SJ series arylphenylamine compound, preparation method and medical use thereof

Abstract

The application discloses an SJ series aryl aniline compound and a preparation method and medical application thereof. The compound has a structure of formula I, and the compound is a stereoisomer, a tautomer, a solvate or a pharmaceutically acceptable salt thereof. The compound or the pharmaceutically acceptable salt thereof can be applied to the field of pharmacy. Animal model experiments show that the compound can significantly improve cognitive impairment of a subject, reduce pathological indexes of Alzheimer's disease, and has the effects of treating and / or preventing Alzheimer's disease.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to a SJ series aryl aniline compound and a preparation method and medical use thereof. BACKGROUND

[0002] Alzheimer's disease (AD) is a neurodegenerative disease caused by multiple factors, and its pathological features include the deposition of senile plaques formed by the aggregation of amyloid β-protein (Aβ) and neuroinflammation tangles (NFT) formed by the over-phosphorylation of Tau (p-tau) protein, and the clinical manifestations include cognitive dysfunction, mental and behavioral abnormalities, and impaired daily living skills. China has entered a rapid development stage of population aging, and the number of elderly population continues to rise. As of the end of 2018, the number of elderly population aged 60 and above in China reached 249 million, accounting for 17.9% of the total population. According to statistics, the prevalence rate of Alzheimer's disease among people aged 65 and above in China is 5.56%. There are currently about 10 million Alzheimer's disease patients in China, and it is estimated that by 2050, there will be more than 40 million Alzheimer's disease patients in China. China has the largest number of AD patients and the fastest growth rate in the world, which brings a heavy burden to patients, families, society and healthcare.

[0003] At present, the common drugs for treating AD are symptomatic treatment drugs, including donepezil, rivastigmine, galantamine and memantine, which are used to improve the cognitive function of patients with mild to moderate Alzheimer's disease. They mainly improve symptoms by inhibiting acetylcholinesterase (AchE) or antagonizing glutamate receptor (NMDAR). However, years of research by scientists around the world have only produced drugs that improve symptoms in certain cases, and through combination therapy, symptoms can be relieved to some extent, but none of them can delay the course of the disease and multiple adverse reactions occur. In recent years, the sugar multi-target drug GV-971 and the antibody biological drug targeting Aβ "Aducan" have been approved for listing, but their clinical efficacy is still controversial. Due to the complexity of AD, its pathogenesis is still unclear. Therefore, the development of effective drugs for preventing and treating AD is still a difficult problem to be solved at present. SUMMARY

[0004] The purpose of the present application is to provide SJ series compounds or pharmaceutically acceptable salts thereof with drug value and a preparation method thereof.

[0005] The present invention also provides a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt thereof as an active ingredient or a major active ingredient, supplemented by a pharmaceutically acceptable carrier.

[0006] A further object of the present invention is to disclose the use of the compound and pharmaceutical composition in the preparation of medicaments for the treatment and / or prevention of Alzheimer's disease.

[0007] The objective of this invention can be achieved through the following technical solutions:

[0008] Compounds having the structure of Formula I, their stereoisomers, tautomers, solvates, or pharmaceutically acceptable salts thereof:

[0009]

[0010] in:

[0011] R1-R5 are independently selected from unsubstituted C. 1-4 Alkyl, hydroxyalkyl, halogen, C 1-3 alkyl halogenates;

[0012] R6-R7 are independently selected from substituted or unsubstituted alkyl, hydroxyalkyl, and halogen groups, wherein the substituent is a halogen.

[0013] R8 is selected from C 1-6 alkyl, C 1-6 cycloalkyl, C 1-6 Heterocyclic and aryl groups.

[0014] As a preferred embodiment of the present invention, the compound, its stereoisomers, tautomers, its solvates, or its pharmaceutically acceptable salts include:

[0015] R1-R5 are independently selected from methyl, halogen, C 1-3 alkyl halogenates;

[0016] R6-R7 are independently selected from methyl, ethyl, and halogen-substituted methyl groups;

[0017] R8 is selected from C 3-6 cycloalkyl, phenyl, C 1-6 Alkyl, tetrahydrofuranyl.

[0018] As a further preferred embodiment of the present invention, the compound, its stereoisomers, tautomers, its solvates, or its pharmaceutically acceptable salts, are characterized by:

[0019] R1-R5 are independently selected from methyl, F, Cl, -CH2F, and CF3;

[0020] R6-R7 are independently selected from methyl, ethyl, -CH2F, and CF3;

[0021] R8 is selected from C 3-6 cycloalkyl, phenyl, C 1-6 Alkyl, tetrahydrofuranyl.

[0022] As a further preferred embodiment of the invention, the compound, its stereoisomers, tautomers, its solvates, or its pharmaceutically acceptable salts, is characterized by being selected from:

[0023]

[0024] In some embodiments of the present invention, the compound having the structure of Formula I is selected from any of the following:

[0025]

[0026] The preparation method of the compound SJ-301, its stereoisomers, tautomers, its solvates, or its pharmaceutically acceptable salts according to the present invention includes the following steps:

[0027]

[0028]

[0029] A pharmaceutical composition comprising the compound of the present invention, its stereoisomers, tautomers, its solvates or pharmaceutically acceptable salts thereof as the active ingredient or the main active ingredient, supplemented by a pharmaceutically acceptable carrier.

[0030] As a preferred embodiment of the present invention, the composition is prepared into any pharmaceutically permissible dosage form; the dosage form includes, but is not limited to, preparations administered by any route of oral, parenteral, intraperitoneal, intravenous, intra-arterial, topical, transdermal, sublingual, intramuscular, rectal, buccal, intranasal, inhalation, vaginal, intraocular, local, subcutaneous, intrafacial, intra-articular, intraperitoneal, or intrathecal administration; the dosage form is preferably an ointment, liniment, lotion, spray, tablet, granule, oral liquid, capsule, drop pill, enema, film, or injection.

[0031] The use of the compounds, stereoisomers, tautomers, solvates, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions thereof, in the preparation of medicaments for the treatment and / or prevention of neurological diseases characterized at least in part by β-amyloid deposition.

[0032] As a preferred embodiment of the invention, the neurological disease characterized at least in part by β-amyloid deposition is a neurological disease characterized at least in part by the accumulation of β-amyloid in the brain.

[0033] As a further preference of the present application, the neurological disease characterized at least in part by beta-amyloid deposition is selected from the group consisting of Alzheimer's disease, Down's syndrome, traumatic brain injury, cerebrovascular disease, or a combination thereof; preferably Alzheimer's disease.

[0034] In some embodiments, administering to the subject an effective amount of the compound comprises administering to the subject a pharmaceutical composition comprising an effective amount of the compound to the subject and at least one pharmaceutically acceptable carrier. In certain embodiments, the subject has or is suspected of having a neurological disease characterized at least in part by accumulation of beta-amyloid in the brain. These neurological diseases can be Alzheimer's disease, Down's syndrome, traumatic brain injury, cerebrovascular disease, or a combination thereof. In any embodiment, administering to the subject an effective amount of the compound can reduce the concentration of beta-amyloid in the brain of the subject.

[0035] The term "or" as used herein, unless otherwise indicated, refers to a single element of the stated alternative, or to a combination of two or more elements of the stated alternative.

[0036] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0037] Unless otherwise indicated, numerical ranges are to be understood as referring to each individual number within the range, including the end points. Unless otherwise indicated, all numbers used herein to describe dimensions, amounts, weight percentages, temperatures, times, and so forth, are to be understood as modified by the term "about." It is also to be understood that the

[0038] While alternative to various components, parameters, operating conditions, and the like are set forth herein, this does not mean that such alternatives are necessarily equivalent and / or equally good performance. Unless otherwise indicated, this also does not mean that the alternatives are listed in a preferred order.

[0039] Definitions of common terms in chemistry can be found in Richard J. Lewis, Sr. (ed.), Hawley's Condensed Chemical Dictionary, published by John Wiley & Sons, Inc., 2016 (ISBN 978-1-118-13515-0).

[0040] To facilitate review of the various embodiments of the present application, the following explanations of specific terms are provided:

[0041] Activator: A drug, medicament, therapeutic agent, nutraceutical, or other compound used to treat, ameliorate, or prevent a disease or disorder or at least one symptom associated therewith. The term activator also includes biologically active agents such as proteins, antibodies, antibody fragments, peptides, oligonucleotides, vaccines, and various derivatives of these materials.

[0042] Aryl: Unless otherwise indicated, aryl is a monovalent aromatic carbocyclic group having from 6 to 15 carbon atoms, having a single ring (e.g., phenyl) or multiple condensed rings wherein at least one ring is aromatic (e.g., quinoline, indole, benzodiazepine, etc.), so long as the point of attachment is through an atom of the aromatic moiety of the aryl group and the aromatic moiety at the point of attachment contains only carbon atoms of the aromatic ring. If any ring moiety of the aryl group contains a heteroatom, the group is a heteroaryl rather than an aryl. The aryl group can be monocyclic, bicyclic, tricyclic, or tetracyclic.

[0043] Combination therapy: Combination therapy refers to administration of a compound of the present application with at least one other active agent within the same general time period such that both compounds exert an effect on the individual, and it does not require that the compounds be administered at the same time (although combination therapy does include administration at the same time point). Thus, combination therapy can be on the same day or different days, or the same week or different weeks.

[0044] Effective dose or therapeutically effective dose: The amount of a compound that elicits a beneficial or therapeutic effect in some percentage of subjects in the statistical population.

[0045] Excipient: A physiologically inert substance used as an additive in a pharmaceutical composition. As used herein, an excipient can be incorporated into the particles of a pharmaceutical composition or can be physically mixed with the particles of a pharmaceutical composition. Excipients can be used, for example, to dilute an active agent and / or to alter the properties of a pharmaceutical composition. Excipients can include, but are not limited to, antiadherents, binders, coatings, enteric coatings, disintegrants, flavoring agents, sweeteners, colorants, lubricants, glidants, adsorbents, preservatives, adjuvants, carriers, or vehicles. Excipients can be starches and modified starches, celluloses and cellulose derivatives, sugar classes and their derivatives such as disaccharides, polysaccharides, and sugar alcohols, proteins, synthetic polymers, cross-linked polymers, antioxidants, amino acids, or preservatives. Exemplary excipients include, but are not limited to, magnesium stearate, stearic acid, phytosterol, sucrose, lactose, starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose, xylitol, sorbitol, maltitol, gelatin, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), tocopheryl polyethylene glycol 1000 succinate (also known as vitamin E TPGS or TPGS), carboxymethylcellulose, dipalmitoyl phosphatidyl choline (DPPC), vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methyl paraben, propyl paraben, sugar, silicon dioxide, talc, magnesium carbonate, sodium starch glycolate, tartrazine, aspartame, benzalkonium chloride, sesame oil, propyl gallate, sodium metabisulfite, or lanolin.

[0046] Haloalkyl: An alkyl group substituted with one or more of the same or different halogen atoms, such as -CH2CI, -CF3, -CH2CF3, -CF2CF3, -CH2CCI3, and the like.

[0047] Heterocycle: Refers to a closed ring compound, or a group that is attached to another group, particularly an organic group, that has at least one atom in its ring structure that is not carbon, usually oxygen, sulfur, and / or nitrogen.

[0048] Pharmaceutically acceptable: A material that does not produce an appreciable adverse toxicological effect on the subject to which it is administered. “Pharmaceutically acceptable form” refers to any pharmaceutically acceptable derivative or variant, such as stereoisomers, mixtures of stereoisomers, enantiomers, solvates, hydrates, isomorphs, crystal forms, pseudomorphs, neutral forms, salt forms, and prodrug formulations.

[0049] Pharmaceutically acceptable carrier: The pharmaceutically acceptable carriers (carriers) used in the present application are conventional. Remington: The Science and Practice of Pharmacy, The University of the Sciences in Philadelphia, Editor, Lippincott, Williams, & Wilkins, Philadelphia, PA, 21st Edition (2005), describes compositions and formulations suitable for drug delivery of one or more compounds of Formula I disclosed herein.

[0050] In general, the nature of the carrier will depend on the particular mode of administration being employed. For example, parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle. In some embodiments, the pharmaceutically acceptable carrier can be sterile to be suitable for administration to a subject (e.g., by parenteral, intramuscular, or subcutaneous injection). In addition to biologically neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, preservatives, and pH buffering agents, and the like, e.g., sodium acetate or sorbitan monolaurate.

[0051] Pharmaceutically acceptable salt: A biologically compatible salt of a disclosed compound derived from various organic and inorganic counter ions known in the art, including, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and salts of organic or inorganic acids, for example hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, and the like, as the molecule contains a basic functional group. Pharmaceutically acceptable acid addition salt refers to those salts of the free base which are formed with acids which are biologically or otherwise tolerable, such as hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, and other inorganic acids such as adipic, aspartic, citric, glutamic, hydroxymaleic, maleic, malic, succinic, tartaric, and other organic acids such as acetic, trifluoroacetic, propionic, glycolic, pyruvic, oxalic, maleic, malonic, succinic, fumaric, tartaric, citric, benzoic, cinnamic, mandelic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, salicylic, and other organic acids. Pharmaceutically acceptable base addition salts include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Typical salts are the ammonium, potassium, sodium, calcium, and magnesium salts. From pharmaceutically acceptable organic non-toxic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrazine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Typical organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine (see, e.g., S. M. Berge et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19, incorporated herein by reference).

[0052] Subject: An animal (human or non-human) that receives treatment, observation, or experimentation.

[0053] The above definitions and the following formulae do not include impermissible substitution patterns (e.g., methyl substituted with 5 fluorine groups). One of ordinary skill in the art will readily recognize such impermissible substitution patterns.

[0054] Any group mentioned herein can be optionally substituted with at least one, and possibly two or more, substituents defined herein. That is, unless otherwise specified in the context in which it occurs, a substituent group has at least one, and possibly two or more substituents, replaceable hydrogens defined herein. Those of ordinary skill in the art will appreciate that compounds can exhibit tautomeric, conformational, geometric, and / or optical isomerism phenomena. For example, certain disclosed compounds can include one or more chiral centers and / or double bonds and, therefore, can exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers), enantiomers, diastereomers, and mixtures of such isomers, e.g., racemic mixtures. As another example, certain disclosed compounds can exist in several tautomeric forms including enol forms, keto forms, and mixtures of such forms. Since the various compound names, chemical formulas, and compound depictions in the specification and claims can represent only one of the possible tautomeric, conformational, optical, or geometric isomeric forms, those of ordinary skill in the art will appreciate that the disclosed compounds include any and all tautomeric, conformational, optical, and / or geometric isomeric forms of the compounds described herein, as well as mixtures of different isomeric forms. In the case of limited rotation, such as around an amide bond or between two directly connected rings like a pyrazolyl and pyridyl ring, atropisomers are also possible and are specifically included in the compounds of the present application.

[0055] In any embodiment, any or all hydrogens present in a compound or in a particular group or moiety in a compound can be replaced by deuterium or tritium. Thus, recitations of alkyl groups include deuterium alkyl groups in which from one up to the maximum number of hydrogens can be replaced by deuterium. For example, an ethyl group can be C2H5or C2H5in which 1-5 hydrogens are replaced by deuterium, such as C2D5. x H5-x.

[0056] In some embodiments, administering an effective amount of a compound to a subject comprises administering to the subject a pharmaceutical composition comprising an effective amount of the compound and at least one pharmaceutically acceptable carrier. The mode of administration can be any suitable route, including but not limited to intravenous, oral, intraperitoneal, subcutaneous, rectal, or buccal administration.

[0057] In some embodiments, the subject has or is suspected of having a neurological disease characterized at least in part by amyloid beta protein accumulation in the brain. Neurological diseases characterized at least in part by beta-amyloid protein accumulation in the brain include, but are not limited to, Alzheimer’s disease, Down’s syndrome, traumatic brain injury, cerebrovascular disease, and combinations thereof. In certain embodiments, the subject has or is suspected of having Alzheimer’s disease.

[0058] Actual dosage will vary according to factors such as the neurological disease and particular state of the subject (e.g., age, size, severity of symptoms, susceptibility factors, etc.), time and route of administration, other drugs or treatments being administered concurrently, and the specific pharmacology of the compound for eliciting the desired activity or biological response in the subject. Dosage regimens can be adjusted to provide the optimum prophylactic or therapeutic response. An effective amount also refers to an amount of the compound that is nontoxic and / or otherwise acceptable to the subject.

[0059] Alternatively, an in vitro model can be used to determine an effective dose. Using such a model, only routine calculation and adjustment are needed to determine the appropriate concentration and dosage of the compound to administer a therapeutically effective amount (e.g., an amount effective to reduce the concentration of beta-amyloid in the brain or to reduce the rate of beta-amyloid accumulation).

[0060] In the methods and formulations of the present application, an effective amount of the compound according to Formula I is in the non-limiting range of 0.0001 gram to 100 grams, e.g., 0.001 gram to 50 grams, 0.01 gram to 25 grams, or 0.1 gram to 10 grams, for an adult human. In some embodiments, the effective amount is in the range of 0.001 mg / kg body weight to 100 mg / kg body weight, e.g., 0.01 mg / kg body weight to 20 mg / kg body weight, 0.01 mg / kg body weight to 10 mg / kg body weight, 0.05 mg / kg body weight to 5 mg / kg body weight, or 0.1 mg / kg body weight to 2 mg / kg body weight. The effective amount can be administered in a single dose, or in two or more doses. In some embodiments, the effective amount of the compound or pharmaceutical composition comprising the compound is administered to the subject in a dosage regimen of once per day.

[0061] The attending physician can vary the dosage regimen to accommodate the needs of the subject. The dose can be increased or decreased, as necessary, to maintain the desired concentration at the target site (e.g., systemic circulation). Higher or lower concentrations can be selected depending on the mode of administration for example, oral administration versus intravenous or subcutaneous administration. The dose can also be adjusted according to the release rate of the administered formulation, for example, oral sustained release versus injected particulate or rectal suppository, etc.

[0062] In some embodiments, a second active agent can be co-administered with the compound. The compound and the second active agent can be administered separately or together in a single composition. The second active agent can be administered by the same route or by a different route. The compound and the second agent can be administered simultaneously or at different times. Separate administration can be in any order. If administered simultaneously, the compound and the second active agent can be combined in a single pharmaceutical composition, or can be administered concurrently as two separate pharmaceutical compositions. The second active agent can be, for example, a cholinesterase inhibitor, an anti-hypertensive agent, an anti-beta-amyloid antibody, a diuretic, an anticonvulsant, an antidepressant, an antipsychotic, an analgesic, a motor system drug, a memory or cognition drug, and combinations thereof.

[0063] The compound of the present application or a pharmaceutically acceptable salt thereof can be applied in the field of pharmacy, and animal model experiments show that the compound can significantly improve the cognitive impairment of the subject, reduce the pathological indicators of Alzheimer's disease, and has the effect of treating and / or preventing Alzheimer's disease. BRIEF DESCRIPTION OF DRAWINGS

[0064] Figure 1 is the high-resolution mass spectrum of the compound SJ-301 obtained in Example 1.

[0065] Figure 2 is a statistical fold line graph of the latency of two groups of mice in Example 5 in the water maze training period for five days.

[0066] Figure 3 is a statistical column chart of the number of times of crossing the platform of two groups of mice in Example 5 during the water maze test.

[0067] Figure 4 is a statistical column chart of the time of activity in the platform quadrant of two groups of mice in Example 5 during the water maze test.

[0068] Figure 5 is a statistical column chart of the proportion of the number of times of entering the new arm of two groups of mice in Example 5 during the Y maze test.

[0069] Figure 6 is an immunofluorescence contrast chart of the Aβ plaque of the dentate gyrus region of the brain frozen section of two groups of mice in Example 5, with a scale of 50 μm.

[0070] Figure 7 is an immunofluorescence contrast chart of the Aβ plaque of the cortex region of the brain frozen section of two groups of mice in Example 5, with a scale of 50 μm.

[0071] Figure 8 is an immunofluorescence contrast chart of the astrocyte of the dentate gyrus region of the brain frozen section of two groups of mice in Example 5, with a scale of 50 μm.

[0072] Figure 9 is an immunofluorescence contrast chart of the astrocyte of the cortex region of the brain frozen section of two groups of mice in Example 5, with a scale of 50 μm.

[0073] Figure 10 is an immunofluorescence contrast chart of the microglia of the dentate gyrus region of the brain frozen section of two groups of mice in Example 5, with a scale of 50 μm.

[0074] Figure 11 is an immunofluorescence contrast chart of the microglia of the cortex region of the brain frozen section of two groups of mice in Example 5, with a scale of 50 μm.

[0075] Figure 12 Figure 6 is a bar graph of the total number of Aβ plaques in brain cryosections from two groups of mice in Example 5.

[0076] Figure 13 Figure 7 is a bar graph of the total number of aggregated astrocytes in brain cryosections from two groups of mice in Example 5.

[0077] Figure 14 Figure 8 is a bar graph of the total number of aggregated microglia in brain cryosections from two groups of mice in Example 5.

[0078] Figure 15 Figure 9 is a bar graph of the number of Aβ plaques in brain cryosections from two groups of mice in Example 5, classified by size.

[0079] Figure 16 Figure 10 is a bar graph of the number of aggregated astrocytes in brain cryosections from two groups of mice in Example 5, classified by size.

[0080] Figure 17 Figure 11 is a bar graph of the number of aggregated microglia in brain cryosections from two groups of mice in Example 5, classified by size. DETAILED DESCRIPTION

[0081] The application is further described in connection with the following examples, which are not intended to limit the application. The examples disclosed herein can be used in subjects having or suspected of having a neurological disease characterized at least in part by β-amyloid accumulation in the brain. In some embodiments, administration of an effective amount of an inhibitor to a subject can reduce the concentration of β-amyloid in the subject's brain.

[0082] Materials and Methods:

[0083] (1) Antibodies:

[0084] Antibodies used in this study and commercial cell lines are listed here (Table 1).

[0085] Table 1. Antibodies used in the Examples and their sources

[0086]

[0087] (2) Animals: Healthy 5×FAD strain mice, 12 weeks old, weighing approximately 20-25g, were purchased from Jiangsu Airingfei Biotechnology Co., Ltd. The mice were housed in an environment with constant and suitable humidity (40-70%), temperature (18-26℃), and normal diurnal rhythm, with normal food and water supply. All mice underwent a one-week acclimatization period before the experiment.

[0088] (3) Reagent: Triton X-100, purchased from Guangzhou Saiguo Biotechnology Co., Ltd. Contains DAPI as an anti-fluorescence quencher. Purchased from Southern Biotech, USA. Bovine Serum Albumin (BSA) was purchased from Sangon Biotech (Shanghai) Co., Ltd. Paraformaldehyde powder was purchased from Sinopharm Chemical Reagent Co., Ltd. The embedding agent was purchased from Japan. Sakura Corporation. Microscope coverslips and slides were purchased from Jiangsu Shitai Experimental Equipment Co., Ltd. PBS buffer (NaCl 8g: purchased from Xilong Scientific, KH2PO4 0.2g: purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd., Na2HPO4·H2O 3.49g: purchased from Nanjing Chemical Reagent Co., Ltd., KCl 0.2g: purchased from Nanjing Chemical Reagent Co., Ltd., 1L distilled water). 0.9% sodium chloride injection (physiological saline): purchased from Chenxin Pharmaceutical Co., Ltd. Isoflurane inhalation anesthetic: purchased from RWD. Sucrose: purchased from Sinopharm Chemical Reagent Co., Ltd. Sodium carboxymethyl cellulose: purchased from Sinopharm Chemical Reagent Co., Ltd. Antibacterial agent ProClin TM 300, purchased from Sigma-Aldrich (48912-U). Compound coloring emulsifier, purchased from Jiangsu Hushen Titanium Dioxide Technology Co., Ltd.

[0089] (4) Instruments: IX73 fluorescence inverted microscope: purchased from Olympus, Japan. Pipettes: purchased from Eppendorf, Germany. Electronic balance: purchased from Shanghai Licheng Bangxi Instrument Technology Co., Ltd. Electronic analytical balance: purchased from Mettler Toledo Instruments (Shanghai) Co., Ltd. 902 ultra-low temperature freezer (-80℃): purchased from Thermo Fisher Scientific, USA. CM1950 cryostat: purchased from Leica, Germany. pH meter: purchased from Shanghai Instrument & Electronics Scientific Instruments Co., Ltd. Water maze video analysis system (Zhongshidichuang), Y-maze video analysis system hardware (Zhongshidichuang), Any-maze analysis software 6.0.

[0090] Example 1: Modification and Synthesis of Compound SJ-301

[0091] Step 1 : Synthesis of 4-amino- 1 -cyclopropylpiperidine:

[0092]

[0093] To a round bottom flask was added 4-tert-butoxycarbonylaminopiperidine (1 g, 1 eq), sodium cyanoborohydride (0.9426 g, 3 eq) in 20 ml methanol, followed by (1-ethoxycyclopropoxy)trimethylsilane (0.9599 g, 1.1 eq) and catalytic amount of acetic acid. The reaction was carried out at 50 °C for 13 h. TLC (V DCM :V MeOH = 10:1) indicated the completion of the reaction. The solvent was removed and washed with 1 M aqueous NaOH (20 ml), saturated NaCl (20 ml), saturated NH4Cl (20 ml) and extracted with ethyl acetate (30 ml x 3). The solvent was removed to obtain a white solid (1.1076 g).

[0094] To a round bottom flask was added the white solid (0.65 g, 1 eq) in dichloromethane (8 ml) and trifluoroacetic acid (0.85 ml, 5 eq) was added dropwise at -10 °C. The reaction was carried out at -10 °C for 6 h. TLC (V DCM :V MeOH = 5:1) indicated the completion of the reaction. The reaction was basified with 10% NaOH (W / V) aqueous solution (20 ml) and extracted with dichloromethane / methanol (V DCM :V MeOH = 10:1) (50 ml x 3). The solvent was removed to obtain a pale yellow oily product (0.33 g, 87% yield).

[0095] Step 2: Synthesis of N-(l-(3-chloro-4-fluorophenyl)-3,5-dimethyl-lH-pyrazol-4-yl)methyl- cyclopropylpiperidin-4-amine:

[0096]

[0097] To a round bottom flask was added 3-chloro-4-fluorobenzhydrazine hydrochloride (0.985 g, 1 eq), acetoacetone (0.57 ml, 1.1 eq), p-toluenesulfonic acid (0.043 g, 0.05 eq) in 10 ml ethanol and the reaction was carried out at room temperature for 14 h. TLC (V Pe :V EA = 3:1) indicated the completion of the reaction. The solvent was removed and washed with saturated NaHCO3 (30 ml) and extracted with ethyl acetate (30 ml x 3). The solvent was evaporated under reduced pressure and purified on a silica gel column (V Pe :V EA = 3:1) to obtain a yellow oily material (1.090 g).

[0098] Under nitrogen atmosphere, dry N,N-dimethylformamide (5 ml) was added to a round bottom flask, and phosphorus oxychloride (0.91 ml, 2 eq) was added dropwise. After the ice-salt bath was removed and the mixture was stirred for 15 min to allow it to warm to room temperature, the resulting yellow oily material (1.090 g, 1 eq) was dissolved in dry N,N-dimethylformamide (5 ml) and added to the flask. The reaction was carried out at 100 °C for 7 h. TLC (V Pe :V EA = 3:1) indicated that the reaction was complete. The reaction was quenched by the addition of 10% NaOH (W / V) aqueous solution (20 ml) under ice-salt bath, and the solvent was evaporated under reduced pressure after extraction with ethyl acetate (30 ml x 3). The residue was purified by silica gel column chromatography (V Pe :V EA = 5:1-V Pe :V EA = 3:1) to give a white solid (0.933 g).

[0099] The resulting white solid (0.933 g, 1 eq), 4-amino-l-cyclopropylpiperidine (0.330 g, 1.2 eq), and acetic acid (0.12 ml, 1 eq) were dissolved in tetrahydrofuran and added to a round bottom flask under nitrogen atmosphere. After the mixture was stirred for 30 min, sodium triacetoxyborohydride (1.24 g, 3 eq) was added, and the reaction was carried out at room temperature for 14 h. TLC (V DCM :V MeOH = 5:1) indicated that the reaction was complete. The reaction was quenched by the addition of 1 M NaOH aqueous solution (20 ml), and the solvent was evaporated under reduced pressure after extraction with ethyl acetate (30 ml x 3). The residue was purified by silica gel column chromatography (V DCM :V MeOH = 10:1-V DCM :V MeOH :V Et3N = 40:2:5) to give a yellow oily product, which was named SJ-301 (0.578 g, 78% yield).

[0100] 1H NMR (300 MHz, Chloroform-d) δ 7.50 (dd, J = 6.5, 2.5 Hz, 1H), 7.32 - 7.24 (m, 1H), 7.19 (t, J = 8.6 Hz, 1H), 3.65 (s, 2H), 3.05 - 2.89 (m, 2H), 2.62 (ddt, J = 14.7, 10.2, 4.0 Hz, 1H), 2.30 (d, J = 2.0 Hz, 6H), 2.21 (td, J = 11.7, 2.6 Hz, 2H), 2.00 - 1.88 (m, 2H), 1.54 (td, J = 6.8, 3.4 Hz, 1H), 1.44 (dt, J = 11.5, 5.8 Hz, 2H), 0.48 - 0.29 (m, 4H).

[0101] 13 C NMR (75 MHz, Chloroform-d) δ 158.73, 155.42, 148.89, 138.14, 136.54, 127.15, 124.38 (d, J = 7.5 Hz), 121.49 (d, J = 19.0 Hz), 116.81 - 115.31 (m), 54.86, 52.27, 39.38, 38.26, 31.53, 11.91, 11.12, 5.91.

[0102] HRMS (ESI) calculated for C20H27ClFN4 [M+H]+m / z 377.19028, found 377.19012. Figure 1 )

[0103] Effect of Example 2 compound SJ-301 on Alzheimer's disease model in mice

[0104] 1.1 Method for preparing therapeutic drugs

[0105] The excipient is sodium carboxymethyl cellulose, which is prepared into a 0.5% concentration with triple distilled water. The SJ-301 drug is precisely weighed into the 0.5% sodium carboxymethyl cellulose to a final concentration of 0.5 mg / mL, and is thoroughly ground in a mortar to uniformly distribute the compound powder. The body weight of the mice is weighed daily, and is gavaged at a dosage volume of 10 mL / kg, so that the dosage concentration of each mouse is 5 mg / kg.

[0106] 1.2 Grouping of animals

[0107] Randomly divide 5xFAD mice into two groups, one group is solvent control group, one group is drug experimental group, of which 10 mice in drug experimental group, 7 mice in solvent control group. Drug experimental group is given SJ-301 treatment drug 5mg / kg by gavage every day, and solvent control group is given control solvent 0.5% sodium carboxymethyl cellulose by gavage every day according to the drug volume of 10mL / kg. The drug administration period is 8 weeks, starting from the age of 8 weeks to the 16th week.

[0108] 1.3 Observation index

[0109] 1.3.1 Morris water maze and Y maze to determine the cognitive differences between the two groups of mice

[0110] After 8 weeks of drug administration in mice, behavioral tests were conducted on the two groups of mice.

[0111] (1) Morris water maze experiment: It is a kind of experiment that forces experimental animals (rats, mice) to swim and learn to find a hidden platform in water, mainly used to test the learning and memory ability of experimental animals to space location and direction (spatial orientation). All mice in the two groups were moved to the water maze experiment room for feeding 1 week before the formal experiment to adapt to the environment.

[0112] The water maze experiment is divided into training period and test period, the training period is also called positioning navigation test, which lasts for 5 days, and the platform is hidden under the water surface 0.5-1cm. Each mouse is put into the water from four water entry points respectively each day, and the time for finding the hidden platform under the water within 60s (escape latency) is recorded. The test period is one day after the training period, also known as spatial exploration test. Remove the platform and put the mouse into the pool from the opposite side of the platform as the water entry point. Record the swimming track of the mouse within 60s, the number of times crossing the original platform position, and investigate the memory of the mouse to the original platform.

[0113] (2) Y maze can be used to evaluate the short-term memory of mice. By placing the experimental mice in a Y maze with one arm closed for 15 minutes during training. After a 1-hour interval, open the arm and let the mouse move freely in the three arms, and record the number of times the mouse enters each arm. Mice with better spatial reference memory should remember the arm they have not explored before and will visit the previously closed arm more frequently.

[0114] 1.3.2 Immunofluorescence detection of AD pathological indicators in mouse brain frozen sections

[0115] After 8 weeks of drug administration in mice, and after the completion of behavioral tests, the mice were anesthetized according to the grouping, and after anesthesia, the mice were heart perfused with normal saline, and the brain was fixed in 4% PFA at 4°C overnight, and dehydrated with gradient sucrose solution. After dehydration was completed, the embedding agent was used for embedding, and the mouse brain was stored in the refrigerator at -80°C. The coronal sections of the hippocampus of the mouse were prepared by using a freezing microtome, and the thickness was 30 μm, and about 35 sections were cut for each mouse brain. After the brain slices of the mouse hippocampus were cut, they were placed in a six-well plate containing 30% sucrose, and then the subsequent related immunofluorescence experiments were carried out.

[0116] The brain slices were selected, washed with PBS at room temperature for 3 times, each time for 5 min. Then the brain slices were permeabilized with 0.2% Triton (prepared in PBS) at room temperature for 25 min, and then washed with PBS for 3 times, each time for 7 min; blocked with blocking solution (2.5% BSA + 0.3% Triton + 0.05% bacteriostatic agent, prepared in PBS) at room temperature for 1 h. After diluting the corresponding primary antibody to the working concentration with the blocking solution, the brain slices were incubated in the primary antibody at 4°C for 12 h or more. After the incubation of the primary antibody was completed, the brain slices were washed with PBS for 3 times, each time for 7 min. The working concentration of the fluorescent secondary antibody prepared in PBS was placed in the brain slices, and incubated in the dark for 1-2 h. After the completion of the secondary antibody, the brain slices were washed with PBS for 3 times, each time for 7 min. The brain slices were attached to the glass slide by using the slide method, and an appropriate amount of DAPI-containing anti-fluorescence quencher was dropped, and a cover glass was covered, and the edges were coated with nail polish. After air-drying in the dark, the expression of Aβ, GFAP, Iba1 and other proteins on each brain slice was observed under a fluorescence microscope, and the size of Aβ plaque, the diameter and number of aggregated forms of astrocytes (GFAP marker) and microglia (Iba1 marker) were counted.

[0117] 1.4 Statistical method

[0118] All data statistical analysis and curve fitting analysis were performed by using GraphPad Prism 6.0 software. The experimental data were expressed by mean and standard error of mean (Mean ± SEM). The statistical software GraphPad Prism was used to perform variance analysis and Student’s t-test on the data to test the differences between groups. *P<0.05 indicates that the experimental data of two groups have statistical difference, **P<0.01 indicates that the experimental data of two groups have significant statistical difference, ***P<0.001 indicates that the experimental data of two groups have extremely significant statistical difference, and ns indicates that the experimental data of two groups have no statistical difference.

[0119] 1.5 Experimental results

[0120] 1.5.1 Animal behavioral experiment

[0121] (1) Morris water maze experiment

[0122] The results of the five-day training are shown in Table 2:

[0123] Table 2 Effects of SJ-301 on escape latency of 5xFAD five-transgenic mice (±SEM)

[0124]

[0125]

[0126] Compared with the control group, *P<0.05, **P<0.01, ***P<0.001.

[0127] As can be seen from Table 2:

[0128] Compared with the solvent control group, the compound of the drug experimental group can significantly shorten the escape latency time of 5xFAD five-transgenic mice in the training period. The mice in the drug experimental group can find the platform location more quickly after training. From the second day, the escape latency of the mice in the drug experimental group began to be significantly different from that of the control group, and the difference between the two groups became more and more significant as the training days increased. This phenomenon shows that the mice in the drug experimental group learn and remember the platform location more quickly during the training process, reflecting the significant improvement of the spatial learning and memory ability of the mice in this group. Figure 2

[0129] The results of the test (spatial exploration experiment) are shown in Table 3:

[0130] Table 3 Platform crossing times and time spent in the quadrant where the platform was located during the test period (±SEM)

[0131]

[0132] Compared with the control group, *P<0.05, **P<0.01, ***P<0.001.

[0133] As can be seen from Table 3, the mice in the drug experimental group have a significant increase in the number of times of crossing the original platform location and the time spent in the quadrant where the original platform was located during the test period after removing the platform compared with the solvent control group. These two indicators can measure the degree of memory of the mice for the platform location, reflecting whether the mice remember and can explore the platform location after five days of training. Therefore, the spatial learning and memory of the mice in the drug experimental group have been significantly improved. Figures 3-4

[0134] (2) Y maze experiment

[0135] The Y maze method used in the present application measures the spatial reference memory of mice, and the standard indicator is the proportion of the number of times of entering the new arm during the test period, and the calculation formula is:

[0136] ​​The ratio of the number of times of entering the new arm = the number of times of entering the new arm / the total number of times of entering the three arms x 100%

[0137] The experimental results of the 5-minute test of the two groups of mice are shown in Table 4:

[0138] Table 4: The ratio of the number of times of entering the new arm of each group of mice in the Y maze test period (±SEM)

[0139]

[0140]

[0141] *P<0.05, **P<0.01, ***P<0.001 compared with the control group.

[0142] According to the data in Table 4 and Figure 5 Statistical data, under the same training conditions, the test performance of the drug experimental group of mice in the Y maze was significantly better than that of the control experimental group. The drug experimental group of mice entered the new arm more times and had stronger exploration ability for the new arm, reflecting that the mice in this group had stronger spatial reference memory. Therefore, it can be concluded that giving the compound drug can improve the spatial reference memory ability of mice.

[0143] 1.5.2 Immunofluorescence experiment of mouse brain frozen sections

[0144] Both groups of mice were perfused after the end of the behavior test, and brain frozen sections were prepared to observe the deposition of Aβ plaques in the brain of mice, as well as the number and size of aggregated astrocytes and microglia. These three indicators are the standard for measuring AD. The more and larger the Aβ plaques, aggregated astrocytes and microglia in the brain, the more severe the AD pathology of the mice.

[0145] The specific results are shown in Figures 6-11 .

[0146] Figure 6 The left side vehicle is the solvent control group, and the right side SJ-301 is the drug experimental group. As shown in Figure 6 , in the dentate gyrus region of the mouse hippocampus, the drug experimental group of mice has reduced Aβ deposition density and reduced Aβ plaque size.

[0147] Figure 7 The left side vehicle is the solvent control group, and the right side SJ-301 is the drug experimental group. As shown in Figure 7 , in the cortical region of the mouse hippocampus, the drug experimental group of mice has reduced Aβ deposition density and reduced Aβ plaque size.

[0148] Figure 8 The left side vehicle is the solvent control group, and the right side SJ-301 is the drug experimental group. As shown inFigure 8 As shown in the dentate gyrus region of the mouse hippocampus, the size and density of the aggregated astrocytes of the drug experimental group mice were reduced.

[0149] Figure 9 The left vehicle is the solvent control group, and the right SJ-301 is the drug experimental group. As shown in the dentate gyrus region of the mouse hippocampus, Figure 9 As shown in the cortex region of the mouse hippocampus, the size and density of the aggregated astrocytes of the drug experimental group mice were reduced.

[0150] Figure 10 The left vehicle is the solvent control group, and the right SJ-301 is the drug experimental group. As shown in the dentate gyrus region of the mouse hippocampus, Figure 10 As shown in the cortex region of the mouse hippocampus, the size and density of the aggregated astrocytes of the drug experimental group mice were reduced.

[0151] Figure 11 The left vehicle is the solvent control group, and the right SJ-301 is the drug experimental group. As shown in the dentate gyrus region of the mouse hippocampus, Figure 11 As shown in the cortex region of the mouse hippocampus, the size and density of the aggregated astrocytes of the drug experimental group mice were reduced.

[0152] At the same time, through statistical analysis of the immunofluorescence results of multiple brain slices of the two groups, the size and number were taken as indicators, and the results are shown in Tables 5-7:

[0153] Table 5 Number of Aβ plaques in the brain frozen sections of the solvent control group and the drug experimental group mice (±SEM)

[0154]

[0155]

[0156] Table 6 Number of aggregated astrocyte proliferation in the brain frozen sections of the solvent control group and the drug experimental group mice (±SEM)

[0157]

[0158] Table 7 Number of aggregated microglial proliferation in the brain frozen sections of the solvent control group and the drug experimental group mice (±SEM)

[0159]

[0160] Compared with the control group, *P<0.05, **P<0.01, ***P<0.001.

[0161] According to Tables 3-5 and Figures 12-17The total number of Aβ plaque in the brain frozen section of the drug experimental group mice was significantly less than that of the control group, and there were almost no large diameter plaques (>40 μm), medium diameter (20-40 μm) and small diameter (<20 μm) plaques. The number of plaques in the drug experimental group mice was concentrated in the small diameter range, indicating that the intragastric administration of compound SJ-301 could effectively reduce the deposition of Aβ plaques in the brain of Alzheimer's disease model mice (5xFAD five-transgenic mice). Compared with the control group, the total number of aggregated astrocyte proliferation in the drug experimental group mice was also significantly reduced, and the number of large diameter, medium diameter and small diameter aggregated astrocytes was also significantly reduced. For the aggregation of microglial cells, the total number of aggregated microglial cells in the brain frozen section of the drug experimental group mice was significantly less than that of the control group, and although the number of small diameter aggregated microglial cells was more than that of the control group, the medium diameter and large diameter microglial cell proliferation was almost none, indicating that the microglial cell proliferation was concentrated in the small diameter range, and the inflammatory response in the brain of the mice in this group was significantly improved.

[0162] From the above data, compared with the solvent control group, the pathological indicators in the brain of Alzheimer's disease model mice administered with compound SJ-301 by intragastric administration were significantly improved.

[0163] Combining the behavior and pathology experiments, it can be concluded that compound SJ-301 has a therapeutic effect on Alzheimer's disease and can significantly improve the AD pathology and learning and cognitive impairment of mice.

[0164] Considering the many possible embodiments that can apply the principles of the disclosed patent, it should be recognized that the examples shown are merely preferred examples of the patent and should not be considered limiting the scope of the patent. On the contrary, the scope of the patent is defined by the following statements. Therefore, we claim all within the scope of these statements and the core idea is our patent.

Claims

1. An aryl aniline compound, a tautomer thereof, or a pharmaceutically acceptable salt thereof, characterized by selected from the group consisting of: 。 2. A pharmaceutical composition, characterized by, The aryl aniline compound according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt thereof is used as an active ingredient or a main active ingredient, in combination with a pharmaceutically acceptable carrier.

3. The pharmaceutical composition of claim 2, wherein, The composition is prepared in any dosage form permitted in pharmacy; the dosage form is selected from preparations for oral, parenteral, intraperitoneal, intravenous, intraarterial, external use on skin, transdermal, sublingual, intramuscular, rectal, transbuccal, intranasal, inhalation, vaginal, intraocular, topical, subcutaneous, intralipid, intraarticular, intraperitoneal or intrathecal administration.

4. The pharmaceutical composition of claim 3, wherein, The dosage form is selected from the group consisting of ointment, liniment, lotion, spray, tablet, bolus, oral liquid, capsule, dripping pill, enema, film or injection.

5. Use of the compound according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to any one of claims 2 to 4 in the manufacture of a medicament for the treatment or / and prevention of a neurological disease characterized by β-amyloid deposition, selected from the group consisting of Alzheimer's disease, Down's syndrome, traumatic brain injury, cerebrovascular disease or a combination thereof.

6. Use according to claim 5, characterized in that The neurological disease characterized by β-amyloid deposition is Alzheimer's disease.

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