml / min. The combined pure fractions were freeze dried to give the product 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-(3-hydroxybenzyl)piperidin-1-carbonate (0.02 g). MS (ESI): m / z 381.38 [M] + . 1 H NMR (500MHz, DMSO-d6) δppm 1.11 (3H, br s), 1.31-1.62 (2H, m), 1.76-2.25 (12H, m), 2.93-3.13 (1H, m), 3.43-3.54 (3H, m), 3.60-3.76 (1 H, m), 3.92-4.17 (2H, m), 4.21-4.39 (1H, m), 5.29-5.63 (1H, m), 6.67-6.85 (4H, m), 6.91 (2H, br s), 7.19 (1H, br t, J=7.78Hz).

[0443] 30. Synthesis of 1-benzyl-1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)azepine-1-bromide Object:

[0444]

[0445] ●Synthetic intermediate 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)butanamide.

[0446] The intermediate 2-bromo-N-(2,6-dimethylphenyl)butyramide was synthesized according to the method for synthesizing compound 6 described above. K2CO3 (0.393g, 2.85mmol, 1.5eq) and nitrogen mustard (0.376g, 3.8mmol, 2.0eq) were added to a solution of 2-bromo-N-(2,6-dimethylphenyl)butyramide (0.5g, 1.9mmol, 1.0eq) stirred in ACN (10mL), and the reaction mixture was stirred at 75°C for 16 hours and the reaction progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; ultraviolet observation). The crude reaction mixture was annealed with water (10mL) and extracted with EtOAc (2x15mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to produce the intermediate 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)butyramide (0.20g). MS (ESI): m / z 289.33 [M+H] + . 1 H NMR (400MHz, CDCl3-d) δppm1.00-1.16 (3H, m), 1.57-1.86 (12H, m), 1.95-2.13 (2H, m), 2.18-2.29 (8H, m), 2.72-2.97 (4H, m), 3.24 (1H, br t, J=6.72Hz), 3.46-3.55 (2H, m), 7.03-7.15 (4H, m), 8.75 (1H, br s).

[0447] ●Synthesis of 1-benzyl-1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)azepine-1-bromide.

[0448] To a stirred solution of 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)butanamide (0.2 g, 0.7 mmol, 1.0 eq) in ACN (5 mL) was added benzyl bromide (0.459 g, 2.85 mmol, 1.5 eq) and the reaction mixture was stirred at 75° C. for 16 hours while the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM; UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product which was triturated with ethyl acetate (20 mL) and dried under vacuum to give 1-benzyl-1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)azepine-1-bromide (0.118 g). MS (ESI): m / z 379.52[M]+.1HNMR (500MHz, DMSO-d6) δppm 1.16 (3H, t, J=7.32Hz), 1.26-1.51 (4H, m), 1.60 (2H, br s), 1.77-1.94 (2H, m), 2.10-2.31 (7H, m), 2.34-2.48 (1H, m), 3.56-3.81 (4H, m), 4.26 (1H, br d, J=10.68Hz), 4.66 (1H, d, J=13.12Hz), 5.21 (1H, d, J=13.12Hz), 7.06-7.22 (3H, m), 7.45-7.61 (3H, m), 7.61-7.69 (2H, m), 10.42 (1H, s).

[0449] 31. Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)azepine-1-bromide

[0450]

[0451] Synthetic intermediate 2-(azo-1-yl)-N-(2,6-dimethylphenyl)acetamide:

[0452] To a solution of 2-bromo-N-(2,6-dimethylphenyl)acetamide (500 mg, 2.065 mmol) stirred in ACN (5.0 mL), KCO (713 mg, 5.159 mmol) was added, followed by azo (467 mg, 4.125 mmol), and the reaction mixture was stirred at 80 ° C for 16 hours while monitoring the reaction progress of the reaction mixture by TLC (mobile phase: 5% methanol in DCM; ultraviolet observation). The resulting reaction mixture crude product was poured into ice water (40 mL) and extracted with ethyl acetate EtOAc (2x50 mL). The combined organic extracts (50 mL) were washed with saline solution, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to produce 2-(azo-1-yl)-N-(2,6-dimethylphenyl)acetamide (480 mg) as an off-white solid. Mass spectrum (ESI): m / z 275.1[M+H]+.1H NMR (400MHz, CHLOROFORM-d) δppm 8.74(br s, 1H), 7.02-7.16 (m, 3H), 3.34 (s, 2H), 2.74-2.88 (m, 4H), 2.25 (s, 6H), 1.53-1.80 (m, 10H).

[0453] · Synthesis of 1-Benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)azo-1-bromide

[0454] To a stirred solution of 2-(azo-1-yl)-N-(2,6-dimethylphenyl)acetamide (200 mg, 0.728 mmol) in ACN (2.0 mL) was added benzyl bromide (240 mg, 1.403 mmol) and the resulting reaction mixture was stirred at 90°C for 16 hours while the progress of the reaction mixture was monitored by TLC (mobile phase: 5% methanol in DCM; UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was triturated with a 1:1 mixture of EtOAc:Et2O (3 x 50 mL) to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)azo-1-bromide as a light brown solid. Mass spectrum (ESI): m / z 365.54 [M] +. 1H NMR (400 MHz, DMSO-d6) δ ppm 10.02 (s, 1H), 7.46-7.70 (m, 5H), 7.05-7.22 (m, 3H), 4.89 (s, 2H), 4.07 (s, 2H), 3.64-3.76 (m, 2H), 3.48-3.62 (m, 2H), 2.23 (s, 6H), 1.94-2.14 (m, 4H), 1.50-1.83 (m, 6H).

[0455] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3,3-difluoropiperidin-1-bromide Chemicals:

[0456]

[0457] Synthesis of 2-(3,3-difluoropiperidin-1-yl)-N-(2,6-dimethylphenyl)acetamide:

[0458] To a solution of 3,3-difluoropiperidine hydrochloride (0.371 g, 0.309 mmol) stirred in ACN (25 mL) was added TEA (0.626 g, 6.195 mmol) at 0°C. After stirring at 0°C for 10 minutes, 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.5 g, 2.065 mmol) was added at 0°C. The resulting reaction mixture was stirred at room temperature for 16 hours while the reaction progress of the reaction mixture was monitored by TLC (mobile phase: 30% ethyl acetate in petroleum ether; ultraviolet observation). The reaction mixture was diluted with water (10 mL) and extracted with ETOAc (2x25 mL). The combined organic layers were concentrated under reduced pressure to produce a crude product, which was purified by normal phase flash chromatography (5% methanol in DCM) to produce 2-(3,3-difluoropiperidin-1-yl)-N-(2,6-dimethylphenyl)acetamide (0.4 g) as a white solid. MS (ESI): m / z 283.24 [M+H] + . 1 HNMR (400MHz, CHLOROFORM-d) δppm 8.58 (s, 1H), 7.07-7.13 (m, 3H), 3.27 (s, 2H), 2.86 (t, 2H), 2.50 (d, 2H), 2.23 (s, 6H), 1.86-2.04 (m, 4H).

[0459] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3,3-difluoropiperidin-1-bromide:

[0460] At room temperature, in a sealed test tube, benzyl bromide (0.145 g, 0.850 mmol) was added to a solution of 2-(3,3-difluoropiperidin-1-yl)-N-(2,6-dimethylphenyl)acetamide (0.2 g, 0.708 mmol) stirred in ACN (10 mL). The resulting reaction mixture was heated to 75 ° C and maintained for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (5% methanol in DCM, ultraviolet observation). The reaction mixture was diluted with water (25 mL) and extracted with 10% methanol in DCM (2x100 mL). The combined organic layer was concentrated under reduced pressure to produce a crude product, which was ground with ethyl acetate (10 mL) to produce 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3,3-difluoropiperidin-1-bromide (0.060 g) as an off-white solid. MS (ESI): m / z 373.1 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.10(s, 1H), 7.56-7.62(m, 5H), 7.07-7.18(m, 3H), 5.16(d, 1H), 5.01(d, 1H) , 4.31-4.40(m, 2H), 3.98-4.18(m, 2H), 3.61-3.68(m, 2H), 2.11-2.48(m, 10H).

[0461] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-((phenyl-d5)methyl)azepine-1-chloro :

[0462]

[0463] ●Synthesis of 1-(chloromethyl)benzene-2,3,4,5,6-d5:

[0464] To a solution of benzene-d6 (5 g, 59.41 mmol) in concentrated hydrochloric acid (15 mL) was added paraformaldehyde (5.3522 g, 178.23 mmol) at room temperature, and the reaction mixture was stirred in a sealed container at 80° C. for 48 hours, and the progress of the reaction mixture was monitored by 1H NMR. The reaction mixture was extracted with Et2O (2×25 mL) and the combined organic extracts were washed with saturated aqueous sodium bicarbonate solution (50 mL), dried over anhydrous sodium sulfate and concentrated at 25° C. under reduced pressure to yield 1-(chloromethyl)benzene-2,3,4,5,6-d5 (1.6 g) as a colorless liquid. 1 H NMR (400MHz, CDCl3) δppm 4.6 (s, 2H).

[0465] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-((phenyl-d5)methyl)azepine-1-chloride

[0466] To a solution of 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)acetamide (1.5 g, 5.76 mmol) stirred in ACN (15 mL) was added 1-(chloromethyl)benzene-2,3,4,5,6-d5 (1.592 g, 12.1 mmol), and the resulting reaction mixture was stirred in a sealed test tube at 80°C for 16 hours, and the progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was cooled to room temperature, filtered and the precipitated solid was washed with ethyl acetate to give 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-((phenyl-d5)methyl)azepine-1-chloride (1.67 g) as a white solid. MS (ESI): m / z 356.36 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.55(s, 1H), 7.1-7.18(m, 3H), 4.95(s, 2H), 4.2(s, 2H), 3.72-3.81(m, 1H), 3.5-3.6(m, 1H), 2.25(s, 6H), 1.9-2.1(m, 4H), 1.1-1.2(m, 4H).

[0467] Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-((1-methyl-1H-pyrazole-4- 1-(2-(4-(2-methyl)piperidin-1-yl)methyl)piperidin-1 :

[0468]

[0469] Synthesis of 4-(bromomethyl)-1-methyl-1H-pyrazole hydrobromide:

[0470] At room temperature, to a solution of (1-methyl-1H-pyrazol-4-yl)methanol (1.0 g, 8.918 mmol) in AcOH (5 mL) was added a solution of 33% HBr in AcOH (12 mL). The reaction mixture was heated to 100° C. and maintained for 16 hours, and the progress of the reaction mixture was monitored using TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to produce 4-(bromomethyl)-1-methyl-1H-pyrazole hydrobromide (1.5 g) as a light brown solid. 1 H NMR (400MHz, DMSO-d6) δppm 8.01 (s, 1H), 7.50 (s, 1H), 5.51 (s, 2H), 3.81 (s, 3H).

[0471] Synthesis of 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-((1-methyl-1H-pyrazol-4-yl)methyl)piperidin-1-bromide:

[0472] To a solution of 4-(bromomethyl)-1-methyl-1H-pyrazole bromate (0.5 g, 1.953 mmol) stirred in DMF (5 mL), N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)butyramide (0.783 g, 2.853 mmol) and potassium carbonate (0.473 g, 3.427 mmol) were added. The reaction mixture was heated to 100 ° C and maintained for 24 hours, and the reaction progress of the reaction mixture was monitored using TLC (5% methanol in DCM, ultraviolet observation). The reaction mixture was diluted with water (25 mL) and extracted with 10% methanol in DCM (2x100 ml). The combined organic layers were concentrated under reduced pressure to give the crude product which was purified by normal phase flash chromatography (5% methanol in DCM) to give 1-(1-((2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)-1-((1-methyl-1H-pyrazol-4-yl)methyl)piperidin-1-bromide (0.120 g) as an off-white solid. MS (ESI): m / z 369.48 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.4 (s, 1H), 8.01 (s, 1H), 7.64 (s, 1H), 7.10-7.14 (m, 3H), 5.17 (d, 1H), 4.48 (d, 1H), 4.1-4.3 (m, 1H), 3.90 (s, 3H), 3. 8-3.9 (m, 1H), 3.20-3.45 (m, 2H), 2.30-2.36 (m, 1H), 2.16 (s, 6H), 1.92-2.08 (m, 6H), 1.54 (d, 2H), 1.11-1.17 (m, 3H).

[0473] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(3-(methoxycarbonyl)benzyl)piperidin Pyridine-1-bromide :

[0474]

[0475] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(3-(methoxycarbonyl)benzyl)piperidin-1-bromide:

[0476] To a stirred solution of N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)acetamide (200 mg, 0.812 mmol) in ACN (3 mL) was added methyl 3-(bromomethyl)benzoate (241.6 mg, 1.055 mmol) at room temperature. The resulting reaction mixture was stirred at 90°C for 16 hours and the reaction was monitored by TLC (5% methanol in DCM, UV observation). The resulting reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product which was triturated with ethyl acetate (3X10 mL) to give pure 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(3-(methoxycarbonyl)benzyl)piperidin-1-bromide (57.5 mg) as an off-white solid. MS (ESI): m / z 395.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.09(s, 1H), 8.18(s, 1H), 8.13(d, 1H), 7.83-7.85(m, 1H), 7.69-7.72(m, 1H), 7.12-7.17(m, 3H), 5.06(s, 2H), 4.22(s, 2H) , 3.88(s, 3H), 3.52-3.65(m, 2H), 3.47-3.50(m, 2H), 2.21(s, 6H), 1.90-1.96(m, 4H), 1.71-1.74(m, 1H), 1.56-1.57(m, 1H).

[0477] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(4-(methoxycarbonyl)benzyl)piperidin Pyridine-1-bromide :

[0478]

[0479] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(4-(methoxycarbonyl)benzyl)piperidin-1-bromide:

[0480] To a stirred solution of N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)acetamide (0.4 g, 1.623 mmol) in ACN (5 mL) was added methyl 4-(bromomethyl)benzoate (0.743 g, 3.247 mmol) and the reaction mixture was stirred at 80° C. for 16 hours and the reaction was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product which was further purified with ethyl acetate (30) by refluxing at 70° C. for 2 hours. The refluxed product was filtered and washed with hot ethyl acetate (30 mL) to give (1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(4-(methoxycarbonyl)benzyl)piperidin-1-bromide (43 mg) as a white solid. Mass spectrum (ESI): m / z 395.3 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.06 (s, 1H), 8.09 (d, 2H), 7.72 (d, 2H), 7.17-7.12 (m, 3H), 5.03 (s, 2H), 4.22 (s, 2H), 3.89 (s, 3H), 3.6 7-3.64(m, 2H), 3.54-3.49(m, 2H), 2.20(s, 6H), 2.0-1.97(m, 4H), 1.72-1.69(m, 1H), 1.58-1.53(m, 1H).

[0481] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(2-(methoxycarbonyl)benzyl)piperidin Pyridine-1-bromide:

[0482]

[0483] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(2-(methoxycarbonyl)benzyl)piperidin-1-bromide:

[0484] To a solution of N-(2,6-dimethylphenyl)-2-(piperidin-1-yl)acetamide (0.500 g, 2.0296 mmol) in ACN (5.0 mL) was added methyl 2-(bromomethyl)benzoate (0.929 g, 4.0592 mmol) and the resulting reaction mixture was heated to 80°C for 16 hours and the reaction progress was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude compound which was triturated with 3x10 mL of ethyl acetate followed by 2x10 mL of diethyl ether to give pure 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1-(2-(methoxycarbonyl)benzyl)piperidin-1-bromide (250 mg) as an off-white solid. MS (ESI): m / z 395.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.11(s, 1H), 8.01(d, 1H), 7.69-7.76(m, 3H), 7.11-7.17(m, 3H), 5.37(s, 2H), 4.43-4.46(m, 2H), 3.85(s, 3H), 3. 75(d, 2H), 3.17(t, 2H), 2.20(s, 6H), 1.96-2.07(m, 2H), 1.79-1.83(m, 2H), 1.66-1.70(m, 1H), 1.37-1.40(m, 1H).

[0485] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)(methyl)amino)-2-oxoethyl)azepine-1-carboxylic acid Salt:

[0486]

[0487] Synthesis of 2-iodo-N-(2,6-dimethylphenyl)-N-methylacetamide:

[0488] At 0 ℃, to 2-chloro-N-(2,6-dimethylphenyl) acetamide (2.0g, 10.118mmol) in THF (16mL) after stirring, add NaH (60%) (0.607g, 25.295mmol), and stir the reaction mixture at 0 ℃ for 20 minutes.Add iodomethane (0.5ml, 8.031mmol) at 0 ℃ and stir the obtained reaction mixture at room temperature for 16 hours.Use TLC (10% ethyl acetate solution in Pet, ultraviolet observation) to monitor the reaction process of the reaction mixture.The reaction mixture is poured into ice water (50mL) and extracted with ethyl acetate (2x50mL).Wash the combined organic extract with saline solution (25mL), dry with anhydrous sodium sulfate and concentrate under reduced pressure to produce crude product, which is purified by column chromatography (eluted with 20-30% ethyl acetate solution in petroleum ether). The collected pure fractions were concentrated under reduced pressure to give N-(2,6-dimethylphenyl)-2-iodo-N-methylacetamide (0.55 g) as a yellow liquid. Mass spectrum (ESI): m / z 304.03 [M+H] + .

[0489] Synthesis of 2-(aza-1-yl)-N-(2,6-dimethylphenyl)-N-methylacetamide:

[0490] Under room temperature, K2CO3 (0.569g, 4.123mmol) and azo (0.327ml, 3.298mmol) were added to a solution of N-(2,6-dimethylphenyl)-2-iodo-N-methylacetamide (0.5g, 1.649mmol) stirred in ACN (5.0mL). The reaction mixture was stirred at 90°C for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, ultraviolet observation). The reaction mixture was poured into ice water (25mL) and extracted with ethyl acetate (2x25mL). The combined organic extracts were washed with brine solution (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude compound which was purified by normal phase flash chromatography (eluting with 10%-50% ethyl acetate in petroleum ether) and the collected pure fractions were concentrated under reduced pressure to give 2-(aza-1-yl)-N-(2,6-dimethylphenyl)-N-methylacetamide (0.23 g). MS (ESI): m / z 275.49 [M+H] + .

[0491] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)(methyl)amino)-2-oxoethyl)azepine-1-carboxylate

[0492] To a stirred solution of 2-(azepine-1-yl)-N-(2,6-dimethylphenyl)-N-methylacetamide (0.1 g, 0.364 mmol) in acetonitrile (1.0 mL) was added (bromomethyl)benzene (0.124 g, 0.728 mmol). The reaction mixture was stirred at 90° C. for 16 hours and the progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was purified by reverse phase preparative HPLC (column: X-selective C18 (250*19) mm, 5u, mobile phase A: 0.1% FA in water, mobile phase B: ACN:MeOH, flow rate: 13 ml / min, solubility: water+THF+ACN, method (T% of B): 0 / 10, 2 / 10, 10 / 50, 13 / 50, 13.2 / 98, 17 / 98, 17.2 / 10, 20 / 10, temperature: room temperature).

[0493] The collected pure fractions were freeze-dried to give the product (1-benzyl-1-(2-((2,6-dimethylphenyl)(methyl)amino)-2-oxoethyl)azepine-1-carboxylate (70 mg) as a white gum. MS (ESI): m / z 365.3 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 8.43(s, 1H), 7.56-7.42(m, 5H), 7.26-7.17(m, 3H), 4.95(s, 2H), 4.41(s, 1H), 3.91-3.85(m, 1H), 3. 62-3.50(m, 3H), 3.36(s, 1H), 3.14(d, 3H), 2.32-2.05(s, 7H), 1.95-1.65(m, 2H), 1.46-1.23(m, 5H).

[0494] Synthesis: Synthesis of 1-benzyl-1-(1-(mesitylamino)-1-oxobutyl-2-yl)piperidin-1-bromide :

[0495]

[0496] Synthesis of 2-bromo-N-mesitylene butyramide:

[0497] To a stirred solution of 2,4,6-trimethylaniline (2.0 g, 14.791 mmol) in DCM (20 mL) was added DIPEA (5.735 g, 44.373 mmol) at room temperature, followed by the dropwise addition of 2-bromobutyryl chloride (3.291 g, 17.749 mmol) at 0°C, and the resulting reaction mixture was stirred for 3 hours, the progress of the reaction mixture was monitored using TLC (20% ethyl acetate in petroleum ether, UV observation). The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 100 mL). The combined organic extracts were washed with brine solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product, which was purified by column chromatography (eluted with 10%-50% ethyl acetate in petroleum ether), and the pure fractions were combined and concentrated to give the desired compound 2-bromo-N-mesitylenebutanamide (1.5 g). MS (ESI): m / z 286.11 [M+2] + . 1 H NMR (400MHz, CDCl3) δppm 7.60 (s, 1H), 6.95 (s, 2H), 4.45 (t, 1H), 2.25 (s, 3H), 2.2 (s, 6H), 1.60-1.40 (m, 2H), 1.10 (t, 3H).

[0498] Synthesis of N-mesitylene-2-(piperidin-1-yl)butyramide:

[0499] Potassium carbonate (2.18 g, 15.834 mmol) and piperidine (0.943 g, 11.083 mmol) were added to a stirred solution of 2-bromo-N-mesitylene butyramide (1.5 g, 5.278 mmol) in acetonitrile (30 ml) at room temperature. The resulting reaction was heated to reflux for 16 hours and the progress of the reaction mixture was monitored by TLC (50% EtOAc / hexane, UV observation). The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3x20 mL). The combined organic extracts were washed with brine solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product, which was triturated with ethyl acetate (40 mL) to give the pure product N-mesitylene-2-(piperidin-1-yl) butyramide (760 mg). MS (ESI): m / z 289.43 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δppm 9.07(s, 1H), 6.86(s, 2H), 3.06-3.01(t, 1H), 2.60-2.48(m, 4H), 2.21(s, 3H), 2.06(s, 6H), 1.80-1.25(t, 8H), 0.95(t, 3H).

[0500] Synthesis of 1-benzyl-1-(1-(mesitylamino)-1-oxobutyl-2-yl)piperidin-1-bromide:

[0501] To a stirred solution of N-mesitylene-2-(piperidin-1-yl)butyramide (0.15 g, 0.52 mmol) in acetonitrile (1 mL) was added benzyl bromide (0.177 g, 1.04 mmol). The reaction mixture was stirred at 80°C for 20 hours and the progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was triturated with ethyl acetate (20 mL) to give the desired product 1-benzyl-1-(1-(mesityleneamino)-1-oxobutyl-2-yl)piperidin-1-bromide (194 mg) as a white solid. Mass spectrum (ESI): m / z 379.3 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.31(s, 1H), 7.62-7.50(m, 5H), 6.93(s, 2H), 5.42-5.39(m, 1H), 4.63-4.20(m, 2H), 4.05-3.55(m, 2H), 3.30-3.10( m, 2H), 2.41-2.31 (m, 1H), 2.23 (s, 9H), 2.15-2.06 (m, 2H), 2.05-1.80 (m, 3H), 1.57-1.43 (m, 2H), 1.23-1.07 (m, 3H).

[0502] Synthesis of 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl) Aza-1-bromide :

[0503]

[0504] Synthesis of 2-bromo-N-(4-nitrophenyl)acetamide:

[0505] At 0°C, bromoacetyl bromide (29.226 g, 144.790 mmol) was added to a solution of 4-nitroaniline (10 g, 72.395 mmol) stirred in H2O (100 mL), and the resulting reaction mixture was stirred at room temperature for 16 hours, and the progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, UV observation). The resulting mixture was alkalized with saturated sodium carbonate (100 mL), and the resulting precipitate was filtered and dried. The resulting crude product was ground with diethyl ether (2X100 mL) to produce 2-bromo-N-(4-nitrophenyl)acetamide (8 g) as a yellow solid. Mass spectrum (ESI): m / z 259.17 [M+1]+ . 1 H NMR (400MHz, DMSO-d6) δppm 10.97 (s, 1H), 8.25 (d, 2H), 7.83 (d, 2H), 4.11 (s, 2H).

[0506] Synthesis of 2-(aza-1-yl)-N-(4-nitrophenyl)acetamide:

[0507] To a solution of 2-bromo-N-(4-nitrophenyl)acetamide (5 g, 19.300 mmol) stirred in ACN (50 ml), K2CO3 (8 g, 57.900 mmol) was added, followed by aza (3.8 g, 38.600 mmol, 2 eq), and the reaction mixture was heated to 90°C in a sealed container and maintained for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature, diluted with water (200 mL) and extracted with ethyl acetate (2×200 mL). The combined organic extracts (100 mL) were washed with brine solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product, which was triturated with diethyl ether (2×50 mL) to give pure 2-(aza-1-yl)-N-(4-nitrophenyl)acetamide (2.5 g,). Mass spectrum (ESI): m / z 278.27[M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.26 (s, 1H), 8.21 (d, 1H), 7.91 (d, 2H), 3.33-3.4 (m, 2H), 2.72-2.81 (m, 4H), 1.56-1.62 (m, 8H).

[0508] Synthesis of N-(4-aminophenyl)-2-(azepine-1-yl)acetamide:

[0509] To a stirred solution of 2-(aza-1-yl)-N-(4-nitrophenyl)acetamide (2.5 g, 9.014 mmol) in MeOH (25 mL) was added 10% Pd / C (2.5 g) at room temperature. The resulting reaction mixture was stirred at room temperature for 16 hours under hydrogen (balloon pressure). The progress of the reaction mixture was monitored by TLC (50% EtOAc / pet-ether, UV observation). The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give pure N-(4-aminophenyl)-2-(aza-1-yl)acetamide (1.5 g). Mass spectrum (ESI): m / z 248.30 [M+H]. 1H NMR (400MHz, DMSO-d6) δppm 9.20 (s, 1H), 7.22 (d, 2H), 6.49 (d, 2H), 4.85 (s, 1H), 3.16-3.19 (m, 2H), 2.68-2.72 (m, 4H), 1.57-1.62 (m, 8H).

[0510] Synthesis of 2-(azepine-1-yl)-N-(4-(butylamino)phenyl)acetamide:

[0511] To a stirred solution of N-(4-aminophenyl)-2-(azepin-1-yl)acetamide (1.8 g, 7.277 mmol) and butyronitrile (2.51 g, 36.385 mmol) in methanol (20 mL) was added 10% Pd / C (1.8 g). The reaction mixture was stirred at room temperature under hydrogen (balloon) for 16 hours. The progress of the reaction mixture was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give the desired product 2-(azepin-1-yl)-N-(4-(butylamino)phenyl)acetamide (1 g). Mass spectrum (ESI): m / z 304.43 [M+1] + .

[0512] Synthesis of tert-butyl (4-(2-(azepine-1-yl)acetamido)phenyl)(butyl)carbamate

[0513] To a stirred solution of 2-(azepine-1-yl)-N-(4-(butylamino)phenyl)acetamide (1.4 g, 4.613 mmol) in ACN (20 mL) was added triethylamine (1.39 g, 13.839 mmol) followed by di-tert-butyl bicarbonate (1.5 g, 6.919 mmol) and the reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, UV observation). After consumption of the starting material as determined by TLC, the crude compound was diluted with water (100 mL), extracted with EtOAc (2×120 mL) and the combined organic extracts (70 mL) were washed with brine solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified using column chromatography (eluting with 10%-20% ethyl acetate in petroleum ether) to give the product tert-butyl (4-(2-(azepin-1-yl)acetamido)phenyl)(butyl)carbamate (1 g,). Mass spectrum (ESI): m / z 404.58 [M+H]. 1H NMR (400MHz, DMSO-d6) δppm 9.65(s, 1H), 7.59(d, 2H), 7.13(d, 2H), 3.53(t, 2H), 3.24(s, 2H), 2.70-2.75(m, 4H), 1.61-1.86(m, 8H), 1.35-1.44(m, 9H), 1.24-1.32(m, 3H), 1.17-1.22(m, 3H).

[0514] Synthesis of 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl)azepine-1-bromide:

[0515] To a stirred solution of tert-butyl(4-(2-(azepin-1-yl)acetamido)phenyl)(butyl)carbamate (200 mg, 0.495 mmol) in ACN (2 mL) at room temperature was added benzyl bromide (127 mg, 0.742 mmol) and the reaction mixture was stirred at 90°C for 16 hours. The progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure and the crude compound was triturated with diethyl ether (2x20 mL) and EtOAc (2x20 mL) to give 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl)azepin-1-bromide (80 mg). Mass spectrum (ESI): m / z 494.56 [M] + . 1 H NMR (500MHz, DMSO-d6) δppm10.63 (s, 1H), 7.66 (d, 2H), 7.50-7.60 (m, 5H), 7.24 (d, 2H), 4.91 (s, 2H), 4 .00(s, 2H), 3.52-3.76(m, 6H), 1.90-1.98(m, 4H), 1.63-1.76(m, 4H), 1.37-1.41(m, 13H), 0.85(t, 3H).

[0516] Synthesis of 1-benzyl-1-(2-((4-(butylamino)phenyl)amino)-2-oxoethyl)azepine-1-carboxylate:

[0517]

[0518] Synthesis of 1-benzyl-1-(2-((4-(butylamino)phenyl)amino)-2-oxoethyl)azepine-1-carboxylate:

[0519] A reaction mixture of 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl)azepine-1-bromide (50 mg, 0.087 mmol) in 47% Aq.HBr (1 mL) was stirred at 80° C. for 16 hours and the progress of the reaction mixture was monitored using LCMS. The reaction mixture was concentrated under reduced pressure to give a crude product which was purified by reverse phase preparative HPLC (column: X-selective C18 (19×250) mm 5u, mobile phase: 0.1% formic acid in water: ACN, flow rate: 18 ml / min, gradient method: 0 / 51, 7.1 / 50, 7.2 / 99, 9.2 / 99, 9.3 / 5, 12 / 5). The resulting pure fractions were collected and freeze-dried to give 1-benzyl-1-(2-((4-(butylamino)phenyl)amino)-2-oxoethyl)azepine-1-carboxylate (25 mg). Mass spectrum (ESI): m / z 394.53 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.95(s, 1H), 8.56(s, 1H), 7.68(d, 2H), 7.49-7.54(m, 3H), 7.36(d, 2H), 6.54(d, 2H), 5.52(t, 1H), 4.89(s, 2H), 4.01(s, 2H ), 3.67-3.69(m, 2H), 3.51-3.53(m, 2H), 2.97(t, 2H), 1.91-2.01(m, 4H), 1.49-1.62(m, 2H), 1.35-1.40(m, 2H), 1.36(t, 3H).

[0520] Synthesis of 1-Benzyl-1-(2-((4-(butylamino)phenyl)amino)-2-oxoethyl)piperidine-1-carbonate :

[0521]

[0522] Synthesis of N-(4-nitrophenyl)-2-(piperidin-1-yl)acetamide:

[0523] To 2-bromo-N-(4-nitrophenyl)acetamide (3.0 g, 11.580 mmol) in ACN (30.0 mL) stirred solution, potassium carbonate (4.801 g, 34.74 mmol) and piperidine (1.972 g, 23.16 mmol) were added and the resulting reaction mixture was stirred at 85 ° C for 1 hour, using TLC (50% ethyl acetate in petroleum ether, ultraviolet observation) to monitor the reaction progress of the reaction mixture. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to produce a crude residue. The crude residue was diluted with water (150 mL), extracted with EtOAc (3 x 100 mL), the combined organic extracts were washed with brine solution (1 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was purified by column chromatography (eluting with 5% ethyl acetate in petroleum ether) to give the product N-(4-nitrophenyl)-2-(piperidin-1-yl)acetamide (1.40 g) as a light yellow solid. MS (ESI): m / z 264.26 [M+H + ]. 1 H NMR (400MHz, DMSO-d6) δppm 10.28 (s, 1H), 8.20-8.23 (m, 2H), 7.89-7.93 (m, 2H), 3.14 (s, 2H), 2.44-2.51 (m, 4H), 1.53-1.59 (m, 4H), 1.40 (t, 2H).

[0524] Synthesis of N-(4-aminophenyl)-2-(piperidin-1-yl)acetamide:

[0525] To a stirred solution of N-(4-nitrophenyl)-2-(piperidin-1-yl)acetamide (1.4 g, 5.3171 mmol) in MeOH (15.0 ml) was added 10% Pd / C (1.4 g) and the resulting reaction mixture was stirred under hydrogen atmosphere (balloon pressure) at room temperature for 16 hours and the progress of the reaction mixture was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was filtered through celite and washed with excess methanol (2 x 20 ml) and the combined filtrate was concentrated under reduced pressure to give the crude product which was triturated with diethyl ether (3 x 20 mL) to give N-(4-aminophenyl)-2-(piperidin-1-yl)acetamide (1.20 g). MS (ESI): m / z 234.29 [M+]. 1H NMR (500 MHz, DMSO-d6) δ ppm 10.25 (s, 1H), 8.22 (d, 2H), 7.92 (t, 2H), 3.32 (d, 2H), 2.72-2.85 (m, 4H), 1.52-1.62 (m, 8H). Analytical data are attached below.

[0526] Synthesis of N-(4-(butylamino)phenyl)-2-(piperidin-1-yl)acetamide:

[0527] To a solution of N-(4-aminophenyl)-2-(piperidin-1-yl)acetamide (1.2 g, 5.1431 mmol) in methanol (15.0 mL) under nitrogen was added butyronitrile (1.777 g, 25.7157 mmol) and 10% Pd / C (1.2 g). The resulting reaction mixture was stirred under hydrogen atmosphere (balloon pressure) at room temperature for 16 hours and the progress of the reaction mixture was monitored by TLC (ethyl acetate in petroleum ether, UV observation). The reaction mixture was filtered through celite and washed with excess methanol (2x20 ml) and concentrated to give a crude product which was purified by column chromatography (eluted with 20% ethyl acetate in petroleum ether) to give pure N-(4-(butylamino)phenyl)-2-(piperidin-1-yl)acetamide (1.10 g). MS (ESI): m / z 290.42 [M+H]. 1 H NMR (400MHz, DMSO-d6) δppm 9.21(s, 1H), 7.27(d, 2H), 6.48(d, 2H), 5.33(t, 2H), 2.88-2.95(m, 4H) , 2.49-2.51(m, 4H), 1.47-1.58(m, 6H), 1.34-1.41(m, 4H), 0.90(t, 3H).

[0528] Synthesis of tert-butylbutyl (4-(2-(piperidin-1-yl)acetamide)phenyl)carbamate:

[0529] At 0 ° C, to N- (4- (butylamino) phenyl) -2- (piperidin-1-yl) acetamide (0.956g, 3.30mmol) in DCM (10.0mL) after stirring, TEA (1.672g, 16.527mmol) and di-tert-butyl bicarbonate (1.803g, 8.2635mmol) were added, and the resulting reaction mixture was allowed to stir at room temperature for 16 hours, using TLC (100% EtOAc, UV observation) to monitor the reaction progress of the reaction mixture. The reaction mixture was concentrated under reduced pressure, diluted with water (50mL) and extracted with DCM (3x50mL). The combined organic extracts were washed with brine solution (1 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude compound which was purified by normal phase flash chromatography (eluting with 30% solution in petroleum ether). The collected pure fractions were concentrated under reduced pressure to give the product tert-butylbutyl (4-(2-(piperidin-1-yl)acetamido)phenyl)carbamate (1.15 g). MS (ESI): m / z 390.57 [M+H] + .

[0530] Synthesis of 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl)piperidin-1-bromide:

[0531] To a stirred solution of tert-butylbutyl(4-(2-(piperidin-1-yl)acetamido)phenyl)carbamate (0.225 g, 0.5776 mmol) in acetonitrile (3 mL) was added benzyl bromide (0.197 g, 1.1552 mmol) in a microwave tube. The reaction mixture was stirred at 100°C in a microwave (CEM instrument) for 1 hour and the progress of the reaction mixture was monitored using LCMS and TLC (5% methanol in DCM, detection: UV). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude compound, which was triturated with diethyl ether (5X5 ml) to give the product 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl)piperidin-1-bromide (110 mg). MS (ESI): m / z 480.58 [M]+ . .

[0532] Synthesis of 1-benzyl-1-(2-((4-(butylamino)phenyl)amino)-2-oxoethyl)piperidin-1-carbonate

[0533] A solution of 1-benzyl-1-(2-((4-((tert-butoxycarbonyl)(butyl)amino)phenyl)amino)-2-oxoethyl)piperidin-1-bromide (105 mg, 0.2184 mmol) in aqueous hydrogen bromide (48%) (1 mL) was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored using LCMS and TLC (80% ethyl acetate in petroleum ether, detection: UV). The reaction mixture was freeze-dried to give a crude product which was purified using reverse phase preparative HPLC (column: X-selective C18 (19*250) 5u, mobile phase (A): 10 mM ammonium bicarbonate, mobile phase (B): 100% ACN method: 0 / 3, 2 / 40, 20 / 40, 20.50 / 100, 30 / 35, flow rate: 18 ml / min, solubility: ACN+THF+MeOH). The pure fractions were combined and freeze-dried to give 1-benzyl-1-(2-((4-(butylamino)phenyl)amino)-2-oxoethyl)piperidine-1-carbonate (15.1 mg). MS (ESI): m / z 380.2 [M] +. 1 H NMR (400MHz, DMSO-d6) δppm 11.10 (br s, 1H), 7.54-7.72 (m, 5H), 7.34 (d, 2H), 6.52 (d, 2H), 5.44 (t, 1H), 4.94 (s, 2H), 4.09 (s, 2H), 3.65 (d, 2H), 3.33-3 .5(m, 2H), 2.97-3.01(m, 2H), 1.92-1.97(m, 4H), 1.63(q, 1H), 1.48-1.55(m, 3H), 1.34-1.42(m, 2H), 0.91(t, 3H).

[0534] Synthesis of 1-benzyl-1-(1-((4-(butylamino)-2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl) Piperidine-1-carbonate :

[0535]

[0536] Synthesis of N-(2,6-dimethylphenyl)-4-methylbenzenesulfonamide:

[0537] To a stirred solution of 2,6-dimethylaniline (50 g, 413 mmol) in pyridine (1.2 L) was added tosyl chloride (94.396 g, 495.131 mmol) and the reaction mixture was heated to reflux at 115° C. for 4 hours, and the progress of the reaction mixture was monitored using TLC (30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude residue, which was diluted with water (500 mL), the pH was adjusted to 6 with 2N HCl (500 mL), and extracted with ethyl acetate (2×1 L). The combined organic extracts were washed with brine solution (1.0 L), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product, which was triturated with petroleum ether to give N-(2,6-dimethylphenyl)-4-methylbenzenesulfonamide (90 g) as an off-white solid. MS (ESI): m / z 276.23 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δppm 9.231 (s, 1H), 7.530-7.551 (d, 2H), 7.354-7.375 (d, 2H), 6.985-7.069 (m, 3H), 2.384 (s, 3H), 1.940 (s, 6H).

[0538] Synthesis of N-(2,6-dimethyl-4-nitrophenyl)-4-methylbenzenesulfonamide:

[0539] To N-(2,6-dimethylphenyl)-4-methylbenzenesulfonamide (90g, 326.8mmol) in AcOH (675mL) and water (450mL) after stirring, NaNO2 (45.103g, 653.666mmol) was added, followed by dropwise addition of concentrated HNO3 (41.181g, 653.666mmol) at room temperature over a period of 55 minutes. The resulting reaction mixture was heated to 110°C and maintained for 5 hours, and the reaction progress of the reaction mixture was monitored by TLC (30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was diluted with ice water (500mL), alkalized with 1N NaOH solution (800mL) and extracted with ethyl acetate (2x1L). The combined organic extracts were washed with brine (1 L), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product which was triturated with petroleum ether (500 mL) to give the product N-(2,6-dimethyl-4-nitrophenyl)-4-methylbenzenesulfonamide (40 g) as an off-white solid. MS (ESI): m / z 321.18 [M+H] + . 1H NMR (400MHz, DMSO-d6) δppm 9.72 (s, 1H), 7.925 (s, 2H), 7.562-7.583 (d, 2H), 7.390-7.409 (d, 2H), 2.400 (s, 3H), 2.073 (s, 6H).

[0540] Synthesis of 2,6-dimethyl-4-nitroaniline:

[0541] To a stirred solution of N-(2,6-dimethyl-4-nitrophenyl)-4-methylbenzenesulfonamide (25 g, 78.037 mmol) in AcOH (125 ml) was added HClO4 (250 ml) at room temperature and the reaction mixture was heated to 100°C for 3 h. The progress of the reaction mixture was monitored by TLC (30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was poured into crushed ice, basified with aqueous amine solution (pH ~ 11) and extracted with ethyl acetate (2 x 1 L). The combined organic extracts were washed with brine solution (300 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product which was purified by column chromatography (eluted with 20% ethyl acetate in petroleum ether) to give 2,6-dimethyl-4-nitroaniline (10 g) as a yellow solid. MS (ESI): m / z 167.03 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.787 (s, 2H), 6.142 (s, 2H), 2.155 (s, 6H).

[0542] Synthesis of 2-bromo-N-(2,6-dimethyl-4-nitrophenyl)butyramide:

[0543] To a frozen solution (0°C) of 2,6-dimethyl-4-nitroaniline (3 g, 18.05 mmol) in DCM (45 mL) was added pyridine (1.713 g, 21.66 mmol) followed by 2-bromobutyryl chloride (4.017 g, 21.66 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was filtered and the isolated solid was washed with DCM (20 mL) and dried under high vacuum to yield 2-bromo-N-(2,6-dimethyl-4-nitrophenyl)butyramide (2 g) as an off-white solid. MS (ESI): m / z 315.26 [M] + . 1H NMR (400MHz, DMSO-d6) δppm 10.05 (s, 1H), 8.0 (s, 2H), 4.55 (t, 1H), 2.279 (s, 6H), 1.940-2.143 (m, 2H), 0.99 (t, 3H).

[0544] Synthesis of N-(2,6-dimethyl-4-nitrophenyl)-2-(piperidin-1-yl)butyramide:

[0545] To a stirred solution of 2-bromo-N-(2,6-dimethyl-4-nitrophenyl)butyramide (2 g, 6.345 mmol) in ACN (30 mL) was added K2CO3 (2.627 g, 19.03 mmol) and piperidine (1.080 g, 12.690 mmol). The resulting reaction mixture was heated to 80°C and maintained for 16 hours, and the progress of the reaction was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was concentrated, diluted with ethyl acetate (150 mL) and washed with water (50 mL x 3) and brine solution (60 mL). The combined organic extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce a crude product, which was purified by column chromatography (eluted with 50% ethyl acetate in petroleum ether) to produce N-(2,6-dimethyl-4-nitrophenyl)-2-(piperidin-1-yl)butyramide (1.70 g). MS (ESI): m / z 320.42 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δppm 9.578 (s, 1H), 7.992 (s, 2H), 3.12 (t, 1H), 2.55-2.62 (m, 4H), 2.28 (s, 6H), 1.60-1.81 (m, 2H), 1.35-1.56 (m, 6H), 0.93 (t, 3H).

[0546] Synthesis of N-(4-amino-2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide:

[0547] To a stirred solution of N-(2,6-dimethyl-4-nitrophenyl)-2-(piperidin-1-yl)butanamide (1.7 g, 5.322 mmol) in EtOH (40 mL) was added 10% Pd-C (0.900 g) and the resulting reaction mixture was stirred under hydrogen atmosphere (balloon pressure) at room temperature for 16 hours and the progress of the reaction was monitored by TLC (100% ethyl acetate, UV observation). The mixture was filtered through celite, the filter bed was washed with methanol (200 ml) and the filtrate was concentrated under reduced pressure to give the crude compound which was purified by normal phase flash chromatography (eluting with EtOAc) to give N-(4-amino-2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide (1.0 g). MS (ESI): m / z 290.46 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δppm 8.78 (s, 1H), 6.23 (s, 2H), 4.82 (s, 2H), 2.95 (t, 1H), 2.45-2.6 (m, 4H), 1.2-1.6 (m, 8H), 0.910 (t, 3H).

[0548] Synthesis of N-(4-(butylamino)-2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide:

[0549] To a stirred solution of N-(4-amino-2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide (1.0 g, 3.455 mmol) in MeOH (20 mL) was added butyronitrile (2.387 g, 34.551 mmol) and 10% Pd / C (1 g) and the resulting reaction mixture was stirred under hydrogen atmosphere (balloon pressure) at room temperature for 16 hours and the progress of the reaction was monitored by TLC (ethyl acetate, UV observation). The mixture was filtered through a celite filter bed, the filter bed was washed with methanol (150 ml) and the filtrate was concentrated under reduced pressure to give the crude compound which was purified by normal phase flash chromatography (eluting with ethyl acetate) to give N-(4-(butylamino)-2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide (0.500 g). MS (ESI): m / z 346.51 [M+H] + . 1H NMR (500MHz, DMSO-d6) δppm 8.789(s, 1H), 6.242(s, 2H), 2.96(t, 3H), 2.50-2.57(m, 3H), 2.027(s, 6H), 1.49 8-1.518(m, 2H), 1.475-1.488(m, 6H), 1.22-1.387(m, 6H), 0.885-0.918(m, 6H).

[0550] Synthesis of tert-butylbutyl(3,5-dimethyl-4-(2-(piperidin-1-yl)butylamino)phenyl)carbamate

[0551] To a stirred solution of N-(4-(butylamino)-2,6-dimethylphenyl)-2-(piperidin-1-yl)butanamide (0.400 g, 1.157 mmol) in EtOH (10 mL) at 0°C, DIPEA (0.179 g, 1.389 mmol) and di-tert-butyl bicarbonate (0.758 g, 3.473 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was concentrated under vacuum to give a crude product, which was diluted with ethyl acetate (80 ml) and washed with water (40 ml) and brine solution (40 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product which was purified by normal phase flash chromatography (eluting with 40% ethyl acetate in petroleum ether) to give tert-butylbutyl(3,5-dimethyl-4-(2-(piperidin-1-yl)butylamino)phenyl)carbamate (0.300 g). MS (ESI): m / z 446.66 [M+H] + . 1 H NMR data showed that the product was accompanied by aliphatic impurities. Data analysis is shown in the attachment.

[0552] Synthesis of 1-benzyl-1-(1-((4-((tert-butoxycarbonyl)(butyl)amino)-2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidin-1-bromide:

[0553] To a stirred solution of tert-butylbutyl(3,5-dimethyl-4-(2-(piperidin-1-yl)butylamino)phenyl)carbamate (0.250 g, 0.561 mmol) in ACN (5 mL) was added benzyl bromide (0.191 g, 1.116 mmol) and the resulting reaction mixture was heated at 85°C for 24 hours and the progress of the reaction was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under vacuum to give a crude product which was triturated with diethyl ether (20 ml) and ethyl acetate (10 ml) to give 1-benzyl-1-(1-((4-((tert-butoxycarbonyl)(butyl)amino)-2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidin-1-bromide (0.200 g). MS (ESI): m / z 536.61 [M] + . 1 HNMR (500MHz, DMSO-d6) δppm 10.317(s, 1H), 7.547(s, 5H), 7.002(s, 2H), 5.41(d, 1H), 4.62(d, 1H), 3.6-3.9(m, 3H), 3. 1-3.4(m, 2H), 2.4-2.5(m, 2H), 1.953-2.184(m, 12H), 1.163-1.620(m, 18H), 0.857(s, 3H).

[0554] Synthesis of 1-benzyl-1-(1-((4-(butylamino)-2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidine-1-carbonate:

[0555] 47% aqueous HBr solution (8 ml) was added to 1-benzyl-1-(1-((4-((tert-butoxycarbonyl)(butyl)amino)-2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidin-1-bromide (0.150 g, 0.243 mmol) at 0°C and the resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure and washed with diethyl ether (5 ml) to give a crude product which was purified by reverse phase preparative HPLC (column: kromasil C18 (25X150) mm 10u, mobile phase-10 mM ammonium bicarbonate in H2O:CAN, flow rate: 25 ml / min, gradient method: 0 / 40, 10 / 82, 10.1 / 99, 12 / 99, 12.1 / 40, 14 / 40). The resulting pure fractions were collected and freeze-dried to give 1-benzyl-1-(1-((4-(butylamino)-2,6-dimethylphenyl)amino)-1-oxobutyl-2-yl)piperidine-1-carbonate (0.050 g). MS (ESI): m / z 436.61 [M] + . 1 H NMR (500MHz, DMSO-d6) δppm 10.826(s, 1H), 7.54(t, 5H), 6.271(s, 2H), 5.44(t, 2H), 4.60(d, 2H), 3.708(s, 2H), 2.96(q, 2H), 2.362(s, 2H).2. 027-2.066(m, 8H), 1.838-1.99(m, 2H), 1.492-1.549(m, 6H), 1.32-1.45(m, 3H), 1.12-1.25(m, 3H), 0.922(t, 3H).

[0556] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-4-(methoxycarbonyl)piperidin Pyridine-1-bromide :

[0557]

[0558] Synthesis of methyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-4-carboxylate:

[0559] To 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.500g, 2.065mmol) and methyl piperidine-4-carboxylate (0.443g, 3.097mmol) in ACN (10ml) after stirring, K2CO3 (0.712g, 5.162mmol) was added. The resulting reaction mixture was heated to 90°C for 16 hours, and TLC (50% ethyl acetate solution in petroleum ether, ultraviolet observation) monitored the reaction progress of the reaction mixture. The reaction mixture was concentrated under reduced pressure, diluted with EtOAc (100ml) and washed with water (50ml x 3) and saline solution (50ml). The organic extract was dried over anhydrous sodium sulfate and concentrated to produce a crude product, which was purified by normal phase flash chromatography (10%-30% of ethyl acetate gradient solution in petroleum ether). The collected pure fractions were concentrated under reduced pressure to give the desired product, methyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-4-carboxylate (0.400 g). MS (ESI): m / z 305.36 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δppm 9.171(s, 1H), 7.060-7.09(m, 3H), 3.607(s, 3H), 3.089(s, 2H), 2.879-2.908(m, 2H), 2.316- 2.371(m,1H),2.198-2.261(m,2H),2.129(s,6H),1.816-1.856(m,2H).1.680-1.778(m,2H).

[0560] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-4-(methoxycarbonyl)piperidin-1-bromide:

[0561] To a stirred solution of methyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-4-carboxylate (0.200 g, 0.657 mmol) in ACN (5 ml) was added benzyl bromide (0.224 g, 1.314 mmol) and the resulting reaction mixture was heated to 90° C. for 24 hours. The progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated to give a crude product, which was triturated with ethyl acetate (30 ml) and n-pentane (20 ml) to give pure 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-4-(methoxycarbonyl)piperidine-1-bromide (0.080 g) as a white solid. MS (ESI): m / z 395.3 [M] +. 1 H NMR (400MHz, DMSO-d6) δppm 10.048(s, 1H), 7.551-7.591(m, 5H), 7.144-7.159(m, 3H), 4.947(s, 2H), 4.237(s, 2H), 3.671-3.766(m, 5H), 3.541-3.549(m, 2H), 2.732-2.788(m, 1H), 2.128-2.212(m, 10H).

[0562] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(ethoxycarbonyl)piperidin Pyridine-1-bromide :

[0563]

[0564] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylic acid ethyl ester

[0565] To a solution of 2-chloro-N-(2,6-dimethylphenyl)acetamide (5 g, 25.295 mmol) stirred in ACN (100 ml) was added KCO (8.726 g, 63.237 mmol) and ethylpiperidine-3-carboxylate (5.964 g, 37.942 mmol) at room temperature. The resulting reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored by TLC (30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was filtered to remove inorganic salts and the filtrate was concentrated under reduced pressure to produce a crude product, which was diluted with ethyl acetate (250 ml) and washed with water (80 ml x 3) and a saline solution (80 ml). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce ethyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (6.5 g). MS (ESI): m / z 319.01 [M+H] + . 1 HNMR(400MHz, CDCl3)δppm 8.751(s, 1H), 7.066-7.101(m, 3H), 4.075-4.168(m, 2H), 3.154-3.249(m, 2H), 2.927-2.949(m, 1H), 2.761-2 .817(m, 2H), 2.613-2.664(m, 1H), 2.45-2.49(m, 1H), 2.23(s, 6H), 1.506-2.045(m, 4H), 1.209-1.277(m, 3H).

[0566] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(ethoxycarbonyl)piperidin-1-bromide:

[0567] To a stirred solution of ethyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (0.300 g, 0.942 mmol) in ACN (10 ml) was added benzyl bromide (0.322 g, 1.884 mmol) and the resulting reaction mixture was heated to 90° C. in a sealed container for 16 hours and the progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was purified by normal phase flash chromatography (eluted with 2% methanol in DCM). The collected pure fractions were concentrated under reduced pressure to give the desired product as a colorless gum (LCMS-85%), which was diluted with ethyl acetate (15 ml) and stirred for 1 hour, then filtered and washed with ethyl acetate (15 ml x 3) to give pure 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(ethoxycarbonyl)piperidin-1-bromide (70 mg) as an off-white solid. MS (ESI): m / z 409.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.05(s, 1H), 7.545-7.585(m, 5H), 7.125-7.178(m, 3H), 4.945-5.060(m, 2H), 4.109-4.342(m, 4H), 3.904-3.934(m, 1H), 3.644 -3.677(m, 1H), 3.433-3.569(m, 2H), 3.230-3.262(m, 1H), 2.213(s, 6H), 1.948-2.195(m, 3H), 1.553-1.620(m, 1H), 1.24(t, 3H).

[0568] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(isopropoxycarbonyl)piperidin Pyridine-1-trifluoroacetate

[0569]

[0570] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylic acid:

[0571] A stirred solution of ethyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (5 g, 15.702 mmol) in concentrated hydrochloric acid (100 ml) was heated to 100° C. for 16 hours and the progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was concentrated under reduced pressure to give 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylic acid (4.2 g). MS (ESI): m / z 291.13 [M+H] + .

[0572] Synthesis of isopropyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate:

[0573] To a stirred solution of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylic acid (0.5 g, 1.722 mmol) in isopropanol (10 mL) at 0°C was added thionyl chloride (5 mL) dropwise. The resulting reaction mixture was stirred at 90°C for 16 hours and the progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, UV observation). The resulting reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product which was quenched with ice-cold saturated aqueous sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (2X 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude compound which was purified by normal phase flash chromatography (eluting with 20%-30% of EtOAc / petroleum ether). The collected pure fractions were concentrated under reduced pressure to give 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (0.54 g) as a pale yellow solid. MS (ESI): m / z 332.27 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.5 (br s, 1H), 7.06-7.11 (m, 3H), 4.97-5.03 (m, 1H), 3.19-3.24 (m, 2H), 2.93-2.95 (m, 1H), 2.59- 2.48(m, 2H), 2.48-2.40(m, 2H), 2.18-2.20(m, 5H), 1.41-1.65(m, 3H), 1.18-1.24(m, 6H).

[0574] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(isopropoxycarbonyl)piperidine-1-trifluoroacetate:

[0575] To a solution of isopropyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (0.4 g, 1.203 mmol) stirred in ACN (10 mL) was added benzyl bromide (0.6173 g, 3.609 mmol) at room temperature. The resulting reaction mixture was stirred at 90° C. in a sealed test tube for 16 hours, and the progress of the reaction mixture was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give 430 mg (LCMS 51%) of crude compound which was further purified by reverse phase preparative HPLC (column: X-selective csh C18 (250*19) mm, 5u; mobile phase A: 0.1% TFA (Aq.) in water, mobile phase B: acetonitrile, flow rate: 22 ml / min, method (T / % of B): 0 / 10, 2 / 10, 10 / 50, 13 / 50, 13.1 / 100, 16 / 100, 16.1 / 10, 19 / 20, solubility: ACN+H2O+THF, temperature: room temperature). The combined pure fractions were freeze dried to give pure 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(isopropoxycarbonyl)piperidine-1-trifluoroacetate (168 mg). LCMS: 99.37% (77.95% + 21.42%, isomer mixture), MS (ESI): m / z 423.2, [M] + . 1 HNMR (400MHz, DMSO-d6) δppm 10.01-10.05(m, 1H), 7.57-7.61(m, 5H), 7.12-7.17(m, 3H), 4.92-5.06(m, 3H), 4.14-4.22(m, 2H), 3.87- 3.67(dd, 1H), 3.46-3.55(m, 3H), 3.15-3.21(m, 1H), 2.208(d, 6H), 1.85-2.07(m, 3H), 1.19-1.23(m, 6H).

[0576] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(propyloxycarbonyl)piperidin- 1-Bromide

[0577]

[0578] Synthesis of propylpiperidinium-3-carboxylate:

[0579] To a stirred solution of piperidine-3-carboxylic acid (2.0 g, 15.484 mmol) in propan-1-ol (20.0 ml) was added SOCl2 (11.05 g, 92.908 mmol) at 0-5°C. The resulting reaction mixture was stirred at 80°C for 16 hours and the progress of the reaction mixture was monitored using TLC (30% ethyl acetate / petroleum ether, observation: ninhydrin). The reaction mixture was concentrated under reduced pressure to give a crude compound which was dissolved in ethyl acetate (100 ml) and washed with saturated aqueous sodium bicarbonate solution (1 x 100 ml) and brine solution (1 x 50 ml). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give propylpiperidine-3-carboxylate (2.0 g) as a colorless liquid. The crude product was directly carried forward to the next step without further purification and analysis.

[0580] Synthesis of propyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate:

[0581] Under room temperature, add K2CO3 (4.14g, 29.95mmol) and propyl piperidine-3-carboxylate (3.47g, 20.28mmol) to the solution after stirring in ACN (20ml) of 2-chloro-N-(2,6-dimethylphenyl) acetamide (2.0g, 10.15mmol). Stir the reaction mixture at 80 ℃ for 16 hours. Use TLC (50% ethyl acetate / petroleum ether, ultraviolet observation) to monitor the reaction process of the reaction mixture. After completing the reaction on TLC, concentrate the reaction mixture under reduced pressure to produce crude compound, dilute the crude compound with water (30ml) and extract with ethyl acetate (2X100 ml). Wash the combined organic extract with saline (50.0ml), dry with anhydrous sodium sulfate and concentrate under reduced pressure to produce crude compound, which is purified by column chromatography (eluted with 20%-80% ethyl acetate / petroleum ether). The collected pure fractions were concentrated under reduced pressure to give propyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (1.0 g) as a colloidal compound. MS (ESI): m / z 333.10 [M+H] + .

[0582] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(propyloxycarbonyl)piperidin-1-bromide:

[0583] To a solution of propyl 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxylate (0.3 g, 0.9030 mmol) in acetonitrile (10 ml) was added benzyl bromide (0.3113 g, 1.806 mmol) at room temperature. The resulting reaction mixture was stirred at 90°C for 16 hours and the progress of the reaction mixture was monitored using TLC (10% MeOH / DCM, UV observation). After complete consumption of the starting material as observed by TLC, the reaction mixture was directly concentrated under reduced pressure to give the crude compound as a pale yellow semi-solid, which was purified by normal phase flash chromatography (eluted with 0%-30% MeOH / DCM). The collected pure fractions were concentrated under reduced pressure to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(propyloxycarbonyl)piperidine-1-bromide (70 mg) as an off-white solid. MS (ESI): m / z 423.3 [M] + . 1 HNMR (400MHz, DMSO-d6) δppm 10.30 (s, 1H), 7.55-7.60 (m, 5H), 7.11-7.18 (m, 3H), 4.97-5.15 (m, 2H), 4.33 (dd, 2H) 4.05 (t, 2H), 3.88 (d, 1H), 3.66(d, 1H), 3.31-3.54(m, 3H), 2.09-2.21(m, 8H), 1.96(d, 2H), 1.55-1.64(m, 3H), 0.87-0.93(m, 3H).

[0584] The following examples were synthesized from piperidine-3-carboxylic acid, 2-chloro-N-(2,6-dimethylphenyl)acetamide and benzyl bromide according to the synthetic procedure described for the synthesis of compound 47A.

[0585]

[0586] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(propionyloxy)piperidin- 1-Bromide

[0587]

[0588] Synthesis of tert-butyl 3-(propionyloxy)piperidine-1-carboxylate:

[0589] To a solution of tert-butyl 3-hydroxypiperidine-1-carboxylate (2 g, 9.936 mmol) and pyridine (2.357 g, 29.808 mmol) stirred in DCM (30 ml) was added propionyl chloride (1.103 g, 11.923 mmol) at 0°C, and the resulting reaction mixture was stirred at room temperature for 24 hours, and the progress of the reaction mixture was monitored by TLC (20% ethyl acetate in petroleum ether, UV observation). The reactant was diluted with DCM (120 mL) and washed twice with water (50 ml), dried over sodium sulfate and concentrated under reduced pressure to produce tert-butyl 3-(propionyloxy)piperidine-1-carboxylate (1.34 g) as an off-white solid. 1 HNMR (400MHz, CDCl3) δppm 4.85-4.75(m, 1H), 3.51-3.43(m, 3H), 3.3-3.29(m, 1H), 2.41-2.39(m, 2H), 1.85-1.72(m, 3H), 1.45(s, 9H), 1.18-1.12(m, 4H).

[0590] ●Synthesis of piperidin-3-yl propionate:

[0591] To a solution of tert-butyl 3-(propionyloxy)piperidine-1-carboxylate (1.3 g, 5.05 mmol) stirred in DCM (20 ml) was added TFA (5 ml) at 0°C, and the resulting reaction mixture was stirred at room temperature for 16 hours, and the progress of the reaction was monitored using TLC (50% ethyl acetate in petroleum ether, observation: ninhydrin). The reaction mixture was concentrated under reduced pressure to give a crude compound, which was distilled with toluene (2 x 15 mL) to give piperidin-3-yl propionate TFA salt (1.34 g) as a bright yellow oil. 1 H NMR (400MHz, CDCl3) δppm 5.16(s, 1H), 3.41(d, 2H), 3.24(t, 1H), 3.10(d, 1H), 2.42-2.36(m, 2H), 2.13-2.10(m, 2H), 2.02-2.01(m, 2H), 1.88-1.84(m, 3H).

[0592] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-3-yl propionate:

[0593] At room temperature, DIPEA (0.5884g, 4.551mmol) and piperidin-3-yl propionate TFA salt (0.4937g, 1.8204mmol) were added to the solution of 2-chloro-N-(2,6-dimethylphenyl) acetamide (0.3g, 1.517mmol) in acetonitrile (15mL) after stirring. The obtained reaction mixture was stirred in a sealed test tube at 80°C for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (50% ethyl acetate solution in petroleum ether UV observation). The reaction mixture was concentrated under reduced pressure to produce a residue, which was diluted with water (50ml) and extracted with ethyl acetate (2x25ml). The combined organic extract was dried with sodium sulfate and concentrated under reduced pressure to produce a crude compound, which was purified by normal phase flash chromatography (eluted with 25%-30% EtOAc / petroleum ether). The collected pure fractions were concentrated under reduced pressure to give 2-(aza-1-yl)-N-(3-methyl-[1,1'-diphenyl]-2-yl)acetamide (310 mg). Mass spectrum (ESI): 319.26 m / z, [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.67(s, 1H), 7.12-7.06(m, 3H), 4.95-4.92(m, 1H), 3.20(s, 2H), 2.95-2.89(m, 1H), 2.73-2.56( m, 1H), 2.59-2.54 (m, 2H), 2.31-2.23 (m, 8H), 1.88-1.84 (m, 2H), 1.68-1.56 (m, 3H), 1.09 (t, 3H).

[0594] ●Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(propionyloxy)piperidin-1-bromide

[0595] To a solution of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-3-yl propanoate (0.2 g, 0.6281 mmol) stirred in acetonitrile (15 ml) at room temperature was added benzyl bromide (0.4297 g, 2.5124 mmol) in a sealed container, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product, which was triturated with 1:1 EtOAc:Et2O (30:30 ml) to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(propionyloxy)piperidin-1-bromide (90 mg) (mixture of isomers, peak-1: 35.75% + peak-2: 63.34%). Mass spectrum (ESI): m / z 409.03[M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.05(d, 1H), 7.60-7.56(m, 5H), 7.15-7.13(m, 3H), 5.45-5.25(m, 1H), 5.05-4.97(m, 1H), 4.35-4.18(m, 2H), 4. 29(d, 2H), 3.87-3.52(m, 4H), 2.44-2.35(m, 2H), 2.21(m, 6H), 2.20-1.98(m, 3H), 1.80-1.6(m, 1H), 1.04(t, 3H).

[0596] Synthesis of 1-benzyl-3-carbamoyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-1- Bromide :

[0597]

[0598] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxamide

[0599] To a stirred solution of 2-chloro-N-(2,6-dimethylphenyl)acetamide (2.0 g, 10.1183 mmol) in ACN (20.0 ml) was added potassium carbonate (4.195 g, 30.3551 mmol) at room temperature and the resulting reaction mixture was stirred at room temperature for 10 minutes. The resulting reaction mixture was treated with piperidine-3-carboxamide (2.6081 g, 20.2366 mmol) at room temperature and then heated at 90° C. while stirring for 16 hours, using TLC (10% methanol in DCM. UV observation) to monitor the progress of the reactants. The reaction mixture was cooled to room temperature, diluted with 70 ml of ethyl acetate and washed with 2x50 ml of water, followed by 1x30 ml of brine. The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to produce 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxamide (2.5 g) as an off-white solid. MS (ESI): m / z 290.25 [M+H] + . 1 H NMR (400MHz, chloroFORM-d) δppm 8.57 (s, 1H), 7.06-7.09 (m, 3H), 5.95 (br s, 1H), 5.45 (br s, 1H), 3.15-3.30 (m, 2H), 2.70-2.90 (m, 3H), 2.45-2.60 (m, 2H), 2.15-2.30 (m, 6), 1.81-1.86 (m, 2H), 1.60-1.72 (m, 2H).

[0600] Synthesis of 1-benzyl-3-carbamoyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidin-1-bromide:

[0601] To a stirred solution of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-3-carboxamide (0.250 g, 0.865 mmol) in ACN (3.0 ml) was added benzyl bromide (0.295 g, 1.730 mmol) and the resulting reaction mixture was heated at 90°C for 16 hours. The progress of the reaction mixture was monitored using TLC (10% methanol in DCM, UV observation). The reaction mixture was directly concentrated under reduced pressure to give a crude compound, which was triturated with 3x10 ml of ethyl acetate to give 1-benzyl-3-carbamoyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)piperidine-1-bromide (130 mg) as an off-white solid (33.18% + 65.55%, isomer mixture). MS (ESI): m / z 380.2 [M] + . 1H NMR(400MH-DMSO-d6)δppm 9.92-10.15(m, 1H), 7.45-7.60(m, 6H), 7.16-7.22(m, 4H), 4.94-5.06(m, 2H), 4.05-4.29(m, 2 H), 3.41-3.83(m, 4H), 2.92(t, 1H), 2.19-2.21(m, 6H), 1.98-2.08(m, 3H), 1.50-1.54(m, 1H).

[0602] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(ethylcarbamoyl)piperidin Pyridine-1-bromide

[0603]

[0604]

[0605] Synthesis of 3-(ethylcarbamoyl)piperidine-1-carboxylate

[0606] To a solution of 1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (2 g, 8.722 mmol) stirred in THF (20 mL), Et3N (2.64 g, 26.166 mmol) and ethylamine hydrochloride (1.42 g, 17.444 mmol) were added, followed by HATU (4.97 g, 13.083 mmol), and the resulting reaction mixture was stirred at room temperature for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (50% EtOAc-petroleum ether, observation: ninhydrin). After the reaction was completed, the reaction mixture was concentrated under reduced pressure, quenched with 2N HCl and extracted with ethyl acetate (2X200 mL). The combined organic phases were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude compound (2.2 g), which was purified by column chromatography (eluted with 40% ethyl acetate in petroleum ether) to give pure tert-butyl 3-(ethylcarbamoyl)piperidine-1-carboxylate (2.1 g) as a light brown liquid. 1 H NMR (400MHz, DMSO-d6) δppm 6.12 (br s, 1H), 3.83 (br s, 2H), 2.80 (S, 5H), 2.26-2.28 (m, 1H), 2.04-1.05 (m, 2H), 1.84-1.85 (m, 1H), 1.46-1.48 (m, 11H), 1.13 (t, 3H).

[0607] Synthesis of N-ethylpiperidine-3-carboxamide

[0608] To a stirred solution of tert-butyl 3-(ethylcarbamoyl)piperidine-1-carboxylate (2.5 g, 9.752 mmol) in DCM (20 mL) was added TFA (10 mL) at 0°C. The resulting reaction mixture was stirred at room temperature for 3 hours. The progress of the reaction mixture was monitored by TLC (10% MeOH-DCM, observation: ninhydrin). The reaction mixture was concentrated under reduced pressure to give pure N-ethylpiperidine-3-carboxamide TFA salt (2.4 g) as a light brown liquid. LCMS purity: 99.89%, mass spectrum (ESI): m / z 157.09 [M+H] + .

[0609] Synthesis of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-N-ethylpiperidine-3-carboxamide

[0610] To a stirred solution of N-ethylpiperidine-3-carboxamide TFA salt (1 g, 3.96 mmol) in ACN (10 mL) was added DIPEA (1.53 g, 11.89 mmol), followed by 2-bromo-N-(2,6-dimethylphenyl)acetamide (960 mg, 3.96 mmol), and the resulting reaction mixture was stirred in a sealed test tube at 90° C. for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (10% MeOH-DCM, UV observation). The reaction mixture was concentrated under reduced pressure to produce a crude compound (1.5 g), which was further purified by column chromatography (eluted with 2% methanol in DCM) to produce the product 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-N-ethylpiperidine-3-carboxamide (1 g,) as a semisolid. LCMS purity: 39%, mass spectrum (ESI): m / z 318.12 [M+H] + The product was taken to the next step without further purification.

[0611] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(ethylcarbamoyl)piperidin-1-bromide

[0612] To a stirred solution of 1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-N-ethylpiperidine-3-carboxamide (0.3 g, 0.945 mmol) in acetonitrile (2 ml) was added benzyl bromide (0.808 g, 4.724 mmol) and the resulting reaction mixture was stirred at 90° C. for 16 hours. The progress of the reaction mixture was monitored using TLC (10% methanol in DCM. UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product which was purified by normal phase flash chromatography (eluting with 10%-15% methanol in DCM) to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-3-(ethylcarbamoyl)piperidine-1-bromide (100 mg) as a white solid. MS (ESI): m / z 408.19 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.15(s, 1H), 7.98(t, 1H), 7.60-7.53(m, 5H), 7.18-7.12(m, 3H), 5.01-5.00(m, 2H), 4.22(s, 2H), 3.73-3.66(m, 2H), 3 .51-3.42(m, 2H), 3.13-3.04(m, 2H), 2.9-2.78(m, 1H), 2.21(s, 6H), 2.02-1.98(m, 3H), 1.54-1.51(m, 1H), 0.99(t, 3H).

[0613] The following examples were synthesized from 1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid, 2-bromo-N-(2,6-dimethylphenyl)acetamide and benzyl bromide as described for the synthesis of compound 54A. The products were purified by normal phase flash chromatography or reverse phase preparative HPLC.

[0614]

[0615]

[0616]

[0617] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-4-(propyloxycarbonyl)-1,4- Diaza-1-bromide

[0618]

[0619] Synthesis of 1-(tert-butyl)4-propyl 1,4-diaza-1,4-dicarboxylate

[0620] At 0°C, carbonyl diimidazole (1.21 g, 7.488 mmol, 3.0 eq) and TEA (0.758 g, 7.488 mmol) were added to a solution of tert-butyl 1,4-diaza-1-carboxylate (0.5 g, 2.496 mmol) stirred in THF (5 ml), followed by 1-propanol (0.449 g, 7.488 mmol). The resulting reaction mixture was stirred at 80°C for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (30% EtOAc-hexane, observation: PMA,). The reaction mixture was concentrated under reduced pressure to produce a crude product, which was diluted with ethyl acetate (150 ml) and washed with water (3 x 30 ml). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce 1-(tert-butyl) 4-propyl 1,4-diaza-1,4-dicarboxylate (0.655 g). MS (ESI): m / z 287.19 [M+H] + .

[0621] ·Synthesis of propyl 1,4-diaza-1-carboxylate

[0622] 1-(tert-butyl)-4-propyl-1,4-diaza-1,4-dicarboxylate (0.65 g, 2.269 mmol, 1.0 eq) was added to 4M dioxane hydrochloride (10 ml) at 0°C and the resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (10% methanol in DCM, observation: PMA). The reaction mixture was concentrated under reduced pressure to give propyl-1,4-diaza-1-carboxylate hydrochloride (0.41 g) as a light yellow gummy solid. MS (ESI): m / z 187.12 [M+H] + .

[0623] ●Synthesis of propyl 4-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1,4-diaza-1-carboxylate

[0624] To a stirred solution of 2-chloro-N-(2,6-dimethylphenyl)acetamide (0.4 g, 2.023 mmol) in ACN (10 ml) at 0°C, DIPEA (0.522 g, 4.046 mmol) and propyl 1,4-diaza-1-carboxylic acid hydrochloride (0.376 g, 2.023 mmol) were added. The resulting reaction mixture was stirred at 90°C for 16 hours and the progress of the reaction mixture was monitored by TLC (50% EtOAc-hexane, UV observation). The reaction mixture was concentrated under reduced pressure to give a residue, which was diluted with EtOAc (100 ml) and washed with water (60 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude product was purified by normal phase flash chromatography (eluting with 10%-50% ethyl acetate in petroleum ether) to give propyl 4-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1,4-diaza-1-carboxylate (0.25 g) as a white solid. MS (ESI): m / z 348.69 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δppm 9.19(s, 1H), 7.00(s, 3H), 3.96-3.91(m, 2H), 3.50-3.43(m, 4H), 3.25(s, 2H), 2.80- 2.65(m, 4H), 2.13(s, 6H), 1.84-1.81(m, 2H), 1.60-1.53(m, 2H), 0.90-0.85(m, 3H).

[0625] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-4-(propyloxycarbonyl)-1,4-diazepine-1-bromide

[0626] To a stirred solution of propyl 4-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-1,4-diazepine-1-carboxylate (0.25 g, 0.719 mmol) in ACN (5 ml) was added benzyl bromide (0.419 g, 2.876 mmol) at room temperature. The reaction mixture was stirred at 90°C for 16 hours. The progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product, which was triturated with ethyl acetate (40 ml) to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-4-(propyloxycarbonyl)-1,4-diazepine-1-bromide (122.1 mg) as a white solid. Mass spectrum (ESI): m / z 438.2 [M] + . 1H NMR (400MHz, DMSO-d6) δppm 9.99(s, 1H), 7.61-7.53(m, 5H), 7.18-7.11(m, 3H), 5.00(s, 2H), 4.29-4.16(m, 2H), 4.0 1-3.56(m, 10H), 2.32-2.30(m, 2H), 2.21(s, 6H), 1.62-1.57(m, 2H), 0.92-0.88(m, 3H).

[0627] Synthesis of 1-Benzyl-1-(2-((2-fluoro-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide

[0628]

[0629] Synthesis of 2-bromo-N-(2-fluoro-6-methylphenyl)acetamide

[0630] A solution of 2-fluoro-6-methylaniline (3.0 g, 23.980 mmol) in water (30.0 ml) was cooled to 0°C and bromoacetyl bromide (29 g, 143.884 mmol) was added. The resulting reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored using TLC (50% ethyl acetate in petroleum ether, UV observation). The resulting reaction mixture was basified with NaCO3, stirred for 20 minutes, filtered, washed with water (100 ml) and then with petroleum ether (50 mL) to produce the crude compound 2-bromo-N-(2-fluoro-6-methylphenyl)acetamide (2.8 g) as an off-white solid. MS (ESI): m / z 246.08 [M] + . 1 HNMR (400MHz, DMSO-d6) δppm 9.95 (s, 1H), 7.18 (d, 3H), 4.15 (s, 2H), 2.20 (s, 3H).

[0631] Synthesis of 2-(aza-1-yl)-N-(2-fluoro-6-methylphenyl)acetamide

[0632] At room temperature, potassium carbonate (2.243 g, 48.762 mmol) was added to a solution of 2-bromo-N-(2-fluoro-6-methylphenyl)acetamide (4.0 g, 16.254 mmol) in ACN (40 ml), and the reaction mixture was stirred for 10 minutes. Subsequently, azepine (3.225 g, 32.509 mmol) was added and the reaction mixture was stirred for 16 hours at room temperature. The reaction mixture was monitored by TLC (50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was diluted with ethyl acetate (100 ml), washed with water (2x50 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude compound was ground with n-pentane (3x30 ml) to produce 2-(azepine-1-yl)-N-(2-fluoro-6-methylphenyl)acetamide (2.2 g) as an off-white solid. MS (ESI): m / z 264.34 [M] + . 1 H NMR (400 MHz, chloro FORM-d) δ ppm 8.88 (s, 1H), 7.26 (s, 1H), 7.10-7.16 (m, 1H), 6.93-7.03 (m, 1H), 3.31 (s, 2H), 2.82 (t, 4H), 2.27 (s, 3H), 1.58-1.73 (m, 8H).

[0633] Synthesis of 1-Benzyl-1-(2-((2-fluoro-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide

[0634] To a stirred solution of 2-(azepine-1-yl)-N-(2-fluoro-6-methylphenyl)acetamide (0.50 g, 1.89 mmol) in ACN (5.0 mL) was added benzyl bromide (1.9411 g, 11.35 mmol) at room temperature, and the resulting reaction mixture was stirred at 80°C for 16 hours, and the progress of the reaction mixture was monitored using TLC (10% methanol in DCM, UV observation). The resulting reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude compound, which was triturated with 1:1 EtOAc / petroleum ether (3x10 mL) to give the product 1-benzyl-1-(2-((2-fluoro-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide (200 mg). MS (ESI): m / z 355.2 [M] + . 1H NMR (400MHz, DMSO-d6) δppm 10.19(s, 1H), 7.64(t, 2H), 7.51-7.57(m, 3H), 7.27-7.32(m, 1H), 7.17(t, 2H), 4.88(s, 2 H), 4.11(s, 2H), 3.78(q, 2H), 3.51-3.56(m, 2H), 2.27(s, 3H), 1.96(s, 4H), 1.66(s, 4H).

[0635] The following examples were synthesized from bromoacetyl bromide, benzyl bromide, aza and the appropriate aniline according to the procedure used in the synthesis of compound 69A.

[0636]

[0637]

[0638]

[0639] Synthesis of 1-benzyl-1-(2-((2-hydroxy-6-methylphenyl)amino)-2-oxoethyl)

[0640] Aza-1-carboxylate

[0641]

[0642] ·1-Benzyl-1-(2-((2-hydroxy-6-methylphenyl)amino)-2-oxoethyl)aza-1-

[0643] To a stirred solution of 1-benzyl-1-(2-((2-methoxy-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide (1 g, 2.235 mmol) in DCM (25 ml) at 0°C was added BBr3 (2.239 g, 8.94 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored using TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give the crude compound which was purified by normal phase flash chromatography (eluting with a gradient of 6% methanol in DCM). The pure fractions were concentrated under reduced pressure to give 1-benzyl-1-(2-((2-hydroxy-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide (0.9 g) as an off-white solid. MS (ESI): m / z 353.21 [M] + . 1H NMR (300MHz, DMSO-d6) δppm 9.82(s, 1H), 9.59(s, 1H), 7.73-7.71(m, 2H), 7.59-7.49(m, 3H), 7.07-7.02(t, 1H), 6.77-6.70(m, 2H), 4.88( s, 2H), 4.06 (s, 2H), 3.77-3.72 (m, 2H), 3.58-3.53 (m, 2H), 2.17 (s, 3H), 2.05-1.9 (m, 4H), 1.66-1.45 (m, 4H).

[0644] Synthesis of 1-benzyl-1-(2-((2-methyl-6-(propionyloxy)phenyl)amino)-2-oxoethyl)azepine-1-bromo Chemical

[0645]

[0646] Synthesis of 1-Benzyl-1-(2-((2-methyl-6-(propionyloxy)phenyl)amino)-2-oxoethyl)azepine-1-bromide

[0647] Propionic anhydride (5 ml) was added to 1-benzyl-1-(2-((2-hydroxy-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide (0.4 g, 0.922 mmol) at room temperature and the resulting reaction mixture was stirred at 80°C for 16 hours. The progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give the crude compound which was purified by normal phase flash chromatography (eluting with 15% methanol in DCM). Pure fractions were concentrated under reduced pressure to give the product which was triturated with diethyl ether (45 ml) to give 1-benzyl-1-(2-((2-methyl-6-(propionyloxy)phenyl)amino)-2-oxoethyl)azepine-1-bromide (95.6 mg) as an off-white solid. MS (ESI): m / z 409.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 10.16(s, 1H), 7.66-7.64(m, 2H), 7.57-7.51(m, 3H), 7.31-7.27(t, 1H), 7.22-7.21(d, 1H), 7.09-7.07(m, 1H), 4.89(s, 2H), 4. 08(s, 2H), 3.75-3.71(m, 2H), 3.51-3.47(m, 2H), 2.58(q, 2H), 2.27(s, 3H), 2.05-1.85(m, 4H), 1.75-1.55(m, 4H), 1.12(t, 3H).

[0648] Synthesis of 1-Benzyl-1-(2-((2-carbamoyl-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide thing

[0649]

[0650] Synthesis of N, N, 3-trimethyl-2-nitrobenzamide:

[0651] To a stirred solution of 3-methyl-2-nitrifying benzoic acid (3.0 g, 16.56 mmol) in DMF (30 ml) was added DIPEA (6.42 g, 49.68 mmol) and HATU (9.44 g, 24.84 mmol), followed by dimethylamine (2M in THF) (16.56 ml, 33.12 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction mixture was monitored using TLC (30% ethyl acetate / petroleum ether, UV observation). The reaction mixture was poured into ice water and extracted with ethyl acetate (2 x 50 ml). The combined extracts were washed with brine (30 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce a crude product, which was purified by normal phase flash chromatography (using 20%-80% ethyl acetate in petroleum ether) to produce N, N, 3-trimethyl-2-nitrifying benzamide (2.2 g) as a black liquid. Mass spectrum (ESI): m / z 209.09 [M+H] + . 1 HNMR (400MHz, CDCl3) δppm 7.47-7.43 (m, 1H), 7.36-7.33 (m, 1H), 7.22-7.19 (m, 1H), 3.09 (s, 3H), 2.93 (s, 3H), 2.80 (s, 6H), 2.33 (s, 3H).

[0652] ●Synthesis of 2-amino-N, N, 3-trimethylbenzamide:

[0653] At room temperature, to N, N, 3-trimethyl-2-nitrifying benzamide (2g, 9.605mmol) in ethanol (20ml) and H2O (20ml) in the solution after stirring, add Fe (3.75g, 67.235mmol) and NH4Cl (3.59g, 67.235mmol). Stir the obtained reaction mixture under 80 ℃ of conditions for 8 hours, use TLC (10% ethyl acetate solution in petroleum ether, ultraviolet observation) to monitor the reaction process of the reaction mixture. Filter the reaction mixture through diatomaceous earth pad and wash twice with EtOH (2x50ml). Concentrate the filtrate under reduced pressure to produce residue, dilute the residue with water (50ml) and extract (2x50ml) with ethyl acetate. The combined organic extracts were washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product which was purified by column chromatography (eluting with 10%-40% ethyl acetate in Pet) to give 2-amino-N,N,3-trimethylbenzamide (600 mg). LCMS purity: 98.02%, mass spectrum (ESI): m / z 179.12 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.26 (s, 1H), 7.06 (s, 1H), 6.97 (d, 1H), 6.66-6.63 (m, 1H), 4.33 (bs, 2H), 3.05 (s, 6H), 2.17 (s, 3H).

[0654] Synthesis of 2-(2-bromoacetamide)-N,N,3-trimethylbenzamide:

[0655] To a stirred solution of isopropyl 2-amino-N,N,3-trimethylbenzamide (0.6 g, 3.366 mmol) in H2O (6 ml) at 0°C was added 2-bromoacetyl bromide (5.34 g, 26.928 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). The reaction mixture was basified with saturated Na2CO3 at 0°C to produce a solid precipitate, which was filtered, washed with water and dried to produce (2-(2-bromoacetamide)-N,N,3-trimethylbenzamide (650 mg) as a white solid. LCMS purity: 93.98%, mass spectrum (ESI): m / z 301.12 [M+2] + . 1 H NMR (400MHz, CDCl3) δppm 8.84 (s, 1H), 7.26-7.07 (m, 3H), 3.93 (s, 2H) 3.09 (s, 3H), 2.93 (s, 3H), 2.21 (s, 3H).

[0656] ●Synthesis of 2-(2-(aza-1-yl)acetamide)-N,N,3-trimethylbenzamide

[0657] To a solution of 2-(2-bromoacetamide)-N,N,3-trimethylbenzamide (0.6 g, 2.005 mmol) in acetonitrile (6 ml) was added K2CO3 (0.692 g, 5.012 mmol) and aza (0.397 g, 4.01 mmol) at room temperature. The resulting reaction mixture was stirred at 80°C for 16 hours and the progress of the reaction mixture was monitored by TLC (10% methanol in DCM, UV observation). After TLC confirmed the completion of the reaction, the reaction mixture was concentrated under reduced pressure to produce a residue, which was poured into ice water and extracted with ethyl acetate (2x25 ml). The combined organic extracts were washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce (2-(2-(azepine-1-yl)acetamide)-N,N,3-trimethylbenzamide (600 mg). MS (ESI): m / z 318.27 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 9.19 (s, 1H) 7.27-7.25 (m, 1H), 7.20-7.17 (m, 1H), 7.09-7.07 (m, 1H), 3.24 (s, 2 H)3.04(s,3H),2.91(s,3H),2.79-2.76(t,4H),2.26(s,3H)1.71-1.62(m,8H).

[0658] Synthesis of 1-benzyl-1-(2-((2-(dimethylcarbamoyl)-6-methylphenyl)amino)-2-oxoethyl)azepine-1-bromide

[0659] To a stirred solution of 2-(2-(azepin-1-yl)acetamide)-N,N,3-trimethylbenzamide (0.5 g, 1.575 mmol) in acetonitrile (5 ml) was added benzyl bromide (0.538 g, 3.15 mmol) at room temperature. The resulting reaction mixture was stirred at 80°C for 48 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: -10% methanol in DCM, Rf: 0.47, UV observation). After the consumption of the starting material was complete as observed on TLC, the reaction mixture was concentrated under reduced pressure to give a crude product which was purified by column chromatography (eluted with 4%-8% of methanol in DCM) to give (1-benzyl-1-(2-((2-(dimethylcarbamoyl)-6-methylphenyl)amino)-2-oxoethyl)azepin-1-bromide (100 mg) as a brown solid. Mass spectrum (ESI): m / z 408.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm10.31 (s, 1H), 7.70-7.68 (m, 2H), 7.56-7.51 (m, 3H), 7.38-7.36 (m, 1H), 7.32-7.28 (m, 1H), 7.19-7.16 (m, 1H), 4. 85(s, 2H), 4.03(s, 2H), 3.70-3.66(m, 2H), 3.47-3.43(m, 2H), 2.95(s , 3H), 2.87(s, 3H), 2.27(s, 3H), 1.94-1.90(s, 4H), 1.70-1.65(s, 4H).

[0660] Synthesis of 1-benzyl-1-(2-((2,6-dimethylbenzyl)oxy)ethyl)azepine-1-bromide

[0661]

[0662] ●Synthetic intermediate 2-bromoethyl 2,6-dimethylbenzoate:

[0663] 2-Bromoethyl-1-ol (4.16 g, 33.29 mmol) and a catalytic amount of concentrated H2SO4 (0.2 ml) were added to a solution of 2,6-dimethylbenzoic acid (5 g, 33.29 mmol) stirred in toluene (40 ml). The resulting reaction mixture was refluxed for 16 hours using a Dean Stark condenser and the progress of the reaction mixture was monitored using TLC (30% ethyl acetate in petroleum ether, UV observation). After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to produce a residue, which was washed with ice water (250 ml) and extracted with ethyl acetate (3 x 150 ml). The combined organic extracts were washed with saturated sodium bicarbonate aqueous solution (50 ml) and brine (100 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce 2-bromoethyl 2,6-dimethylbenzoate (5.52 g). MS (ESI): m / z 258.82 [M+H+2] +.1 H NMR (400MHz, CDCl3) δppm 7.22-7.18 (m, 1H), 7.0-7.02 (m, 2H), 4.64 (t, 2H), 3.63 (t, 2H), 2.33 (s, 6H).

[0664] ●Synthetic intermediate 2-(aza-1-yl)ethyl 2,6-dimethylbenzoate:

[0665] To a solution of 2-bromoethyl 2,6-dimethylbenzoate (2.0 g, 7.778 mmol) stirred in ACN (30 ml) was added K2CO3 (3.224 g, 23.334 mmol) and aza (1.157 g, 11.667 mmol). The resulting reaction mixture was stirred at 90°C for 16 hours, and the reaction progress of the reaction mixture was monitored by TLC (30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to produce a residue, which was washed with cold water.

[0666] The residue was diluted with ethyl acetate (100 ml) and extracted with ethyl acetate (3 x 50 ml). The combined organic extracts were washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-(azepine-1-yl)ethyl 2,6-dimethylbenzoate (1.80 g) as a brown liquid. MS (ESI): m / z 276.35 [M+H] +.1 H NMR (400MHz, CDCl3) δppm7.19-7.15 (m, 1H), 7.03-7.01 (m, 2H), 4.41 (t, 2H), 2.86 (t, 2H), 2.72-2.69 (mt, 4H), 2.33 (s, 6H), 1.65-1.56 (m, 8H).

[0667] ●Synthesis of 1-benzyl-1-(2-((2,6-dimethylbenzyl)oxy)ethyl)azepine-1-bromide:

[0668] To a stirred solution of 2-(azepine-1-yl)ethyl 2,6-dimethylbenzoate (300 mg, 1.089 mmol) in ACN (5 ml) was added benzyl bromide (279.3 mg, 1.633 mmol) and the resulting reaction mixture was stirred at 90°C for 16 hours. The progress of the reaction mixture was monitored by TLC (methanol in DCM, UV observation). After the consumption of the starting material was complete, the reaction mixture was cooled to room temperature and then concentrated under reduced pressure to give a crude product, which was triturated with a 1:2 mixture of Et2O & EtOAc (60 ml) to give 1-benzyl-1-(2-((2,6-dimethylbenzyl)oxy)ethyl)azepine-1-bromide (117.4 mg,) as an off-white solid. Mass spectrum (ESI): m / z 366.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.61-7.59(m, 2H), 7.54-7.48(m, 3H), 7.29-7.26(m, 1H), 7.13-7.11(d, 2H), 4.89(t, 2 H), 4.67(s, 2H), 3.65-3.45(m, 6H), 2.28(s, 6H), 1.83-.1.77(d, 4H), 1.6-1.5(m, 4H).

[0669] Synthesis of 1-benzyl-1-(2-(2,6-dimethylphenoxy)-2-oxoethyl)azepine-1-2,2,2-trifluoroacetate

[0670]

[0671] Synthetic intermediate 2,6-dimethylphenyl 2-bromoacetate:

[0672] At 0 ℃, to 2,6-dimethylbenzene (0.5g, 4.092mmol) in ACN (5ml) after stirring, add pyridine (0.647g, 8.184mmol) and 2-bromoacetyl bromide (1.23g, 6.138mmol), stir the resulting reaction mixture at 0 ℃ for 15 minutes, use TLC (10% ethyl acetate solution in petroleum ether, ultraviolet observation) to monitor the reaction progress of the reaction mixture. After consuming the starting compound, dilute the reaction mixture with water (20ml) and extract with ethyl acetate (2x25ml). Wash the combined organic extracts with brine (30ml), dry with anhydrous sodium sulfate and concentrate under reduced pressure to produce product 2,6-dimethylphenyl 2-bromoacetate (500mg) as a light yellow liquid.1 H NMR (400MHz, CDCl3) δppm 7.07 (s, 3H), 4.07 (s, 2H), 2.18 (s, 6H).

[0673] Synthetic intermediate 2,6-dimethylphenyl 2-(azepine-1-yl) acetate:

[0674] At room temperature, 2,6-dimethylphenyl 2-bromoacetate (0.5 g,

[0675] To a solution of 2,6-dimethylphenyl 2-(azepine-1-yl)acetate (500 mg) was added KCO (0.71 g, 5.14 mmol) and aza (0.407 g, 4.112 mmol) in ACN (5 ml). The reaction mixture was stirred at 90° C. for 16 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: ethyl acetate, UV observation). After the reaction was completed, the reaction mixture was concentrated under reduced pressure to produce a residue, which was diluted with ice water (25 ml) and extracted with ethyl acetate (2×100 ml). The combined organic extracts were washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce 2,6-dimethylphenyl 2-(azepine-1-yl)acetate (500 mg) as a red liquid. Mass spectrum (ESI): m / z 262.09 [M+H] + .

[0676] Synthesis of 1-benzyl-1-(2-(2,6-dimethylphenoxy)-2-oxoethyl)azepine-1-2,2,2-trifluoroacetate.

[0677] To a stirred solution of 2,6-dimethylphenyl 2-(azepin-1-yl)acetate (0.45 g, 1.721 mmol) in ACN (4.5 ml) was added benzyl bromide (0.588 g, 3.442 mmol) and the reaction mixture was stirred at 90°C for 16 hours. The progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM, UV observation). After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give a crude compound which was purified by reaction preparative HPLC (column: X-selected phenylhexyl C18 (19*250), 5um; mobile phase A: 0.1% TFA (Aq); mobile phase B-acetonitrile; flow rate: 15 ml / min; method: 0 / 20, 2 / 20, 10 / 50, 15 / 75, 15.2 / 98, 19 / 98, 19.2 / 20, 23 / 20; solubility: ACN+water+THF; temperature: room temperature). The collected pure fractions were freeze-dried to give 1-benzyl-1-(2-(2,6-dimethylphenoxy)-2-oxoethyl)azepine-1-2,2,2-trifluoroacetate (56 mg) as a pale yellow solid. MS (ESI): m / z 352.2 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.60-7.53(m, 5H), 7.21-7.16(m, 3H), 4.87(s, 2H), 4.71(s, 2H), 3.90-3.84( m, 2H), 3.66-3.60 (m, 3H), 2.19 (s, 6H), 2.07-1.91 (m, 4H), 1.40-1.20 (s, 4H).

[0678] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)ethyl)azepine-1-trifluoroacetate

[0679]

[0680] Synthetic intermediate 1-(aza-1-yl)-2-chloroethan-1-one:

[0681] To a cooled solution of azepine (10 g, 100.826 mmol) (0°C) in ACN (100 mL) was added TEA (30.6 g, 302.478 mmol) followed by 2-chloroacetyl chloride (13.6 g, 120.98 mmol). The reaction mixture was stirred at room temperature for 3 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: EtOAc, UV observation: ninhydrin). The reaction was quenched with saturated sodium bicarbonate and extracted with ethyl acetate (2X250 mL). The combined organic extracts were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product which was purified by column chromatography (eluting with 30% ethyl acetate in petroleum ether) to give 1-(azepine-1-yl)-2-chloroethane-1-one (8.5 g) as a brown liquid. Mass spectrum (ESI): m / z 175.89 [M+1] + . 1 H NMR (400MHz, CDCl3) δppm 4.08 (s, 2H), 3.49-3.55 (m, 4H), 1.72-1.79 (m, 4H), 1.58-1.63 (d, 4H).

[0682] Synthetic intermediate 1-(aza-1-yl)-2-((2,6-dimethylphenyl)amino)ethane-1-one:

[0683] To a solution of 1-(azepine-1-yl)-2-chloroethane-1-one (5 g, 28.464 mmol) stirred in ACN (40 mL) was added DIPEA (11.03 g, 85.392 mmol), followed by 2,6-dimethylaniline (5.17 g, 42.696 mmol) and the reaction mixture was stirred at 90° C. for 3 days, and the reaction progress of the reaction mixture was monitored using TLC (mobile phase: 30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was quenched with saturated sodium bicarbonate aqueous solution (20 ml) and extracted with ethyl acetate (2×250 ml). The combined organic extracts (200 mL) were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product (8.5 g), which was purified by column chromatography (eluting with 30% ethyl acetate in petroleum ether) to give 1-(azepan-1-yl)-2-((2,6-dimethylphenyl)amino)ethan-1-one (2.7 g, 36%) as a brown solid. Mass spectrum (ESI): m / z 261.18 [M+1] + . 1H NMR (400MHz, cdcl3) δppm 6.97-7.99(m, 2H), 6.76-6.80(m, 1H), 4.78(s, 1H), 3.81(s, 2H), 3.33-3.58( m, 2H), 3.30-3.32 (m, 2H), 2.34 (s, 6H), 1.72-1.73 (m, 4H), 1.55-1.59 (m, 5H).

[0684] ●Synthetic intermediate N-(2-(aza-1-yl)ethyl)-2,6-dimethylaniline

[0685] A solution of 1-(aza-1-yl)-2-((2,6-dimethylphenyl)amino)ethane-1-one (1.5 g, 5.760 mmol) in THF (15 mL) was cooled to 0° and LAH (2M solution in THF, 5.76 mL, 11.521 mmol) was added dropwise while stirring. The resulting reaction mixture was stirred at room temperature for 15 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was quenched with an aqueous solution containing ammonium chloride at 0°C and then stirred at room temperature for 1 hour. The precipitated solid was recovered by filtration and the filtrate was concentrated under reduced pressure to produce N-(2-(aza-1-yl)ethyl)-2,6-dimethylaniline (1.4 g) as a light yellow liquid. Mass spectrum (ESI): m / z 247.23 [M+1] + .

[0686] ●Synthetic intermediate t-butyl (2-(azepine-1-yl) ethyl) (2,6-dimethylphenyl) carbamate:

[0687] To a stirred solution of N-(2-(azepine-1-yl)ethyl)-2,6-dimethylaniline (800 mg, 3.247 mmol) in a 1:1 mixture of 1,4-dioxane and water (10 ml) was added NaOH (260 mg, 6.494 mmol) followed by (Boc)2O (2.12 g, 9.741 mmol) and the resulting reaction mixture was stirred at room temperature for 24 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 30% ethyl acetate in petroleum ether, detection: ninhydrin). The reaction mixture was diluted with water (75 mL), extracted with EtOAc (2×75 mL) and the combined organic extracts (70 mL) were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (eluting with 15% ethyl acetate in petroleum ether) to give tert-butyl (2-(azepin-1-yl)ethyl)(2,6-dimethylphenyl)carbamate (600 mg) as a colorless liquid. Mass spectrum (ESI): m / z 347.24 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.00-7.26 (m, 3H), 3.47-3.57 (m, 2H), 2.60-2.73 (m, 6H), 2.30 (s, 6H), 2.22 (t, 11H), 1.57 (s, 6H).

[0688] Synthetic intermediate 1-benzyl-1-(2-((tert-butoxycarbonyl)(2,6-dimethylphenyl)amino)ethyl)azepine-1-chloride:

[0689] To a stirred solution of tert-butyl (2-(azepin-1-yl)ethyl)(2,6-dimethylphenyl)carbamate (300 mg, 0.865 mmol) in ACN (4 mL) was added benzyl chloride (328 mg, 2.595 mmol). The reaction mixture was stirred at 90° C. for 15 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give the crude product 1-benzyl-1-(2-((tert-butoxycarbonyl)(2,6-dimethylphenyl)amino)ethyl)azepin-1-chloride salt (400 mg) as a colorless gum. Mass spectrum (ESI): m / z 437.30 [M+1] + .

[0690] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)ethyl)azepine-1-trifluoroacetate:

[0691] To a stirred solution of 1-benzyl-1-(2-((tert-butoxycarbonyl)(2,6-dimethylphenyl)amino)ethyl)azepine-1-chloride (360 mg, 0.634 mmol) was added 4M HCl in 1,4-dioxane (3.17 mL, 12.680) at 0° C. The reaction mixture was stirred at room temperature for 15 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was purified by reverse phase preparative HPLC (column: X-bridge C18 (150*25) mm, 10u; mobile phase A: 0.1% TFA in water (Aq); mobile phase B: acetonitrile; flow rate: 14 ml / min; method (T / % of B): 0 / 20, 2 / 30, 10 / 40, 19 / 40, 19.2 / 98, 22 / 98, 22.2 / 20, 26 / 20; solubility: ACN+water+THF; temperature: room temperature). The pure fractions were collected and freeze dried to give 1-benzyl-1-(2-((2,6-dimethylphenyl)amino)ethyl)azepine-1-trifluoroacetate (165 mg) as a light brown solid. Mass spectrum (ESI): m / z 337.30 [M] + .LCMS: 99.72%. 1 H NMR (400MHz, DMSO-d6) δppm 7.41-7.53(m, 5H), 6.96-7.21(m, 2H), 6.83(t, 1H), 4.56(s, 2H), 3.31-3.53(m, 8H), 2.27(s, 6H), 1.82-1.83(m, 4H), 1.58-1.59(m, 4H).

[0692] Synthesis of 1-Benzyl-1-(2-(2,6-dimethylphenoxy)ethyl)azepine-1-bromide

[0693]

[0694] Synthesis of 2-(2-bromoethoxy)-1,3-dimethylbenzene:

[0695] Potassium carbonate (1.698 g, 12.288 mmol) and 1,2-dibromoethane (3.847 g, 20.480 mmol) were added to a solution of 2,6-dimethylbenzyl alcohol (0.500 g, 4.096 mmol) in acetonitrile (8.0 mL) after stirring. The reaction mixture was stirred at 90°C for 16 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: 5% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure, diluted with water (100 mL) and extracted with dichloromethane (3×50 mL), the combined organic extracts were washed with brine solution (1×50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce a crude product, and the crude product was purified by silica gel chromatography (eluted with petroleum ether) to produce pure 2-(2-bromoethoxy)-1,3-dimethylbenzene (0.20 g) as a colorless liquid. 1 H NMR (400MHz, CDCl3) δppm 7.00-7.25 (m, 2H), 6.91-6.95 (m, 1H), 4.01-4.10 (m, 2H), 3.65-3.68 (m, 2H), 2.30 (s, 6H).

[0696] Synthetic intermediate 1-(2-(2,6-dimethylphenoxy)ethyl)azepine:

[0697] To a solution of 2-(2-bromoethoxy)-1,3-dimethylbenzene (200 mg, 0.87 mmol) in acetonitrile (3.0 mL) was added DIPEA (451.2 mg, 3.49 mmol) and aza (120.2 mg, 1.22 mmol). The resulting reaction mixture was stirred at 90° C. for 16 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 10% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure, diluted with water (20 mL), and extracted with dichloromethane (3×20 mL). The combined organic extracts (1×20 ml) were washed with saline solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to produce 1-(2-(2,6-dimethylphenoxy)ethyl)azepine (210 mg). 1 H NMR (400MHz, CDCl3) δppm 6.98-7.00(m, 2H), 6.88-6.92(m, 1H), 3.85-3.88(m, 2H), 2.93-2.96(m, 2H), 2.75-2.77(m, 4H), 2.28(s, 6H), 1.59-1.68(m, 8H).

[0698] Synthesis of 1-benzyl-1-(2-(2,6-dimethylphenoxy)ethyl)azepine-1-bromide:

[0699] To a solution of 1-(2-(2,6-dimethylphenoxy)ethyl)azepine (100 mg, 0.404 mmol) in acetonitrile (1.5 mL) was added benzyl bromide (0.072 ml, 0.606 mmol) and the reaction mixture was stirred at 90° C. in a sealed container for 16 hours. The progress of the reaction mixture was monitored by TLC (mobile phase: 50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude gum, which was triturated with ethyl acetate (3×5 ml) to give 1-benzyl-1-(2-(2,6-dimethylphenoxy)ethyl)azepine-1-bromide (80.5 mg) as an off-white solid. MS (ESI): m / z 338.41 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.63-7.65(m, 2H), 7.49-7.57(m, 3H), 7.04-7.08(m, 2H), 6.96-6.98(m, 1H), 4.72(s, 2 H), 4.29(t, 2H), 3.52-3.69(m, 6H), 2.29(s, 6H), 1.88-1.92(m, 4H), 1.59-1.62(m, 4H).

[0700] Synthesis of N-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-N,N-diethyl-3-phenylpropyl-1-bromide Ammonium

[0701]

[0702] Synthetic intermediate N, N-diethyl-3-phenylpropyl-1-amine:

[0703] To a solution of (3-bromopropyl)benzene (2 g, 10.04 mmol) in acetonitrile (5 ml) was added diethylamine (1.46 g, 19.96 mmol) at room temperature. The resulting reaction mixture was stirred at 75°C for 15 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was cooled to room temperature, quenched with water and extracted with ethyl acetate (2 x 15 ml). The combined organic extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure to yield N, N-diethyl-3-phenylpropyl-1-amine (1.3 g). MS (ESI): m / z 192.25 [M+H] + . 1H NMR (400MHz, CDCl3) δppm 7.3-7.25(m, 2H), 7.22-7.15(m, 3H), 2.6-2.25(m, 2H), 2.55-2.42(m, 6H), 1.81-1.75(m, 2H), 1.0(t, 6H).

[0704] Synthesis of N-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-N,N-diethyl-3-phenylpropyl-1-ammonium bromide:

[0705] To a solution of N,N-diethyl-3-phenylpropyl-1-amine (0.2 g, 1.045 mmol) in toluene (5 ml) was added 2-bromo-N-(2,6-dimethylphenyl)acetamide (0.328 g, 1.354 mmol) and the resulting reaction mixture was heated to reflux for 16 hours, and the progress of the reaction mixture was monitored using TLC (mobile phase: 10% methanol in DCM, UV observation). The resulting reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product, which was triturated with ethyl acetate (20 ml) to give N-(2-((2,6-dimethylphenyl)amino)-2-oxoethyl)-N,N-diethyl-3-phenylpropyl-1-ammonium bromide (95 mg) as an off-white solid. MS (ESI): m / z 353.45 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm9.9 (s, 1H), 7.45-7.3 (m, 5H), 7.15-7.09 (m, 3H), 4.28 (s, 2H), 3. 58(q, 4H), 3.51-3.45(m, 2H), 2.68-2.51(m, 2H), 2.15(s, 6H), 2.08-2.0(m, 2H), 1.28(t, 6H).

[0706] Synthesis of N-Benzyl-2-((2,6-dimethylphenyl)sulfonyl)-N,N-diethylethane-1-ammonium bromide

[0707]

[0708] Synthetic intermediate (2-chloroethyl)(2,6-dimethylphenyl)sulfonamide:

[0709] To a stirred solution of 2,6-dimethylbenzenethiol (2 g, 14.468 mmol, 1 eq) in ethanol (30 ml) was added NaOH (1.736 g, 43.405 mmol) and 1-bromo-2-chloroethane (4.149 g, 28.937 mmol) at 0°C. The resulting reaction mixture was stirred at room temperature for 16 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 100% petroleum ether, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude residue and the residue was diluted with ethyl acetate (80 ml) and washed with water (50 ml x 2) and brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude compound which was purified by flash chromatography (eluted with petroleum ether). The collected pure fractions were concentrated under reduced pressure to give (2-chloroethyl)(2,6-dimethylphenyl)sulfonamide (0.500 g) as a colorless liquid. 1 H NMR (400MHz, CDCl3) δppm 7.085-7.163 (m, 3H), 3.502-3.542 (m, 2H), 2.963-3.003 (m, 2H), 2.544 (s, 6H).

[0710] ●Synthetic intermediate 2-((2-chloroethyl)sulfonyl)-1,3-dimethylbenzene:

[0711] To a stirred solution of (2-chloroethyl)(2,6-dimethylphenyl)sulfonamide (0.500 g, 2.491 mmol) in acetic acid (10 ml) was added hydrogen peroxide (33% aqueous solution) (0.847 g, 24.910 mmol) at room temperature. The resulting reaction mixture was heated to 110°C for 16 h. The progress of the reaction mixture was monitored using TLC (mobile phase: 20% ethyl acetate in petroleum ether, UV observation). The reaction mixture was diluted with water (60 ml) and extracted with ethyl acetate (70 ml x 3). The combined organic extracts were washed with aqueous sodium thiosulfate (50 ml x 2) and concentrated under reduced pressure to give a crude product which was purified by flash chromatography (eluted with 10% ethyl acetate in petroleum ether). The collected pure fractions were concentrated under reduced pressure to give 2-((2-chloroethyl)sulfonyl)-1,3-dimethylbenzene (0.350 g) as an off-white solid. MS (ESI): m / z 232 [M] + . 1 H NMR (400MHz, CDCl3) δppm 7.345-7.383 (m, 1H), 7.176-7.195 (m, 2H), 3.815-3.854 (m, 2H), 3.531-3.569 (m, 2H), 2.714 (s, 6H).

[0712] ●Synthetic intermediate 2-((2,6-dimethylphenyl)sulfonyl)-N,N-diethylethane-1-amine:

[0713] Potassium iodide (0.428 g, 2.578 mmol) and diethylamine (0.283 g, 3.867 mmol) were added to a solution of 2-((2-chloroethyl)sulfonyl)-1,3-dimethylbenzene (0.300 g, 1.289 mmol) in acetonitrile (10 ml) at room temperature. The resulting reaction mixture was heated to 90° C. in a sealed container and maintained for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (mobile phase: 50% ethyl acetate in petroleum ether, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude compound which was diluted with ethyl acetate (60 ml), washed with water (30 ml x 3) and brine (30 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude compound which was purified by flash chromatography (eluted with 40% ethyl acetate in petroleum ether). The collected pure fractions were concentrated under reduced pressure to give the desired product 2-((2,6-dimethylphenyl)sulfonyl)-N,N-diethylethan-1-amine (0.200 g) as an off-white solid. MS (ESI): m / z 270.16 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.308-7.346(m, 1H), 7.153-7.172(m, 2H), 3.244-3.283(m, 2H), 2.947- 2.985(m, 2H), 2.716(s, 6H), 2.437-2.490(m, 4H), 0.937-0.973(m, 6H).

[0714] ●Synthesis of N-benzyl-2-((2,6-dimethylphenyl)sulfonyl)-N,N-diethylethane-1-ammonium bromide:

[0715] To a stirred solution of 2-((2,6-dimethylphenyl)sulfonyl)-N,N-diethylethane-1-amine (0.100 g, 0.371 mmol) in acetonitrile (5 ml) was added benzyl bromide (0.127 g, 0.742 mmol). The resulting reaction mixture was heated to 90°C for 16 hours and the progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product, which was triturated with ethyl acetate (20 ml) and n-pentane (10 ml) to give N-benzyl-2-((2,6-dimethylphenyl)sulfonyl)-N,N-diethylethane-1-ammonium bromide (0.060 mg) as an off-white solid. MS (ESI): m / z 360.0 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm7.475-7.552 (m, 6H), 7.347-7.366 (d, 2H), 4.604 (s, 2H), 3.925- 3.965(m, 2H), 3.520-3.560(m, 2H), 3.243-3.288(m, 4H), 2.676(s, 6H), 1.235-1.312(m, 6H).

[0716] Synthesis of 1-Benzyl-1-(2-((2,6-dimethylphenyl)sulfonyl)ethyl)azepine-1-bromide

[0717]

[0718] Synthetic intermediate (2-((2,6-dimethylphenyl)sulfonyl)ethyl):

[0719] To a solution of 2-((2-chloroethyl)sulfonyl)-1,3-dimethylbenzene (1.0 g, 4.2970 mmol) stirred in ACN (10.0 ml) was added azepine (0.639 g, 6.4455 mmol) and potassium iodide (1.426 g, 8.5940 mmol). The resulting reaction mixture was heated to reflux for 16 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 30% ethyl acetate in petroleum ether, UV observation). The reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (2 x 50 ml) and brine (1 x 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to produce a crude compound, which was ground with 2 x 10 ml of n-pentane to produce (2-((2,6-dimethylphenyl)sulfonyl)ethyl)azepine (180 mg) as an off-white solid. MS (ESI): m / z 295.44 [M+H]. 1H NMR (400MHz, CDCl3) δppm 7.34 (t, 1H), 7.17 (d, 2H), 3.1-3.8 (m.6H), 2.71 (d, 6H), 1.57-1.82 (m, 10H), 2.71 (d, 6H).

[0720] Synthesis of 1-benzyl-1-(2-((2,6-dimethylphenyl)sulfonyl)ethyl)azepine-1-bromide:

[0721] To a stirred solution of 1-(2-((2,6-dimethylphenyl)sulfonyl)ethyl)azepine (0.550 g, 1.864 mmol) in ACN (6.0 ml) was added benzyl bromide (0.637 g, 3.728 mmol) and the reaction mixture was heated to reflux for 48 hours. The progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM, UV observation). The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude compound, which was triturated with 3x10 ml of ethyl acetate to give 1-benzyl-1-(2-((2,6-dimethylphenyl)sulfonyl)ethyl)azepine-1-bromide (80 mg) as an off-white solid. MS (ESI): m / z 386.40 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.5-7.63(m, 6H), 7.44(d, 2H), 4.59-4.65(d, 2H), 4.06-4.15(m, 2H), 3. 49-3.55(m, 6H), 2.696(s, 6H), 1.817-1.95(m, 4H), 1.569-1.65(m, 4H).

[0722] Synthesis of 1-Benzyl-1-(1-(2,6-dimethylphenyl)-2-oxypyrrolidin-3-yl)piperidin-1-bromide

[0723]

[0724] Synthesis of 3-bromo-1-(2,6-dimethylphenyl)pyrrolidin-2-one:

[0725] At 0°C, K3PO4 (1.751 g, 8.252 mmol) and 2,4-dibromobutyryl chloride (2.181 g, 8.252 mmol) were added to a solution of 2,6-dimethylaniline (1.0 g, 8.252 mmol) stirred in ACN (20 ml) for treatment. The reaction mixture was stirred at room temperature for 1 hour. After 1 hour, the reaction mixture was treated with 50% NaOH aqueous solution (1.650 g in 3.32 mL water) and the reaction mixture was stirred for 1 hour at room temperature. TLC (mobile phase: 50% ethyl acetate in petroleum ether, UV observation) was used to monitor the reaction progress of the reaction mixture. After the reaction was completed, the reaction mixture was filtered to remove inorganic salts and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce a colorless colloidal crude product. The crude product was ground with n-pentane (30 ml) to produce 3-bromo-1-(2,6-dimethylphenyl)pyrrolidin-2-one (1.5 g). MS (ESI): m / z 268.03 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.18-7.14(m, 1H), 7.10-7.08(m, 2H), 4.56-4.54(m, 1H), 3.92-3.86(m, 1H), 3.5 1-3.46(m, 1H), 2.84-2.78(m, 1H), 2.52-2.47(m, 1H), 2.29(s, 3H), 2.19(s, 3H).

[0726] ●Synthesis of 1-(2,6-dimethylphenyl)-3-(piperidin-1-yl)pyrrolidin-2-one:

[0727] To a stirred solution of 3-bromo-1-(2,6-dimethylphenyl)pyrrolidin-2-one (0.300 g, 1.118 mmol) in ACN (10 ml) was added piperidine (0.114 g, 1.342 mmol) and DIPEA (0.578 g, 4.475 mmol) at room temperature and the resulting reaction mixture was heated in a sealed container at 90°C for 16 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 10% methanol in DCM, UV observation). The reaction mixture was concentrated under reduced pressure to give a crude product which was diluted with DCM (70 ml) and washed with water (40 ml x 3) and brine (40 ml). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 1-(2,6-dimethylphenyl)-3-(piperidin-1-yl)pyrrolidin-2-one (0.220 g). MS (ESI): m / z 273.35 [M+H] + . 1H NMR (400MHz, CDCl3) δppm 7.14-7.06(m, 3H), 3.70-3.48(m, 3H), 2.80-2.89(m, 2H), 2.60-2.65(m, 2H), 2.3 5-2.28(m, 2H), 2.21(s, 3H), 2.15(s, 3H), 1.67-1.60(m, 4H), 1.50-1.46(m, 2H).

[0728] Synthesis of 1-benzyl-1-(1-(2,6-dimethylphenyl)-2-oxypyrrolidin-3-yl)piperidin-1-bromide:

[0729] To a stirred solution of 1-(2,6-dimethylphenyl)-3-(piperidin-1-yl)pyrrolidin-2-one (0.220 g, 0.807 mmol) in ACN (5 ml) was added benzyl bromide (0.276 g, 1.615 mmol) and the reaction mixture was heated in a sealed container at 90°C for 16 hours. The progress of the reaction mixture was monitored by TLC (mobile phase: 10% methanol in DCM, UV observation). The reaction mixture was cooled to room temperature, diluted with ethyl acetate (20 ml) and stirred for 2 hours to produce a solid, which was filtered and washed with ethyl acetate (40 ml), then washed with n-pentane (20 ml) and dried under high vacuum to produce 1-benzyl-1-(1-(2,6-dimethylphenyl)-2-oxopyrrolidin-3-yl)piperidin-1-bromide (0.090 g) as an off-white solid. MS (ESI): m / z 363.1 [M] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.60-7.52(m, 5H), 7.25-7.16(m, 3H), 5.58-4.02(m, 4H), 3.65-3.57(m, 2H), 3.13-2.92(m, 3H), 2.67-2.66(m, 1H), 2.21-1.94(m, 10H), 1.61-1.23(m, 3H).

[0730] The following examples were prepared according to the procedure described for the synthesis of compound 89A from 3-bromo-1-(2,6-dimethylphenyl)pyrrolidin-2-one, aziridine and benzyl bromide.

[0731]

[0732] Synthesis of 1-Benzyl-1-(1-(2,6-dimethylphenyl)-2-oxypiperidin-3-yl)pyrrolidine-1-bromide

[0733]

[0734] ●Synthesis of 2,5-dibromopentanoyl chloride:

[0735] A stirred suspension of 5-bromovaleryl chloride (5 g, 25.066 mmol) was treated with bromine (2.56 ml, 50.132 mmol) at room temperature and the resulting reaction mixture was stirred at 100°C for 2 hours. After 2 hours, the reaction was cooled to room temperature and concentrated under reduced pressure to give crude 2,5-dibromovaleryl chloride (7.2 g) which was used in the next step without further purification.

[0736] Synthesis of 3-bromo-1-(2,6-dimethylphenyl)piperidin-2-one:

[0737] A solution of 2,6-dimethylaniline (2.0 g, 16.504 mmol) and potassium phosphate (3.503 g, 16.504 mmol) stirred in acetonitrile (50 ml) was cooled to 0°C and 2,5-dibromopentanoyl chloride (4.594 g, 16.504 mmol) was added dropwise. The resulting reaction mixture was stirred at room temperature for 1 hour. After 1 hour, 50% aqueous NaOH (3.3 g, 6.6 ml water) was added to the reaction mixture and the resulting reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction mixture was monitored using TLC (mobile phase: 50% EtOAc: petroleum ether, UV observation). The reaction mixture was filtered to remove inorganic salts and the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure to produce a crude product, which was purified by column chromatography (eluted with 25%-30% ethyl acetate in petroleum ether) to produce 3-bromo-1-(2,6-dimethylphenyl)piperidin-2-one (2.2 g) as an off-white solid. MS (ESI): 282.25 m / z, [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.15-7.08(m, 3H), 4.87(t, 1H), 3.54-3.49(m, 1H), 3.37-3.34(m, 1H), 2.51-2.45(m, 1H), 2.27-2.21(m, 2H), 2.22-2.10(m, 6H), 1.94(m, 1H).

[0738] Synthesis of 1-(2,6-dimethylphenyl)-3-(pyrrolidin-1-yl)piperidin-2-one:

[0739] Pyrrolidine (0.0655 g, 0.921 mmol) was added to a solution of 3-bromo-1-(2,6-dimethylphenyl)piperidin-2-one (0.2 g, 0.709 mmol) and DIPEA (0.366 g, 2.83 mmol) in acetonitrile (10 mL) and stirred, and the resulting reaction mixture was stirred at 90 ° C in a sealed container for 16 hours, and the reaction progress of the reaction mixture was monitored using TLC (mobile phase: 50% ethyl acetate in petroleum ether, UV observation). The reaction was concentrated under reduced pressure to produce a crude product, which was diluted with water (20 mL) and extracted with ethyl acetate (2×50 mL). The combined organic extracts were dried over anhydrous sodium sulfate and concentrated to give the crude compound which was purified by normal phase flash chromatography (eluting with 25%-30% ethyl acetate in petroleum ether) to give 1-(2,6-dimethylphenyl)-3-(pyrrolidin-1-yl)piperidin-2-one (350 mg) as a light yellow gummy oil. LCMS purity: 75.17%, MS (ESI): 273.43 m / z, [M+H] + .

[0740] Synthesis of 1-benzyl-1-(1-(2,6-dimethylphenyl)-2-oxypiperidin-3-yl)pyrrolidine-1-bromide:

[0741] To a solution of 1-(2,6-dimethylphenyl)-3-(pyrrolidin-1-yl)piperidin-2-one (250 mg, 0.9178 mmol) stirred in acetonitrile (10 mL) was added benzyl bromide (0.2041 g, 1.1931 mmol) and the resulting reaction mixture was stirred in a sealed container at 90°C for 16 hours and the progress of the reaction mixture was monitored using TLC (mobile phase: 10% methanol in DCM, UV observation). The resulting reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product which was triturated with ethyl acetate (20 mL x 2) to give 1-benzyl-1-(1-(2,6-dimethylphenyl)-2-oxopiperidin-3-yl)pyrrolidine-1-bromide (105 mg) as an off-white solid. MS (ESI): m / z 363.4 [M] + . 1H NMR (400MHz, DMSO-d6) δppm 7.64-7.62(m, 2H), 7.57-7.48(m, 3H), 7.20-7.13(m, 3H), 4.78(m, 3H), 4.03-4.01(m, 2H), 3.80-3.72(m, 2H), 3.5 5-3.50 (m, 1H), 3.41-3.61 (m, 1H), 2.51-2.49 (m, 1H), 2.39-2.35 (m, 1H), 1.21-1.99 (m, 10H), 1.97-1.90 (m, 2H).

[0742] The following examples were prepared from 3-bromo-1-(2,6-dimethylphenyl)piperidin-2-one, benzyl bromide and the appropriate azacycloalkane using the same procedure as described for the synthesis of compound 91A. The products were purified by trituration or reverse phase preparative HPLC.

[0743]

[0744]

[0745] The following examples were prepared using the same procedure as described for the synthesis of compound 69A from bromoacetyl bromide, benzyl bromide and the appropriate azacycloalkane and substituted aniline.

[0746]

[0747]

[0748]

[0749] Example 2 - Inhibition of Nav1.7 Current

[0750] Representative compounds of the invention were synthesized according to the methods described above and tested for their ability to inhibit voltage-gated sodium channels.

[0751] Manual patch clamp:

[0752] Cell culture

[0753] NaV1.7 was induced with tetracycline. Cells were cultured in DMEM containing 10% dialyzed fetal bovine serum (VWR, Radnor, PA), 1% glutamine (VWR, Radnor, PA), 1% penicillin-streptomycin (VWR, Radnor, PA), 100 mg / L hygromycin (ThermoFisher Scientific, Waltham, MA) and 5 mg / L juvenile cytokines (Alfa Aesar, Haverhill, MA). Cells were grown in a humidified environment at 37°C and kept in air containing 10% carbon dioxide. Cells were separated 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 and placed on 24-well plates. The cells were washed with DPBS (VWR, Radnor, PA), trypsinized, and then triturated five times in 10 mL of growth medium to separate cell aggregates. 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 tetracycline was added. Then 1 mL of mixed medium with cells was added to each well of the 24-well plate, and a 12 mm coverslip was placed in each well. The cells were then cultured overnight at 37°C and 10% CO2.

[0754] Patch clamp solutions and drugs

[0755] The intracellular solution contained the following (in mM): CsCl 135, NaCl 10, EGTA 10, HEPES 10, MgCl2 2, and was adjusted to pH 7.2 with CsOH. The external solution was normal Ringer's solution, containing (in mM) NaCl 155, HEPES 10, glucose 10, KCl 3.5, CaCl2 1.5, MgCl2 1, and was adjusted to pH 7.4 with NaOH. CsCl was from Alpha Essar, Haverhill, MA. All other chemicals were from Sigma-Aldrich, St. Louis, MO. To test the degree of internal blockage of a compound, the compound to be tested was dissolved in the internal solution at the indicated test concentration. In control experiments, the internal solution did not contain any compound. To test the degree of external blockage by a test compound, the compound was dissolved in the external solution at the indicated test concentration.

[0756] Whole-cell patch clamp protocol

[0757] 18-24 hours after induction of cells with tetracycline, coverslips were placed in a chamber filled with normal Ringer's solution at room temperature and the chamber was placed on a microscope. Pipettes were pulled from borosilicate glass on a P97 puller (Sutter Instrument, Novato, CA) and polished with a MF-830 micro glass polisher (Narishige International USA, Inc, Amityville, NY) to a resistance of 1.5-2.5 mΩ when filled with CsCl internal solution at room temperature. Healthy cells (those that were round, translucent and free of visible defects) were selected to form a seal. A seal was formed between the pipette and the cell and a brief pulse of suction was used to "cut in" and establish the entire cell structure. The membrane potential was held at -100 mV before the voltage protocol began. Only cells with a series resistance between 1.5 and 5 MΩ were retained for analysis. The voltage protocol was as follows: the cell was held at -100 mV for 12 ms, followed by a hyperpolarizing step at -105 mV for 12 ms to monitor leakage. The cell was then depolarized to -100 mV for 40 ms. The cell was then depolarized to -20 mV for 10 ms and then back to -100 mV for 26 ms.

[0758] Internal blocking of test compounds

[0759] Once the recording begins, the voltage scheme runs for 5 minutes at 30-second intervals to obtain a stable baseline. Four 30-second 5Hz identical voltage stimulations are subsequently performed, with a 1-minute rest after each voltage stimulation, and then a 0.33Hz stimulation is performed after the last stimulation. Current is recorded using PatchMaster software and Heka EPC10 (Heka Electronics, Lambrecht, Germany). Only cells with inward current amplitudes between -20mV, 400pA and 4nA are accepted. In addition, batteries with leakage currents greater than 10% of their current amplitudes are discarded.

[0760] Data Analysis: Internal Blocking

[0761] Data were plotted using Patchmaster software (HEKA Electronics, Lambrecht, Germany) and analyzed by plotting the minimum current during the voltage step to -20 mV (peak inward current) as a function of time. To determine the extent of depletion during the experiment, the mean peak inward current amplitude before 5 Hz stimulation (2-3 points) was designated as the baseline (I baseline The average peak inward current (I test ). By dividing Itest by I baselineCalculation of the remaining control fraction current. On each recording day, three cells were tested with the control internal solution, and the mean fraction of the remaining current (Ctrl fraction current) was calculated.

[0762] To determine the % block produced by the internally applied test compound, the following procedure was performed. The average peak inward current amplitude (2-3 points) before 5 Hz stimulation was designated as 0% block (I 0％block To correct for current variations under control conditions, I0% block was multiplied by the remaining average Ctrl fractional current to obtain the corrected 0% block current. The average peak inward current during the last 2 seconds of the last 5 Hz stimulation was designated as the unblocked current (I unblocked ). Calculate % blocking using the following equation: (1-I UNBLOCK / (I 0％block *Residual current control fraction) x 100).

[0763] Representative examples of the present invention were tested for intracellular inhibition of NaV 1.7. At 10 μM, the activity range was percent inhibition: "++++" (>95%), "+++" 95-70%, "++" (70-40%) or "+" (<40%). The results are shown below.

[0764]

[0765] Representative examples of the present invention were tested for intracellular inhibition of NaV 1.7. At 3 μM, the activity range is the percentage of inhibition: "++++" (>95%), "+++" 95-70%, "++" (70-40%) or "+" (<40%). The results are shown below.

[0766]

[0767]

[0768] External blocking of test compounds

[0769] Once recording begins, the voltage scheme runs for 5 minutes at 30-second intervals to obtain a stable baseline. The same voltage scheme is then used to stimulate at 5Hz until the end of the experiment. Test compounds are added during the 5Hz stimulation, and wait until the cells show a stable consumption rate before the addition of the compound. Test compounds are added for five minutes, followed by conventional Ringer's solution washout. PatchMaster software and Heka EPC10 (Heka Electronics, Lambrecht, Germany) are used to record current. Only cells with inward current amplitudes between -20mV, 400pA and 4nA are accepted. In addition, batteries with leakage currents greater than 10% of their current amplitudes are discarded.

[0770] Data Analysis: External Blocking

[0771] Data were plotted using Patchmaster software (HEKA Electronics, Lambrecht, Germany) and analyzed by plotting the minimum current during the voltage step to -20 mV (peak inward current) as a function of time. To determine the percentage of block produced by externally applied test compounds, the following was performed. Rate (Rate) was calculated by dividing the change in peak current amplitude by time after a stable current decay rate was established during 5 Hz stimulation. rundown The average peak inward current amplitude before compound addition (2-3 seconds) was used to determine the 0% block (I 0％block ). To correct for the loss, I 0％block Subtract(Rate rundown *5 minutes) to obtain the corrected 0% blocking current. The average peak inward current in the last 2-3 seconds of the 5-minute compound addition period before washing was the unblocked current (I unblocked ). The % blockade was then calculated using the following equation: Current blockade fraction = 1 - I UNBLOCK / (I 0％block -Rate rundown *5 minutes).

[0772] Representative examples of the present invention were tested for extracellular inhibition of NaV 1.7. At 10 μM, the activity range is the percentage of inhibition: "++++" (>90%), "+++" 90-70%, "++" (70-40%) or "+" (<40%). The results are shown below.

[0773] Compound <![CDATA[Na V 1.7 Extracellular Inhibition]]> Compound <![CDATA[Na V 1.7 Extracellular Inhibition]]> 3A ++ 47A ++ 4A + 53A + 6A + 69A + 9A + 70A ++ 33A ++ 76A ++ 35A + 80A + 38A + 81A + 44A + 86A + 46A ++ 90A +

[0774] Automated patch clamp:

[0775] Cell culture

[0776] Tetracycline induced HEK293 cells to express NaV1.7. Cells were cultured in DMEM containing 10% dialyzed fetal bovine serum (VWR, Radnor, PA), 1% glutamine (VWR, Radnor, PA), 1% penicillin-streptomycin (VWR, Radnor, PA), 100 mg / L hygromycin (Thermo Fisher Scientific, Waltham, MA) and 5 mg / L juvenile cytokines (Alfa Aesar, Haverhill, MA). Cells were grown and stored in a humid environment at 37°C and 10% CO2 in the air. Cells were separated 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, IBIScientific, Peosta, IA) one day before recording.

[0777] Prior to the experiment, cells were washed with DPBS (VWR, Radnor, PA), digested with deacetylase (VWR, Radnor, PA), and then triturated 10 times in CHO serum-free medium (VWR, Radnor, PA) to resuspend the cells and dissociate cell aggregates. Cells were counted and the final concentration was set at 2-5 million cells per ml.

[0778] Patch clamp solutions and drugs

[0779] The intracellular solution contained the following: 140 mM CsF, 1 mM / 5 mM EGTA / CsOH, 10 mM HEPES, 10 mM NaCl, pH adjusted to 7.3 with CsOH, and osmolarity adjusted to 320 with sucrose. The external solution contained the following: 145 mM NaCl, 4 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM HEPES, 10 mM glucose, pH adjusted to 7.4 with CsOH, and osmolarity adjusted to 305 with sucrose. All chemicals were from Sigma-Aldrich, St. Louis, MO. To test the extent of internal blocking by the assay, the compound was dissolved in the internal solution at the indicated test concentration. In control experiments, the internal solution did not contain any compound. To test the extent of external blocking by the assay, the compound was dissolved in the external solution at the indicated test concentration.

[0780] Automated patch clamp solution

[0781] Automated patch clamping was performed using multi-well Qchips on a Qube 384 (Sophion Bioscience, Woburn MA) with the temperature set to 22 degrees. Whole cell configurations were formed using default Qube seal and run-in parameters. The membrane potential was held at -100 mV before the voltage protocol began. A two-step voltage protocol was followed.

[0782] Step 1: The cell is held at -100mV and a depolarizing pulse is applied at -20mV for 10ms with the interval set to 5s. By default, the current is corrected by subtracting the leak from each pulse. The duration is set to 5 minutes.

[0783] Step 2: Hold the cell at -100 mV and apply a depolarizing pulse at -20 mV for 10 ms. The frequency is 5 Hz. Correct the current by subtracting the leak calculated before step 2. The duration is set to 4 minutes.

[0784] Internal blocking of test compounds

[0785] After step 2, remove the Qchip from the recording chamber. Replace the internal solution with the solution containing the test compound. Qchips remain at -100 mV in the recording chamber after replacement with no pulses. The total solution switching time is 8 min. After the internal solution is exchanged, record the cell and repeat step 2 for 10 min.

[0786] Data analysis was performed by Sophion analyzer. Cells were filtered with a minimum seal resistance of 50 MOhm and a minimum start-up current of 5 nA. Control cells (non-drug) were used to correct the current drop. The remainder was calculated by averaging the last 3 points at the end of the experiment. The baseline was calculated as the average of the last 3 points in step 2. IC50 curves were drawn using DR graphs / Hill functions (dose-response graphs using Hill fit). Data analysis was performed by Sophion analyzer. Representative embodiments of the present invention were tested for intracellular inhibition of NaV 1.7 in automated patch clamp analysis. The range of activity is expressed as IC 50 : “++++” (<0.3 μM), “+++” (0.3-1 μM), “++” (1-3 μM) or “+” (3-10 μM). The results are shown in the table below.

[0787]

[0788]

[0789] External blocking of test compounds

[0790] After step 2, the external solution was replaced with a solution containing the test compound. Qchips was held at -100 mV with no pulses. The total solution switching time was 8 min. After the external solution exchange, cells were recorded for 10 min using the same procedure as step 2.

[0791] Data analysis was performed by Sophion analyzer. Data were corrected with control cells (non-drug). IC50 data were plotted using DR plot / Hill function (dose-response plot with Hill fit).

[0792] Representative examples of the invention were tested for their extracellular inhibitory effects on NaV1.7 in an automated patch clamp assay. The range of activity is expressed as IC 50 : “++++” (<1 μM), “+++” (1-3 μM), “++” (3-10 μM) or “+” (>10 μM). The results are shown in the table below.

[0793]

[0794] Example 3 - Membrane Permeability

[0795] The ability of the compounds of the invention to pass through artificial lipid membranes by passive diffusion was determined using the PAMPA assay (pION, Inc., Woburn MA). The test compound was dissolved in dimethyl sulfoxide (10 mM) and diluted 200-fold in buffer (pION Inc., pH 7.4) to provide a 50 um stock solution. Buffer (150 μL) was added to the UV blank plate, and the stock solution (150 μL) was transferred to the UV control plate. The blank and reference spectra were read using a spectrophotometer. The stock solution (200 μL) was added to the donor plate of the PAMPA sandwich plate, and a receiving plate (pION Inc, 5 μL) coated with GIT lipids was placed on top. Buffer (200 μL) was added to the acceptor plate, and the PAMPA sandwich plate was incubated for 4 hours. A small portion (150 μL) from the acceptor plate was added to the UV plate and read as the acceptor spectrum. An aliquot (150 μL) of the donor solution was added to the UV analysis plate and read as the donor spectrum. The permeability coefficients of the test compounds were calculated based on the AUCs of the reference plate, donor plate, test plate, and receptor plate using PAMPA Explorer™ software (version 3.5.0.4).

[0796] PAMPA permeability results of representative compounds (10 -6 cm / s) is reported as “+” (<0.1 10 -6 cm / s), "++" (0.1-2.0 10 -6 cm / s), "+++" (2.0-10.010 -6cm / s) or "++++" (>10.0 10 -6 cm / s).

[0797]

[0798]

[0799] The patents and scientific literature referred to herein build upon the knowledge available to one of ordinary skill in the art to which the invention pertains. All U.S. patents and published or unpublished U.S. patent applications cited herein are hereby incorporated by reference in their entirety. All published foreign patents and foreign patent applications cited herein are hereby incorporated by reference in their entirety. All other published references, documents, manuscripts and scientific articles are hereby incorporated by reference in their entirety.

[0800] Although the present invention has been specifically shown and described in its preferred embodiments, it will be understood by those skilled in the art that various changes may be made in form and detail without departing from the scope of the invention as defined by the appended claims. It should also be understood that none of the embodiments described herein are immutable and may be combined with one another in various ways without departing from the scope of the invention as defined by the claims.

[0801] Although the present invention has been specifically shown and described in its preferred embodiments, it will be understood by those skilled in the art that various changes may be made in form and detail without departing from the scope of the invention as defined by the appended claims. It should also be understood that none of the embodiments described herein are immutable and may be combined with one another in various ways without departing from the scope of the invention as defined by the claims.

Claims

1. Compound of formula (I): in, Y - is a pharmaceutically acceptable anion; R A , R B , and R C are independently selected from H, D, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, OR I , CN, NR J R K , NR L C(O)R M ,S(O)R N , S(O)2R N , SO2R O R P , SO2NR Q R R , SO3R S ,CO2R T , C(O)R U , and C(O)NR V R W ; 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 of R is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl; R J and R K or R V and R W or R Q and R R They may also form a substituted or unsubstituted 5-, 6-, 7- or 8-membered ring together with the nitrogen atom to which they are attached; R A , R B and / or R C can form fused bi- or tricyclic ring systems with the benzene rings to which they are attached; X 1 selected from —CR X R Y —, —NR Z C(O)—, —NR Z C(O)CR X R Y ——OC(O)—, —SC(O)—, —C(O)NR 1A —, —C(O)O—, —(O)CS—, —NR 1A S(O)—, —S(O)NR 1A —, —NR 1A C(O)NR 1A —, —S(O)— and —S(O)2—; R X , R Y , R Z , and R 1A Each of which is independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl; R D and R E each independently selected from H, D, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl; or R D and R E Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted C3-C6 cycloalkyl or a substituted or unsubstituted heterocyclyl; or R D and R Z Together with the carbon atom to which they are attached or with -NC(O)-, they form an optionally substituted 5- to 8-membered lactam; R F and R G N + together to form optionally substituted, except N + In addition to the heterocyclic ring, there is one or more nitrogen atoms, or, R F and R G are independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted C3-C6 cycloalkyl; and R H is a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group.

2. The compound according to claim 1, wherein Y - It is iodide, bromide or chloride.

3. The compound according to claim 1, wherein X 1 It is —NHC(O)—.

4. The compound according to claim 1, wherein R H is replaced by C 6-10 Aryl or substituted C 5-10 Heteroaryl, which is optionally substituted by C 1-6 Alkanes, C 3-6 The alkylene group may be substituted with cycloalkyl, heterocyclyl, phenyl, substituted phenyl, heteroaryl, substituted heteroaryl, hydroxy, amide, ester, sulfonamide, urea, nitrile or halogen.

5. The compound according to claim 1, wherein R H It is C 6-10 aryl or 5-membered to 10-membered heteroaryl, which is optionally substituted or unsubstituted C 1-6 The moiety can be substituted by alkyl, halogen, nitrile, hydroxyl and alkoxy.

6. The compound according to claim 1, wherein Each R A and R B are independently selected from H, halogen, C 1-4 Alkyl, and NR J R K ; R J and R K Each of the 1-4 alkyl; and R C is H, halogen, C 1-4 Alkyl, or NR J R K .

7. The compound according to claim 1, wherein R A , R B , and R C Independently selected from H, D, halogen, OR I , substituted or unsubstituted C 1- C4 alkyl, and NR J R K ;in, R I , R J and R K Each of which is independently selected from H and substituted or unsubstituted C 1- C4 alkyl.

8. The compound according to claim 1, wherein R A and R B Each of is CH3, and R C Selected from the group consisting of H, CH3, halogen, nitrile, methoxy, and ethoxy.

9. The compound according to claim 1, wherein R D It is C 1-4 Alkyl, which is optionally substituted by halogen, oxygen, C 3-8 cycloalkyl, aryl or heteroaryl, and wherein R E H, D or C 1-4 alkyl.

10. The compound according to claim 1, wherein R D and R E All are hydrogen.

11. The compound according to claim 1, wherein R D is hydrogen and R E It is an alkyl group.

12. The compound according to claim 1, wherein R D and R E Together with the carbon source to which they are attached they form a C3-C6 cycloalkyl group.

13. The compound according to claim 1, wherein The R F and R G N + Together they form a substituted or unsubstituted five-, six-, seven- or eight-membered heterocyclyl ring.

14. The compound according to claim 1, wherein R F and R G are independently selected from unsubstituted C1-C4 alkyl groups.

15. The compound according to claim 1, wherein The compound is selected from the following table:

16. The compound according to claim 1, wherein The compound is selected from the following table:

17. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

18. The composition according to claim 16, wherein The compositions are formulated for oral, intravenous, intramuscular, rectal, cutaneous, subcutaneous, topical, transdermal, sublingual, nasal, inhalation, vaginal, intrathecal, epidural, or intraocular administration.

19. A method for treating pain, cough, pruritus or a neurogenic inflammatory disorder in a patient, the method comprising administering to the patient an effective amount of a compound of claim 1.

20. The method according to claim 19, wherein: The pain is selected from the group consisting of back and neck pain, lower back pain, cancer pain, gynecological and labor pain, 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 anus syndrome, rectal pain, perianal pain, hemorrhoidal pain, stomach pain, ulcers, inflammatory bowel disease, irritable bowel syndrome, irritable bowel syndrome, oral mucositis, esophagitis, interstitial cystitis, Cystitis, urethritis and other urinary pain, toothache, burning pain, headache, eye irritation, conjunctivitis (such as allergic conjunctivitis), eye redness, dry eyes, dry eyes (chronic eye pain), complex regional pain syndrome, acute postoperative pain, postoperative pain, postoperative eye pain, and procedural pain (i.e. pain associated with injections, abscess drainage, surgery, dental surgery, ocular surgery, eye irritation, conjunctivitis (such as allergic conjunctivitis)), eye redness, dry eyes, arthroscopy and other medical device use, cosmetic surgery, dermatologic surgery, fixation of fractures, biopsies, etc.).

21. The method according to claim 19, wherein: The cough is selected from the group consisting of asthma, COPD, asthma-COPD overlap syndrome (ACOS), interstitial pulmonary fibrosis (IPF), idiopathic pulmonary fibrosis, post-viral cough, post-infectious cough, chronic idiopathic cough and lung cancer.

22. The method according to claim 19, wherein: The pruritus is selected from the group consisting of pruritus, brachioradial pruritus, chronic idiopathic pruritus, genital / anal pruritus, paresthesia, scalp pruritus, allergic dermatitis, contact dermatitis, atopic dermatitis, hand eczema, poison ivy, infection, parasites, insect bites, pregnancy, metabolic disorders, liver or kidney failure, drug reaction, allergic reaction, eczema, genital and anal pruritus, hemorrhoidal pruritus, and cancer.

23. The method according to claim 19, wherein: The neurogenic inflammatory disease 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, eczema, cystitis, gastritis, migraine, rosacea, sunburn, pancreatitis, chronic rhinosinusitis, traumatic brain injury, polymicrobial sepsis, tendinopathy, chronic urticaria, rheumatic disease, acute lung injury, exposure to irritants, inhaled irritants, pollutants, chemical warfare agents, and atopic dermatitis.

24. The method according to claim 19, wherein: The compound of formula (I) is used in combination with one or more exogenous macroporous receptor antagonists.

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