RU2847108C1 - Magl inhibitor, method for obtainment and use thereof - Google Patents

Abstract

FIELD: medicine; pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics and particularly to a MAGL inhibitor, a method for preparing and using it. Disclosed is a compound of formula I or pharmaceutically acceptable salts thereof

where R is H, Ar₁ is selected from diphenyl ether, indolyl and biphenyl, Ar₂ is C₆ aryl, X₁ is selected from one of halogen and C_1-3 alkyl, X₂ is selected from one or two of halogen, C_1-3 alkyl and C_1-3 alkoxy and compound is not N-{1-[(5-chloroindol-2-yl)carbonyl](4-piperidyl)}(4-chlorophenyl)carboxamide, {1-[(5-chloroindol-2-yl)carbonyl](4-piperidyl)}-N-(3-chlorophenyl)carboxamide or {1-[(5-chloroindol-2-yl)carbonyl](4-piperidyl)}-N-(3-fluorophenyl)carboxamide; compound of formula I, where R is H, Ar₁ is phenyl, X₁ is hydroxyl, Ar₂ is phenyl, X₂ is one or two of methoxy, fluorine and chlorine and compound is not

compound of formula I, wherein R is H, Ar₁ is indolyl, X₁ is H, methyl or fluorine, Ar₂ is phenyl, X₂ is chlorine and the compound is not N-(3-chlorophenyl)-1-(1H-indol-2-ylcarbonyl)-4-piperidine carboxamide; method for synthesis thereof; pharmaceutical composition for treating a MAGL-mediated disease or disorder; use of the compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a MAGL-mediated disease or disorder; a method of inhibiting MAGL; and a method of treating a MAGL-mediated disease or condition.

EFFECT: novel compounds which can be used to treat a MAGL-mediated disease or disorder are obtained.

14 cl, 14 tbl, 45 ex

Description

Field of invention

The invention relates to the field of pharmaceuticals and, in particular, to a MAGL inhibitor, a method for its production and use.

State of the art of the invention

Monoacylglycerol lipase (MAGL) is a serine hydrolase that promotes the breakdown of lipids into glycerol and fatty acids. It is highly expressed in aggressive and primary human tumor cells. High concentrations of MAGL can maintain the high pathogenicity of tumor cells by increasing the concentration of free fatty acids. MAGL has become an important target for the development of anticancer drugs.

Disclosure of invention

The present invention provides a compound of formula I or pharmaceutically acceptable salts thereof.

wherein R is H or C _1-5 alkyl, said C _1-5 alkyl being selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, neo-butyl, n-pentyl, iso-pentyl, neopentyl, cyclopentyl or C _1-5 alkyl substituted with halogen, hydroxyl, carboxyl, C _1-3 alkyl, C _1-3 alkoxy, C _3-8 cycloalkyl, C _3-8 heterocyclyl, C _3-8 cycloalkoxy, aryl, heteroaryl; the number of R is 0, 1, 2, 3 or 4;

each Ar ₁ , Ar ₂ is independently selected from 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), (5-10-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-(5-10-membered heteroaryl), (5-10-membered heteroaryl)-O-(5-10-membered heteroaryl), (5-10-membered heteroaryl)-(5-10-membered heteroaryl), the position of the substituents is arbitrary;

each X ₁ , X ₂ is independently selected from one or more of 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 _1-3 alkyl, C _1-3 alkoxy, C _3-8 cycloalkyl, C _3-8 heterocyclyl, C _3-8 cycloalkoxy, aryl, heteroaryl, which are substituted by hydroxyl, carboxyl, halogen, and alkyl, the position of the substituents is arbitrary.

In some examples, each Ar ₁ , Ar ₂ is independently selected from phenyl, C _5-18 aryl, heteroaryl, (C _6-10 aryl)-(C _6-10 aryl), (C 6-10 aryl)-O-(C _6-10 aryl); X ₁ , X ₂ is independently selected from one or more of H, hydroxyl, halogen, acyloxy, C _1-3 alkyl and C _1-3 alkoxy, the position of the substituents is arbitrary.

In some examples, each Ar ₁ , Ar ₂ is independently selected from one or more substituents representing phenyl, naphthyl, diphenyl ether, indole, biphenyl, the position of the substituents is arbitrary.

The above compound of formula I or its pharmaceutically acceptable salts, wherein said R is H, has the structural formula respectively

where each Ar ₁ , Ar ₂ is independently selected from 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), (5-10-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-(5-10-membered heteroaryl), (5-10-membered heteroaryl)-O-(5-10-membered heteroaryl), (5-10-membered heteroaryl)-(5-10-membered heteroaryl), the position of the substituents is arbitrary;

each X ₁ , X ₂ is independently selected from one or more of H, hydroxyl, carboxyl, halogen, C _1-3 alkyl, C _1-3 alkoxy, C _3-8 cycloalkyl, C _3-8 heterocyclyl, C _3-8 cycloalkoxy, aryl, heteroaryl, or C _1-3 alkyl, C _1-3 alkoxy, C _3-8 cycloalkyl, C _3-8 heterocyclyl, C _3-8 cycloalkoxy, aryl, heteroaryl, which are substituted by hydroxyl, carboxyl, halogen and alkyl, the position of the substituents is arbitrary.

More specifically, each Ar ₁ , Ar ₂ is independently selected from 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), (5-10 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-(5-10 membered heteroaryl), (5-10 membered heteroaryl)-O-(5-10 membered heteroaryl), (5-10 membered heteroaryl)-(5-10 membered heteroaryl), the position of the substituents is arbitrary;

each X ₁ , X ₂ is independently selected from one or more of H, hydroxyl, halogen, C _1-3 alkyl, C _1-3 alkoxy, C _3-8 heterocyclyl, C _3-8 cycloalkoxy, and C _1-3 alkyl and C _1-3 alkoxy, which are substituted by hydroxyl, halogen, alkyl, the position of the substituents is arbitrary.

In some examples, Ar ₁ is one or more of phenyl, naphthyl, diphenyl ether, biphenyl, and the position of the substituents is arbitrary;

X ₁ represents one or more of H, methoxy, hydroxyl, fluorine, chlorine, the position of the substituents is arbitrary;

Ar ₂ is benzene;

X ₂ is one or more of H, methyl, dimethyl, methoxy, hydroxyl, fluorine, chlorine, the position of the substituents is arbitrary.

In specific examples, Ar ₁ has a meta-position of substitution.

In some examples, said Ar ₁ is one or more of benzofuranyl, benzimidazolyl, benzoxazolyl, indolyl, benzothiophenyl, quinolinyl, isoquinolinyl, purinyl, the position of the substituents is arbitrary;

X ₁ represents one or more of H, methyl, methoxy, hydroxyl, fluorine, chlorine, bromine, the position of the substituents is arbitrary;

Ar ₂ is phenyl, the position of the substituent is arbitrary.

X ₂ represents one or more of H, methyl, dimethyl, methoxy, hydroxy, fluoro, chlorine, the position of the substituents is arbitrary. In some examples, the above formula I may be limited to formula III, as shown in the following formula:

wherein said R ₁ is selected from hydroxy, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy; R ₂ is selected from hydroxy, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy, phenyl and aryl substituted with hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy. The number of R ₁ and R ₂ is 0, 1, 2, 3 or 4; the mode of binding R ₁ , R ₂ to the parent nucleus is not limited to a single bond connection.

The present invention provides a compound of formula b

,

intended for the synthesis of a compound of formula I, wherein said Ar ₂ , X ₂ are described in formula I,

In some examples, fragment A, outlined by a dotted line ,

replaced or .

The present invention provides a method for synthesizing a compound of formula I comprising the following steps:

b or salt b reacts with , as a result of the reaction, I was obtained; the indicated Ar ₁ , X ₁ are described in formula I.

The specified b or salt b reacts with , as a result of the reaction, I was obtained; The addition of a condensing agent and a base at the reaction stage makes the reaction more favorable, and the condensing agent is selected from HBTU, DMC, HOBT, HOBT/EDCI, HATU, HATU/DIEPA, DCC, CDI, isopropyl chloroformate; the structural formulas are respectively the following:

b or salt b is obtained by deprotection of a

;

the said base is an organic base such as N-diisopropylethylamine (DIPEA), diethylamine (DEA) or triethylamine (TEA).

a or salt a was obtained as a result of the reaction of sm with or its salt;

,

where sm can be obtained according to the prior art or can be synthesized by a person skilled in the art based on the prior art and by a limited number of experiments without any inventive efforts. The addition of a condensing agent and a base during the reaction step makes the reaction more favorable; said condensing agent is selected from HBTU, DMC, HOBT, HOBT/EDCI, HATU, HATU/DIEPA, DCC, CDI, and isopropyl chloroformate.

In some specific examples, the following methods may be used:

wherein said reducing agent is preferably at least one of NaBH ₄ , KBH ₄ , NaBH ₄ /LiCl.

Unless otherwise specifically stated, the solvents selected for the reactions in the above or subsequent steps of the present invention are conventional solvents in the art, and the principle of choosing a solvent is to dissolve the reactants, but not to participate in the reaction, to extract the products or to crystallize the corresponding products therein and to separate them from impurities, such as water, haloalkanes, alkylamines, aliphatic hydrocarbons, esters, alcohols, aromatic hydrocarbons, ethers and heterocyclic solvents; specifically selected from these solvents, but not limited to: methanol, ethanol, propanol, isopropanol, diethyl ether, ethyl acetate, acetic acid, cyclohexane, methylene chloride, trichloromethane, tetrahydrofuran, pyridine, diethylamine, triethylamine, dimethylformamide, toluene and mixtures of at least two of them.

Serial numbers a, b, etc., used in the present invention are ordinal numbers adopted for the convenience of listing general formulas. They may be transformed into other numbers, such as 1, 2, 3, etc., in specific examples. All of them are intended for ease of presentation and do not affect the essence of the structural formulas and reaction equations; they represent an expression of the general formula and its reaction equations. Those skilled in the art understand that the substituents of each intermediate compound in all of the synthetic routes described above are determined by the structure of the target compound.

Chiral isomers or cis-trans isomers and mixtures of isomers in any ratio among the compounds covered by the general formula of the target compound of general formula I and their specific representative substances are also covered by the compounds covered by the general formula of the target compound of general formula I and their specific representative substances.

The term "arbitrary substituent position" refers to any of a number of examples of permissible valence bond positions. For example, a substituent on a benzene ring may be in one or more of the ortho, meta, or para positions; for example, a substituent on an aromatic heterocycle may be in one or more of the 2-, 3-, 4-, 5-, and so on positions. With respect to the selection of substituents, "one or more" and "one or more moieties" refer to the number of substituents selected, and the specific number is 1, 2, 3, 4... specifically, for example, "Ar ₁ is phenyl substituted at any position, X ₁ is one or more of fluorine, chlorine, at any position of substitution" includes phenyl, which may be substituted with one Cl, two Cl, three Cl at the ortho position (two), meta position (two), para position (one), or two meta positions substituted with one Cl and one F, or one Cl at the ortho position and one F at the para position, and so on.

Unless otherwise specifically stated, the solvents selected for the reactions in the above or subsequent steps of the present invention are conventional solvents in the art, and the principle of choosing a solvent is to dissolve the reactants, but not to participate in the reaction, to extract the products or to crystallize the corresponding products therein and to separate them from impurities, such as water, haloalkanes, alkylamines, aliphatic hydrocarbons, esters, alcohols, aromatic hydrocarbons, ethers and heterocyclic solvents; specifically selected from these solvents, but not limited to: methanol, ethanol, propanol, isopropanol, diethyl ether, ethyl acetate, acetic acid, cyclohexane, methylene chloride, trichloromethane, tetrahydrofuran, pyridine, diethylamine, triethylamine, dimethylformamide, toluene and mixtures of at least two of them.

Unless otherwise specifically stated, in each of the above or following reactions of the present invention, when there is an excess amount of reactants, substances capable of reacting with the excess amount of reactants may be added at the end of the reaction to quench the reaction. In some examples, quenching with water or quenching with a saturated ammonium chloride solution may be used.

Unless otherwise specifically stated, in each of the reactions described above or below according to the present invention, the method for purifying the product at each reaction stage is selected from extraction, crystallization, solvent removal, and column chromatography. These operations are conventional in the art and can be performed by those skilled in the art, as appropriate.

The general formulas and methods for synthesizing them according to the present invention make it possible to obtain these specific substances, but are not limited to them, and specific compounds that can be obtained without creative work by specialists in this field under the guidance of the general formulas of the present invention and the general method of synthesis are included in the scope of the present invention.

Although preferred examples have been shown and described herein, it will be apparent to those skilled in the art that such examples are provided by way of example only. Those skilled in the art should understand that any changes, modifications, substitutions, and variations of these examples that do not depart from the principles and spirit of the present invention are also within the scope of protection of the present invention. The appended claims are intended to limit the scope of protection of the present invention and, thus, encompass methods and structures, and their equivalent forms, within the scope of the said claims.

The present invention provides pharmaceutical compositions comprising the above compounds, i.e. compounds covered by the general formula I of the target compound and their specific material representatives, or their pharmaceutically acceptable salts, and one or more pharmaceutically acceptable medicinal additives.

The present application provides pharmaceutical compositions comprising a compound described herein or pharmaceutically acceptable salts thereof and a pharmaceutically acceptable carrier. Said pharmaceutical compositions include, but are not limited to, an oral dosage form, a parenteral dosage form, a topical dosage form, and a rectal dosage form. Said compositions may be in liquid, solid, semi-solid, gel, or aerosol form. In some examples, said pharmaceutical compositions may be tablets, capsules, pills, powders, extended-release preparations, solutions and suspensions for oral use, sterile solutions, suspensions, or emulsions for parenteral administration, ointments or creams for topical use, or suppositories for rectal administration. In other examples, said pharmaceutical compositions are unit dosage forms suitable for single administration of precise doses.

The present invention provides all of the above compounds or pharmaceutically acceptable salts thereof and pharmaceutical compositions, their use as monoacylglycerol lipase inhibitors; or their use in the preparation of drugs for the treatment of diseases or disorders having pathological features of the monoacylglycerol lipase metabolic pathway.

The present invention provides methods of using all of the above compounds or pharmaceutically acceptable salts thereof and pharmaceutical compositions for inhibiting monoacylglycerol lipase. These methods include methods of inhibiting monoacylglycerol lipase in vivo and in vitro. Also provided are methods of using all of the above compounds or pharmaceutically acceptable salts thereof and pharmaceutical compositions for the treatment of diseases or disorders mediated by monoacylglycerol lipase.

The diseases or disorders listed are selected from the following: metabolic disease (e.g., obesity); kidney disease (e.g., acute inflammatory kidney injury and diabetic nephropathy); vomiting or regurgitation (e.g., chemotherapy-induced vomiting); nausea (e.g., refractory nausea or chemotherapy-induced nausea); eating disorder (e.g., anorexia or bulimia); neuropathy (e.g., diabetic neuropathy, pellagra neuropathy, alcoholic neuropathy, beriberi neuropathy); neurodegenerative disease [multiple sclerosis (MS), Parkinson's disease (PD), Huntington's disease, dementia, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), epilepsy, frontotemporal dementia, sleep disorder, Creutzfeldt-Jakob disease (CJD), or prion disease]; schizophrenia, depression, bipolar disorder, tremor, dyskinesia, dystonia, spasticity, Tourette's syndrome, withdrawal syndrome [alcohol withdrawal syndrome, antidepressant withdrawal syndrome, antipsychotic drug withdrawal syndrome, benzodiazepine withdrawal syndrome, marijuana withdrawal syndrome, neonatal abstinence syndrome, nicotine withdrawal syndrome, or opioid withdrawal syndrome]; traumatic brain injury; spinal cord injury; epileptic seizures; a disease involving abnormal growth or proliferation of cells [e.g., benign tumor or cancer, such as benign skin tumor, brain tumor, papilloma, prostate tumor, brain tumor (glioblastoma, medulloepithelioma, medulloblastoma, neuroblastoma, astrocytoma, astroblastoma, ependymoma, oligodendroglioma, plexus tumor, neuroepithelioma, pineal tumor, ependymoblastoma, malignant meningioma, sarcomatosis, melanoma, schwannoma), melanoma, metastatic tumor, kidney cancer, bladder cancer, brain cancer, glioblastoma (GBM), gastrointestinal cancer, leukemia, or blood cell cancer]; inflammatory disease [eg, appendicitis, bursitis, colitis, cystitis, dermatitis, phlebitis, rhinitis, tendinitis, tonsillitis, vasculitis, acne vulgaris, chronic prostatitis, glomerulonephritis, hypersensitivity, CPK, pelvic inflammatory disease, sarcoidosis, HIV encephalitis, rabies, brain abscess, neuroinflammation, central nervous system (CNS) inflammation]; immune system disease (eg, transplant rejection or gluten-sensitive 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 [eg, acute pain; chronic pain; inflammatory pain; visceral pain; postoperative pain; migraine; low back pain; joint pain; abdominal pain; chest pain; postmastectomy pain; menstrual pain; pain due to endometriosis; pain due to physical trauma; headache; sinus headache; tension headache; arachnoiditis; pain due to herpes virus; diabetic pain; pain caused by a condition selected from: rheumatoid arthritis, osteoarthritis, spondylitis, gout, childbirth, musculoskeletal disorder, skin disorder, dental disorder, fever, burn, sunburn, snakebite, venomous snakebite, spider bite, insect bite, neurogenic bladder, interstitial cystitis, urinary tract infection (UTI), rhinitis, contact dermatitis/hypersensitivity, pruritus, eczema, pharyngitis, mucositis, enteritis, irritable bowel syndrome (IBS), cholecystitis, and pancreatitis; neuropathic pain (eg, neuropathic low back pain, complex regional pain syndrome, trigeminal neuralgia, causalgia, toxic neuropathy, reflex sympathetic dystrophy, diabetic neuropathy, chronic chemotherapy-induced neuropathy, or lumbosacral pain)]; demyelinating disease [e.g., multiple sclerosis (MS), Devic disease, CNS neuropathies, central pontine myelinolysis, syphilitic myelopathy, leukoencephalopathy, leukodystrophies, 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 impairment [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 dysfunction (POCD)].

The present invention provides the use of a MAG inhibitor for the preparation of medicaments intended for the treatment of a MAGL-mediated disease or disorder.

Some examples of MAGL inhibitor-associated disorders include pain, migraine, irritable bowel syndrome, or ulcerative colitis.

In the above application examples, pain is defined as traumatic pain or pathological pain. Specifically, pathological pain is inflammatory pain or endogenous pain. More specifically, this pain includes inflammatory pain of biological origin, inflammatory pain of chemical origin, pain of blood origin, pain of immune origin, pain of endocrine origin, pain caused by metabolic disorders, neurogenic pain, or cardiogenic pain.

In the above application examples, the pain referred to is transient pain, acute pain, or chronic pain.

In the above application examples, the pain referred to is pain in the nervous system, pain in the cardiovascular system, pain in the blood system, pain in the respiratory system, pain in the digestive system, pain in the endocrine system, pain in the urinary system, pain in the musculoskeletal system, or pain in the immune system.

In the above application examples, the migraine in question is simple migraine, typical migraine, cluster migraine.

In the above examples of use, the irritable bowel syndrome mentioned is irritable bowel syndrome with constipation, irritable bowel syndrome with diarrhea, irritable bowel syndrome with alternating diarrhea and constipation.

The MAGL inhibitor of the present invention is capable of reducing the duration of writhing in mice induced by acetic acid, prolonging the latency time, and increasing the thermal pain threshold in mice in the hot plate test. It has a pronounced analgesic effect.

In some examples, the MAGL inhibitor of the present invention is able to significantly prolong blood clotting time in migraine models, increase the number of immobility episodes during tail suspension in mice, and significantly increase 5-HT levels in brain tissue. It has a significant therapeutic effect on migraine.

The MAGL inhibitor of the present invention is capable of significantly reducing the AWR score and colonic and serum 5-HT levels in a model of irritable bowel syndrome. It exhibits significant therapeutic efficacy in irritable bowel syndrome. Thus, the compounds have been confirmed to possess biological activity for the prevention and treatment of irritable bowel syndrome.

The MAGL inhibitor of the present invention is able to significantly increase colon length, reduce the total amount of blood in stool, and reduce the level of IL-1 in an ulcerative colitis model.

Some chemical terms

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Unless otherwise defined, all patents, patent applications, and published materials cited herein are hereby incorporated by reference in their entirety.

It should be understood that the foregoing general description and the following detailed description are merely examples and explanations and are not limiting of any of the claimed subjects of the invention. In this application, the use of the singular includes the plural unless expressly stated otherwise. It should be noted that references to the singular in this description and the appended claims include references to the plural unless the context clearly indicates otherwise. It should also be noted that the use of "or" means "and/or" unless otherwise stated. Furthermore, the use of the term "including," as well as its other forms such as "includes," "comprises," and "has," is not limiting.

Definitions of standard chemical terms can be found in works referenced herein, including, but not limited to, Carey and Sundberg, "Advanced Organic Chemistry 4th Ed." Vols. A (2000) and B (2001), Plenum Press, New York. Unless otherwise noted, standard techniques of mass spectroscopy, nuclear magnetic resonance (NMR), high-performance liquid chromatography (HPLC), infrared (IR) spectroscopic analysis and UV/Vis spectroscopy, and pharmacology known in the art are used. Unless specifically defined, the nomenclature used herein is that known in the art. Standard techniques can be used for chemical synthesis, chemical analysis, preparation, manufacture, and delivery of pharmaceuticals, and treatment of patients. Reactions and purification methods can be carried out, for example, using a Reagent Kit according to the manufacturer's instructions, or as commonly performed in the art, or as described herein. The above methods and procedures can generally be performed using conventional methods known in the art and as described in various general and more specific references cited and discussed throughout the description of this invention. Throughout the description, groups and their substituents can be selected by those skilled in the art to produce stable fragments and compounds.

As used herein, the term "alkyl" refers to a saturated hydrocarbon with a straight or branched chain. Examples of alkyls include, but are not limited to, straight or branched hydrocarbons with 1-6, 1-4, or 1-3 carbon atoms, referred to herein as C1-6 alkyl, C1-4 alkyl, and C1-3 alkyl. Examples of 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 2,2-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, tert-butyl, amyl, neo-butyl, neopentyl, hexyl, etc.

The term "C _1-4 alkyl" as used herein includes C _1-3 alkyl, C _1-2 alkyl, C _2-3 alkyl, C _2-4 alkyl, specifically includes, but is not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, neo-butyl as an example.

Alkoxy is an alkyloxy group. As used herein, the term "C _1-3 alkoxy" includes C _1-2 alkoxy and C _2-3 alkoxy, in particular, examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy.

The term "cycloalkyl" as used herein refers to saturated or partially unsaturated hydrocarbons, such as a monocyclic 5-8-membered ring, a bicyclic 8-12-membered ring. _C3-8 cycloalkyl includes 3-8, 3-6, 4-6, 6-8, 3-7 or 5-8 carbons, and is referred to herein as _C3-8 cycloalkyl, _C3-6 cycloalkyl or _C4-6 cycloalkyl, which have different forms but are equivalent in meaning. Specific examples of cycloalkyl groups include, but are not limited to, cyclohexyl, cyclopentyl, cyclopentenyl, cyclobutyl or cyclopropyl.

The term "heterocycle" as used herein means a monocyclic 5-8-membered ring or a bicyclic 8-12-membered ring containing one or more heteroatoms (e.g., one to three heteroatoms such as nitrogen, oxygen, and sulfur). As described herein, _C3-8 heterocycle refers to monocyclic or polycyclic groups having 3-8 atoms, including _C3-6 heterocycle, _C4-8 heterocycle, _C5-8 heterocycle, _C4-7 heterocycle, _C3-7 heterocycle, _C6-8 heterocycle; having at least one ring with one, two, three, or four heteroatoms independently selected from N, O, or S, wherein none of the rings is aromatic. Examples of heterocycles include, but are not limited to, morpholinyl, thiomorpholinyl, furanyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofurfuryl, and 1,3-dioxanyl.

As used herein, the term "C _3-8 cycloalkoxy" means cyclic alkoxy, where cycloalkoxy is analogous to the above interpretation of cycloalkyl.

The term "aryl" as used herein means a monocyclic or polycyclic aromatic carbocyclic group having at least one aromatic ring comprising 6-12 carbon atoms. The term " _C6-10 aryl" as used herein includes _C6-8 aryl, C6-10 aryl, _C6-9 aryl, C7-10 aryl, C8-10 _aryl , C6 _aryl , _C7 aryl, _{C8 aryl} , _C9 aryl, _{C10 aryl} . Specific examples include, but are not limited to, phenyl, mono- or polyalkyl-substituted phenyl, as well as pentalenyl, indenyl, naphthyl and azulenyl. Mono- or polyalkyl-substituted phenyl groups include, but are not limited to, tolyl, ethylphenyl, dimethylphenyl, propylphenyl, isopropylphenyl, methylethylphenyl; trimethylphenyl, butylphenyl, methylpropylphenyl, methylisopropylphenyl, diethylphenyl, dimethylethylphenyl, tetramethylphenyl, tert-butylphenyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, 1H-indenyl.

The term "C _5-18 aryl" used herein includes C _6-10 aryl, C _11-12 aryl, C _13-15 aryl and C _16-18 aryl, such as pentylphenyl, methylbutylphenyl, methylisobutylphenyl, methyl tert-butylphenyl, ethylpropylphenyl, ethyl isopropylphenyl, methylpropylpropylphenyl, methylisopropylphenyl, trimethylethylphenyl, pentamethylphenyl, etc.

The term "heteroaryl" or "heteroaromatic ring" or "heteroaromatic group" as used herein means a monocyclic aromatic 5-8 membered ring or a bicyclic aromatic 8-12 membered ring containing one or more heteroatoms (e.g., 1-3 heteroatoms such as nitrogen, oxygen and sulfur). The term "5-10 membered heteroaryl" as used in the text includes 5-9-membered heteroaryl, 5-8-membered heteroaryl, 5-7-membered heteroaryl, 5-6-membered heteroaryl, 6-10-membered heteroaryl, 6-9-membered heteroaryl, 6-8-membered heteroaryl, 7-10-membered heteroaryl, 7-9-membered heteroaryl, 8-10-membered heteroaryl, 9-10-membered heteroaryl, 5-membered heteroaryl, 6-membered heteroaryl, 7-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, 10-membered heteroaryl. If 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 heteroaryl groups include, but are not limited to: benzofuranyl, iso-benzofuranyl, 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-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl, 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-alpha]pyridyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, pyridyl, benzoisoxazolyl.

The terms "one or more substituents selected from benzene, naphthalene..." or "one or more benzene, naphthalene..." refer to one or two or three selected from phenyl, naphthyl...

The term "acyloxy" refers to the group -OC(=O)-R; wherein R is selected from H and substituted or unsubstituted C _1-5 alkyl, said unsubstituted C _1-5 alkyl is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, neobutyl, n-pentyl, iso-pentyl, neopentyl, cyclopentyl; said substituted C _1-5 alkyl is C _1-5 alkyl substituted with halogen, hydroxyl, amino, carboxyl, C _1-3 alkyl, C _1-3 alkoxy, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, C _3-8 cycloalkoxy, aryl, heteroaryl. In some examples, R is n-propyl or .

The term "halogen" or "halo" as used herein means F, Cl, Br or I.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes the chemically equivalent substituent obtained when the structural formula is written from right to left. For example, CH ₂ O is equivalent to OCH ₂ .

The term "compound" in this application may include all stereoisomers, geometric isomers, tautomers, and isotopes. The compounds of the present application may be asymmetric, for example, having one or more stereoisomers. Unless otherwise specified, all stereoisomers, such as enantiomers and diastereoisomers, are included. The compounds of the present application containing asymmetrically substituted carbon atoms may be isolated in optically active pure form or in racemic form. The optically active pure form may be isolated from a racemic mixture or synthesized using chiral raw materials or chiral reagents. The compounds of the present application also include tautomeric forms. Tautomeric forms arise from the exchange of a single bond with an adjacent double bond along with the migration of a proton. The compounds of the present application also include all isotopes of atoms in the intermediate compound and in the final compound. Isotopes of atoms 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 part or all of the hydrogen (H) is replaced by tritium (T) and/or deuterium (D); also compounds in which part or all of ¹² C is replaced by ¹³ C and/or ¹⁴ C; and compounds in which other atoms (e.g., N, O, P, S) are replaced by other isotopes, such as ¹⁴ N and ¹⁵ N; ¹⁸ O and ¹⁷ O; ³¹ P and ³² P; ³⁵ S and ³⁶ S, etc. The compounds described herein may contain one or more stereocenters, and each center may exist in the R or S configuration or combinations thereof. Similarly, the compounds described herein may contain one or more double bonds, and each may exist in the E (trans) or Z (cis) configuration, or combinations thereof. The representation of one particular stereoisomer, regioisomer, diastereomer, enantiomer, or epimer should be understood to include all possible stereoisomers, regioisomers, diastereomers, enantiomers, or epimers, and mixtures thereof. Thus, the compounds described herein include all individual stereoisomeric, regioisomeric, diastereomeric, enantiomeric, and epimeric forms, as well as corresponding mixtures thereof. Techniques for inverting or leaving unchanged a particular stereocenter and techniques for separating mixtures of stereoisomers are well known in the art, and one skilled in the art will be able to select the appropriate method for a 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.

The term "optional/voluntary" or "optionally/voluntarily" means that the phenomenon or state described below may or may not occur and that the description includes cases where the stated phenomenon or state occurs and cases where it does not occur.

Some pharmacological terms

As used herein, the terms "treat," "treating," or "treatment" and other grammatical equivalents include alleviating, mitigating, or ameliorating the symptoms of a disease or condition, preventing additional symptoms, alleviating or preventing the underlying metabolic causes of symptoms, inhibiting a disease or condition, such as stopping the progression of a disease or condition, alleviating a disease or condition, causing remission of a disease or condition, alleviating a condition caused by a disease or condition, or stopping the symptoms of a disease or condition; these terms include prophylactic use. The terms further include achieving a therapeutic effect and/or a prophylactic effect. A therapeutic effect is understood to mean eliminating or ameliorating the underlying disorder being treated. Furthermore, a therapeutic effect is achieved by eliminating or ameliorating one or more physiological symptoms associated with the underlying disorder, so that the patient experiences an improvement, although the patient may still suffer from the underlying disorder. For a prophylactic effect, the compositions may be administered to a patient at risk of developing a particular disease or to a patient complaining of one or more physiological symptoms of a disease, even if a diagnosis of the disease may not be made.

As used herein, the terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" refer to the amount of one or more active substances (such as a compound of the present application) that, when administered, is sufficient to provide some relief from one or more undesirable physiological changes or disorders being treated. The result may be a reduction and/or remission of the signs, symptoms, or causes of a disease, or any other desired change in a biological system. For example, an "effective amount" for therapeutic purposes is the amount of a composition comprising the compounds disclosed herein necessary to provide clinically significant relief from the condition. An effective amount suitable for use in each particular case can be determined using methods such as dose escalation tests.

As used herein, the terms "administer," "administering," or "administering," and the like, refer to methods that can be used to ensure delivery of compounds or compositions to the desired site of biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, or infusion), topical, and rectal administration. One skilled in the art is familiar with administration techniques that can be used with the compounds and methods described herein, such as those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

As used herein, the term "acceptable" in relation to a composition, formulation, or ingredient means not having a permanent adverse effect on the general health of the subject being treated.

The term "pharmaceutically acceptable" as used herein refers to a substance (such as a carrier or diluent) that does not interfere with the biological activity or properties of the compounds of the present application and is relatively non-toxic, i.e., the substance can be administered to individuals without causing adverse effects of a biological response or without interacting in an undesirable manner with any component contained in the composition.

As used herein, the term "pharmaceutical composition" refers to a biologically active compound mixed with at least one pharmaceutically acceptable chemical component, such as, but not limited to, carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents and/or excipients.

The term "carrier" as used herein refers to a relatively non-toxic substance that facilitates the introduction of the compounds of the present application into cells or tissues.

As used herein, the term "pharmaceutically acceptable salts" refers to salts that retain the biological potency of the free acid and free base of the compound and do not exhibit adverse biological or other effects. The compounds of the present application also include pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to the form in which a basic group in the parent compound is converted to a salt. Pharmaceutically acceptable salts include, but are not limited to, salts of basic groups, such as inorganic or organic salts of the amine (amino) group. Pharmaceutically acceptable salts of the present application can be synthesized from the parent compound, i.e., the basic groups in the parent compound are reacted with 1-4 equivalents of acid in a solvent system. Suitable salts are listed in Remingtong's Pharmaceutical Sciences, 17th ^ed ., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

Unless otherwise specified, salts in this application refer to acid salts formed with organic acids/inorganic acids and base salts formed with organic bases/inorganic bases. In addition, zwitterions (inner salts) are formed when the basic functional group of the compound is pyridine or imidazole (but not limited to pyridine or imidazole) and the acid functional group is a carboxylic acid (but not limited to a carboxylic acid); internal salts are also included in the salts in this application.

Examples of specific implementation

All specific compounds provided by the present invention can be prepared using the synthetic methods disclosed herein. A more detailed description of the present invention is provided below, accompanied by specific examples, but the invention is not limited to them. Furthermore, all other embodiments obtained by those skilled in the art based on the embodiments of the present invention, without creative effort, will fall within the scope of protection of the present invention.

Example 1. Synthesis

Obtaining intermediate product a.

N-Boc-4 piperidinecarboxylic acid (458.6 mg, 2 mmol, 1 equiv) was dissolved in 15 mL of DCM, DIPEA (0.7 mL, 4 mmol, 2 equiv), HATU (912 mg, 2.4 mmol, 1.2 equiv) were added and the mixture was stirred for 10 min at room temperature. Aniline (0.365 mL, 4 mmol, 2 equiv) was added and the mixture was stirred for 12 min at room temperature, washed twice with water and twice with 0.1 M dilute hydrochloric acid, and evaporated under reduced pressure to remove the solvent to give intermediate compound a.

Obtaining intermediate compound b.

Intermediate a was dissolved in 10 ml ethyl acetate, 2 ml HCl/EA solution was added and stirred at room temperature for 24 h. A white solid was isolated, after suction filtration it was washed with ethyl acetate to give intermediate b as a white solid 347.5 mg (72.4%).

Receipt of MAGLZ-II-01,

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

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). The MS results are consistent with the theoretical values.

In examples 2-23, the synthesis was carried out according to the general method similar to the method in example 1,

where, in examples 22 and 23, b was replaced by

And .

The MS results of all target products were consistent with theoretical values, and the NMR results were as follows:

Number Target product number Characteristic data Example 2 ¹ H NMR(CDCl ₃ ) δ (ppm):7.32-7.42(m,4H), 7.11-7.20(m,3H),6.95-7.07(m,4H),6.74(s,1H), 2.79-3.05(m,2H),2.43-2.56(m,1H),2.27(s,6H), 1.71-2.07(m,6H). Example 3 ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 9.92 (s,1H), 7.40-7.47(m,4H), 7.32-7.34 (m,1H), 7.07-7.23 (m,5H), 7.00-7.05 (m,2H), 3.78-4.68 (m,2H), 3.72 (s,3H), 2.78-3.17 (m,2H), 2.57-2.68 (m,1H), 1.74-1.93 (m,2H), 1.53-1.68 (m,2H); Example 4 ¹ H NMR(DMSOd ₆ ) δ(ppm):
9.83(s,1H),9.37(s,1H),7.40-7.46(m,4H),7.17-7.25 (m,2H),7.08-7.13(m,2H),7.00-7.06(m,3H),6.93-6.99 (m,1H),6.38-6.45(m,1H),2.88-2.92(m,1H),2.68-2.75 (m,3H),2.56-2.66(m,1H), 1.75-1.93(m,2H),1.50-1.65(m,2H) Example 5 ¹ H NMR(CDCl ₃ ) δ (ppm): 7.32-7.44(m,5H), 7.17-7.25(m,1H),6.92-6.99(m,1H),6.65-6.71(m,1H),3.80(s,3H),2.73-3.31(m,4H),2.46-2.56(m,1H),1.78-1.97 (m,4H). Example 6 ¹ H NMR(DMSOd ₆ ) δ
(ppm):9.82(s,1H),9.38(s,1H),7.42-7.55(m,4H), 7.16-7.20(m,1H),7.01-7.09(m,1H),6.93-6.99 (m,1H),6.39-6.45(m,1H),2.68-2.91(m,4H), 2.55 -2.64(m,1H),1.80-2.02(m,2H), 1.53-1.65(m,2H). Example 7 ¹ H NMR(CDCl ₃ ) δ
(ppm):7.70(s,1H),7.42-7.52(m,2H),7.21-7.37(m,3H),6.92-6.98(m,2H),3.81(s,3H),2.80-2.99(m,4H), 2.45-2.55(m,1H),1.99-2.12(m,2H), 1.78 -1.89(m,2H). Example 8 ¹ H NMR(DMSOd ₆ ) δ
(ppm):10.08(s,1H),9.68(s,1H),7.60-7.66(m,2H), 7.32-7.38(m,2H),7.20-7.28(m,1H),6.73-6.86(m,3H), 2.67-2.93(m,3H),2.57-2.66(m,1H),1.68-2.00(m,3H), 1.52-1.65(m,2H). Example 9 ¹ H NMR(DMSOd ₆ ) δ
(ppm):10.02(s,1H),9.72(s,1H),7.60-7.66(m,2H),7.31-7.37(m,2H),7.18-7.25(m,1H ),7.08-7.13(m,1H),6.81-6.89(m,2H),3.98-4.13(m,2H),2.85-2.97(m,2H),2.54-2.66 (m,1H),1.75-1.86(m,2H), 1.52-1.66(m,2H). Example 10 ¹ H NMR(DMSOd ₆ ) δ(ppm):
10.10(s,1H),9.87(s,1H),7.61-7.66(m,2H),7.33-7.38 (m,2H),7.23-7.29(m,2H),6.77-6.83(m,2H),2.86-3.02 (m,2H),2.54-2.85(m,3H),1.77-1.87(m,2H),1.52-1.62 (m,2H) Example 11 ¹ H NMR(DMSOd ₆ ) δ(ppm): 10.08(s,1H), 9.62(s,1H), 7.82(t,1H,J=3Hz), 7.44-7.50 (m,1H), 7.32(t,1H, J=6Hz), 7.24 (t,1H,J=6Hz), 7.07-7.12(m,1H), 6.74-6.86(m,3H), 2.89-2.98 (m,2H), 2.60-2.74 (m,1H), 2.49-2.58 (m,2H), 1.82-1.91 (m,2H), 1.52-1.64 (m,2H); Example 12 ¹ H NMR(DMSOd ₆ ) δ(ppm):
10.15(s,1H),9.79(s,1H),7.835(t,1H, J=1.5Hz),7.42-7.48(m,1H),7.32(t,1H,J=6Hz),7.18-7.26(m,1H),7.06-7.13(m,2H),6.80-6.89(m,2H),2.69-2.89 (m,4H),2.55-2.65(m,1H),1.69-1.93(m,2H),1.49-1.67 (m,2H). Example 13 ¹ H NMR(DMSOd ₆ ) δ(ppm):
9.64(s,1H),9.07(s,1H),7.87-8.00(m,1H), 7.24 (t,1H, J=7.8Hz),6.99-7.13(m,2H),6.72-6.94(m,4H),3.83 (s,3H),2.62-3.16(m,4H),1.43-1.95(m,5H). Example 14 ¹ H NMR(DMSOd ₆ ) δ(ppm): 10.12(s,1H), 7.83(t,1H,J=1.92Hz),7.43-7.50(m,1H),7.30-7.39 (m,2H), 7.07-7.11(m,1H),7.00-7.04(m,1H), 6.91-6.96(m,2H),3.79(s,3H),2.67-3.30(m,4H), 2.58-2.67(m,1H),1.76-1.95(m,2H),1.53-1.66(m,2H). Example 15 ¹ H NMR(DMSOd ₆ ) δ(ppm): 9.68(s,1H),
9.23(s,1H),7.32-7.42(m,1H),7.15-7.20(m,1H), 6.90-7.06(m,5H),6.40-6.48(m,1H),3.80(s,3H), 2.68-3.20(m,4H),2.56-2.68(m,1H),1.76-1.79(m,2H), 1.54-1.66(m,2H). Example 16 ¹ H NMR(DMSOd ₆ ) δ(ppm): 10.13(s,1H), 7.79-7.88(m,1H),7.41-7.48(m,5H),7.29-7.36(m,1H), 7.17-7.24(m,1H),7.07-7.12(m,3H),7.01-7.05(m,3H), 2.56-3.29(m,4H), 1.46-1.99(m,5H) Example 17 ¹ H NMR(DMSOd ₆ ) δ(ppm):
10.14(s,1H),7.97-8.04(m,4H),7.81-7.87(m,1H), 7.57-7.62(m,2H),7.44-7.54(m,2H),7.29-7.38 (m,1H),7.06-7.14(m,1H),3.65-4.66(m,2H), 2.88-3.22(m,2H),2.64-2.71(m,1H),1.62-1.97(m,4H). Example 18 ¹ H NMR(DMSOd ₆ ) δ(ppm): 11.58(s,1H),
10.17(s,1H),7.82-7.88(m,1H),7.58-7.65(m,1H), 7.40-7.50(m,2H),7.29-7.37(m,1H),7.15-7.23(m,1H), 7.01-7.13(m,2H),6.78-6.82(m,1H),4.44-4.58(m,2H), 3.00-3.26(m,2H),2.66-2.74(m,1H),1.86-2.00(m,2H), 1.58-1.74(m,2H). Example 19 ¹ H NMR(DMSOd ₆ ) δ(ppm):
10.16(s,1H),7.83-7.86(m,1H),7.69-7.78(m,4H), 7.47-7.52(m,4H),7.37-7.46(m,2H),7.33(t,1H, J=8.04),7.07-7.13(m,1H),3.67-4.61(m,2H), 2.89-3.16(m,2H),2.61-2.69(m,1H),1.82-1.99(m,2H), 1.56-1.69(m,2H). Example 20 [M+H] ⁺ 401.2 Example 21 ¹ H NMR(300 MHz, DMSO-d ₆ ) δ(ppm): 10.17 (s,1H),7.83-7.87(t,1H),7.59-7.65(d,1H),7.50-7.56 (d,1H), 7.45-7.49(m,1H),7.30-7.37(t,1H),7.23-7.30 (t,1H), 7.11-7.14(m,1H),7.07-7.11(m,1H),6.67 (s,1H),3.77 (s,3H), 2.63-2.76(m,2H),1.98-2.06(s,1H), 1.85-1.97(d,2H), 1.57-1.73(m,2H),1.15-1.28(m,2H)
MS:[M+Na] ⁺ :418.2 Example 22 ¹ HNMR (300 MHz, DMSO-d ₆ ) δ(ppm): 9.68 (s,1H), 7.83-7.84(d,1H), 7.73-7.75 (d,1H), 7.40-7.43 (m,1H), 7.21-7.25 (t,1H), 6.81-6.84 (m,1H), 6.78-6.80 (d,1H), 6.75-6.76 (m,1H), 4.40 (d,1H), 3.67 (s,1H), 3.21 (s,2H),3.09-3.15 (m,1H), 2.11-2.15 (m,2H), 1.75-1.78 (m,2H);
[M+Na] ⁺ 379.1 Example 23 ¹ H NMR (300 MHz, DMSO-d ₆ ) δ(ppm): 12.48 (s,1H), 9.73 (s,1H), 7.56-7.62(t,1H), 7.50-7.56 (d,1H), 7.24-7.29 (t,1H), 7.18-7.20 (m,1H), 6.80-6.87 (m,3H), 4.50 (d,1H), 3.71 (d,1H), 3.21 (s,2H), 3.02-3.17 (m,1H),
2.03-2.10 (m,2H), 1.71-1.81 (m,2H);
[M+H] ⁺ 356.1

In Examples 24-30, the description of the synthesis of the target product is based on the following structural formula:

wherein said R ₁ is selected from hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy; R ₂ is selected from hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy, phenyl and aryl substituted by hydroxyl, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, methoxy. The number of R ₁ and R ₂ is 0, 1, 2, 3 or 4; the mode of binding R ₁ , R ₂ to the parent nucleus is not limited to a single bond.

Example Compound R1 R2 Characteristic data Example 24 MAGLZ-II-20 3-C1 4-OH ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.12 (s,1H), 9.83 (s,1H), 7.81-7.84 (m,1H), 7.42-7.49 (m,1H), 7.29-7.36 (m,1H), 7.22-7.28 (m,2H), 7.06-7.12 (m,1H), 6.77-6.83 (m,2H), 2.89-2.90 (m,2H), 2.73-2.74 (m,2H), 2.55-2.67 (m,1H), 1.77-1.89 (m,2H), 1.50-1.65 (m,2H);
[M+H] ⁺ 359.1 Example 25 MAGLZ-II-21 3-C1 4-C1 ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.12 (s,1H), 7.81-7.84 (m,1H), 7.49-7.55 (m,2H), 7.41-7.48 (m,3H), 7.29-7.36 (m,1H), 7.07-7.12 (m,1H), 3.16-3.19 (m,2H), 2.79-3.11 (m,2H), 2.58-2.69 (m,1H), 1.71-1.95 (m,2H), 1.52-1.68 (m,2H);
[M+H] ⁺ 377.08 Example 26 MAGLZ-II-22 3-C1 and 4-C1 3-OH ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.24 (s,1H), 9.67 (s,1H),8.00-8.03 (m,1H), 7.47-7.58 (m,2H), 7.24 (t,1H, J = 7.5 Hz), 6.80-6.86 (m,1H), 6.74-6.80 (m,2H), 2.88-2.91 (m,2H), 2.69-2.74 (m,2H), 2.56-2.67 (m,1H), 1.72-1.93 (m,2H), 1.50-1.63 (m,2H);
[M+H] ⁺ 393.08 Example 27 MAGLZ-II-23 3-C1 and 5-C1 3-OH ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.27 (s,1H), 9.67 (s,1H), 7.94-7.97 (m,1H), 7.67-7.70 (m,2H), 7.25-7.28 (m,1H), 6.72-6.85 (m,3H), 2.88-2.91 (m,2H), 2.73-2.74 (m,2H), 2.56-2.67 (m,1H), 1.73-1.92 (m,2H), 1.48-1.63 (m,2H);
[M+H] ⁺ 393.08 Example 28 MAGLZ-II-24 3-C1 and 4-C1 ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.24 (s,1H), 7.95-8.02 (m,5H), 7.57-7.60 (m,2H), 7.48-7.54 (m,3H), 2.89 (s,2H), 2.73 (s,2H), 2.61-2.65 (m,1H), 1.74-1.90 (m,2H), 1.57-1.67 (m,2H);
[M+H] ⁺ 427.09 Example 29 MAGLZ-II-25 3-C1 and 5-C1 ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.28 (s,1H), 7.95-8.00 (m,4H), 7.68-7.69 (d,2H), 7.57-7.60 (m,2H), 7.49-7.52 (m,1H), 7.25-7.26 (t,1H), 2.89 (s,2H), 2.73 (s,2H), 2.59-2.63 (m,1H), 1.73-1.91 (m,2H), 1.58-1.65 (m,2H);
[M+H] ⁺ 427.09 Example 30 MAGLZ-II-26 3-F 3-OH ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.16 (s,1H), 9.69 (s,1H), 7.58-7.65 (m,1H), 7.27-7.38 (m,2H), 7.24 (t,1H, J = 7.5 Hz), 6.73-6.90 (m,4H), 2.71-3.12 (m,4H), 2.56-2.68 (m,1H), 1.73-1.93 (m,2H), 1.49-1.65 (m,2H);[M+H] ⁺ 343.15 Example 31 MAGLZ-II-27 3-F and 4-C1 3-OH ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.30 (s,1H), 9.76 (s,1H), 7.76-7.81 (m,1H), 7.47-7.53 (t,1H), 7.32-7.36 (m,1H), 7.21-7.27 (t,1H), 6.74-6.85 (m,3H), 2.66-2.89 (m,4H), 2.63-2.73 (m,1H), 1.76-1.85 (m,2H), 1.54-1.57 (m,2H);
[M+H] ⁺ 377.10 Example 32 MAGLZ-II-29 3-C1 ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.13 (s,1H), 9.93 (s,1H), 7.82-7.87 (m,3H), 7.72-7.75 (d,1H), 7.44-7.47 (m,1H), 7.37-7.40(m,1H),7.29-7.34 (t,1H), 7.07-7.15 (m,3H), 2.64-3.02 (m,4H), 2.60-2.64 (m,1H), 1.82-1.85 (m,2H), 1.56-1.68 (m,2H);
[M+Na] ⁺ 431.12 Example 33 MAGLZ-II-11(a) 3-C1 2-F and 5-OH ¹ H NMR(300 MHz, DMSO-d ₆ ) δ (ppm): 10.17 (s,1H), 9.68 (s,1H), 7.82 (s,1H), 7.43-7.45 (d,1H), 7.29-7.35 (t,1H), 7.06-7.09 (m,2H), 6.79-6.82 (m,1H), 6.67 (s,1H), 2.84-2.89 (d,2H), 2.68-2.73 (d,2H), 2.61-2.62 (m,1H), 1.76-1.93 (m,4H);
[M+Na] ⁺ 399.1

Example 34. Preparation of MAGLZ-II-11F.

The starting N-Boc-4-piperidinecarboxylic acid (20) (229.3 mg, 1 mmol) was dissolved in 10 ml CH₂Cl₂, added DIPEA (0.35 ml, 2 mmol) and HATU (456 mg, 1.2 mmol), stirred for 10 min at room temperature, added 3-chloroaniline (0.21 ml, 2 mmol), stirred for 6 h at room temperature, washed with water three times, washed three times with 10% dilute hydrochloric acid, dried with anhydrous Na₂SO₄, and removed CH₂Cl₂by evaporation on a rotary evaporator, after which intermediate compound 21h was obtained without further purification.

Intermediate 21h was dissolved in 10 mL of EtOAc, 4 M HCl/EtOAc solution (2 mL) was added, and the mixture was stirred at room temperature for 12 h. A white solid was isolated. After suction filtration, the filter cake was washed with EtOAc, yielding intermediate 22h as a white solid (188.1 mg, 68.4% yield).

Intermediate 24 (62.5 mg, 0.3 mmol) was dissolved in anhydrous DMF, HATU (136.8 mg, 0.36 mmol), triethylamine (151.8 mg, 1.5 mmol) were added, stirred for 30 min at room temperature, then intermediate 22h (107.33 mg, 0.39 mmol) was added, stirred for 12 h at room temperature, after completion of the reaction, a saturated solution was added, extracted three times with EtOAc, the organic phases were combined, washed three times with saturated sodium chloride solution, dried with anhydrous Na ₂ SO ₄ , after isolation and purification by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain 94.9 mg of yellow solid, the yield was 74.1%.

Characteristic data.

¹ H NMR (300 MHz, CDCl ₃ ) ¹ H NMR (300 MHz, CDCl ₃ ) 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.

Example 35 Synthesis of MAGLZ-II-11J

The starting N-Boc-L-tyrosine (0.5 mmol, 140.5 mg) was dissolved in 10 ml of anhydrous ₂ CHCl ₂ , intermediate compound II-11 (179.5 mg, 0.5 mmol), DMAP (0.1 mmol, 12.2 mg) were added, after the reaction started, it was cooled to 0 °C, DCC (0.55 mmol, 113.4 mg) dissolved in 3 ml of anhydrous CHCl ₂ (113.4 mg) was added, stirred for 10 min, the reaction temperature was brought to room temperature, stirred for 12 h at room temperature, the reaction was not complete, after stopping the reaction, suction filtration was performed, the filtrate was evaporated on a rotary evaporator, 20 ml of CH ₂ Cl ₂ were added, the mixture was washed three times with a saturated sodium chloride solution and dried over anhydrous sodium sulfate. After isolation and purification by column chromatography (dichloromethane:methanol = 100:1), 52.6 mg of a white solid was obtained, the yield was 16.9%.

Intermediate 25 (37 mg) was dissolved in 10 mL EtOAc, 4 M HCl/EtOAc solution (2 mL) was added, and the mixture was stirred at room temperature for 12 h. The resulting white solid was filtered off with suction and washed with EtOAc. The isolated precipitate was treated with 10 mL sodium bicarbonate solution and extracted three times with EtOAc. The organic phases were combined, washed three times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the EtOAc was removed by rotary evaporation to yield 12 mg of a pale white solid; the yield was 40%.

Characteristic data of MAGLZ-II-11J: ¹ H NMR (300 MHz, DMSO-d ₆ ) 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.

Example 36. MAGL enzyme analysis

Using the Cayman MAGL inhibitor screening reagent kit, an ethanol solution of 4-nitrophenylacetic acid was used as the MAGL substrate according to the instructions; the final substrate concentration was 236 μM. 150 μl of 1X assay buffer, 10 μl of recombinant human MAGL protein, and 10 μl of MAGL inhibitor at various concentrations (six points in the range of 1 nM-1000 nM) were added to each well of a 96-well plate. JZL195 (inhibitor, positive control) was used as a positive control well. After incubation for 5 min at room temperature, 10 μl of MAGL substrate was added to each well. The 86 wells were shaken for 10 seconds and left at room temperature for 10 min to determine the absorbance at 410 nm. In addition, 100% inhibition control wells (three replicate wells with 150 µl 1X assay buffer, 10 µl MAGL, and 10 µl DMSO per well) and background wells (three replicate wells with 160 µl 1X assay buffer and 10 µl DMSO per well) were added. IC50 curves for each inhibitor were then calculated using Graphad Prism 5.0 software, and the resulting values are presented in Table 2.

Table 2. Activity data

MAGLX connection number IC50(μM) MAGLZ-II-01 0.045 MAGLZ-II-02 0.027 MAGLZ-II-03 0.749 MAGLZ-II-04 ND MAGLZ-II-05 4.07 MAGLZ-II-06 1.96 MAGLZ-II-07 0.576 MAGLZ-II-08 1.34 MAGLZ-II-09 10.8 MAGLZ-II-10 0.662 MAGLZ-II-11 0.096 MAGLZ-II-12 ND MAGLZ-II-13 0.926 MAGLZ-II-14 0.333 MAGLZ-II-15 ND MAGLZ-II-16 0.018 MAGLZ-II-17 0.015 MAGLZ-II-18 0.670 MAGLZ-II-19 2.82 MAGLZ-II-18 (c) 0.0026 MAGLZ-II-18 (a) 2.15 MAGLZ-II-20 9.92 MAGLZ-II-21 ND MAGLZ-II-22 1.77 MAGLZ-II-23 4.03 MAGLZ-II-24 0.275 MAGLZ-II-25 0.139 MAGLZ-II-26 12.2 MAGLZ-II-27 2.90 MAGLZ-II-29 0.252 MAGLZ-II-30 0.011 MAGLZ-II-31 ND MAGLZ-II-11 (a) 2.15 MAGLZ-II-11F 0.126 MAGLZ-II-11J 1.60

Example 37. Tissue distribution test of the MAGL inhibitor of the present invention

Reagents for detection: MAGLZ-II-11, MAGLZ-II-16, MAGLZ-II-17.

Test object: mice.

Administered dose: 16 mg/kg (drug concentration: 8 mg/ml).

Method of administration: subcutaneous injection.

Sampling time: Samples were taken at 1, 2 and 5 hours after administration.

Tested tissues: brain, heart, liver, spleen, lungs, kidneys, pancreas, blood;

The results of the experiment show that

All three compounds tested were detected in the brain, with MAGLZ-II-16 having the highest abundance in the brain, followed by MAGLZ-II-17, and MAGLZ-II-11 having the lowest abundance (see Table 3).

Table 3. Distribution pattern in tissues

Time/h MAGLZ-II-
01 MAGLZ-II-
02 MAGLZ-II-
11 MAGLZ-II-
16 MAGLZ-II-
17 1.0 0 0 0 0 2.9 2.0 0.6 1.2 1.6 32.8 9.4 5.0 0.9 3.7 1.3 16.9 4.8

Example 38. Effect of the MAGL inhibitor of the present invention on reserpine-induced depression in mice.

1) Basic information

Species: ICR mice

Microbiological class: SPF

Number and sex of acquired animals: 75 males, 75 females

Number and sex of animals used: 75 males, 75 females

Weight: 13-18 g

2) Grouping of animals

Grouping method: A total of 3 batches of animals with 50 animals each were used in the experiment, each batch was randomly divided into 5 groups: low dose, medium dose, high dose of the test compound treatment group, blank control group and model group, and each batch was used to test one test compound.

3) Test methods

The drug was administered subcutaneously once daily at doses of 4, 8, and 16 mg/kg for 6 days. After the final dose, each animal, except for the blank control group, received a subcutaneous injection of 4 mg/kg reserpine. After 2 hours, arousal, finger jamming, fear, head jamming, autonomic activity, upright posture, eyelid closure, and other measures were measured.

4) Statistical analysis

Statistics and data analysis were performed using SPSS version 17.0 and Office 2016 software on a computer. Data statistics were performed using the Kruskal-Wallis test. If the Kruskal-Wallis test showed statistical significance (p ≤ 0.05), further multiple comparison tests were performed using the nonparametric Mann-Whitney U test.

5) The results are presented in Tables 4A, 4B and 4C

Table 4A

Group Awaken-
denie Stuck
fingering Stuck
head twitching Fear Autonomous
activity
nost Straightening Closing the eyelids Empty control group 4±0 4±0 4±0 4±0 4±0 4±0 4±0 Model group 2.2±1.40 ^** 1.2±0.98 ^** 1.4±0.92 ^** 0.2±0.60 ^** 0±0 ^** 0±0 ^** 1.2±1.33 ^** MAGL-II-11-L 3±1.61 1.6±1.50 1±1.34 0±0 0.8±1.60 0.8±1.60 1.2±0.98 MAGL-II-11-M 2.6±1.56 1.8±1.66 1.8±1.40 0±0 0.8±1.60 0.8±1.60 1.4±1.28 MAGL-II-11-H 3.2±1.33 2±1.79 2.6±1.56 0.2±0.60 1.8±1.66 ^** 1±1.61 ^* 2±1.26 Note: when comparing the model group with the blank group: ^* P < 0.05, ^** P <0.01; when comparing the experimental group with the model group, ^* P < 0.05, ^** P < 0.01

Table 4B

Group Awaken-
denie Stuck
fingering Stuck
head twitching Fear Autonomous
activity
nost Straightening Closing the eyelids Empty control group 4±0 4±0 4±0 4±0 4±0 4±0 4±0 Model group 3.5±1.32 ^** 3.5±1.32 ^** 3.58±1.10 ^** 0.25±0.66 ^** 0.58±0.85 ^** 0±0 ^** 1.83±0.44 ^** MAGL-II-17-L 4±0 4±0 3.75±0.66 3.5±1.32 0.5±0.87 0.25±0.66 2.25±0.66 MAGL-II-17-M 4±0 4±0 4±0 0±0 1.5±1.66 1±1.41 1±1.00 MAGL-II-17-H 4±0 4±0 3.75±0.66 0±0 0.25±0.66 0±0 0.25±0.66 Note: when comparing the model group with the blank group: ^* P < 0.05, ^** P <0.01; when comparing the experimental group with the model group ^{: *} P < 0.05, ^** P < 0.01,

Table 4C

Group Awakening Fingers getting stuck Head stuck Fear Autonomous activity Straightening Closing the eyelids Empty control group 4±0 4±0 4±0 4±0 4±0 4±0 4±0 Model group 1.6±1.2 ^** 1.2±0.98 ^** 0±0 ^** 0±0 ^** 0.2±0.6 ^** 0±0 ^** 0±0 ^** MAGL-II-16-L 2±0 1.4±0.92 0.4±0.8 0±0 0±0 0±0 0±0 MAGL-II-16-M 1.8±0.6 1.2±0.98 0.8±0.98 ^* 0±0 0.2±0.6 0±0 0±0 MAGL-II-16-H 2±0 1.6±0.8 0.8±0.98 ^* 0±0 0.6±0.6 ^* 0.4±1.2 0±0 Note: when comparing the model group with the blank group: ^* P < 0.05, ^** P <0.01; when comparing the experimental group with the model group ^{: *} P < 0.05, ^** P < 0.01

1) MAGLZ-II-11,

Nonparametric tests

For each indicator, a statistically significant difference was observed in the Kruskal-Wallis test (P < 0.05).

Comparison of differences between groups

Mann-Whitney U test

The differences between the model group and the blank control group were statistically significant (P < 0.05), indicating good modeling results; The differences between the low/medium/high dose groups and the model group were not statistically significant (P > 0.05); The differences in autonomic activity and straightening between the high dose group and the model control group were statistically significant (P < 0.01, P < 0.05), and the change trend of the indicators was the same as the blank control group.

5.2) MAGLZ-II-17: For each indicator in the Kruskal-Wallis test, a statistically significant difference was observed (p < 0.05);

The differences between the model group and the blank control group were statistically significant (P < 0.05), indicating good modeling results.

The differences between the low dose group and the model group were not found to be statistically significant (P> 0.05).

The differences in eyelid closure between the medium dose group and the model group were statistically significant (P < 0.05), but the mean value was smaller than that of the model group, and other indices did not show statistical significance (P > 0.05).

The differences in eyelid closure between the high dose group and the model group were statistically significant (P < 0.01), but the mean value was smaller than that of the model group, and other indices did not show statistical significance (P > 0.05).

5.3) MAGLZ-II-16: For each indicator in the Kruskal-Wallis test, a statistically significant difference was observed (p < 0.05); Mann-Whitney U-test

The differences between the model group and the blank control group were statistically significant (P < 0.05), indicating good modeling results.

There was no statistical significance between the low dose group and the model group (P> 0.05).

The differences in head sticking between the medium dose group and the model group were statistically significant (P < 0.05), but 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; other indicators did not show statistical significance (P > 0.05).

The differences in head stuckness and autonomic activity between the high dose group and the model group were statistically significant (P < 0.05), but the mean was greater than that of the model group, and the trend change was the same as that of the blank control group; other indices did not show statistical significance (P > 0.05).

6) Conclusion

6.1) Under the conditions of this test, MAGLZ-II-11 had a moderate effect on reserpine-induced depression in mice.

6.2) Under the conditions of this test, MAGLZ-II-16 had a moderate effect on reserpine-induced depression in mice.

6.3) Under the conditions of this test, MAGLZ-II-17 may have a potentiating effect on reserpine-induced depression in mice.

Example 39, Effect of the MAGL inhibitor of the present invention on the forced swimming model in mice and the tail suspension model in mice.

1. Materials and methods

1.1 Experimental animals

ICR mice, male, 18-22 g. The light regimen was 12 hours of light and 12 hours of darkness. The air temperature was maintained at 22±1°C, and the relative humidity was 55±5%.

1.2. Experimental method

1.2.1 Animal grouping and administration route

The experiment involved 7 groups of mice, 10 in each, all animals were divided into the following groups: control group (saline group), positive control group (fluoxetine hydrochloride capsule, 5 mg/kg), 5 groups of treatment with MAGLZ-II series compounds: MAGLZ-II-06, MAGLZ-II-07, MAGLZ-II-11, MAGLZ-II-17, MAGLZ-II-18, drug concentration upon administration was 5 mg/kg.

All drugs were administered intragastrically at a dose of 20 ml/kg body weight, once daily for 10 days. Behavioral testing began 1 hour after the last drug administration.

1.2.2. Mouse forced swimming test

The forced swim test in mice was conducted using the method developed by Porsalt (Porsalt RD. Behavioural 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 in diameter, 25 cm in height) with a water temperature of 25 ± 1°C and a water depth of 19 cm. The total time during which each animal remained motionless for 6 minutes was recorded as the immobility time (in seconds). Each mouse was considered immobile when it stopped struggling and remained suspended in the water, making only the movements necessary to keep its head above the surface. A reduction in immobility time indicates an antidepressant effect.

1.2.3. Tail suspension test

The tail suspension test (TST) was performed according to the method developed 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 clamp, mice were suspended 10 mm from the tail tip of the chamber (250 mm × 250 mm × 300 mm) and 5 cm from the bottom near the head. Noise tests were performed in a dark room with minimal background noise, and the cumulative duration of immobility in the final 4-minute interval was recorded for 6 minutes. The evaluation criterion was cessation of struggling and complete immobility of the mice.

1.2.4. Data processing

All data are expressed as Differences between groups were analyzed using one-way ANOVA.

2. Results

2.1 Effect of MAGLZ-II series compounds on forced swim test behavior in mice

The experiments showed that the positive control drug fluoxetine hydrochloride and MAGLZ-II series compounds could significantly reduce the total immobility time in forced swimming in mice (P < 0.05) (Table 5).

Table 5. Effect of 5MAGLZ-II series compounds on behavior in the forced swim test in mice

Group Dose (mg/kg) Number of animals (pieces) Accumulated time of immobility (s) Control group - 10 136.84±46.69 Fluoxetine hydrochloride capsules 5 10 92.64±29.17 ^** MAGLZ-II-06 5 10 89.23±46.37 ^* MAGLZ-II-07 5 10 91.31±41.85 ^* MAGLZ-II-11 5 10 73.57±27.28 ^** MAGLZ-II-17 5 10 79.38±41.67 ^** MAGLZ-II-18 5 10 81.71±38.12 ^** Note: ^* P < 0.05, ^** P < 0.01 when compared with the control group.

2.2 Effect of MAGLZ-II series compounds on tail-suspension behavior in mice

Differences with the control group showed that MAGLZ-II series compounds significantly reduced the total duration of motionless hovering (P < 0.05) (Table 6).

Table 6. Effect of 6MAGLZ-II series compounds on tail-suspension behavior in mice

Group Dose (mg/kg) Number of animals (pieces) Accumulated time of immobility (s) Control group - 10 132.38±44.51 Fluoxetine hydrochloride capsules 5 10 92.19±23.63 ^** MAGLZ-II-06 5 10 90.73±30.51 ^* MAGLZ-II-07 5 10 89.66±31.52 ^* MAGLZ-II-11 5 10 72.81±23.49 ^** MAGLZ-II-17 5 10 89.87±29.68 ^** MAGLZ-II-18 5 10 79.68±25.72 ^** Note: when compared with the control group ^* P < 0.05, ^** P < 0.01,

3. Discussion

Forced swim and tail suspension are the most commonly used classic animal models of depression. The hopelessness and behavioral distortions exhibited by mice in these models are hallmarks of in vitro depression models. They are also frequently used as antidepressant screening models due to their sensitivity to antidepressants and ease of use. In this study, the antidepressant activity of MAGLZ-II compounds was further evaluated and confirmed in these two classic animal models of depression. The results showed that MAGLZ-II compounds significantly reduced the total duration of immobility during forced swim and tail suspension, indicating that MAGLZ-II compounds possess potent antidepressant activity.

Example 40. Effect of the MAGL inhibitor of the present invention on the number of acetic acid-induced writhings in mice

1. Grouping of animals and route of administration

200 male ICR mice, (20±2) g, were randomly divided into 20 groups according to body weight, 10 pieces in each, all animals were divided into the following groups: model group: administered 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Positive control group: diclofenac sodium, 10 mg/kg, intragastrically

Low dose group MAGLZ-II-11: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-11a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18c: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18c: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-10: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-10: 15 mg/kg, intragastrically

High dose group MAGLZ-II-10: 30 mg/kg, intragastrically

Each compound was prepared as a suspension in 0.5% sodium carboxymethylcellulose solution respectively, then prepared according to the prescribed dose, and each treatment group was administered intragastrically; the model group was administered 0.5% sodium carboxymethylcellulose intragastrically in the same volume.

2. Experimental method and data processing

Mice were given acetic acid by intraperitoneal injection to produce larger and more persistent pain stimuli deep in the abdominal cavity, eliciting a writhing response. One hour after the injection, each treatment group received an intraperitoneal injection of 0.1 ml/10 g of 0.7% acetic acid saline.

The time at which mice began to writhe after intraperitoneal injection of acetic acid was recorded.

The number of acetic acid-induced writhing movements in each mouse was recorded for 20 min after acetic acid administration, and the writhing inhibition ratio was calculated in each treatment group.

Writhing inhibition ratio = [(number of writhings in the control group - number of writhings in the treatment group)/number of writhings in the control group] × 100%

The experimental data were expressed as , analysis of variance was performed using SPSS15.0 software.

3. Experimental results

The results of the experiment are shown in Table 7.

Table 7. Effect of the MAGL inhibitor of the present invention on the number of acetic acid-induced writhings in mice

Group Number of writhings Inhibition coefficient (%) Latent time (s) Model group 39.3±12.9 - 226.5±38.2 Positive control group 24.5±12.9 ^* 37.7 358.5±172.7 ^* Low-dose MAGLZ-II-18a group 24.2±8.5 ^** 38.4 355.5±136.5 ^* Group with medium dose of MAGLZ-II-18a 20.8±12.5 ^** 47.1 380.7±130.7 ^** High-dose MAGLZ-II-18a group 17.8±11.1 ^*** 54.7 422.4±137.9 ^*** Low dose MAGLZ-II-11 group 23.4±11.7 ^** 40.5 379.7±129.3 ^** Group with medium dose of MAGLZ-II-11 17.5±9.6 ^*** 55.5 408.4±128.2 ^*** High-dose MAGLZ-II-11 group 13.6±13.1 ^*** 65.4 467.8±175.6 ^*** Low-dose 11MAGLZ-II-a group 22.6±12.0 ^** 42.5 401.2±168.3 ^** Group with medium dose of MAGLZ-II-1a1 18.6±14.6 ^** 52.7 438.9±230.0 ^** High-dose 11MAGLZ-II-a group 12.5±9.4 ^*** 68.2 488.8±172.0 ^*** Low-dose MAGLZ-II-18 group 25.0±13.5 ^* 36.4 318.7±120.2 ^* Group with the average dose of MAGLZ-II-18 21.8±12.4 ^** 44.5 381.4±127.1 ^** High dose MAGLZ-II-18c group 15.8±15.3 ^** 59.8 410.9±186.5 ^** Low dose MAGLZ-II-18c group 28.3±11.9 28.0 263.5±77.7 Group with the average dose of MAGLZ-II-18c 24.9±12.2 ^* 36.6 312.7±90.9 ^* High dose MAGLZ-II-18c group 19.9±12.2 ^** 49.4 373.6±114.0 ^** Low-dose MAGLZ-II-10 group 34.0±9.1 13.5 235.4±59.6 Group with the average dose of MAGLZ-II-10 32.8±11.6 16.5 246.2±69.5 High-dose MAGLZ-II-10 group 24.8±11.1 ^* 36.9 306.4±90.2 ^{* *} P < 0.05, ^** P < 0.01, ^*** P < 0.001 when compared with the model group.

From the experimental results shown in Table 1, it can be seen that, when compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a and MAGLZ-II-18c of the present invention were able to significantly reduce the number of acetic acid-induced writhings in mice and prolong the latency time in mice.

Example 41. Effect of the MAGL inhibitor of the present invention on the pain threshold in mice under thermal stimulation using the hot plate method

Female ICR mice (20 ± 2 g) were placed on a hot plate apparatus at 55 ± 0.5°C. The latency (s) between the contact of the mouse's lower paw with the hot plate and the onset of the hind paw licking response was recorded as an index of pain threshold. Mice with a response latency of < 5 s or > 30 s, or jumping mice, were excluded.

The grouping and routes of administration were the same as in Example 40.

The mice were continuously administered the drug intragastrically for 7 consecutive days; the pain threshold of mice in each group was measured once at 30, 60, 90 and 120 min after the last administration of the drug; the pain threshold exceeding 60 s was calculated at 60 s.

The results of the experiment are shown in Table 8.

Table 8. Effect of the MAGL inhibitor of the present invention on the pain threshold in mice under thermal stimulation using the hot plate method

Pain threshold (s) in mice at different time periods after drug administration Group 30 min 60 min 90 min 120 min Model group 15.63±7.32 16.85±7.46 15.50±6.11 15.38±6.93 Positive control group 22.32±6.76 ^* 24.34±8.19 ^* 22.64±6.48 ^* 22.52±6.91 ^* Low dose MAGLZ-II-11 group 37.92±8.83 ^*** 44.14±7.56 ^*** 45.93±8.38 ^*** 43.80±6.12 ^*** Group with medium dose of MAGLZ-II-11 40.33±8.25 ^*** 44.87±5.67 ^*** 46.41±5.36 ^*** 43.44±5.91 ^*** High-dose MAGLZ-II-11 group 41.43±5.15 ^*** 46.16±6.63 ^*** 47.22±5.98 ^*** 45.49±6.31 ^*** Low-dose 11MAGLZ-II-a group 36.18±8.50 ^*** 43.17±7.43 ^*** 42.83±7.34 ^*** 42.16±7.05 ^*** Group with medium dose of MAGLZ-II-11a 38.16±7.00 ^*** 45.47±7.03 ^*** 43.71±5.87 ^*** 42.34±7.04 ^*** High-dose 11MAGLZ-II-a group 39.45±6.75 ^*** 46.56±6.35 ^*** 45.42±6.84 ^*** 43.73±5.68 ^*** Low-dose MAGLZ-II-18a group 34.62±6.65 ^*** 40.29±6.73 ^*** 43.98±6.55 ^*** 43.56±6.01 ^*** Group with medium dose of MAGLZ-II-18a 37.51±6.07 ^*** 42.42±5.90 ^*** 45.86±7.09 ^*** 44.38±5.44 ^*** High-dose MAGLZ-II-18a group 40.86±7.17 ^*** 46.29±5.27 ^*** 48.70±5.73 ^*** 47.04±5.54 ^*** Low-dose MAGLZ-II-18 group 26.33±7.52 ^** 26.74±7.05 ^** 27.16±7.32 ^** 26.64±10.03 ^** Group with the average dose of MAGLZ-II-18 26.41±7.53 ^** 27.09±7.53 ^** 27.58±7.71 ^** 27.38±9.97 ^** High dose MAGLZ-II-18c group 26.73±7.37 ^** 27.22±7.29 ^** 27.86±7.93 ^** 27.62±10.01 ^** Low dose MAGLZ-II-18c group 23.98±6.89 ^* 24.75±6.91 ^* 25.16±8.87 ^* 24.28±7.17 ^* Group with the average dose of MAGLZ-II-18c 26.82±7.99 ^** 28.52±6.18 ^** 28.66±8.73 ^** 27.88±7.74 ^** High dose MAGLZ-II-18c group 29.23±7.95 ^** 30.24±7.80 ^** 30.66±8.35 ^** 29.87±7.43 ^** Low-dose MAGLZ-II-10 group 16.37±6.00 17.93±5.75 19.24±4.72 19.14±4.63 Group with the average dose of MAGLZ-II-10 16.67±5.97 18.79±5.95 22.43±5.31 20.60±4.64 High-dose MAGLZ-II-10 group 17.73±6.20 19.77±6.73 25.25±7.01 ^* 24.90±5.73 ^{* *} P < 0.05, ^** P < 0.01, ^*** P < 0.001 when compared with the model group.

When 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 were able to significantly prolong the pain threshold of mice receiving a thermal stimulus.

Example 42. Effect of the MAGL inhibitor of the present invention on the pain threshold in mice with inflammatory pain

Male SD rats (180 ± 20) g were injected with 50 μl of complete Freund's adjuvant into the right hind paw, while control mice were injected with the same volume of 0.5% sodium carboxymethylcellulose to create a model of inflammatory pain with paw edema. Following the modeling, the mice were continuously administered the drug intragastrically once daily for 7 days. The pain threshold (sec) of the mice was measured before drug administration and on days 1, 3, and 7 after the modeling at different time points.

The grouping and routes of administration were the same as in Example 40.

Table 9. Effect of the MAGL inhibitor of the present invention on pain threshold in mice with inflammatory pain

Group Pain threshold (s) in mice at different time periods after drug administration 0d 1d 3D 7d Control group 12.67±2.52 13.02±3.03 12.91±2.81 12.88±2.88 Model group 12.56±2.50 5.49±0.94 ^### 7.06±0.63 ^### 8.27±0.57 ^### Positive control group 12.71±2.55 6.38±0.91 ^* 8.05±1.23 ^* 9.00±0.911 ^* Low dose MAGLZ-II-11 group 12.94±2.41 7.55±1.29 ^*** 9.00±0.73 ^*** 10.26±1.08 ^*** Group with medium dose of MAGLZ-II-11 12.80±2.27 7.73±1.10 ^*** 9.47±0.79 ^*** 10.93±1.62 ^*** High-dose MAGLZ-II-11 group 12.88±2.27 7.99±0.90 ^*** 9.88±1.34 ^*** 11.26±1.82 ^*** Low-dose MAGLZ-II-11a group 12.96±2.44 7.38±1.19 ^*** 8.87±0.74 ^*** 9.91±1.05 ^*** Group with medium dose of MAGLZ-II-11a 13.03±2.35 7.58±0.84 ^*** 9.30±0.75 ^*** 10.66±1.30 ^*** High-dose MAGLZ-II-11a group 12.97±2.32 7.83±1.01 ^*** 9.55±1.58 ^*** 11.00±161 ^*** Low-dose MAGLZ-II-18a group 12.73±2.43 6.97±0.87 ^** 8.44±1.03 ^** 9.56±0.92 ^** Group with medium dose of MAGLZ-II-18a 12.86±2.25 7.26±0.70 ^*** 9.01±0.68 ^*** 10.37±1.49 ^*** High-dose MAGLZ-II-18a group 12.85±2.26 7.49±0.79 ^*** 9.35±1.2 ^*** 10.37±1.59 ^*** Low-dose MAGLZ-II-18 group 12.73±2.43 ^* 6.39±0.91 ^* 8.12±1.29 ^* 9.16±1.17 ^* Group with the average dose of MAGLZ-II-18 12.86±2.25 ^** 6.62±0.69 ^** 8.24±0.73 ^** 9.51±1.15 ^** High dose MAGLZ-II-18c group 12.85±2.26 ^** 6.95±0.85 ^** 8.49±1.15 ^** 9.62±0.93 ^** Low dose MAGLZ-II-18c group 12.75±2.46 ^* 6.36±0.89 ^* 7.98±1.21 ^* 8.95±0.67 ^* Group with the average dose of MAGLZ-II-18c 12.86±2.07 ^* 6.41±0.91 ^* 8.10±1.30 ^* 8.99±0.92 ^* High dose MAGLZ-II-18c group 12.85±2.09 ^* 6.44±0.92 ^* 8.17±1.14 ^* 9.05±0.84 ^* Low-dose MAGLZ-II-10 group 12.71±2.47 5.59±0.89 7.15±0.59 8.33±0.57 Group with the average dose of MAGLZ-II-10 12.77±2.48 5.62±0.84 7.18±0.55 8.36±0.59 High-dose MAGLZ-II-10 group 12.93±2.26 5.67±0.93 7.95±0.66 ^* 8.93±0.61 ^{* *} P < 0.05, ^** P < 0.01, ^*** P < 0.001 when compared with the model group.

MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, MAGLZ-II-18c significantly increased the pain threshold in mice with inflammatory pain when compared with the model group.

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

Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder characterized by chronic abdominal pain, abdominal discomfort, and altered bowel habits without overt intestinal lesions. This experiment is primarily used to discuss the therapeutic effect of the MAGL inhibitor of the present invention on rats with irritable bowel syndrome induced by early maternal weaning.

1. Grouping of animals and route of administration

200 rats SD (20±2) g were divided into 20 groups randomly according to body weight, 10 pieces in each, all animals were divided into the following groups:

Empty group: 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Model group: 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Low dose group MAGLZ-II-11: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-11a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18c: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18c: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-10: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-10: 15 mg/kg, intragastrically

High dose group MAGLZ-II-10: 30 mg/kg, intragastrically

Each compound was prepared as a suspension in 0.5% sodium carboxymethylcellulose solution respectively, then prepared according to the prescribed dose, and each treatment group was administered intragastrically; the model group was administered 0.5% sodium carboxymethylcellulose intragastrically in the same volume.

2. Experimental method and data processing

Newborn male SD rats were separated from lactating mothers for 3 h every day from day 2 to day 21 after birth of the newborn SD rats, namely, the lactating mother rat was taken out of the cage at 8:30 a.m. In the morning, the newborn rats were removed from the original cage and placed in another separate cage, then the cage was moved to the next room, then the newborn rats were returned to the original cage after 3 h, namely at 11:30 a.m., so that the newborn rats and the mother rat were together; newborn rats were weaned on day 22 after birth, separated from the original cage on day 30, grouped on day 60, then administered drugs for 20 days.

Intragastric administration was carried out during the administration period once a day with continuous administration for 20 days.

Experiment endpoint: The autopsy was performed after completion of the relevant test parameters of the experiment.

Test parameters: Abdominal wall reconstruction reflex (AWR) assessment, test method: The rat was fasted for 18 hours before the experiment, water was drunk freely, the rat was anesthetized with ether, an air balloon covered with paraffin oil was inserted into the colon of the rat by 4.0 cm, a catheter outside the anus was fixed to the root of the rat's tail with adhesive tape, the catheter was connected to a syringe and blood pressure monitor through a T-piece. The rats were placed in a plexiglass chamber (20 cm × 12 cm × 9 cm) for observation, and the experiment was started 30 minutes after the rats woke up and fully adapted to the environment. The AWR assessment was repeated 3 times under a pressure of 20, 40, 60 mmHg, the interval between stimuli was 4 min. The AWR assessment was conducted in a blinded manner according to the following criteria: 0 point: no obvious behavioral changes; 1 point: only simple head movement; 2 points: abdominal muscles began to contract but did not rise off the table; 3 points: abdominal muscles clearly contracted to the point of flattening or the lower abdominal wall rose off the table; 4 points: Bending or arching of the trunk or pelvis.

After the experiment, the animals were necropsied, weighed, anesthetized, blood was collected, and serum 5-HT levels were analyzed using high-performance liquid chromatography. Brains were collected on ice, pre-chilled, washed with 0.5% sodium carboxymethylcellulose, dried on filter paper, then placed in lyophilization tubes and stored at -20°C. Brain tissue was analyzed for 5-HT levels using high-performance liquid chromatography.

Histological examination of the colon: After the colon dilation experiment, the distal colon of the rat (5-6 cm from the anus) was collected and subjected to routine HE staining to observe the inflammation and damage of the intestinal wall.

Data are presented as mean ±SD, and differences between groups were calculated using t-tests or one-way ANOVA, with P < 0.05 indicating a statistically significant difference.

3. Experimental results

The results of the experiment are shown in Table 10.

Table 10. Effect of the MAGL inhibitor of the present invention on AWR and 5-HT in rats with IBS

Group Quantity AWR indicator Colon (mcg/mg) Serum (µg/ml) 20 mmHg 40 mmHg 60 mmHg 5-HT 5-HT Empty group 10 1.93±0.43 2.78±0.35 3.29±0.25 11.78±1.78 28.55±4.03 Model group 10 2.85±0.28 ^### 3.53±0.22 ^### 3.86±0.13 ^### 19.76±2.52 ^### 38.73±3.42 ^### Low dose group 10 2.28±0.58 ^* 3.20±0.43 ^* 3.61±0.24 ^** 16.84±2.87 ^* 34.2±5.37 ^* MAGLZ-II-18c
Medium dose group 10 1.79±0.23 ^*** 2.87±0.65 ^** 3.27±0.51 ^** 14.99±2.90 ^** 31.88±4.40 ^** MAGLZ-II-18c
High dose group 10 1.38±0.51 ^*** 2.20±0.57 ^*** 2.96±0.70 ^*** 14.34±2.40 ^*** 29.03±5.29 ^*** MAGLZ-II-18c
Low dose group 10 2.12±0.36 ^*** 3.02±0.53 ^* 3.58±0.26 ^** 16.33±2.78 ^** 31.76±5.42 ^** MAGLZ-II-11a
Medium dose group 10 1.51±0.43 ^*** 2.52±0.38 ^*** 3.27±0.48 ^** 14.79±2.73 ^*** 29.89±5.34 ^*** MAGLZ-II-11a
High dose group 10 1.23±0.50 ^*** 2.13±0.48 ^*** 2.87±0.46 ^*** 12.65±3.36 ^*** 24.05±6.17 ^*** MAGLZ-II-11a
Low dose group 10 2.87±0.39 3.43±0.30 3.79±0.20 21.44±2.72 36.32±3.14 MAGLZ-II-10
Medium dose group 10 2.83±0.47 3.56±0.22 3.80±0.17 20.97±2.24 38.65±2.68 MAGLZ-II-10
High dose group 10 2.89±0.46 3.21±0.35 ^* 3.65±0.17 ^* 18.66±2.38 ^* 33.09±2.31 ^* MAGLZ-II-10
Low dose group 10 2.50±0.38 ^* 3.00±0.49 ^** 3.60±0.22 ^** 16.24±2.34 ^** 31.75±5.33 ^** MAGLZ-II-11
Medium dose group 10 2.02±0.57 ^** 2.42±0.41 ^*** 3.03±0.37 ^*** 14.33±2.10 ^*** 29.47±4.91 ^*** MAGLZ-II-11
High dose group 10 1.39±0.30 ^*** 1.92±0.33 ^*** 2.86±0.26 ^*** 12.00±2.93 ^*** 23.82±6.34 ^*** MAGLZ-II-11
Low dose group 10 2.59±0.33 3.26±0.39 3.65±0.23 ^* 21.91±2.46 35.73±3.32 MAGLZ-II-18
Medium dose group 10 2.29±0.36 ^** 3.01±0.40 ^** 3.3±0.45 ^** 14.89±3.19 ^** 29.12±7.05 ^** MAGLZ-II-18
High dose group 10 1.87±0.36 ^*** 2.6±0.54 ^*** 3.09±0.34 ^*** 14.33±2.10 ^*** 30.86±5.28 ^*** MAGLZ-II-18
Low dose group 10 2.58±0.26 ^* 3.06±0.38 ^** 3.50±0.29 ^** 16.43±2.15 ^** 32.51±5.01 ^** MAGLZ-II-18a
Medium dose group
MAGLZ-II-18a 10 2.35±0.40 ^** 3.02±0.51 ^** 3.39±0.34 ^*** 14.64±2.17 ^*** 30.20±5.59 ^*** High dose group
MAGLZ-II-18a 10 1.96±0.46 ^*** 2.60±0.58 ^*** 3.00±0.54 ^*** 13.90±1.13 ^*** 29.44±4.37 ^{*** #} P < 0.05, ^## P < 0.01, ^### P < 0.001 when compared with the empty group.
^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 when compared with the model group.

From the experimental results presented in Table 10, it can be seen that

When compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18, MAGLZ-II-18c significantly reduced the AWR score and decreased the 5-HT level in the colon and serum.

Example 44. Effect of the MAGL inhibitor of the present invention on reserpine-induced migraine in mice

1. Grouping of animals and route of administration

210 male ICR mice, (20±2) g, were randomly divided into 21 groups according to body weight, 10 pieces in each, all animals were divided into the following groups: model group: administered 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Model group: 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Positive control group: zolmitriptan 0.5 mg/kg, intragastrically

Low dose group MAGLZ-II-11: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-11a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18c: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18c: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-10: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-10: 15 mg/kg, intragastrically

High dose group MAGLZ-II-10: 30 mg/kg, intragastrically

Each compound was prepared as a suspension in 0.5% sodium carboxymethylcellulose solution respectively, then prepared according to the prescribed dose, and each treatment group was administered intragastrically; the model group was administered 0.5% sodium carboxymethylcellulose intragastrically in the same volume.

2. Experimental method and data processing

All groups were subcutaneously injected with reserpine solution (0.2 mg/kg), except for the blank control group, which was subcutaneously injected with saline, for 10 days; if after the administration of reserpine, the mice showed closing of the eyes, squatting and decreased activity, diarrhea, decreased food intake, arching of the back and other typical reserpine-induced symptoms, the modeling was considered successful.

Mice in the blank control group and model group were administered the solvent intragastrically from the 5th day after the modeling, and the other groups of mice were respectively administered the corresponding drugs at the corresponding dose for 10 consecutive days; 1 hour after the last administration, the mice were weighed; blood was collected from the internal circuit to determine the blood clotting time; whole blood was collected, and the brain was placed on ice and stored at -20°C for subsequent experiments.

The end point of the experiment: the modeling administration continued for 10 days, the test and blood sampling were performed 1 hour after the last administration on the 10th day, thus the experiment was completed.

Test parameters: mice were weighed once every 2 days after the start of the experiment;

Behavioral test: tail suspension was performed; on the 9th day of the experiment, mice were attached with adhesive tape 1 cm from the tail, a steel needle was passed through the tape, and the mice were suspended in a dark box so that they hung upside down. The number of struggle episodes (the total number of attempts to turn over while suspended by the tail) was observed for 2 minutes.

Mice were weighed 1 hour after completion of drug administration; blood was collected from the inner part of the canthus through a capillary to determine blood clotting time.

Blood was collected for serum 5-HT determination using high-performance liquid chromatography. Brains were collected on ice, pre-chilled, lightly rinsed with 0.5% sodium carboxymethylcellulose, then dried with filter paper, placed in lyophilization tubes, and stored at -20°C for further use. Brain tissue was then analyzed for 5-HT content using high-performance liquid chromatography.

The 5-HT content analysis was carried out as follows: the sample was precipitated with 10% perchloric acid solution at a ratio of 1:1, then centrifuged to collect the supernatant for analysis. Chromatographic conditions: the chromatographic column is a SHIMADZU VP-ODS C18 column (250*4.6 mm, 5 μm), the mobile phase is methanol-0.01 mol/L potassium acetate buffer solution (10:90, V/V, pH adjusted to 4.00 with 0.2 mol/L citric acid) for gradient elution; column temperature is 25°C, flow rate is 1.00 mL/min, wavelength is 275 nm.

Data are presented as mean ±SD, and differences between groups were calculated using t-tests or one-way ANOVA, with P < 0.05 considered statistically significant.

3. Experimental results

The results of the experiment are presented in Tables 11-13 below.

Table 11. Effect of the MAGL inhibitor of the present invention on blood clotting time

Grouping Dosage Clotting time (s) Normal group - 352.50±37.48 Model group - 115.38±24.05 ^*** Zolmitriptan 0.5 mg/kg 185.12±38.86 ^### Low-dose MAGLZ-II-18a group 7.5 mg/kg 137.25±67.22 Group with medium dose of MAGLZ-II-18a 15 mg/kg 191.50±63.66 ^## High-dose MAGLZ-II-18a group 30 mg/kg 252.25±30.99 ^### Low dose MAGLZ-II-11 group 7.5 mg/kg 222.5±35.52 ^### Group with medium dose of MAGLZ-II-11 15 mg/kg 326.88±45.9 ^### High-dose MAGLZ-II-11 group 30 mg/kg 336.88±33.09 ^### Low dose MAGLZ-II-10 group 7.5 mg/kg 113.25±23.31 Group with the average dose of MAGLZ-II-10 15 mg/kg 125.25±19.31 High-dose MAGLZ-II-10 group 30 mg/kg 148.00±30.23 ^# Low-dose MAGLZ-II-11a group 7.5 mg/kg 136.75±33.22 Group with medium dose of MAGLZ-II-11a 15 mg/kg 256.88±42.93 ^### High-dose MAGLZ-II-11a group 30 mg/kg 300.63±45.26 ^### Low dose MAGLZ-II-18c group 7.5 mg/kg 148.00±30.23 ^# Group with the average dose of MAGLZ-II-18c 15 mg/kg 198.63±21.36 ^### High dose MAGLZ-II-18c group 30 mg/kg 237.13±28.75 ^### Low-dose MAGLZ-II-18 group 7.5 mg/kg 186.5±51.71 ^## Group with the average dose of MAGLZ-II-18 15 mg/kg 212.63±46.02 ^### High dose MAGLZ-II-18c group 30 mg/kg 241.5±41.63 ^### When compared with the model group, #P < 0.05, ##P < 0.01, ###P < 0.001.

From the experimental results shown in Table 11, it can be seen that, when 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 blood clotting time.

Table 12. Effect of the MAGL inhibitor of the present invention on the number of episodes of immobility

Grouping Dosage Number of times Normal group - 22.38±3.38 Model group - 8.38±2.62 ^*** Zolmitriptan 0.5 mg/kg 18.13±3.94 ^### Low-dose MAGLZ-II-18a group 7.5 mg/kg 11.38±2.67 ^# Group with medium dose of MAGLZ-II-18a 15 mg/kg 13.38±2.67 ^## High-dose MAGLZ-II-18a group 30 mg/kg 18.50±2.00 ^### Low-dose MAGLZ-II-10 group 7.5 mg/kg 9.25±2.66 Group with the average dose of MAGLZ-II-10 15 mg/kg 10.63±1.19 ^# High-dose MAGLZ-II-10 group 30 mg/kg 11.63±1.85 ^# Low dose MAGLZ-II-11 group 7.5 mg/kg 14.13±1.89 ^### Group with medium dose of MAGLZ-II-11 15 mg/kg 16.13±1.73 ^### High-dose MAGLZ-II-11 group 30 mg/kg 23.00±3.59 ^### Low-dose MAGLZ-II-11a group 7.5 mg/kg 16.13±3.68 ^### Group with medium dose of MAGLZ-II-11a 15 mg/kg 19.88±4.09 ^### High-dose MAGLZ-II-11a group 30 mg/kg 20.75±4.98 ^### Low dose MAGLZ-II-18c group 7.5 mg/kg 10.13±2.3 ^## Group with the average dose of MAGLZ-II-18c 15 mg/kg 12.75±2.6 ^## High dose MAGLZ-II-18c group 30 mg/kg 15.88±3.8 ^### Low-dose MAGLZ-II-18 group 7.5 mg/kg 11.88±4.02 ^### Group with the average dose of MAGLZ-II-18 15 mg/kg 13.75±4.56 ^### High dose MAGLZ-II-18c group 30 mg/kg 16.51±4.38 ^{### #} P < 0.05, ^## P < 0.01, ^### P < 0.001 when compared with the model group.

From the experimental results presented in Table 12, it can be seen that MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18 and MAGLZ-II-18c of the present invention were able to significantly increase the number of immobility episodes during tail suspension when compared with the model group.

Table 13. Effect of the MAGL inhibitor of the present invention on the content of 5-hydroxytryptamine in brain tissue

Grouping Dosage 5-HT (nmol/g) Normal group - 3.36±0.4 Model group - 0.88±0.23 ^*** Zolmitriptan 0.5 mg/kg 1.93±0.37 ^### Low-dose MAGLZ-II-18a group 7.5 mg/kg 1.21±0.17 ^## Group with medium dose of MAGLZ-II-18a 15 mg/kg 1.65±0.18 ^### High-dose MAGLZ-II-18a group 30 mg/kg 2.76±0.34 ^### Low-dose MAGLZ-II-10 group 7.5 mg/kg 0.85±0.20 Group with the average dose of MAGLZ-II-10 15 mg/kg 1.05±0.14 High-dose MAGLZ-II-10 group 30 mg/kg 0.90±0.21 ^# Low dose MAGLZ-II-11 group 7.5 mg/kg 1.54±0.27 ^### Group with medium dose of MAGLZ-II-11 15 mg/kg 2.10±0.22 ^### High-dose MAGLZ-II-11 group 30 mg/kg 3.63±5.34 ^### Low-dose MAGLZ-II-11a group 7.5 mg/kg 1.35±0.38 ^## Group with medium dose of MAGLZ-II-11a 15 mg/kg 1.86±0.29 ^### High-dose MAGLZ-II-11a group 30 mg/kg 2.97±0.28 ^### Low dose MAGLZ-II-18c group 7.5 mg/kg 1.14±0.22 ^# Group with the average dose of MAGLZ-II-18c 15 mg/kg 1.37±0.34 ^## High dose MAGLZ-II-18c group 30 mg/kg 1.79±0.32 ^### Low-dose MAGLZ-II-18 group 7.5 mg/kg 1.13±0.45 ^# Group with the average dose of MAGLZ-II-18 15 mg/kg 1.54±0.54 ^### High dose MAGLZ-II-18c group 30 mg/kg 2.08±0.59 ^{### #} P < 0.05, ^## P < 0.01, ^### P < 0.001 when compared with the model group.

From the experimental results presented in Table 13, it can be seen that when compared with the model group, MAGLZ-II-11, MAGLZ-II-11a, MAGLZ-II-18a, MAGLZ-II-18 and MAGLZ-II-18c were able to significantly increase the 5-HT level in the brain tissues of reserpine-induced low 5-HT mice.

Example 45. Studies of the effect of a MAG inhibitor on ulcerative colitis in mice

Animal grouping and route of administration

Healthy male BALB/c mice of SPF class, 8 weeks old, body weight (20±2) g.

The mice were quarantined for 7 days, and healthy male mice were selected as test animals. Key checks during the quarantine period include, but are not limited to, the following: whether the animals' vital signs meet the required values; general condition; and body weight. Animals that failed the above-mentioned quarantine were not included in this test.

Modeling: Mice were randomly divided into 21 groups after quarantine, with 10 mice in each. Mice in the normal groups drank plain water freely, while mice in the other groups freely drank a 1.0%-1.5% DSS modeling solution. Seven days after modeling, mice in the model group drank plain water freely for seven days. Day 14 served as the modeling period for four consecutive cycles. Mice in the model group were administered the drug simultaneously.

Empty group: 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Model group: 0.5% sodium carboxymethylcellulose in the same volume, intragastrically

Positive control group: mesalazine (100 mg/kg), intragastrically

Low dose group MAGLZ-II-11: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-11a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-11a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-11a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18a: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18a: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18a: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-18c: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-18c: 15 mg/kg, intragastrically

High dose group MAGLZ-II-18c: 30 mg/kg, intragastrically

Low dose group MAGLZ-II-10: 7.5 mg/kg, intragastrically

Medium dose group MAGLZ-II-10: 15 mg/kg, intragastrically

High dose group MAGLZ-II-10: 30 mg/kg, intragastrically

Each compound was prepared as a suspension in 0.5% sodium carboxymethylcellulose solution respectively, then prepared according to the prescribed dose, and each treatment group was administered intragastrically; the model group was administered 0.5% sodium carboxymethylcellulose intragastrically in the same volume.

2. Test process and method

Experimental endpoint: The experimental administration was carried out for four cycles, and the autopsy was performed on the 3rd and 4th days of the recovery period of the 4th cycle.

Test parameters: mice were weighed twice a week after the start of drug administration; diarrhea and hematochezia of mice were observed, and the time of hematochezia was recorded.

At the end of the experiment, the animals were necropsied, weighed, anesthetized, blood was collected, and the serum was stored at -80°C. The spleen was removed, weighed, and the splenic index was calculated. An ELISA was performed to determine the amount of IL-1β in the serum. The colon was dissected, its length was measured, fixed with formaldehyde, stained with HE, and pathological changes were identified.

Data are presented as mean ±SD, and differences between groups were calculated using t-tests or one-way ANOVA, with P < 0.05 considered statistically significant.

3. Results and discussion

The results of the experiment are shown in Table 14.

Table 14. Effect of the MAGL inhibitor of the present invention on colon length, number of hematochezia episodes and IL-1 content in mice with ulcerative colitis

Group Number Length of the colon (cm) Total number of hematochezia episodes over four cycles IL-1 level (pg/ml) Empty group 10 8.22±1.45 0±0 35.36±5.92 Model group 10 5.70±0.78 ^### 38.60±2.80 ^### 55.53±3.62 ^### Mesalazine (100 mg/kg) 10 6.66±0.68 ^** 32.80±4.96 ^** 48.33±6.49 ^** Low dose MAGLZ-II-11 group 10 6.88±0.89 ^** 32.00±4.94 ^** 44.57±10.32 ^** Group with medium dose of MAGLZ-II-11 10 7.26±0.67 ^*** 27.20±5.39 ^*** 41.40±9.50 ^*** High-dose MAGLZ-II-11 group 10 7.68±0.75 ^*** 24.80±5.43 ^*** 33.83±7.14 ^*** Low-dose MAGLZ-II-18a group 10 6.51±0.90 ^* 33.60±5.83 ^** 47.32±9.06 ^* Group with medium dose of MAGLZ-II-18a 10 7.06±1.20 ^** 29.90±8.95 ^** 43.77±10.69 ^** High-dose MAGLZ-II-18a group 10 7.72±0.92 ^*** 27.90±7.28 ^*** 36.22±4.31 ^*** Low dose MAGLZ-II-18c group 10 6.25±1.17 43.10±7.59 48.08±13.18 Group with the average dose of MAGLZ-II-18c 10 6.47±0.71 ^* 43.20±5.31 ^* 44.09±9.13 ^** High dose MAGLZ-II-18c group 10 6.65±0.67 ^** 31.80±6.6 ^** 45.10±8.88 ^** Low-dose MAGLZ-II-11a group 10 7.21±1.10 ^** 28.80±7.04 ^** 42.01±13.92 ^** Group with medium dose of MAGLZ-II-11a 10 7.20±0.97 ^** 29.10±7.14 ^** 40.70±8.50 ^*** High-dose MAGLZ-II-11a group 10 7.56±0.74 ^*** 27.90±3.73 ^*** 38.61±10.98 ^*** Low-dose MAGLZ-II-10 group 10 5.92±0.66 38.40±5.02 51.81±7.86 Group with the average dose of MAGLZ-II-10 10 5.72±0.60 38.20±7.83 50.83±7.79 High-dose MAGLZ-II-10 group 10 6.35±0.61 ^* 35.15±4.52 ^* 47.25±6.37 ^* Low-dose MAGLZ-II-18 group 10 6.44±0.64 ^* 32.80±4.64 ^** 46.15±9.65 ^* Group with the average dose of MAGLZ-II-18 10 6.37±0.39 ^* 33.90±4.77 ^* 49.01±5.65 ^* High dose MAGLZ-II-18c group 10 6.73±0.63 ^** 32.80±4.37 ^** 46.98±7.56 ^{** #} P < 0.05, ^## P < 0.01, ^### P < 0.001 when compared with the empty group.
^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 when compared with the model group.

When 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 were able to significantly increase the length of the colon, reduce the total number of hematochezia episodes and reduce the level of IL-1.

The invention has been described with the aid of the above examples, but any equivalent substitutions are obvious and are within the scope of the present invention.

Claims (56)

1. A compound of formula I or a pharmaceutically acceptable salt thereof where R is H; Ar ₁ is selected from diphenyl ether, indolyl and biphenyl; Ar ₂ is C ₆ aryl; X ₁ is selected from one of halogen and C _1-3 alkyl; X ₂ is selected from one or two of halogen, C _1-3 alkyl and C _1-3 alkoxy and the connection does not represent N-{1-[(5-chloroindol-2-yl)carbonyl](4-piperidyl)}(4-chlorophenyl)carboxamide, {1-[(5-chloroindol-2-yl)carbonyl](4-piperidyl)}-N-(3-chlorophenyl)carboxamide or {1-[(5-chloroindol-2-yl)carbonyl](4-piperidyl)}-N-(3-fluorophenyl)carboxamide. 2. A compound of formula I or a pharmaceutically acceptable salt thereof, where R is H; Ar ₁ is phenyl; X ₁ is hydroxyl; Ar ₂ is phenyl; and X ₂ represents one or two of methoxy, fluorine and chlorine and the connection does not represent 3. The compound or pharmaceutically acceptable salt thereof according to claim 2, characterized in that said Ar ₁ has a meta-position of substitution. 4. A compound of formula I or a pharmaceutically acceptable salt thereof, where R is H; Ar ₁ is indolyl; X ₁ is H, methyl, or fluorine; Ar ₂ is phenyl; X ₂ is chlorine and the connection does not represent N-(3-chlorophenyl)-1-(1H-indol-2-ylcarbonyl)-4-piperidinecarboxamide. 5. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 6. Compound of formula III or its pharmaceutically acceptable salt, wherein said R ₁ is selected from hydroxy, fluorine, chlorine and methoxy; R ₂ is selected from hydroxy, Cl and methoxy; the number of R ₁ is 1 or 2; and the quantity R ₂ is 1 and the connection does not represent 7. A method for synthesizing a compound of formula I, comprising the following steps: where b or salt b reacts with with the receipt of I; the said X ₁ , X ₂ , Ar ₁ , Ar ₂ are as described in paragraphs 1, 2 or 4; The addition of a condensing agent and a base at this stage makes the reaction more favorable, said condensing agent being selected from HBTU, DMC, HOBT, HOBT/EDCI, HATU, HATU/DIEPA, DCC, CDI, isopropyl chloroformate. 8. The method of synthesis according to claim 7, characterized in that b or salt b is obtained by removing the protection from a: 9. The method of synthesis according to claim 7, characterized in that a or salt a is obtained as a result of the reaction of sm with or its salt: 10. A pharmaceutical composition for the treatment of a MAGL-mediated disease or disorder, comprising a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier. 11. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment of a MAGL-mediated disease or disorder. 12. The use according to claim 11, characterized in that said disorder is one or more of the following disorders: metabolic disease, nephropathy, vomiting or regurgitation or nausea, eating disorders, neuropathy, schizophrenia, depression, bipolar disorder, tremor, movement disorders, withdrawal syndromes, traumatic brain injury, non-traumatic brain injury, spinal cord injury, seizures, conditions associated with abnormal growth or proliferation of cells [eg, benign tumors or cancer], inflammatory diseases, conditions of the immune system, 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 impairment. 13. A method for inhibiting MAGL, comprising contacting MAGL with a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6. 14. A method of treating a MAGL-mediated disease or condition in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6.