MAGL inhibitors, methods for producing them and their uses

Abstract

The present invention is in the field of medicine and relates to the compound of formula I as a MAGL inhibitor JPEG2022546958000066.jpg40170 and pharmaceutically acceptable salts thereof, as well as methods for producing them, uses thereof, and intermediates for producing them.

Description

[Technical Field]

[0001]

[0001] The present invention relates to the field of medicines, in particular to MAGL inhibitors, and their preparation methods and uses. [Background technology]

[0002]

[0002] Monoacylglycerol lipase (MAGL), a serine hydrolase that promotes the breakdown of lipids into glycerol and fatty acids, is highly expressed in human invasive and primary tumor cells. High levels of MAGL can maintain the high pathogenicity of tumor cells by increasing the levels of free fatty acids. MAGL has become an important target for the development of antitumor drugs. Summary of the Invention

[0003] The present invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof: JPEG0007749540000001.jpg43170, where R is H or C 1-5 and the C 1-5 The alkyl in the formula is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, neobutyl, n-pentyl, isopentyl, neopentyl, cyclopentyl, or halogen, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 3-8 Cycloalkyl, C 3-8 Heterocyclyl, C 3-8 C substituted with cycloalkoxy, aryl, or heteroaryl 1-5 wherein the number of R is 0, 1, 2, 3, or 4; Ar1 and Ar2 are each C 5-18 aryl, heteroaryl, (C 6-10 Aryl)-C 1-4 Alkyl, (C 6-10 Aryl)-C1-4 Alkyl-(C 6-10 (aryl), (5- to 10-membered heteroaryl)-C 1-4 Alkyl-(C 6-10 aryl), (C 6-10 Aryl)-(C 6-10 aryl), (C 6-10 aryl)-O-(C 6-10 aryl), (C 6-10 (aryl)-O-(5- to 10-membered heteroaryl), (5- to 10-membered heteroaryl)-O-(5- to 10-membered heteroaryl), (5- to 10-membered heteroaryl)-(5- to 10-membered heteroaryl), and the substitution position is optional; X1 and X2 are H, hydroxyl, carboxyl, halogen, acyloxy, C 1-3 Alkyl, C 1-3 Alkoxy, C 3-8 Cycloalkyl, C 3-8 Heterocyclyl, C 3-8 Cycloalkoxy, aryl, heteroaryl, or C substituted with hydroxy, carboxyl, halogen, or alkyl 1-3 Alkyl, C 1-3 Alkoxy, C 3-8 Cycloalkyl, C 3-8 Heterocyclyl, C 3-8 The substituent is independently selected from one or more of cycloalkoxy, aryl, and heteroaryl, and the position of the substituent is optional.

[0004] In some embodiments, Ar and Ar are each phenyl, C 5-18 aryl, heteroaryl, (C 6-10 Aryl)-(C 6-10 aryl), (C 6-10 aryl)-O-(C 6-10 aryl), and X1 and X2 are independently selected from H, hydroxyl, halogen, C 1-3 Alkyl, C 1-3 The substituents are independently selected from one or more of alkoxy, and the positions of the substituents are arbitrary.

[0005] In some embodiments, Ar1 and Ar2 are each independently selected from one or more substituents of phenyl, naphthyl, diphenylether, indole, and biphenyl, at any position.

[0006] A compound of formula I above, or a pharmaceutically acceptable salt thereof, wherein R is H and the corresponding structural formula is JPEG0007749540000002.jpg44170, where Ar1 and Ar2 are C 5-18 aryl, heteroaryl, (C 6-10 Aryl)-C 1-4 Alkyl, (C 6-10 Aryl)-C 1-4 Alkyl-(C 6-10 (aryl), (5- to 10-membered heteroaryl)-C 1-4 Alkyl-(C 6-10 aryl), (C 6-10 Aryl)-(C 6-10 aryl), (C 6-10 aryl)-O-(C 6-10 aryl), (C 6-10 (aryl)-O-(5- to 10-membered heteroaryl), (5- to 10-membered heteroaryl)-O-(5- to 10-membered heteroaryl), (5- to 10-membered heteroaryl)-(5- to 10-membered heteroaryl), and the substitution position is optional; X1 and X2 are H, hydroxyl, carboxyl, halogen, acyloxy, C 1-3 Alkyl, C 1-3 Alkoxy, C 3-8 Cycloalkyl, C 3-8 Heterocyclyl, C 3-8 Cycloalkoxy, aryl, heteroaryl, or C substituted with hydroxy, carboxyl, halogen, or alkyl 1-3 Alkyl, C 1-3 Alkoxy, C 3-8 Cycloalkyl, C 3-8 Heterocyclyl, C 3-8 The substituent is independently selected from one or more of cycloalkoxy, aryl, and heteroaryl, and the position of the substituent is optional.

[0007] Specifically, Ar1 and Ar2 are each C 5-18 aryl, heteroaryl, (C 6-10 Aryl)-C 1-4 Alkyl, (C 6-10 Aryl)-C 1-4 Alkyl-(C 6-10 (aryl), (5- to 10-membered heteroaryl)-C 1-4 Alkyl-(C 6-10 aryl), (C 6-10 Aryl)-(C 6-10 aryl), (C 6-10 aryl)-O-(C 6-10 aryl), (C 6-10 (aryl)-O-(5- to 10-membered heteroaryl), (5- to 10-membered heteroaryl)-O-(5- to 10-membered heteroaryl), (5- to 10-membered heteroaryl)-(5- to 10-membered heteroaryl), and the substitution position is optional; X1 and X2 are H, hydroxyl, halogen, or C, respectively. 1-3 Alkyl, C 1-3 Alkoxy, C 3-8 Heterocyclyl, C 3-8 Cycloalkoxy or C substituted with hydroxyl, carboxyl, halogen, or alkyl 1-3 Alkyl, C 1-3 The substituents are independently selected from one or more of alkoxy, and the positions of the substituents are arbitrary.

[0008] In some embodiments, Ar is one or more of phenyl, naphthyl, diphenylether, biphenyl, optionally substituted; X1 is one or more of H, methoxy, hydroxyl, fluorine, and chlorine, and the substituents may be positioned arbitrarily; Ar2 is phenyl; X2 is one or more of H, methyl, dimethyl, methoxy, hydroxyl, fluorine, and chlorine, and the position of the substituent is optional.

[0009] In a particular embodiment, the substitution position of Ar1 is at the meta position.

[0010] In some embodiments, Ar1 is one or more of benzofuran, benzimidazolyl, benzoxazolyl, indolyl, benzothienyl, quinolinyl, isoquinolinyl, and purinyl, optionally substituted; X1 is one or more of H, methyl, methoxy, hydroxyl, fluorine, chlorine, and bromine, and the substituents may be in any position; Ar2 is phenyl, and the substituents are optionally positioned; X2 is one or more of H, methyl, dimethyl, methoxy, hydroxyl, fluorine, and chlorine, and the position of the substituent is optional.

[0011] In some embodiments, Formula 1 above may be limited to Formula III, As shown in JPEG0007749540000003.jpg48170, wherein R1 is selected from hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, and methoxy, and R2 is selected from hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy, phenyl, and aryl groups substituted with hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, and methoxy. The numbers of R1 and R2 are 0, 1, 2, 3, or 4, and the bonding mode between R1 and R2 and the mother nucleus is not limited to a single bond connection.

[0012] The present invention relates to a compound of formula b JPEG0007749540000004.jpg36170, which is used to synthesize a compound of Formula I, wherein Ar2 and X2 are as described in Formula 1.

[0013] In some embodiments, Segment A within the dashed line in JPEG0007749540000005.jpg40170 is Replaced by JPEG0007749540000006.jpg21170.

[0014] The present invention provides a method for synthesizing a compound of formula I, the specific steps of which are: JPEG0007749540000007.jpg44170, and the salt of b or b JPEG0007749540000008.jpg16170 to obtain I, wherein Ar1 and X1 are as described in Formula 1.

[0015] The b or salt of b JPEG0007749540000009.jpg16170 to obtain I, and adding a condensing agent and a base in this step of the reaction helps the reaction, and the condensing agent is selected from HBTU, DMC, HOBT, HOBT / EDCI, HATU, HATU / DIEPA, DCC, CDI, and isopropyl chloroformate, whose structural formulas are as follows:

[0016] The b or salt of b is obtained by deprotection of a, JPEG0007749540000010.jpg51170. The base is an organic base such as N,N-diisopropylethylamine (DIPEA), diethylamine (DEA), or triethylamine (TEA).

[0017] The a or salt of a is sm JPEG0007749540000011.jpg16170 or a salt thereof, JPEG0007749540000012.jpg51170.

[0018] Here, the preparation of sm can be obtained by the prior art, or can be synthesized by a person skilled in the art through a limited number of experiments based on the prior art without creative efforts. In the reaction of this step, adding a condensing agent and a base helps the reaction, and the condensing agent is selected from HBTU, DMC, HOBT, HOBT / EDCI, HATU, HATU / DIEPA, DCC, CDI, and isopropyl chloroformate.

[0019] In some particular embodiments, the following method can be used.

[0020] JPEG0007749540000013.jpg80170

[0021] Here, the reducing agent is preferably at least one of NaBH4, KBH4, and NaBH4 / LiCl.

[0022] Unless otherwise specified, the solvents selected for the reactions in the above or following steps of the present invention are conventional solvents in this field. The principle of selection is to dissolve the reactants but not participate in the reaction, extract the product, or crystallize the corresponding product and separate impurities from it. Examples include water, halogenated alkanes, alkylamines, aliphatic hydrocarbons, esters, alcohols, aromatic hydrocarbons, ethers, and heterocyclic solvents. These solvents are particularly selected from, but not limited to, methanol, ethanol, propanol, isopropanol, diethyl ether, ethyl acetate, acetic acid, cyclohexane, dichloromethane, chloroform, tetrahydrofuran, pyridine, diethylamine, triethylamine, dimethylformamide, toluene, and mixtures of at least two thereof.

[0023] The numbers a, b, etc. used in the present invention are numbers used for conveniently explaining the general formula, and they can be converted to other numbers such as 1, 2, 3, etc. in specific examples. These are all for the convenience of explanation, do not affect the structural formula and its reaction scheme, and essentially belong to the expression of the general formula and general reaction scheme. Those skilled in the art can determine that the substituents of each intermediate in all the above synthetic routes are all determined according to the structure of the target compound.

[0024] Chiral isomers or cis-trans isomers and mixtures of isomers in any ratio among the compounds included in the general formula of the target compound of general formula I and representatives of specific substances thereof are also included within the scope of the compounds included in the general formula of the target compound of general formula I and representatives of specific substances thereof.

[0025] Regarding "any substitution position," it is understood that any selection can be made when the valence bond is appropriate. Substitution on the benzene ring may be at one or more of the peri-, meta-, and para-positions, and substitution in the aromatic heterocycle may be at one or more of the 2-, 3-, 4-, 5-, etc. positions. With regard to the selection of substituents, "one or more" and "one or more" all refer to the number of selected substituents, specific numbers being 1, 2, 3, 4, etc. Specifically, as in "Ar1 is phenyl, the substitution position is optional, and X1 is one or more of fluorine and chlorine, and the substitution position is optional," this includes the peri- (two), meta- (two), and para- (one) positions of phenyl being substituted with one Cl, two Cl, and three Cl, respectively, or the two meta positions being substituted with one Cl and one F, respectively, or the para position being substituted with one Cl and one F, and similar inferences are included.

[0026] Unless otherwise specified, the solvents selected for the reactions in the above or following steps of the present invention are conventional solvents in this field. The principle of selection is to dissolve the reactants but not participate in the reaction, extract the product, or crystallize the corresponding product and separate impurities from it. Examples include water, halogenated alkanes, alkylamines, aliphatic hydrocarbons, esters, alcohols, aromatic hydrocarbons, ethers, and heterocyclic solvents. These solvents are particularly selected from, but not limited to, methanol, ethanol, propanol, isopropanol, diethyl ether, ethyl acetate, acetic acid, cyclohexane, dichloromethane, chloroform, tetrahydrofuran, pyridine, diethylamine, triethylamine, dimethylformamide, toluene, and mixtures of at least two thereof.

[0027] Unless otherwise specified, in each of the reactions described above or below in the present invention, if there is an excess of reactant, the reaction can be quenched by adding a substance capable of reacting with the excess reactant. For example, in some embodiments, water quenching or saturated ammonium chloride quenching can be used.

[0028] Unless otherwise specified, in the above or below reactions of the present invention, the purification method of the product in each step of the reaction is selected from extraction, crystallization, solvent removal, and column chromatography, all of which operations are conventional techniques in the art and can be handled by those skilled in the art according to specific circumstances.

[0029] The general formula and synthetic method of the general formula of the present invention can derive those that are not limited to these specific substances, and under the guidance of the general formula and synthetic method of the general formula of the present invention, those skilled in the art can obtain specific compounds without creative efforts, all of which are within the scope of the present invention.

[0030] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the described embodiments of the invention may be used in practicing the invention. The following claims are intended to define the scope of the invention, and methods and structures within the scope of these claims and their equivalents are intended to be covered thereby.

[0031] The present invention provides pharmaceutical compositions comprising the above compounds, i.e., compounds covered by the general formula of the I target compounds and their specific substance representatives, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable pharmaceutical excipients.

[0032] The present application provides a pharmaceutical composition comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The pharmaceutical composition may be in the form of an oral dosage form, a parenteral dosage form, an external dosage form, or a rectal dosage form, but is not limited to these. The composition may be in the form of a liquid, solid, semisolid, gel, or aerosol. In some embodiments, the pharmaceutical composition may be an oral tablet, capsule, pill, powder, sustained-release formulation, solution or suspension, a sterile solution, suspension, or emulsion for parenteral injection, an ointment or cream for external use, or a suppository for rectal administration. In other embodiments, the pharmaceutical composition is in a unit dosage form suitable for single administration of a precise dose.

[0033] The present invention provides all of the above compounds or pharmaceutically acceptable salts thereof and pharmaceutical compositions thereof, as well as their use as monoacylglycerol esterase inhibitors or for the manufacture of a medicament for the treatment of a disease or disorder having a pathology characteristic of the monoacylglycerol esterase metabolic pathway.

[0034] The present invention provides a method for inhibiting monoacylglycerol esterase using any of the above compounds or their pharmaceutically acceptable salts and pharmaceutical compositions thereof. The method includes in vivo and in vitro methods for inhibiting monoacylglycerol esterase. Furthermore, the present invention also provides a method for treating diseases or disorders mediated by monoacylglycerol esterase using any of the above compounds or their pharmaceutically acceptable salts and pharmaceutical compositions thereof.

[0035] The disorders include metabolic disorders (e.g., obesity), kidney diseases (e.g., acute inflammatory kidney disease and diabetic kidney disease), vomiting or vomiting (e.g., chemically induced vomiting), nausea (e.g., refractory nausea or chemically induced nausea), eating disorders (e.g., anorexia or bulimia), neuropathy (e.g., diabetic neuropathy, pellagranulopathy, alcoholic neuropathy, velibellineuropathy), neurodegenerative disorders (e.g., multiple sclerosis (MS), Kinson's disease (PD), Huntington's disease, dementia, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), etc. epilepsy, frontotemporal dementia, sleep disorders, Creutzfeldt-Jakob disease (CJD) or prion disease], schizophrenia, depression, bipolar disorder, tremors, movement disorders, dystonia, convulsions, Tourette's syndrome, withdrawal syndromes [alcohol withdrawal syndrome, antidepressant discontinuation syndrome, antipsychotic discontinuation syndrome, benzodiazepine withdrawal syndrome, marijuana withdrawal, neonatal withdrawal, nicotine withdrawal, or opioid withdrawal], traumatic brain injury, non-traumatic brain injury, spinal cord injury, seizures, disorders associated with abnormal cell growth or proliferation [e.g., benign tumors or cancers, e.g., benign skin tumors, brain tumors, papillomas, prostate tumor, brain tumor (glioblastoma, medulloepithelioma, medulloblastoma, neuroblastoma, astrocytoma, astroblastoma, ependymoma, oligodendroglioma, plexus tumor, neuroepithelial tumor, epiphyseal tumor, ependymoblastoma, malignant meningioma, sarcomatosis, melanoma, schwannoma), melanoma, metastatic tumor, kidney cancer, bladder cancer, brain cancer, glioblastoma (GBM), gastrointestinal cancer, leukemia, or blood cancer), inflammatory diseases (e.g., appendicitis, bursitis, colitis, irritable bowel syndrome, ulcerative colitis, cystitis, dermatitis, phlebitis, rhinitis, tendonitis, tonsillitis, vasculitis, prurigo vulgaris, chronic prostatitis, glomerulonephritis, hypersensitivity, IBS, pelvic inflammatory diseases, sarcoidosis, HIV encephalitis, rabies, brain abscess, neuroinflammation, central nervous system (CNS) inflammation], immune system disorders (e.g., transplant rejection or celiac disease), post-traumatic stress disorder (PTSD), acute stress disorder, panic disorder, substance-induced anxiety, obsessive-compulsive disorder (OCD), agoraphobia, specific phobia, social phobia, anxiety disorder, attention deficit disorder (ADD), attention deficit hyperactivity disorder (ADHD), pain [e.g., acute pain, chronic pain, inflammatory pain, visceral pain, post-operative pain, migraine, back pain, joint pain, abdominal pain, chest pain, post-mastectomy pain syndrome, menstrual pain,Pain caused by disorders selected from endometriosis pain, pain due to physical trauma, headache, sinusitis, tension headache, arachnoiditis, herpes virus pain, diabetic pain, osteoarthritis, rheumatoid arthritis, spondylitis, gout, labor pain, musculoskeletal disorders, skin diseases, toothache, carditis, burns, sunburn, snakebite, venomous snakebite, spider bite, insect bite, neurogenic bladder, interstitial cystitis, urinary tract infection (UTI), rhinitis, contact dermatitis / hypersensitivity, itch, eczema, pharyngitis, mucositis, enteritis, irritable bowel syndrome (IBS), cholecystitis, and pancreatitis, neuropathic pain (e.g., neuropathic lower back pain, complex regional pain syndrome, columnar trigeminal neuralgia, causalgia, toxic neuropathy, reflex sympathetic dystrophy, diabetic neuropathy, chemosis, etc.) chronic neuropathy or sciatica due to chemotherapy), demyelinating diseases [e.g., multiple sclerosis (MS), Duke's disease, CNS neuropathy, central pontine myelinolysis, syphilitic myelopathy, leukoencephalopathy, leukodystrophy, Guillain-Barré syndrome, chronic inflammatory demyelinating polyneuropathy, anti-myelin-associated glycoprotein (MAG) peripheral neuropathy, Charcot-Marie-Tooth disease, peripheral neuropathy, myelopathy, optic neuropathy, progressive inflammatory neuropathy, optic neuritis, transverse myelitis], and cognitive disorders [e.g., cognitive impairment associated with Down syndrome, cognitive impairment associated with Alzheimer's disease, cognitive impairment associated with PD, mild cognitive impairment (MCI), dementia, post-chemotherapy cognitive impairment (PCCI), post-operative cognitive impairment (POCD)].

[0036] The present invention provides the preparation of MAGL inhibitors and the use of the drugs to treat diseases associated with MAGL inhibitors.

[0037] In some embodiments, the disorder associated with a MAGL inhibitor is pain, migraine, irritable bowel syndrome, or ulcerative colitis.

[0038] In the above applications, the pain is traumatic pain or pathological pain. Furthermore, the pathological pain is inflammatory pain or endogenous pain. Furthermore, it is biologically derived inflammatory pain, chemically derived inflammatory pain, hematogenous pain, immune derived pain, endocrine derived pain, pain caused by metabolic pathology, neurogenic pain, or cardiac pain.

[0039] In the above applications, the pain may be transient pain, acute pain, or chronic pain.

[0040] In the above applications, the pain is neurological pain, cardiovascular pain, hematological pain, respiratory pain, digestive pain, endocrine pain, urinary pain, locomotor pain, or immune pain.

[0041] For the above applications, the migraine headache is a common migraine headache, a typical migraine headache, or a cluster migraine headache.

[0042] In the above applications, the irritable bowel syndrome is constipation-predominant irritable bowel syndrome, diarrhea-predominant irritable bowel syndrome, and constipation-diarrhea irritable bowel syndrome.

[0043] The MAGL inhibitor of the present invention can reduce the number of body twists induced by acetic acid in mice, prolong the latency time, and increase the pain threshold of mice subjected to thermal stimulation of the pain response using a hot plate, demonstrating significant analgesic effects.

[0044] In some embodiments, the MAGL inhibitor of the present invention can significantly prolong the blood clotting time in a migraine model, increase the number of tail-suspension exercises, and significantly increase the level of 5-HT in brain tissue, thereby having a significant therapeutic effect on migraine.

[0045] The MAGL inhibitor of the present invention can significantly reduce the AWR score of irritable bowel syndrome model, and reduce the level of 5-HT in colon and serum.It is shown to have significant therapeutic effect on irritable bowel syndrome.Therefore, it is confirmed that this compound has the biological activity of preventing and treating irritable bowel syndrome.

[0046] The MAGL inhibitors of the present invention can significantly increase colon length, reduce the total number of bloody stools, and reduce the content of IL-1 in an ulcerative colitis model.

[0047] Specific Chemical Terms

[0048] Unless otherwise defined, all technical and scientific terms herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. Unless otherwise stated, all patents, patent applications, and published materials cited throughout this specification are incorporated herein by reference.

[0049] It is to be understood that the foregoing brief description and the following detailed description are exemplary and explanatory only and are not intended to limit the subject matter of the present application in any way. In this application, the use of the singular includes the plural unless otherwise stated. It should also be noted that the use of "or" means "and / or" unless otherwise stated. Additionally, the use of the term "comprises" and other forms such as "comprises," "includes," and "containing" are open-ended descriptions.

[0050] Definitions for standard chemical terms are given in the reference (Carey and Sundberg, "ADVANCED ORGANIC CHEMISTRY 4 TH ed." Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise explained, conventional methods within the skill of the art, such as mass spectrometry, NMR, IR and UV / Vis spectroscopy, and pharmacological methods, are used. Unless a specific definition is provided, terms used herein are known in the art, such as analytical chemistry, synthetic organic chemistry, and related descriptions of drugs and medicinal chemistry. Standard techniques are used for chemical syntheses, chemical analyses, drug production, formulation and delivery, and treatment of patients. For example, reactions and purifications are performed using instructions provided with manufacturer's kits or according to methods known in the art or described herein. The techniques and methods described above are generally performed according to conventional methods known in the art, based on the descriptions in several general and more specific references cited and discussed herein. As used herein, groups and substituents thereof may be selected by one skilled in the art to provide stable structural moieties and compounds.

[0051] The term "alkyl," as used herein, refers to a saturated straight- or branched-chain hydrocarbon. Exemplary alkyls include, but are not limited to, straight- or branched-chain hydrocarbons of 1 to 6, 1 to 4, or 1 to 3 carbon atoms, referred to herein as C alkyl, C alkyl, and C alkyl, respectively. Exemplary alkyls include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-butyl, 3-methyl-2-butyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, and the like.

[0052] C as described herein 1-4 Alkyl is C 1-3 Alkyl, C 1-2 Alkyl, C 2-3 Alkyl, and C 2-4 Specific examples include, but are not limited to, alkyl, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, and neobutyl.

[0053] Alkoxy is alkyloxy. 1-3 Alkoxy is C 1-2 Alkoxy and C 2-3 Specific examples include, but are not limited to, alkoxy, methoxy, ethoxy, propoxy, and isopropoxy.

[0054] The term "cycloalkyl" as used herein refers to a saturated or partially unsaturated hydrocarbon such as a monocyclic 5- to 8-membered ring system or a bicyclic 8- to 12-membered ring system. 3-8 Cycloalkyl contains 3 to 8, 3 to 6, 4 to 6, 6 to 8, 3 to 7, or 5 to 8 carbons, and is defined herein as C 3-8 Cycloalkyl, C 3-6 Cycloalkyl, or C 4-6and are simply different forms, but have the same meaning. Exemplary cycloalkyls include, but are not limited to, cyclohexyl, cyclopentyl, cyclopentenyl, cyclobutyl, or cyclopropyl.

[0055] The term "heterocyclyl" as used herein refers to a monocyclic 5-8 membered ring system or a bicyclic 8-12 membered ring system containing one or more heteroatoms (e.g., 1-3 heteroatoms, e.g., nitrogen, oxygen, and sulfur). 3-8 Heterocyclyl represents a monocyclic or polycyclic group having 3 to 8 atoms. 3-6 Heterocyclyl, C 4-8 Heterocyclyl, C 5-8 Heterocyclyl, C 4-7 Heterocyclyl, C 3-7 Heterocyclyl, C 6-8 Heterocyclyl includes at least one ring containing 1, 2, 3, or 4 ring heteroatoms selected from N, O, or S, and none of the rings is aromatic. Examples of heterocyclyl include, but are not limited to, morpholinyl, thiomorpholinyl, furanyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuran, and 1,3-dioxanyl.

[0056] C 3-8 Cycloalkoxy is cycloalkyloxy, wherein the cycloalkyl has the same meaning as cycloalkyl given above.

[0057] The term "aryl" as used herein refers to an aromatic mono- or polycarbocyclic group of 6 to 12 carbon atoms having at least one aromatic ring. 6-10 Aryl is C6-8 aryl, C 6-10 Aryl, C 6-9 Aryl, C 7-10 Aryl, C 8-10 Aryl, C6 Aryl, C7 Aryl, C8 Aryl, C9 Aryl, C 10aryl. Specific examples include, but are not limited to, phenyl, mono- or polyhydrocarbyl-substituted phenyl, cyclopentadienyl, indenyl, naphthyl, and chamomyl cyclohydrocarbyl. Here, the mono- or polyhydrocarbyl-substituted phenyl includes, but is not limited to, methylphenyl, ethylphenyl, dimethylphenyl, propylphenyl, isopropylphenyl, methylethylphenyl, trimethylphenyl, butylphenyl, methylpropylphenyl, methylisopropylphenyl, diethylphenyl, dimethylethylphenyl, tetramethylphenyl, tert-butylphenyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H-indenyl.

[0058] C as described herein 5-18 Aryl is C 6-10 Aryl, C 11-12 Aryl, C 13-15 Aryl, and C 16-18 Aryl includes, for example, pentylphenyl, methylbutylphenyl, methylisobutylphenyl, methyltert-butylphenyl, ethylpropylphenyl, ethylisopropylphenyl, methylmethylpropylphenyl, methylmethylisopropylphenyl, trimethylethylphenyl, pentamethylphenyl, and the like.

[0059] As used herein, a heteroaryl or heteroaromatic ring or "heteroaromatic group" refers to a monocyclic aromatic 5- to 8-membered ring system or a bicyclic aromatic 8- to 12-membered ring system containing one or more heteroatoms (e.g., 1 to 3 heteroatoms, e.g., nitrogen, oxygen, and sulfur). A 5- to 10-membered heteroaryl as used herein includes a 5- to 9-membered heteroaryl, a 5- to 8-membered heteroaryl, a 5- to 7-membered heteroaryl, a 5- to 6-membered heteroaryl, a 6- to 10-membered heteroaryl, a 6- to 9-membered heteroaryl, a 6- to 8-membered heteroaryl, a 7- to 9-membered heteroaryl, an 8- to 10-membered heteroaryl, a 9- to 10-membered heteroaryl, a 5-membered heteroaryl, a 6-membered heteroaryl, a 7-membered heteroaryl, an 8-membered heteroaryl, a 9-membered heteroaryl, and a 10-membered heteroaryl. Where possible, the heteroaryl ring can be attached to an adjacent group through a carbon or nitrogen atom. Examples include, but are not limited to, furanyl, thienyl, 1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, pyrimidinyl, pyrazinyl, 1,3,5-triazinyl, 1,2,4-triazinyl, 1,2,3-triazinyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl.Examples of 8-12 membered bicyclic heteroaryls are benzofuranyl, isobenzofuranyl, benzo[b]thienyl, benzo[c]thienyl, isoindolyl, benzo[d]isoxazolyl, benzo[c]isoxazolyl, benzo[d]oxazolyl, benzo[d]isothiazolyl, benzo[c]isothiazolyl, benzo[d]thiazolyl, benzo[d]imidazolyl, benzo[d]imidazolyl, and benzo[d][1,2,3]triazolyl, oxazolyl, thiazolyl, imidazolyl, pyrrolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,2,4-oxadiazol-5-yl, 1,2,4-oxadiazol-3-yl, 1,3 ,4-oxadiazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,2,4-thiadiazol-5-yl, 1,2,4-thiadiazol-3-yl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl, tetrazolyl, indolyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzisothiazolyl and imidazo[1,2-α]pyridyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, pyridyl, benzisoxazolyl.

[0060] Any reference herein to "one or more substituents of benzene, naphthalene, etc." or "one or more of benzene, naphthalene, etc." refers to one, two, or three of phenyl, naphthyl, etc.

[0061] The term "acyloxy" or "acyloxy" refers to the group -OC(=O)-R, where R is H, and substituted or unsubstituted C 1-5 alkyl, wherein the unsubstituted C 1-5 The alkyl in the substituted C is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, neobutyl, n-pentyl, isopentyl, neopentyl, and cyclopentyl. 1-5 Alkyl is halogen, hydroxyl, amino, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C3-8 Cycloalkyl, C 3-8 Heterocyclyl, C 3-8 C substituted with cycloalkoxy, aryl, or heteroaryl 1-5 In some embodiments, R is n-propyl or JPEG0007749540000014.jpg19170.

[0062] The term "halo" or "halogen" as used herein with respect to halogen refers to F, Cl, Br, or I.

[0063] Where a substituent is depicted in a conventional chemical formula written from left to right, the substituent also includes the chemically equivalent substituent that results when the structural formula is written from right to left, e.g., CHO is equivalent to OCH.

[0064] The term "compound" as used herein is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes. The compounds of the present application may be asymmetric, e.g., have one or more stereoisomers. Unless otherwise specified, all stereoisomers include enantiomers, diastereomers, and the like. Compounds of the present application containing asymmetrically substituted carbon atoms may be isolated in optically pure or racemic form. Optically pure forms may be isolated from racemic mixtures or synthesized using chiral starting materials or chiral reagents. The compounds of the present application also include tautomeric forms. Tautomeric forms result from the swapping of one single bond with the adjacent double bond, involving the migration of one proton. The compounds of the present application also include all isotopes of atoms, whether intermediates or final compounds. Atomic isotopes have the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. That is, the compounds of the present application include compounds in which some or all of the hydrogen (H) atoms are replaced with tritium (T) and / or deuterium (D), 12 Part or all of C 13 C and / or 14 C, and 14 N and 15 N, 18 O and 17 O.31 P and 32 P, 35 S and 36 The compounds described herein include compounds in which other isotopes (e.g., N, O, P, S) are substituted, such as S. The compounds described herein have one or more stereoisomeric centers, and each isomeric center can exist in the R or S configuration, or a combination thereof. Similarly, the compounds described herein have one or more double bonds, and each double bond can exist in the E (trans) or Z (cis) configuration, or a combination thereof. A particular stereoisomer, regioisomer, diastereomer, enantiomer, or epimer should be understood to include all possible isomers, such as stereoisomers, regioisomers, diastereomers, enantiomers, epimers, and mixtures thereof. Thus, the compounds described herein include all structurally distinct stereoisomeric, regioisomeric, diastereomeric, enantiomeric, or epimeric forms, and their corresponding mixtures. Techniques for converting specific stereoisomers or maintaining specific stereoisomers as they are, and for resolving mixtures of stereoisomers, are well known to those skilled in the art, and those skilled in the art can select an appropriate method depending on the particular situation. See, for example, Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; and Heller, Acc. Chem. Res. 1990, 23, 128.

[0065] The terms "optionally / optionally" or "optionally / optionally" refer to the occurrence or non-occurrence of a subsequently described event or circumstance, and the description includes both the occurrence and non-occurrence of said event or circumstance.

[0066] Specific pharmaceutical terms

[0067] As used herein, the term "treatment" and other similar synonyms include alleviating, alleviating, or ameliorating the symptoms of a disease or disorder, such as halting the onset of a disease or disorder, alleviating a disease or disorder, ameliorating a disease or disorder, alleviating symptoms caused by a disease or disorder, or halting symptoms of a disease or disorder, preventing other symptoms, or improving or preventing the underlying metabolic causes of the symptoms. Furthermore, the term also includes prophylactic purposes. The term also includes achieving a therapeutic effect and / or a prophylactic effect. The therapeutic effect refers to curing or ameliorating the underlying disease being treated. Furthermore, curing or ameliorating one or more physiological symptoms associated with an underlying disease is also a therapeutic effect, for example, an improvement in the patient's condition is observed, although the patient may still be affected by the underlying disease. From the perspective of a prophylactic effect, the composition can be administered to a patient at risk of developing a particular disease, or to a patient who exhibits one or more physiological symptoms of the disease, even if the disease has not been diagnosed.

[0068] As used herein, the terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" refer to an amount of at least one active agent (e.g., a compound of the present application) sufficient to alleviate to some extent one or more symptoms of the disease or disorder being treated after administration. The result may be a reduction and / or alleviation of signs, symptoms, pathogenesis, or any other desired change in a biological system. For example, a therapeutically "effective amount" refers to the amount of a composition comprising a compound disclosed herein required to provide a clinically significant disorder-alleviating effect. An appropriate effective amount for any individual case may be determined using techniques such as a dose escalation study.

[0069] As used herein, the terms "dosing," "administration," "administering," and the like refer to the method that can deliver a compound or composition to the desired site for biological action.These methods include, but are not limited to, oral route, duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical application, and rectal administration.Those skilled in the art are familiar with the administration techniques that can be used in the compounds and methods described herein, and are described, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current edition; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

[0070] The term "acceptable" as used herein with respect to a formulation, composition, or ingredient means that there are no long-term adverse effects on the general well-being of the subject being treated.

[0071] As used herein, the term "pharmaceutically acceptable" refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compound of the present application and is non-toxic, i.e., the substance can be administered to an individual and does not produce an adverse biological response or interact in an undesirable way with any of the components included in the composition.

[0072] As used herein, the term "pharmaceutical composition" refers to a mixture of a compound of the present application and at least one pharmaceutically acceptable substance, including, but not limited to, a carrier, stabilizer, diluent, dispersant, suspending agent, thickener, and / or excipient.

[0073] As used herein, the term "carrier" refers to a non-toxic substance, which helps to introduce the compounds of the present application into cells or tissues.

[0074] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acid and free base of the specified compound and does not cause any adverse biological or other effects. The compounds of the present application also include pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to a form in which a basic group in the parent compound is converted into a salt. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of a basic group, such as an amine (amino) group. The pharmaceutically acceptable salts of the present application are synthesized from the parent compound, i.e., by reacting the basic group in the parent compound with 1 to 4 equivalents of an acid in a solvent system. Suitable salts are described in Remington's Pharmaceutical Sciences, 17 th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0075] Unless otherwise specified, the term "salt" in this application refers to acidic salts formed with organic acids / inorganic acids and basic salts formed with organic bases / inorganic bases. In addition, when the basic functional group of the compound of the general formula is pyridine or imidazole (but is not limited to pyridine or imidazole) and the acidic functional group is carboxylic acid (but is not limited to carboxylic acid), a zwitterion (inner salt) is formed, and the inner salt is also included in the salt in this application. DETAILED DESCRIPTION OF THE INVENTION

[0004]

[0076] The specific compounds represented by the present invention are all synthesized by the synthetic methods of the general formulas disclosed in the present invention. The present invention will be further described below with reference to specific embodiments, but the present invention is not limited thereto. Furthermore, under the guidance of the general formulas, synthetic methods of the general formulas, and specific embodiments of the present invention, those skilled in the art can obtain other specific compounds without creative effort, all of which are within the scope of the present invention.

[0077] Example 1 Composition of JPEG0007749540000015.jpg40170

[0078] Preparation of intermediate a:

[0079] N-Boc-4-piperidinecarboxylic acid (458.6 mg, 2 mmol, 1 eq) was dissolved in 15 mL of DCM, DIPEA (0.7 mL, 4 mmol, 2 eq) was added, and HATU (912 mg, 2.4 mmol, 1.2 eq) was added. The mixture was stirred at room temperature for 10 min. Aniline (0.365 mL, 4 mmol, 2 eq) was added, and the mixture was stirred at room temperature for 12 h. The mixture was washed twice with water and twice with 0.1 M dilute hydrochloric acid. The solvent was removed by distillation under reduced pressure to give intermediate a without further purification.

[0080] Preparation of intermediate b:

[0081] Dissolve intermediate a in 10 ml of ethyl acetate, add 2 ml of HCl / EA solution, stir at room temperature for 24 h, precipitate a white solid, filter with suction, and wash with ethyl acetate to obtain 347.5 mg of white solid as intermediate b (72.4%).

[0082] MAGLZ-II-01 Production:

[0083] Intermediate b (120 mg, 0.5 mmol, 1.5 eq) was dissolved in 10 ml of DMF, TEA (0.23 ml, 1.65 mmol, 5 eq) was added, and the mixture was stirred at room temperature for 30 min. HATU (152 mg, 0.4 mmol, 1.2 eq) and 4-phenoxybenzoic acid (71.4 mg, 0.33 mmol, 1 eq) were added, and the mixture was stirred at room temperature for 2 h. After the reaction was complete, saturated sodium chloride solution was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed three times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether / ethyl acetate = 1:1) to obtain 102.4 mg of the target compound as a pale yellow solid (76.8%).

[0084] 1 HNMR (CDCl3) δ (ppm): 7.80-7.88 (m, 1H), 7.47-7.54 (m, 2H), 7.29-7.39 (m, 5H), 6.96-7.19 (m, 6H), 2.79-3.05 (m, 2H), 2.45-2.58 (m, 1H), 1.65-2.06 (m, 6H). MS results correspond to theoretical values.

[0085] The preparation processes in Examples 2 to 23 are similar to those in Example 1, based on the synthesis method of the general formula, where b in Examples 22 and 23 is The MS results for each target product are all equivalent to the theoretical values, and the NMR results are as follows:

[0086] JPEG0007749540000017.jpg214170JPEG0007749540000018.jpg213170JPEG0007749540000019.jpg225170 JPEG0007749540000020.jpg249170JPEG0007749540000021.jpg193170JPEG0007749540000022.jpg140170

[0087] The description of the synthetic target products in Examples 24 to 30 is based on the following structural formulas:

[0088] JPEG0007749540000023.jpg35170, wherein R1 is selected from hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, and methoxy, and R2 is selected from hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy, phenyl, and aryl groups substituted with hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, and methoxy. The numbers of R1 and R2 are 0, 1, 2, 3, or 4, and the bonding mode between R1 and R2 and the mother nucleus is not limited to a single bond.

[0089] JPEG0007749540000024.jpg244170JPEG0007749540000025.jpg248170JPEG0007749540000026.jpg181170

[0090] Example 34 Preparation of MAGLZ-II-11F: JPEG0007749540000027.jpg35170

[0091] The raw material N-Boc-4-piperidinecarboxylic acid (20) (229.3 mg, 1 mmol) was dissolved in 10 mL of CHCl, DIPEA (0.35 mL, 2 mmol) and HATU (456 mg, 1.2 mmol) were added, and the mixture was stirred at room temperature for 10 min. 3-Chloroaniline (0.21 mL, 2 mmol) was added, and the mixture was stirred at room temperature for 6 h. The mixture was washed three times with water and three times with 10% dilute hydrochloric acid, dried over anhydrous NaSO, and rotary evaporated to remove CHCl to obtain intermediate 21h without further purification.

[0092] Intermediate 21h was dissolved in 10 mL of EtOAc, and 4 M HCl / EtOAc solution (2 mL) was added. The mixture was stirred at room temperature for 12 h. A white solid precipitated and was filtered off with suction. The filter cake was washed with EtOAc to give 188.1 mg of a white solid as intermediate 22, with a yield of 68.4%.

[0093] JPEG0007749540000028.jpg30170

[0094] Intermediate 24 (62.5 mg, 0.3 mmol) was dissolved in anhydrous DMF, and HATU (136.8 mg, 0.36 mmol) and TEA (151.8 mg, 1.5 mmol) were added. The mixture was stirred at room temperature for 30 min. Then, hydrochloride intermediate 22h (107.33 mg, 0.39 mmol) was added and the mixture was stirred at room temperature for 12 h. The reaction was quenched by adding saturated brine and extracting with EtOAc three times. The combined organic phases were washed with saturated brine three times, dried over anhydrous Na2SO4, and purified by column chromatography (petroleum ether:ethyl acetate=1:1) to obtain 94.9 mg of a yellow solid, with a yield of 74.1%.

[0095] Characterization data:

[0096] 1H NMR(300MHz,CDCl3)δ(ppm):7.65(s,1H),7.40-7.45(t,1H),7.34-7.36(d,2H),7.23-7.28(d,1H),7.14-7.16(m,2H),7.07-7.10(m,1H) ,2.96(s,1H),2.88(s,1H),2.52-2.58(t,2H),2.04(s,1H),1.74-1.86(m,2H),1.59(s,4H),1.24-1.28(m,3H),1.02-1.07(t,3H);[M+Na] + :451.2.

[0097] Example 35 Synthesis of MAGLZ-II-11J JPEG0007749540000029.jpg53170

[0098] The starting material, N-Boc-L-tyrosine (0.5 mmol, 140.5 mg), was dissolved in 10 mL of anhydrous CHCl. ​​Intermediate II-11 (179.5 mg, 0.5 mmol) and DMAP (0.1 mmol, 12.2 mg) were added. The reaction mixture was cooled to 0 °C, DCC (0.55 mmol, 113.4 mg) dissolved in 3 mL of anhydrous CHCl was added, and the mixture was stirred for 10 min. The reaction temperature was raised to room temperature, and the mixture was stirred at room temperature for 12 h. If the reaction was incomplete, the reaction was stopped by suction filtration, the filtrate was spun down, and the mixture was washed three times with saturated brine and dried over anhydrous sodium sulfate. The mixture was purified by column chromatography (dichloromethane:methanol = 100:1) to give 52.6 mg of intermediate 25 as a white solid, in a 16.9% yield. JPEG0007749540000030.jpg29170

[0099] Intermediate 25 (37 mg) was dissolved in 10 mL of EtOAc, and 4 M HCl / EtOAc solution (2 mL) was added. The mixture was stirred at room temperature for 12 h. A white solid precipitated, which was suction filtered and washed with EtOAc to obtain a filter cake. 10 mL of sodium bicarbonate was added to the filter cake, and extracted three times with EtOAc. The organic phases were combined, washed three times with saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated to remove EtOAc to obtain 12 mg of a pale white solid, with a yield of 40%.

[0100] MAGLZ-II-11J's special features: 1 H NMR(300MHz,DMSO-d6)δ(ppm):11.95(s,1H),10.14(s,1H),7.83(s,1H),7.54-7.56(d,1H ),7.48-7.51(t,1H),7.44-7.46(d,1H),7.28-7.34(m,2H),7.10-7.12(m,2H),7.04-7.07( d,2H),6.69-6.71(d,2H),4.40-4.49(m,1H),4.26-4.30(t,1H),4.00-4.06(m,2H),3.61( s,1H),3.07-3.14(d,2H),2.89-2.98(d,2H),2.60-2.65(m,1H),1.58-1.78(m,4H);[M+Na] + 544.17.

[0101] Example 36: MAGL enzyme analysis

[0102] Cayman's MAGL inhibitor reagent cartridge was selected according to the manufacturer's instructions. An ethanolic solution of 4-nitrophenylacetic acid was used as the MAGL substrate, with a final substrate concentration of 236 μM. 150 μL of 1X assay buffer, 10 μL of recombinant human MAGL protein, and 10 μL of different concentrations of MAGL inhibitor (six concentrations ranging from 1 nM to 1000 nM) were added to each well of a 96-well plate. JZL195 (positive control inhibitor) was used as a positive control well. After incubating at room temperature for 5 minutes, 10 μL of MAGL substrate was added to each well. The 86-well plate was shaken for 10 seconds, left at room temperature for 10 minutes, and the absorbance at 410 nm was measured. Additionally, a 100% inhibition control well (150 μL of 1X assay buffer, 10 μL of MAGL, and 10 μL of DMSO were added to each well of a 96-well plate, with three replicate wells) and a background well (160 μL of 1X assay buffer and 10 μL of DMSO were added to each well, with three replicate wells) were set up. Next, Graphad Prism 5.0 was used to calculate and fit the IC50 curves for each inhibitor, and the specific values ​​are shown in Table 2.

[0103] Table 2 Activity data JPEG0007749540000031.jpg255170JPEG0007749540000032.jpg255170JPEG0007749540000033.jpg148170

[0104] Example 37: Tissue distribution study of MAGL inhibitors of the present invention

[0105] Detection reagents: MAGLZ-II-11, MAGLZ-II-16, MAGLZ-II-17

[0106] Test subjects: mice

[0107] Dosage: 16 mg / kg (concentration of preparation: 8 mg / ml);

[0108] Administration method: Subcutaneous injection

[0109] Sampling time: 1, 2, and 5 hours after administration

[0110] Detected tissues: brain, heart, liver, spleen, lungs, kidneys, pancreas, blood

[0111] Test Results:

[0112] The distribution of three test substances in the brain was detected, among which MAGLZ-II-16 had the highest content in the brain, followed by MAGLZ-II-17, and MAGLZ-II-11 was the lowest (see Table 3).

[0113] Table 3. Tissue distribution map JPEG0007749540000034.jpg34170

[0114] Example 38 Effect of MAGL inhibitors of the present invention on reserpine-induced depression in mice

[0115] 1)Basic information

[0116] Type: ICR mouse

[0117] Microbial level: SPF level

[0118] Number and sex of animals purchased: Male: 75, Female: 75

[0119] Number and sex of animals used: Male: 75, Female: 75

[0120] Weight: 13-18g upon receipt

[0121] 2) Grouping of animals

[0122] Group setup: each batch contains 50 animals, totaling three batches, and the animals in each batch are randomly divided into five groups according to their body weight. One test article is tested in each batch for low, medium, and high dose groups of the test article, a blank control group, and a model group.

[0123] 3) Test method

[0124] 3.1 The animals were administered subcutaneously once a day at doses of 4, 8, and 16 mg / kg for a total of 6 days. Two hours after the last administration, each animal was subcutaneously injected with 4 mg / kg of reserpine, except for the blank control group. Then, two hours later, the following indicators were measured: alertness, finger touch, fear, head touch, locomotor activity, upright position, and eyelid closure.

[0125] 4) Statistical analysis

[0126] Data statistics and analysis were performed on a computer using SPSS 17.0 software and Office 2016. Data statistics were performed using the Kruskal-Wallis test. If the Kruskal-Wallis test result was significant (P ≤ 0.05), further multiple comparison tests were performed using the Mann-Whitney U nonparametric test.

[0127] 5) The results are shown in Tables 4A, 4B, and 4C.

[0128] Table 4A JPEG0007749540000035.jpg90170

[0129] Note: Comparison of model and blank groups: * P<0.05, ** P<0.01, comparing the treatment group and the model group: * P<0.05, ** P<0.01.

[0130] Table 4B JPEG0007749540000036.jpg85170

[0131] Note: Comparison of model and blank groups: * P<0.05, ** P<0.01, comparing the treatment group and the model group: * P<0.05, ** P<0.01.

[0132] Table 4C JPEG0007749540000037.jpg70170

[0133] Note: Comparison of model and blank groups: * P<0.05, ** P<0.01, comparing the treatment group and the model group: * P<0.05, ** P<0.01.

[0134] 5.1) MAGLZ-II-11:

[0135] Nonparametric tests

[0136] There is statistical difference between each index by Kruskal-Wallis test (P<0.05).

[0137] Differences between comparison groups

[0138] Mann-Whitney U test

[0139] There was a statistical difference between the model group and the blank control group (P<0.05), indicating good modeling results. There was no statistical difference between the low-dose group, the medium-dose group, and the model group (P>0.05). Comparing the high-dose group and the model control group, there was a statistical difference in locomotor activity and standing (P<0.01, P<0.05), and the change trends of the indexes were consistent with those of the blank control group.

[0140] 5.2) MAGLZ-II-17: There is a statistical difference between the indices of the Kruskal-Wallis test (P<0.05). Mann-Whitney U test

[0141] There was a statistical difference between the model group and the blank control group (P<0.05), indicating that the modeling results were good.

[0142] There was no statistical difference between the low-dose group and the model group (P>0.05).

[0143] Comparing the medium dose group with the model group, there is a statistical difference in the eyelids (P<0.05), but the average value is smaller than that of the model group, and there is no statistical difference in other indicators (P>0.05).

[0144] Comparing the high-dose group with the model group, there is a statistical difference in the eyelids (P<0.01), but the average value is smaller than that of the model group, and there is no statistical difference in other indexes (P>0.05).

[0145] 5.3) MAGLZ-II-16: There is a statistical difference between the indices of the Kruskal-Wallis test (P<0.05). Mann-Whitney U test

[0146] There was a statistical difference between the model group and the blank control group (P<0.05), indicating that the modeling results were good.

[0147] There was no statistical difference between the low-dose group and the model group (P>0.05).

[0148] Comparing the medium dose group and the model group, there was a statistical difference in head contact (P<0.05), the mean value was larger than that of the model group, and the trend change was the same as that of the blank control group, and there was no statistical difference in other indicators (P>0.05).

[0149] Comparing the high-dose group and the model group, there was a statistical difference in head contact and locomotor activity (P<0.05), the mean value was greater than that of the model group, and the trend change was the same as that of the blank control group, and there was no statistical difference in other indexes (P>0.05).

[0150] 6) Conclusion

[0151] 6.1) Under the conditions of this study, MAGLZ-II-11 has a certain influence and effect on reserpine-induced depression.

[0152] 6.2) Under the conditions of this study, MAGLZ-II-16 exerts a certain influence and effect on reserpine-induced depression.

[0153] 6.3) Under the conditions of this study, MAGLZ-II-17 may have a stronger effect and efficacy on reserpine-induced depression.

[0154] Example 39 Effects of MAGL inhibitors of the present invention on mouse forced swimming and mouse tail suspension models

[0155] 1. Materials and Methods

[0156] 1.1 Experimental animals

[0157] Male ICR mice weighing 18-22 g were used. Animals were exercised on a normal 12-h light / 12-h dark schedule. Ambient temperature and relative humidity were maintained at 22 ± 1°C and 55 ± 5%, respectively.

[0158] 1.2 Experimental Method

[0159] 1.2.1 Animal grouping and administration method

[0160] Seven groups of experimental mice, each with 10 mice, were set up as a control group (saline group), a positive control group (fluoxetine hydrochloride capsules, 5 mg / kg), and five MAGLZ-II series treatment groups (MAGLZ-II-06, MAGLZ-II-07, MAGLZ-II-11, MAGLZ-II-17, and MAGLZ-II-18, respectively), each administered at a concentration of 5 mg / kg.

[0161] All drugs were administered by oral gavage at a dose of 20 mL / kg body weight once daily for 10 days. Ethological observations were initiated 1 h after the last administration.

[0162] 1.2.2 Forced swimming test in mice

[0163] The forced swimming experiment in mice was performed according to the method established by Porsalt (Porsalt RD. Behavioral despair in mice: A primary screening test for antidepressants [J]. Arch. Int. Pharmacolodyn. 1977, 229). Mice were forced to swim alone in an open cylindrical container (10 cm diameter, 25 cm height). The water temperature in the container was 25±1°C and the water depth was 19 cm. The total time each animal remained immobile over a 6-minute period was recorded as the immobility time (in seconds). Each mouse was considered immobile when it stopped struggling and remained immobile in the water, making only the movements necessary to keep its head above water. A reduction in immobility time indicated an antidepressant effect.

[0164] 1.2.3 Mouse tail suspension test

[0165] The tail suspension test (TST) was performed according to the method established by Porsolt et al. (Porsolt RD, Le Pichon M, Jalfre M. Depression: a new animal model sensitive to antidepressant treatments [J]. Nature, 1977, 266(5604):730-732). Using a tail suspension clip, each mouse was suspended by its tail in a box (250 mm x 250 mm x 300 mm) with its head 5 cm from the bottom and 10 mm from the tail. The test was conducted in a dark room with minimal background noise. Each mouse was suspended for 6 minutes, with the final immobility time recorded over 4-minute intervals. The criterion for success was when the mouse stopped struggling and remained completely motionless.

[0166] 1.2.4 Data Processing

[0167] All data is JPEG0007749540000038.jpg8170, and comparisons between groups were performed using single-factor analysis of variance.

[0168] 2 Results

[0169] 2.1 Effects of MAGLZ-II series compounds on forced swimming behavior in mice

[0170] The effect of MAGLZ-II series compounds on forced swimming behavior in mice showed that the positive control drug fluoxetine hydrochloride and the MAGLZ-II series compounds were both able to significantly reduce the cumulative immobility time during forced swimming in mice (P<0.05) (Table 5).

[0171] Table 5. Effects of MAGLZ-II series compounds on forced swimming behavior in mice JPEG0007749540000039.jpg87170

[0172] Note: Compared to the control group, * P<0.05, ** P<0.01.

[0173] 2.2 Effects of MAGLZ-II series compounds on tail-suspension behavior in mice

[0174] Compared with the control group, each compound group in the MAGLZ-II series could significantly shorten the cumulative immobility time of mice in tail suspension (P<0.05) (Table 6).

[0175] Table 6. Effects of MAGLZ-II series compounds on tail-suspended behavior in mice JPEG0007749540000040.jpg87170Note: Compared to the control group, * P<0.05, ** P<0.01.

[0176] 3. Consideration

[0177] Mouse forced swimming and mouse tail suspension are two of the more commonly used and typical animal models of depression. The despair and behavioral distortions reflected by these models are key features of in vitro simulated depression. At the same time, they are sensitive to antidepressants and easy to manipulate, making them commonly used screening models for antidepressants. In this study, we used these two typical animal models of depression to further evaluate and validate the antidepressant activity of MAGLZ-II series compounds. The results showed that MAGLZ-II series compounds significantly reduced the cumulative immobility time in mouse forced swimming and mouse tail suspension, demonstrating their distinct antidepressant effects.

[0178] Example 40 Effect of MAGL inhibitors of the present invention on the number of body twists induced by acetic acid in mice

[0179] 1. Animal Grouping and Administration

[0180] There were 200 male ICR mice, weighing (20±2) g, randomly divided into 20 groups according to body weight, with 10 mice in each group. The specific groupings were as follows: Model group: an equal volume of 0.5% sodium carboxymethylcellulose, i.g. Positive control group: diclofenac sodium 10 mg / kg, ig MAGLZ-II-11 low dose group: 7.5mg / kg, ig MAGLZ-II-11 medium dose group: 15mg / kg, ig MAGLZ-II-11 high dose group: 30mg / kg, ig MAGLZ-II-11a low dose group: 7.5mg / kg, ig MAGLZ-II-11a intermediate dose group: 15mg / kg, ig MAGLZ-II-11a high dose group: 30mg / kg, ig MAGLZ-II-18a low dose group: 7.5mg / kg, ig MAGLZ-II-18a intermediate dose group: 15mg / kg, ig MAGLZ-II-18a high dose group: 30mg / kg, ig MAGLZ-II-18 low dose group: 7.5mg / kg, ig MAGLZ-II-18 medium dose group: 15mg / kg, ig MAGLZ-II-18 high dose group: 30mg / kg, ig MAGLZ-II-18c low dose group: 7.5mg / kg, ig MAGLZ-II-18c intermediate dose group: 15mg / kg, ig MAGLZ-II-18c high dose group: 30mg / kg, ig MAGLZ-II-10 low dose group: 7.5mg / kg, ig MAGLZ-II-10 medium dose group: 15mg / kg, ig MAGLZ-II-10 high dose group: 30mg / kg, ig Each compound was prepared into a suspension using 0.5% sodium carboxymethylcellulose, and then dosed according to the dosage. The suspension was administered orally to each treatment group, and an equal volume of 0.5% sodium carboxymethylcellulose was administered orally to the model group.

[0181] 2. Experimental Methods and Data Processing

[0182] Acetic acid was injected intraperitoneally into the mice, causing widespread, persistent pain deep within the abdominal cavity and resulting in a twisting reaction. One hour after administration, each group was injected intraperitoneally with 0.1 ml / 10 g of 0.7% acetic acid in saline.

[0183] The time from intraperitoneal injection of acetic acid to the onset of body twisting (pain latency) is recorded.

[0184] After injecting acetic acid to induce pain, the number of body twisting reactions of each mouse within 20 minutes is recorded, and the body twisting inhibition rate of each administration group is calculated.

[0185] Inhibition rate of body twisting = [(number of body twisting in the control group - number of body twisting in the drug group) / number of body twisting in the control group] × 100%

[0186] The experimental data The data are represented as JPEG0007749540000041.jpg7170 and analysis of variance is performed using SPSS15.0 software.

[0187] 3. Experimental Results

[0188] The experimental results are shown in Table 7.

[0189] Table 7. Effect of MAGL inhibitors of the present invention on the number of acetic acid-induced body twists in mice. JPEG0007749540000042.jpg230170Compared to the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0190] As can be seen from the experimental results shown in Table 1:

[0191] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, and MAGLZ-II-18c of the present invention can significantly reduce the number of body twists induced by acetic acid in mice and prolong the latency time.

[0192] Example 41 Effect of the MAGL inhibitor of the present invention on hot plate-induced pain response in mice

[0193] A female ICR mouse (20 ± 2 g) was placed on an intelligent hot plate device at 55 ± 0.5 °C. The latency period (s) from when the mouse's paw contacted the hot plate to when it licked its hind paw was recorded as an index of pain threshold. Mice with a latency period of <5 s or >30 s, or that jumped, were eliminated.

[0194] Grouping and administration were the same as in Example 40.

[0195] Mice are subjected to oral gavage for 7 consecutive days, and the pain threshold of each mouse in each administration group is measured once at 30, 60, 90, and 120 minutes after the last administration. Pain thresholds of mice exceeding 60 seconds are calculated as 60 seconds.

[0196] The experimental results are shown in Table 8.

[0197] Table 8. Effect of MAGL inhibitors of the present invention on the pain threshold of hot plate-induced pain in mice JPEG0007749540000043.jpg235170Compared to the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0198] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention can significantly prolong the pain threshold of thermally stimulated mice.

[0199] Example 42 Effect of the MAGL inhibitor of the present invention on the pain threshold of mice with inflammatory pain

[0200] Male SD mice (180±20 g) were injected with 50 μL of complete Freund's adjuvant into the right hind paw pad, and a control group was injected with a corresponding volume of 0.5% sodium carboxymethylcellulose to establish the inflammatory pain model of paw swelling. After modeling, mice were subjected to oral gavage once daily for 7 consecutive days. The pain thresholds of the mice were measured before and after modeling on days 1, 3, and 7, respectively.

[0201] Grouping and administration were the same as in Example 40.

[0202] Table 9. Effect of MAGL inhibitors of the present invention on pain threshold in mice with inflammatory pain. JPEG0007749540000044.jpg246170Compared to the model group, * P<0.05, ** P<0.01,*** P<0.001.

[0203] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention can significantly prolong the pain threshold of mice with inflammatory pain.

[0204] Example 43 Therapeutic effect of the MAGL inhibitor of the present invention on rats with irritable bowel syndrome induced by early maternal separation

[0205] Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder characterized by chronic abdominal pain, abdominal discomfort, and altered bowel habits without obvious intestinal lesions. This study primarily investigated the therapeutic effects of MAGL inhibitors on rats with IBS induced by early maternal separation.

[0206] 1. Animal Grouping and Administration

[0207] There were 200 SD rats, each weighing (20±2) g, randomly divided into 20 groups according to their body weight, with 10 mice in each group. The specific groupings were as follows: Blank group: an equal volume of 0.5% sodium carboxymethylcellulose, i.g. Model group: equal volume of 0.5% sodium carboxymethylcellulose, i.g. MAGLZ-II-11 low dose group: 7.5mg / kg, ig MAGLZ-II-11 medium dose group: 15mg / kg, ig MAGLZ-II-11 high dose group: 30mg / kg, ig MAGLZ-II-11a low dose group: 7.5mg / kg, ig MAGLZ-II-11a intermediate dose group: 15mg / kg, ig MAGLZ-II-11a high dose group: 30mg / kg, ig MAGLZ-II-18a low dose group: 7.5mg / kg, ig MAGLZ-II-18a intermediate dose group: 15mg / kg, ig MAGLZ-II-18a high dose group: 30mg / kg, ig MAGLZ-II-18 low dose group: 7.5mg / kg, ig MAGLZ-II-18 medium dose group: 15mg / kg, ig MAGLZ-II-18 high dose group: 30mg / kg, ig MAGLZ-II-18c low dose group: 7.5mg / kg, ig MAGLZ-II-18c intermediate dose group: 15mg / kg, ig MAGLZ-II-18c high dose group: 30mg / kg, ig MAGLZ-II-10 low dose group: 7.5mg / kg, ig MAGLZ-II-10 medium dose group: 15mg / kg, ig MAGLZ-II-10 high dose group: 30mg / kg, ig

[0208] Each compound was prepared into a suspension using 0.5% sodium carboxymethylcellulose, and then dosed according to the dosage. The suspension was administered orally to each treatment group, and an equal volume of 0.5% sodium carboxymethylcellulose was administered orally to the model group.

[0209] 2. Experimental Methods and Data Processing

[0210] After the birth of the SD newborn rats, from the 2nd to the 21st day of life, the male newborn SD rats were separated from the lactating female rats for 3 hours every day. That is, from 8:30 a.m., the lactating female rats were removed from their cages, and then the newborn rats were removed from their cages and placed in separate cages. This cage was then moved to the adjacent room. After 3 hours, that is, at 11:30 a.m., the newborn rats were returned to their original cages and allowed to mingle. Weaning began on the 22nd day of life, and the rats were divided into cages on the 30th day and grouped on the 60th day and administered drugs, for a total of 20 days.

[0211] During the administration period, oral administration is performed once a day for 20 days.

[0212] Experimental endpoint: Dissection is performed after the corresponding detection indicator of the experiment is completed.

[0213] Detection index: Abdominal Nociceptive Reflex Score (AWR). Detection method: Before the experiment, rats were fasted for 18 h, allowed free access to water, and anesthetized with ether. A paraffin oil-coated balloon was inserted 4.0 cm into the rat's colon and rectum. An external anal catheter was fixed to the base of the rat's tail with adhesive tape and connected to a syringe and a blood pressure monitor via a three-way connection. The rat was placed in a Plexiglas observation box (20 cm x 12 cm x 9 cm) for observation. The experiment began 30 min after the rat had woken up and fully acclimatized to the environment. Pressures of 20, 40, and 60 mmHg were applied, with each distention lasting 20 seconds and a 4-minute interval between stimulations. AWR scoring was performed three times for each pressure. The AWR score uses a single-blind method, and the scoring criteria are as follows: 0 points: no obvious behavioral changes; 1 point: only simple head movements; 2 points: abdominal muscles begin to contract but do not lift off the table; 3 points: abdominal muscles contract significantly and flatten or the lower abdominal wall lifts off the table; 4 points: abdominal wall arches or the body and pelvis arch.

[0214] After the experiment, the animals were dissected and weighed, anesthetized, and blood was collected. The 5-HT content in the serum was detected by high-performance liquid chromatography. The brains were collected on ice, washed with pre-cooled 0.5% sodium carboxymethylcellulose, blotted dry with filter paper, and stored in cryopreservation tubes at -20°C for future use. The 5-HT content in the brain tissue was detected by high-performance liquid chromatography.

[0215] Histological observation of the colon: After the colon distension experiment, the distal colon (5-6 cm from the anus) of the rat was removed and subjected to conventional HE staining to observe the inflammation and damage status of the intestinal wall.

[0216] Data were expressed as mean ± SD, and differences between groups were statistically analyzed by t-test or one-way analysis of variance, with P<0.05 considered to indicate a significant difference.

[0217] 3. Experimental Results

[0218] The experimental results are shown in Table 10.

[0219] Table 10. Effects of MAGL inhibitors of the present invention on AWR score and 5-HT in rats with irritable bowel syndrome. JPEG0007749540000045.jpg243170JPEG0007749540000046.jpg99170Compared to the blank group, # P<0.05, ## P<0.01, ### P<0.001.

[0220] Compared with the models, * P<0.05, ** P<0.01, *** P<0.001.

[0221] As can be seen from the experimental results shown in Table 10:

[0222] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention can significantly reduce the AWR score and lower the 5-HT levels in the colon and serum.

[0223] Example 44 Effect of MAGL inhibitors of the present invention on reserpine-induced migraine in mice

[0224] 1. Animal Grouping and Administration

[0225] There were 210 male ICR mice, weighing (20±2) g, randomly divided into 21 groups according to body weight, with 10 mice in each group. The specific groupings were as follows: blank group: an equal volume of 0.5% sodium carboxymethylcellulose, i.g. Model group: equal volume of 0.5% sodium carboxymethylcellulose, i.g. Positive control group: zolmitriptan 0.5 mg / kg, ig MAGLZ-II-11 low dose group: 7.5mg / kg, ig MAGLZ-II-11 medium dose group: 15mg / kg, ig MAGLZ-II-11 high dose group: 30mg / kg, ig MAGLZ-II-11a low dose group: 7.5mg / kg, ig MAGLZ-II-11a intermediate dose group: 15mg / kg, ig MAGLZ-II-11a high dose group: 30mg / kg, ig MAGLZ-II-18a low dose group: 7.5mg / kg, ig MAGLZ-II-18a intermediate dose group: 15mg / kg, ig MAGLZ-II-18a high dose group: 30mg / kg, ig MAGLZ-II-18 low dose group: 7.5mg / kg, ig MAGLZ-II-18 medium dose group: 15mg / kg, ig MAGLZ-II-18 high dose group: 30mg / kg, ig MAGLZ-II-18c low dose group: 7.5mg / kg, ig MAGLZ-II-18c intermediate dose group: 15mg / kg, ig MAGLZ-II-18c high dose group: 30mg / kg, ig MAGLZ-II-10 low dose group: 7.5mg / kg, ig MAGLZ-II-10 medium dose group: 15mg / kg, ig MAGLZ-II-10 high dose group: 30mg / kg, ig

[0226] Each compound was prepared into a suspension using 0.5% sodium carboxymethylcellulose, and then dosed according to the dosage. The suspension was administered orally to each treatment group, and an equal volume of 0.5% sodium carboxymethylcellulose was administered orally to the model group.

[0227] 2. Experimental Methods and Data Processing

[0228] Except for the blank control group, which was subcutaneously injected with saline, all other groups were subcutaneously injected with reserpine solution (0.2 mg / kg) for a total of 10 days. After the reserpine injection, the mice were considered to have been successfully modeled if they showed typical reserpineization symptoms such as closed eyes, squatting and reduced movement, diarrhea, reduced food intake, and hunched posture.

[0229] Starting on the fifth day after modeling, the blank control group and model group were given a vehicle gavage orally, while the other groups were given the corresponding drug and dose for 10 consecutive days. One hour after the last dose, the animals were weighed and blood was collected from the medial canthus of the eye to measure blood clotting time. Whole blood was collected, and the brains were collected on ice and stored at -20°C for subsequent experiments.

[0230] Experimental endpoint: Modeling administration time is 10 days, and on the 10th day, the experiment will be terminated by detection and blood sampling 1 hour after the last administration.

[0231] Detection parameters: After the experiment begins, the weight of the mice is measured every two days.

[0232] Behavioral detection: Tail suspension movement. On the 9th day of the experiment, adhesive tape was attached to the site 1 cm away from the tail of the mouse, a steel needle was passed through the adhesive tape, and the mouse was hung upside down in a dark box. The number of times the mouse struggled within 2 minutes (the total number of times the mouse bent upward after being hung upside down) was observed.

[0233] One hour after the last administration, the body weight is measured and blood is collected from the medial canthus with a capillary to detect the blood clotting time.

[0234] Blood samples were collected and serum 5-HT content was determined by high-performance liquid chromatography. Brains were collected on ice, washed with pre-cooled 0.5% sodium carboxymethylcellulose, blotted dry with filter paper, and stored in cryopreservation tubes at -20°C for future use. Brain tissue 5-HT content was determined by high-performance liquid chromatography.

[0235] The 5-HT detection method is as follows: the sample is precipitated with 10% perchloric acid solution (1:1), centrifuged, and the supernatant is collected for detection. Chromatography conditions: The chromatography column is SHIMADZU VP-ODS C 18 Column (250 *The column was 4.6 mm, 5 μm, and the mobile phase was methanol-0.01 mol / L potassium acetate buffer (10:90, V / V, pH adjusted to 4.00 with 0.2 mol / L citric acid) for isocratic elution. The column temperature was 25°C, the flow rate was 1.00 ml / min, and the detection wavelength was 275 nm.

[0236] Data were expressed as mean ± SD, and differences between groups were statistically analyzed by t-test or one-way analysis of variance, with P<0.05 considered to indicate a significant difference.

[0237] 3. Experimental Results

[0238] The experimental results are shown in Tables 11 to 13 below.

[0239] Table 11. Effect of MAGL inhibitors of the present invention on blood clotting time JPEG0007749540000047.jpg201170Compared to the model group, # P<0.05, ## P<0.01, ### P<0.001.

[0240] As can be seen from the experimental results shown in Table 11:

[0241] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention can significantly prolong the blood clotting time.

[0242] Table 12. Effect of MAGL inhibitors of the present invention on the number of tail suspensions JPEG0007749540000048.jpg220170Compared to the model group, # P<0.05, ## P<0.01, ### P<0.001.

[0243] As can be seen from the experimental results shown in Table 12:

[0244] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention can significantly increase the number of tail-suspension movements.

[0245] Table 13. Effect of MAGL inhibitors of the present invention on serotonin in brain tissue JPEG0007749540000049.jpg140170Compared to the model group, # P<0.05, ## P<0.01, ### P<0.001.

[0246] As can be seen from the experimental results shown in Table 13:

[0247] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention can significantly increase the level of 5-HT in the brain tissue of mice in the reserpine-induced hypo-5-HT model.

[0248] Example 45 Study on the effect of MAGL inhibitors on ulcerative colitis in mice

[0249] 1. Animal Grouping and Administration

[0250] Healthy male SPF grade BALB / c mice, 8 weeks old and weighing (20±2) g.

[0251] After entering the laboratory, the mice will be quarantined for 7 days, and healthy male mice will be selected as test animals. During the quarantine period, the main inspection items include, but are not limited to, whether the number of animals meets the requirements and quality indicators at the time of application, their general condition, and weight. Any animals that do not pass the quarantine will not be included in the study.

[0252] Modeling: After quarantine, the mice were randomly divided into 21 groups with 10 mice in each group. The normal group was given free access to normal water, while the other groups were given free access to 1.0%-1.5% DSS solution for modeling. After 7 days of modeling, the modeling group was given free access to normal water for 7 days. Thus, 14 days constituted one modeling cycle, with four consecutive cycles administered simultaneously with modeling.

[0253] Blank group: an equal volume of 0.5% sodium carboxymethylcellulose, i.g. Model group: equal volume of 0.5% sodium carboxymethylcellulose, i.g. Positive control group: mesalamine (100 mg / kg), i.g. MAGLZ-II-11 low dose group: 7.5mg / kg, ig MAGLZ-II-11 medium dose group: 15mg / kg, ig MAGLZ-II-11 high dose group: 30mg / kg, ig MAGLZ-II-11a low dose group: 7.5mg / kg, ig MAGLZ-II-11a intermediate dose group: 15mg / kg, ig MAGLZ-II-11a high dose group: 30mg / kg, ig MAGLZ-II-18a low dose group: 7.5mg / kg, ig MAGLZ-II-18a intermediate dose group: 15mg / kg, ig MAGLZ-II-18a high dose group: 30mg / kg, ig MAGLZ-II-18 low dose group: 7.5mg / kg, ig MAGLZ-II-18 medium dose group: 15mg / kg, ig MAGLZ-II-18 high dose group: 30mg / kg, ig MAGLZ-II-18c low dose group: 7.5mg / kg, ig MAGLZ-II-18c intermediate dose group: 15mg / kg, ig MAGLZ-II-18c high dose group: 30mg / kg, ig MAGLZ-II-10 low dose group: 7.5mg / kg, ig MAGLZ-II-10 medium dose group: 15mg / kg, ig MAGLZ-II-10 high dose group: 30mg / kg, ig

[0254] Each compound was prepared into a suspension using 0.5% sodium carboxymethylcellulose, and then dosed according to the dosage. The suspension was administered orally to each treatment group, and an equal volume of 0.5% sodium carboxymethylcellulose was administered orally to the model group.

[0255] 2. Testing process and methods

[0256] Experimental endpoint: The experiment involves four cycles of drug administration, with dissections performed on days 3 and 4 of the recovery period of the fourth cycle.

[0257] Detection parameters: After the start of administration, the mice were weighed and observed twice a week for diarrhea and bloody stool, and the number of bloody stools was recorded.

[0258] After the experiment, the animals were dissected, weighed, anesthetized, bled, and the serum was stored at -80°C. The spleens were removed and weighed to calculate the spleen index. The IL-1β content in the serum was detected by ELISA. The colon and rectum were cut and their length was measured. They were then fixed with formaldehyde and stained with HE to detect pathological changes.

[0259] Data were expressed as mean ± SD, and differences between groups were statistically analyzed by t-test or one-way analysis of variance, with P<0.05 considered to indicate a significant difference.

[0260] 3. Results and Discussion

[0261] The experimental results are shown in Table 14.

[0262] Table 14. Effect of MAGL inhibitors of the present invention on colon length, frequency of bloody stools, and IL-1 content in mice with ulcerative colitis. JPEG0007749540000050.jpg245170Compared to the blank group, # P<0.05, ## P<0.01, ### P<0.001.

[0263] Compared with the models, * P<0.05, ** P<0.01, *** P<0.001.

[0264] Compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, and MAGLZ-II-18c of the present invention significantly increase the length of the colon, reduce the total number of bloody stools, and reduce the content of IL-1.

[0265] Since the present invention has been described by the above examples, any equivalent substitutions are obvious to and encompassed by the present invention.

Claims

1. Formula I or a pharmaceutically acceptable salt thereof, where: R is H, Ar １ is phenyl, diphenyl ether or biphenyl, X １ is hydroxy, Ar ２ is phenyl, X ２ is one or more of H, methyl, dimethyl, methoxy, hydroxy, F and Cl; A compound or a pharmaceutically acceptable salt thereof, wherein the substitution position of Ar 1 is at the meta position.

2. A compound selected from the following or a pharmaceutically acceptable salt thereof.

3. 10. A pharmaceutical composition for treating a MAGL-mediated disease or condition, comprising a compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

4. 10. Use of a compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a MAGL-mediated disease or condition.

5. 5. The use of claim 4, wherein the condition is selected from metabolic disorders, kidney disease, vomiting or nausea, eating disorders, neurological disorders, schizophrenia, depression, bipolar disorder, tremors, movement disorders, withdrawal syndromes, traumatic brain injury, non-traumatic brain injury, spinal cord injury, epileptic seizures, disorders associated with abnormal cell growth or proliferation (e.g., benign tumors or cancer), inflammatory disorders, immune system disorders, irritable bowel syndrome, ulcerative colitis, acute stress disorder, substance-induced anxiety, obsessive-compulsive disorder, anxiety disorders, attention deficit disorder, attention deficit hyperactivity disorder, pain, migraine, demyelinating diseases, and cognitive disorders.

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