WO2020237747A1

WO2020237747A1 - Applications of long-acting compound in preparing medicament

Info

Publication number: WO2020237747A1
Application number: PCT/CN2019/091695
Authority: WO
Inventors: 周强; 叶涛; 李书鹏
Original assignee: 北京大学深圳研究生院
Priority date: 2019-05-24
Filing date: 2019-06-18
Publication date: 2020-12-03
Also published as: CN110538169B; CN110538169A

Abstract

Uses of a compound having long-acting therapeutic effects in preparing a medicament. The compound is applicable in preparing a medicament for combating depression, anesthesia, pain relief, improving cognitive function, pulmonary protection, amyotrophic lateral sclerosis, and complex regional pain syndrome.

Description

Application of a long-acting compound in preparing medicine

Technical field

This application relates to the field of medicine, and specifically relates to the application of a compound for treating depression, neuropathic and chronic pain, including complex regional pain syndrome (CRPS), in the preparation of medicines.

Background technique

Ketamine is a representative of intravenous anesthetics of phencyclidine commonly used in clinical practice, and it is one of the rapid development of clinical and basic research in recent years. In clinical practice, it is often used to meet the anesthesia needs of pediatrics, obstetrics, perioperative patients and patients with special diseases because of its characteristics such as rapid induction, shorter action time, faster awakening, and less impact on the respiratory and circulatory systems.

Ketamine was first synthesized in 1962, used in humans in 1965, and was officially approved by the FDA for clinical use in 1970. Its typical "separation anesthesia" and short-acting precise analgesia made it a smash hit, but then it was discovered that mental side effects and the rapid development of other intravenous anesthetics have greatly reduced the clinical use of ketamine. In the past 10 years, with the research on the usage and dosage of ketamine, and its anti-inflammatory, antidepressant, neuroprotective, analgesic and other effects have been discovered, the medical community has renewed interest in ketamine.

For a long time, ketamine has strong analgesia, forgetfulness, while retaining spontaneous breathing and airway protective reflex, maintaining hemodynamic stability, etc., so that the role of ketamine in prehospital anesthesia and analgesia cannot be ignored. Ketamine has both neurotoxic and neuroprotective effects. The huge surgical stimulation may theoretically balance the damage of anesthesia, while ketamine can reduce the neurocognitive damage caused by surgical injury stimulation. Does ketamine cause neuroprotection or nerve damage and the dosage of ketamine? There is a direct correlation.

Ketamine has an effect on postoperative cognitive function. Some researchers tested 50 children receiving ketamine anesthesia and found that general ketamine anesthesia can reduce the cognitive function of children 6 hours after surgery, but it has no effect on the cognitive function after 24 hours. Hudetz et al. found that giving 0.5 mg/kg ketamine during general anesthesia induction can reduce the incidence of postoperative cognitive dysfunction one week after cardiac surgery, and this effect may be related to the anti-inflammatory effect of ketamine. In recent years, numerous clinical trials have confirmed that a single small dose of ketamine during surgery can reduce the incidence of postoperative cognitive dysfunction (POCD).

Ketamine has an analgesic effect. Sub-anaesthetic doses of ketamine are often used for the treatment of hyperalgesia and acute and chronic pain. Studies have confirmed that gargle with ketamine salt water mixture before induction of anesthesia can significantly reduce the incidence and severity of postoperative sore throat caused by general anesthesia tracheal intubation. The use of opioids during surgery increases the amount of opioid analgesia after surgery. This effect is called opioid tolerance. It has been clinically found that the use of ketamine can prevent opioid tolerance and can also reverse morphine tolerance and enhance the analgesic effect of morphine. Studies have also confirmed that the intraoperative application of low-dose ketamine can prevent remifentanil-induced postoperative hyperalgesia. Cagla et al. found that after intravenous injection of ketamine 0.15 mg/kg in patients undergoing knee arthroscopy, ketamine can significantly improve the satisfaction of postoperative analgesia, and the sedation score is lower than that of the ketamine combined midazolam group.

Ketamine has lung protection. In recent years, ketamine has been found to have significant lung protection. Clinical experiments have confirmed that both intravenous and aerosolization before one-lung ventilation in thoracic surgery can reduce the level of inflammatory factors in the blood, and aerosol inhalation is more beneficial to the cardiovascular system and airway pressure, while the effect of aerosolization on the side of lung ventilation Better than double lung nebulization. Ketamine is also commonly used clinically in the rescue of fatal asthma attacks when conventional treatment is ineffective, and it is recognized that its use can improve the prognosis.

Ketamine has an antidepressant effect. In 2000, Berman et al. reported for the first time that a single intravenous injection of sub-anaesthetic dose of ketamine (0.5 mg/kg, over 40 minutes intravenously) within 72 hours of more than 50% of patients with Hamilton Depression Scale score reduced by more than 50% (see: Berman RM, Cappiello A, Anand A, et al. Antidepressant effects of ketamine in depressed patients [J]. Biol Psychiatry, 2000, 47(4): 351-354). PaulR et al. found that compared with racemic ketamine, S-ketamine has similar antidepressant effects, but has fewer mental side effects (see: Paul R, Schaaff N, Padberg F, et al. Comparison of racemic ketamine and s-ketamine in treatment-resistant major depression: report of two cases[J]. World j biological psychiatry,2009,10(3):241-244). In recent years, more animal and clinical studies have further confirmed the antidepressant effect of ketamine. Ketamine is also used as anesthesia for electroconvulsive therapy in depressed patients.

In 2012, the U.S. Department of Health and Human Services, Cooper Health System and Stanford Research Institute jointly applied for application number CN 201280062294X, and the invention name is (2R,6R)-hydroxynorketamine, (S)-dehydronormethyl Patent application for the application of ketamine and other stereoisomeric dehydrogenation and hydroxylation metabolites of (R,S)-ketamine in the treatment of depression and neuropathic pain. CNS (central nervous system) side effects are related to the activity of (R,S)-ketamine on NMDA receptors. In this application, based on ketamine, (2R,6R; 2S,6S)-hydroxynorketamine has been studied and synthesized (HNK), the compound has no activity on NMDA receptors, thereby avoiding possible side effects. At the same time, the compound is said to have treatment for bipolar depression, major depression, Alzheimer’s dementia, amyotrophic lateral sclerosis, complex The role of regional sexual pain syndrome (CRPS), chronic pain, or neuropathic pain.

We found in animal experiments that (2R, 6R; 2S, 6S)-hydroxynorketamine (HNK) does not last for a long time after administration, and it loses efficacy within 1 week, which severely limits the treatment The long-lasting effect desired in depression. Therefore, how to modify the structure of (2R, 6R; 2S, 6S)-hydroxynorketamine (HNK) to obtain a longer-acting drug will have great therapeutic potential.

Summary of the invention

The application relates to the application of a compound with antidepressant, anesthesia, analgesic, cognitive function improvement, lung protection, prevention or treatment of amyotrophic lateral sclerosis, or prevention or treatment of complex regional pain syndrome in the preparation of medicines.

Compared with the existing known HNK compounds, the compound of the present application has a longer drug effect time, which is specifically manifested in that HNK is metabolized within one week and loses efficacy, while the drug effect time of the compound of the present application can last for more than one week.

This application provides a compound represented by the following formula:

Where m is an integer of 0-3, and n is an integer of 0-4;

R ₁ and R _{2 are} independently selected from H, halogen, hydroxyl, amino, cyano, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, mono- and di-C ₁ -C ₄ alkylamino , ₁ or more of C ₁ -C ₂ haloalkyl, C ₁ -C ₂ haloalkoxy, C ₆ -C ₁₀ aryl, or monocyclic or polycyclic heteroaryl;

R _{2 is} independently selected from

R _{3 is} independently selected from C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₁ -C ₈ acyl;

R _{4 is} independently selected from C ₁ -C ₈ acyl, aryl acyl or heteroaryl acyl;

Or salts, stereoisomers, or tautomers of the above compounds.

This application provides a compound represented by the following formula:

among them

R _{3 is} independently selected from C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₁ -C ₈ acyl, C ₆ -C ₁₀ aryl or monocyclic or polycyclic Heteroaryl

Or salts, stereoisomers, or tautomers of the above compounds.

This application provides the following compounds:

Where m is an integer of 0-3, and n is an integer of 0-4;

Or salts, stereoisomers, or tautomers of the above compounds.

This application provides a compound represented by the following formula:

among them

Or salts, stereoisomers, or tautomers of the above compounds.

The aforementioned compound is characterized by the following compounds:

The aforementioned compound is characterized by the following compounds:

Formula 6 (referred to as CF3 compound in this application).

The preparation method of the compound as described above is characterized in that the preparation route is as follows:

The use of any of the aforementioned compounds in the preparation of drugs for anesthesia, analgesia, cognitive function improvement, lung protection, antidepressant, amyotrophic lateral sclerosis, and complex regional pain syndrome.

The pain includes: chronic pain or neuropathic pain; depression includes: bipolar depression, major depression, depression with neurodegenerative diseases; improvement of cognitive function includes prevention or treatment of Alzheimer’s dementia, Pa Jin Sen et al.

All the diseases mentioned above also cover preventive or therapeutic effects.

The application also provides intermediate compounds represented by the following formula:

Where m is an integer of 0-3, and n is an integer of 0-4;

R ₁ and R _{2 are} independently selected from H, halogen, hydroxyl, amino, cyano, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, mono- and di-C ₁ -C ₄ alkylamino , One or more of C ₁ -C ₂ haloalkyl, C ₁ -C ₂ haloalkoxy, C ₆ -C ₁₀ aryl, or monocyclic or polycyclic heteroaryl;

R ₅ and R ₆ are protecting groups;

Or salts, stereoisomers, or tautomers of the above compounds.

Wherein R5 and R6 are H or a protecting group;

Or salts, stereoisomers, or tautomers of the above compounds.

The stereoisomers of all the above compounds include enantiomers and diastereomers.

When all the above-mentioned compounds exist in different tautomeric forms, the present invention is not limited to any specific tautomer, but includes all tautomeric forms.

All the above-mentioned compounds include compounds having all possible isotopes of the atoms present in the compound. Isotopes include those atoms that have the same atomic number but different mass numbers. By way of general example, without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include ¹¹ C, ¹³ C, and ¹⁴ C.

The application also provides a pharmaceutical composition. The compound disclosed herein can be administered as a pure chemical, but is preferably administered as a pharmaceutical composition. Therefore, the present disclosure provides a pharmaceutical composition including a compound or a pharmaceutically acceptable salt together with at least one pharmaceutically acceptable carrier. The pharmaceutical composition may include a compound or salt as the sole active agent, but preferably includes at least one other active agent. In certain embodiments, the pharmaceutical composition contains about 0.1 mg to about 1000 mg, about 1 mg to about 500 mg, or about 10 mg to about 200 mg of the compound of formula I and optionally about 0.1 mg to about 2000 mg in a unit dosage form , About 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of other active agents.

The compounds disclosed herein can be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, by oral cavity, rectal, as an ophthalmic solution, or by other means in dosage unit formulations containing conventional pharmaceutical carriers Come and give. The pharmaceutical composition can be formulated into any medicinal form, such as aerosol, cream, gel, pill, capsule, tablet, syrup, transdermal patch, or ophthalmic solution. Some dosage forms such as tablets and capsules can be subdivided into appropriate dosage unit dosage forms containing appropriate amounts of active ingredients such as effective amounts for the desired purpose.

Carriers include excipients and diluents, and must have sufficiently high purity and very low toxicity to make them suitable for administration to the patient to be treated. The carrier may be inert or it may have medicinal benefits by itself.

The types of carriers include, but are not limited to: binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavoring agents, glidants, lubricants, preservatives, stabilizers, surfactants, tableting Agent, and wetting agent. Some carriers can be listed in more than one category. For example, vegetable oils can be used as lubricants in some formulations and as diluents in others. Exemplary pharmaceutical carriers include sugar, starch, cellulose, powdered tragacanth, malt, gelatin, talc, and vegetable oils. An optional active agent may be included in the pharmaceutical composition, which does not substantially affect the biological function of the compound of the present invention.

The compound or salt of the application may be the only active agent administered or may be administered in conjunction with other active agents. For example, the compound of the application can be administered together with another active agent selected from any of the following:

Antidepressants : edipram oxalate, felocetine, paroxetine, duloxetine, sertraline, citalopram, bupropion, venlafaxine, duloxetine, naltrexone, rice Zapine, venlafaxine, atomoxetine, bupropion, doxepine, amitriptyline, clomipramine, nortriptyline, buspirone, aripiprazole, chlorazide Ping, kesepin, olanzapine, quetiapine, risperidone, ziprasidone, carbamazepine, gabapentin, lamotrigine, phenytoin, pregabalin, donepezil, galantamine, memantine, Rivastigmine, tramiprosate, or their pharmaceutically active salts or prodrugs, or a combination of the above;

Schizophrenia drugs : aripiprazole, lurasidone, asenapine, clozapine, ziprasidone, risperidone, quetiapine, triflurazine, olanzapine, crexepin, Flupentixol (flupentioxol), perphenazine, haloperidin, chlorpromazine, fluphenazine, fluphenazine, paliperidone;

Alzheimer's dementia drugs: donepezil, rivastigmine, galantamine, memantine;

ALS drug : riluzole;

Pain drugs : acetaminophen, aspirin, NSAIDS, including: diclofenac, difluorobenzene salicylic acid, etodolac, fenoprofen, flurbiprofen, ibuprofen, indoxine, ketoprofen , Ketorolac, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, phenylbutazone, piroxicam, sulindac, Tolmetinopiods, Cox -2 inhibitors such as celecoxib, and anesthetic pain drugs such as: buprenorphine, butorphanol, codeine, dihydrocodeinone, hydromorphone, hydroxymethyllevormorphan, pethidine , Methadone, morphine, nalbuphine, oxycodone, oxymorphone, analgesic new, propoxyphene, central analgesic tramadol.

The foregoing list of other active agents is exemplary rather than comprehensive. Other active agents not included in the above list can be administered in combination with the compound of formula I. Although in some embodiments, the other active agent will be administered at a dose less than generally prescribed and in some cases less than the minimum approved dose, the other active agent may be administered according to its approved prescribed information.

The present disclosure includes methods for treating depression, especially bipolar depression and major depression, especially methods for treating treatment-resistant depression, wherein the effective amount of the compound is the lowest that is effective in alleviating the symptoms of depression Dose, where the relief of depressive symptoms is achieved by achieving a 50% or more reduction in the Depressive Symptom Rating Scale score, or a score less than or equal to 7 on HRSD ₁₇ , or less than or equal to 5 on QID-SR ₁₆ , or Less than or equal to 10 on MADRS.

The present disclosure provides an amount effective to reduce pain (or analgesia) symptoms; wherein, the reduction of pain symptoms is to achieve a 50% or more reduction in pain symptoms on a pain rating scale.

Terminology convention:

"Stereoisomers" are compounds that have the same chemical composition but differ in the arrangement of atoms or groups in space.

"Diastereoisomers" are stereoisomers that have two or more centers of chirality and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral characteristics and reactivity. In the presence of a resolving agent or chromatography, a chiral HPLC column can be used to separate a mixture of diastereomers under high-resolution analysis steps such as electrophoresis and crystallization.

"Enantiomers" refer to two stereoisomers of a compound that have no overlapping mirror images of each other. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which can appear in a chemical reaction or process when there is no stereoselectivity or stereospecificity.

"Alkyl" includes both branched and straight chain saturated aliphatic hydrocarbon groups, and has a specified number of carbon atoms, generally 1 to about 12 carbon atoms. The term C ₁ -C ₆ alkyl as used herein means an alkyl group having 1 to about 6 carbon atoms. When a C ₀ -C _n alkyl group is used in conjunction with another group in this text, take (phenyl) C ₀ -C ₄ alkyl as an example, the designated group, in this case, the phenyl group is passed through a single co- The valence bond (C ₀ ) is directly bonded or connected through an alkyl chain having the specified number of carbon atoms (in this case, 1 to about 4 carbon atoms). Examples of alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl, and sec-pentyl.

"Alkenyl" refers to straight and branched hydrocarbon chains that include one or more unsaturated carbon-carbon bonds, which can occur at any stable point along the chain. The alkenyl groups described herein generally have 2 to about 12 carbon atoms. Preferably, the alkenyl groups are lower alkenyl groups, those alkenyl groups having 2 to about 8 carbon atoms, such as: C ₂ -C ₈ , C ₂ -C ₆ , and C ₂ -C ₄ alkenyl groups. Examples of alkenyl include vinyl, propenyl, and butenyl.

"Alkoxy" refers to an alkyl group as defined above having the specified number of carbon atoms connected by an oxygen bridge. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, 3-hexyloxy, and 3-methylpentyloxy.

The term "heterocyclic ring" means a 5- to 8-membered saturated ring, a partially unsaturated ring, or an aromatic ring containing 1 to about 4 heteroatoms selected from N, O, and S and the remaining ring atoms are carbon, or It is a 7 to 11 membered saturated ring, partially unsaturated ring, or aromatic heterocyclic ring system and a 10 to 15-membered tricyclic ring system, which contains at least 1 heteroatom selected from the group consisting of N, O and S polycyclic ring systems And each ring in the polycyclic system contains up to about 4 heteroatoms independently selected from N, O, and S. Unless otherwise specified, the heterocyclic ring can be attached to a group where it is substituted at any heteroatom and carbon atom and results in a stable structure. When indicated, the heterocyclic ring described herein may be substituted on a carbon or nitrogen atom as long as the resulting compound is stable. The nitrogen atom in the heterocyclic ring can optionally be quaternized. Preferably, the total number of heteroatoms in the heterocyclic group is not more than 4 and preferably the total number of S and O atoms in the heterocyclic group is not more than 2, more preferably not more than 1. Examples of heterocyclic groups include: pyridyl, indolyl, pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, furyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl Azolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, benzo[b]phenylthio (benz[b]thiophenyl), isoquinolinyl, quinazolinyl, quinoxalinyl, Thienyl, isoindolyl, dihydroisoindolyl, 5,6,7,8-tetrahydroisoquinoline, pyridyl, pyrimidinyl, furyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidine Group, morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl.

"Aryl or heteroaryl" means a stable 5- or 6-membered monocyclic or polycyclic ring containing 1 to 4, or preferably 1 to 3 heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. ring. When the total number of S and O atoms in the heteroaryl group exceeds 1, these heteroatoms are not adjacent to each other. Preferably, the total number of S and O atoms in the heteroaryl group is not more than 2. It is especially preferred that the total number of S and O atoms in the heteroaryl group is not more than one. The nitrogen atom in the heterocyclic ring can optionally be quaternized. When indicated, these heteroaryl groups may also be substituted with carbon or non-carbon atoms or groups. Such substitution may include fusion with a 5- to 7-membered saturated cyclic group optionally containing 1 or 2 heteroatoms independently selected from N, O, and S, thereby forming, for example, [1,3]dioxin Azolo[4,5-c]pyridyl. Examples of heteroaryl groups include, but are not limited to: pyridyl, indolyl, pyrimidinyl, pyridazinyl, pyrazinyl, imidazolyl, oxazolyl, furyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl Azolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, benzo[b]phenylthio, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl, isoindolyl , And 5,6,7,8-tetrahydroisoquinoline.

"Depression" includes low mood, decreased interest in activities, slowed or irritable mental activities, changes in appetite, inattention or indecision, excessive guilt or low self-esteem, and in depression, bipolar depression, and other diseases or conditions caused by Suicidal ideation may occur in the case of mood disorders, substance-induced mood disorders and other unexplained mood disorders, and can also be associated with various other mental illnesses (including but not limited to mental disorders, cognitive disorders, eating disorders, anxiety Syndrome and personality disorder) coexist. The progression of the disease (longitudinal course), medical history, type of symptoms, and etiology help to distinguish the various forms of affective diseases from each other.

"Salts of compounds" are derivatives of the disclosed compounds, wherein the parent compound is modified by preparing a non-toxic acid or its base addition salt, and also refers to these compounds and pharmaceutically acceptable solvates of these salts, including hydrates . Examples of pharmaceutically acceptable salts include, but are not limited to: inorganic or organic acid addition salts of basic residues such as amines; alkali or organic addition salts of acidic residues such as carboxylic acids; etc., and include one or more A combination of the above salts. Pharmaceutical salts include non-toxic salts and quaternary ammonium salts such as parent compounds formed from non-toxic inorganic or organic acids. For example, non-toxic acid salts include those derived from inorganic acids, such as: hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, etc.; other acceptable inorganic salts include metal salts such as sodium salt, potassium salt, Cesium salt, etc.; alkaline earth metal salt such as calcium salt, magnesium salt, etc., and a combination including one or more of the above-mentioned salts.

The organic salt of the compound includes compounds such as acetic acid, trifluoroacetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, hexanoic acid, maleic acid, hydroxymaleic acid , Phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, p-toluenesulfonic acid, methane Salts prepared from organic acids such as sulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH ₂ ) _n -COOH (where n is 0 to 4); organic amine salts, such as: triethylamine Salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.; and amino acid salt, such as arginine salt, aspartame Amino acid salts, glutamate salts, etc., and combinations including one or more of the foregoing salts.

Description of the drawings

Figure 1 Depression-like behavior test results in mice;

Figure 2 Differential protein analysis;

Figure 3 Enrichment analysis of the biological processes and molecular functions of differential proteins;

Figure 4 Enrichment analysis of signal transduction pathways of differentially expressed proteins;

Figure 5-Figure 10 NMR spectrum of the compound.

Detailed ways

Example 1: Compound preparation method

Step 1: Starting from o-trifluoromethyl benzonitrile, compound D _{1 is} obtained according to the classic ketamine drug synthesis method Calvin Stevens method (shown in the above road map);

Step 2: Add compound D ₁ (2.57g, 10mmol) to 60ml THF, add triethylamine (2.7mL, 20mmol) and Boc ₂ O (3.32g, 15mmol), reflux for 6h, cool, spin dry, pass through a silica gel column 3.25 g of compound E ₁ was obtained with a yield of 91%. ¹ H NMR (300MHz, CDCl ₃ ): δ 7.80 (d, J = 8.1 Hz, 1H), 7.66 (d, J = 7.8 Hz, 1H), 7.49 (t, J = 7.5 Hz, 1H), 7.22 ( t,J=7.5Hz,1H),5.76(s, 1H), 4.30(s,1H),3.46-3.42(m,1H), 2.45–2.36(m,1H),2.35-2.22(m,1H) ,1.76–1.60(m,6H),1.21(s,9H). ¹³ C NMR(75MHz,CDCl ₃ ): δ208.9,151.7,149.0,148.6,136.1,133.8,132.6,130.5,129.0,128.1,128.0,127.9 ,127.8,121.8,121.7,88.9,80.8,73.7,40.0,29.4,28.3,27.2,23.2;

Step 3: Add compound E ₁ (2.14g, 6mmol) to dry 50ml THF, under the protection of argon, cool to -78℃, add 4ml HMPA, and then slowly add 2M LDA in THF solution ( 8ml, 16mmol), stirred for 30-40min, then slowly heated to -30°C and stirred for 1h, then cooled to -78°C and added trimethylchlorosilane TMSCl (1.73g, 16mmol), slowly heated to -50°C, stirred 3h, pour into saturated ammonium chloride solution and return to room temperature, concentrate the solvent THF and add EA for extraction, dry the organic phase with anhydrous Na ₂ SO ₄ , spin off the solvent and vacuum dry, add 100ml of anhydrous DCM to the obtained oil Dissolve, cool to -15°C, add mCPBA (1.75g, 7.7mmol) under argon protection, stir for 1h and then warm to room temperature, add 50ml DCM and stir for 1h, then pour into saturated sodium thiosulfate and sodium bicarbonate The solution (1:1) was extracted with DCM, spinned to dry the solvent and dried in vacuo, the resulting oil was dissolved in 100ml THF, then cooled to -5°C, tetrabutylammonium fluoride (3g, 11.4mmol) was added and stirred for 30min , Adding saturated NaHCO ₃ solution, extracting with EA, spin-drying the solvent and vacuum drying, passing through a silica gel column to obtain 1.34 g of compound F _{1 with a} yield of 60%. ¹ H NMR (300MHz, CDCl ₃ ): δ7.95 (d, J = 7.8 Hz, 1H), 7.68 (d, J = 7.5 Hz, 1H), 7.58 (t, J = 7.5 Hz, 1H), 7.40 ( t,J=7.5Hz,1H),6.44(s,1H),4.12(dd,J=11.7,6.8Hz,1H), 3.87(d,J=14.4Hz,1H), 3.38(m,1H), 2.36(m,1H),1.74(m,2H),1.72–1.63(m,6H),1.30(s,9H). ¹³ C NMR(75MHz,CDCl ₃ ): 208.2, 153.2, 131.8, 131.5, 128.7, 128.3, 79.4, 72.8, 66.3, 60.4, 40.2, 28.2, 27.9, 19.6;

Step 4: Dissolve compound F ₁ (650 mg, 1.74 mmol) in 10 mL of dry THF, add gaseous HCl to saturation at room temperature and stir for 4 h, add 20 mL of dry ether, crystals precipitate out, filter to obtain compound T ₁ cis- The hydrochloride salt of 6-hydroxydesmethyltrifluoromethanone T ₁ was 490 mg, and the yield was 90%. ¹ H NMR (400MHz, CD ₃ OD): δ8.12 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.97 (t, J = 8.0 Hz, 1H), 7.85 (t,J=8.0Hz,1H), 4.28(dd,J ₁ =11.6Hz,J ₂ ＝6.8Hz,1H), 3.46(dd,J ₁ ＝14.4Hz,J ₂ ＝2.8Hz,1H),2.07 (m,1H),2.01–1.72(m,4H). ¹³ C NMR (100MHz, CD ₃ OD): 205,133.7,131.1,130.9,128.9,128.8,127.6,73.1,67.0,60.7,44.2,38.6,38.3, 29.4, 28.9, 18.6;

The compound T ₁ prepared above is the compound CF3 in the activity test.

Example 2: Activity test method:

1. Forced swimming experiment

The mice were transferred to the laboratory 1 hour before the forced swim test (FST). The test is carried out under normal lighting conditions and monitored by a digital video camera. During the test, the mice were put into transparent glass cylinders (28.5 cm high and 14 cm in diameter) containing 20 cm of water (23 ± 1°C). On the first day, the mice were trained for 6 minutes and then removed from the cylinder. On the second day, the mice were given a solvent (normal saline) control group, HNK, CF3, and then tested their immobile time after 1 hour and 7 days, where the immobile time refers to passive floating without other actions. In the last 4 minutes of the entire 6-minute swimming test, the immobile time was recorded through the EthoVision XT (Noldus, Netherlands) of the Nodus system (Castagné et al., 2011, Hajmirzaian et al., 2014, Porsolt et al., 1977). After every two to three tests, the water in the cylinder should be replaced. After the swimming test, the mice were taken out of the water and dried under infrared light.

2. Protein extraction and digestion

The hippocampal tissue samples from all mouse groups were separated, frozen in liquid nitrogen and stored at -80°C for use. The sample was suspended in 8M urea (containing PBS (pH 8.0), 1x protease and phosphatase inhibitor mixture), and then sonicated with Sonics VCX-150 (Newtown, CT, USA). Subsequently, 14,000 g of the homogenate was centrifuged at 4°C for 30 minutes to remove cell debris. The supernatant was then collected in a new 1.5ml tube. The protein concentration was determined by Nanodrop 2000 (Thermo Scientific, USA). According to the results of protein quantification, the concentration of all samples was adjusted to 1 μg/μl. The 100 μg protein of each sample from the same group was pooled, and a system with a total of 100 μg protein from each group was obtained. The four combined samples were treated with 10mM DTT at 55°C for 60 minutes, and then treated with 25mM IAA in the dark at room temperature for 60 minutes. Each combined sample was digested with 4 μg sequencing grade modified trypsin at 37°C for 1 hour, and then diluted with PBS (pH 8.0) to reach the final 1.0M urea concentration. Then, the sample continued to be digested overnight at 37°C. After digestion, the peptide was treated with 100% FA, and then desalted with peptide desalting spin columns (Waters, MC, USA). The peptide was dried with a vacuum concentrator, and finally dissolved in 200mM TEAB, and labeled with TMT working solution.

3.Tandem mass tag (TMT) labeling (tandem tag (TMT))

Re-dissolve each vial of TMT with 40 μL 99.9% acetonitrile (ACN) to obtain a TMT working solution. The peptides were then labeled with TMT working solution for 1 hour at room temperature. Different groups were labeled with different TMT: the control group was labeled with TMT-127, the HNK group was labeled with TMT-128, and the CF3 group was labeled with TMT-131. After labeling, all peptides from the three groups were mixed, desalted, and dried as previously described.

4. Use high pH reverse phase separation for peptide fractionation

According to the high pH reverse phase protocol, the TMT-labeled peptides were separated according to the established components. In short, due to different peptide retention behaviors, different sets of elution solutions were used for TMT-labeled samples. The TMT-labeled peptide was separated in 300 μL of 0.1% formic acid, and then loaded on a spin column for reversed-phase fractionation. The loaded peptide was eluted in 8 fractions with an ACN gradient buffer solution at pH 8. The separated components were dried on a high-speed vacuum concentrator and stored at -80°C, waiting for LC-MS analysis.

5.NanoLC-MS/MS and database search

The labeled separated fractions were reconstituted in 20 μL of 0.1% FA. Then, when equipped with C18 resin (

Peptides were separated on an UltiMate 300 RSLC nano System (Thermo Scientific, USA) of 5 μm; Varian, Lexington, MA) and a silica capillary column (75 μmID, 150 mm length, Upchurch, Oak Harbor, WA). A gradient with 0.1% FA and 5% ACN was run for 120 minutes at a fixed flow rate of 0.3 μL/min for relative quantification and target analysis. The ionized peptides were analyzed on a quadrupole-orbitrap mass spectrometer (Q-Exactive, Thermo Scientific, USA). Proteome Discover 2.1 software (Thermo Scientific, USA) uses the peak analysis and data processing of the Mus musculus database, in which the MS/MS spectrum of each nanoLC-MS/MS run is analyzed. For protein identification, the parameters are set as follows: full trypsin specificity, allowing no more than two missed cuts, carbamoyl methylation (C) and MT plex (K and peptide N-terminus) as static modifications, and oxidation ( M) As a dynamic modification. For all MS data obtained using the Orbitrap mass analyzer, the precursor ion mass tolerance is set to 20 ppm, and for all MS/MS spectra obtained, the fragment ion mass tolerance is set to 20 mmu. Quantitative accuracy is reflected in the variability of protein ratios. The fold change was calculated by the ratio of proteins labeled with TMT-128/TMT-127, TMT-131/TMT-128, and TMT-131/TMT-127. The up-regulation and down-regulation thresholds were set to 1.2 (or 1.5) and 0.83 (or 0.67), respectively.

6. Bioinformatics analysis

Analyze proteomics results through various methods and methods. DAVID version 6.7 (https://david.ncifcrf.gov/) is used to classify the functional categories of differentially expressed proteins and gene ontology (gene ontology, GO) annotation enrichment analysis. Venny 2.1 (http://bioinfogp.cnb.csic.es/tools/venny/index.html) was used to analyze the hippocampal proteome of two model mice. Use STRING version 10.5 (https://string-db.org/) for protein-protein interaction network analysis. The network and wiki paths generated by STRING are visualized and edited in Cytoscape 3.6.1.

7. Statistical analysis

All results are expressed as mean±SEM. The number of N is given in the legend. Use Prism (version 5.0, Graphpad software) and SPSS 19.0 to complete the statistical analysis of the data. All experiments used Bonferroni's hoc test for parameter testing (t test, full two-tailed) or one-way analysis of variance (ANOVA). A value of p<0.05 is considered to be statistically significant.

Example 3: Experimental results

1. Test results of depression-like behavior in mice:

As shown in Figure 1:

A forced swimming test is used to measure depression-like behavior. Mice received 10 mg of HNK, CF3 by intragastric administration, and the immobility time was measured after 1 hour and 7 days. The percentage of immobility time is expressed as: mean ±SEM*p<0.05, **p<0.0 compared with pre-activation. Each group N=8 saline: saline mice; HNK: 2R, 6R-hydroxynorchloramine treated mice; CF3, CF3 treated mice.

2. Differential protein analysis:

as shown in picture 2:

A. Taking 0.833>Abundance Ratio>1.2 as the critical point, a Ratio value higher than 1.2 is defined as an up-regulated protein, and a Ratio value lower than 0.833 is defined as a down-regulated protein. Compared with the control group, there were 101 differentially expressed proteins in the CF3 treatment group, of which 39 differentially expressed proteins were co-expressed with the HNK treatment group. B, Compared with the HNK treatment group, the CF3 treatment group has 243 differentially expressed proteins, of which 60 differentially expressed proteins are co-expressed with the CF3 treatment group. C, the expression abundance of 39 co-expressed differentially expressed proteins among the groups. D, the expression abundance of 60 co-expressed differentially expressed proteins among the groups.

3. Enrichment analysis of differential protein biological processes and molecular functions

As shown in Figure 3:

A, Compared with the control group, the enrichment analysis of the differentially expressed protein in the CF3 treatment group in the biological process. B, Compared with the control group, the molecular function enrichment analysis of differentially expressed proteins in the CF3 treatment group. C, Compared with the HNK treatment group, the enrichment analysis of the differentially expressed protein in the CF3 treatment group in the biological process. D, Enrichment analysis of the molecular functions of differentially expressed proteins in the CF3 treatment group.

4. Pathway enrichment analysis of differentially expressed proteins

As shown in Figure 4:

A, Compared with the control group, the pathway enrichment analysis of differentially expressed proteins caused by CF3 treatment. B, Pathway analysis of differentially expressed protein caused by CF3 treatment compared with HNK treatment group.

The above results can be seen:

1. Compared with HNK compound, the short-term effect of CF3 compound is equivalent to that of HNK, but HNK does not have long-term effect, it becomes invalid within 1 week and no longer has therapeutic effect. The CF3 compound of the present application still has an antidepressant effect after 7 days, that is, it has a long-term effect.

2. The results of venny analysis indicated that compared with the control group, there were 101 differentially expressed proteins in the CF3 treatment group, of which 39 differentially expressed proteins were co-expressed with the HNK treatment group; compared with the HNK treatment group, the CF3 treatment group had a total of 243 Among them, 60 differentially expressed proteins were co-expressed with the CF3 treatment group.

3. GO analysis results show that, compared with the control group, the differential proteins produced by CF3 treatment are mainly enriched in lens development in camera-type eye, camera-type eye development, visual perception and other biological processes, as well as structural constituent of eye lens , Protein binding, poly(A)RNA binding and other molecular functions; compared with the HNK treatment group, the differential proteins produced by CF3 treatment are mainly enriched in biological processes such as transport, protein transport, lens development in camera-type eye, and Structural Consituent of Eye Lens, Protein Domain Specific Binding, Protein Binding and other molecular functions.

4. Pathway analysis results indicate that, compared with the control group, the differentially expressed protein produced by CF3 treatment is mainly enriched in spliceosome and mRNA splicing; compared with the HNK treatment group, the differentially expressed protein produced by CF3 treatment is mainly enriched in oxidative stress ,wnt signaling pathway, and long-term potentiation.

In short, CF3 has a better long-term antidepressant effect than HNK. CF3 treatment can cause the differential expression of a variety of proteins in the hippocampus of mice. The main pathway for its effect lies in spliceosome and mRNA splicing, compared to HNK. In the treatment group, the differentially expressed proteins produced by CF3 mainly focused on oxidative stress, long-term potentiation, and mitochondrial related effects. These results comprehensively indicate that CF3 may affect oxidative stress, long-term potentiation, and the mitochondrial electron transport chain to play an antidepressant effect in the body through the role of spliceosome.

Claims

The application of a long-acting compound in the preparation of antidepressant, anesthesia, analgesic, cognitive function improvement, lung protection, amyotrophic lateral sclerosis or complex regional pain syndrome drugs, characterized in that: the compound structural formula is as follows Shown:

Where m is an integer of 0-3, and n is an integer of 0-4;

R 1 and R 2 are independently selected from H, halogen, hydroxyl, amino, cyano, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, mono- and di-C 1 -C 4 alkylamino , 1 or more of C 1 -C 2 haloalkyl, C 1 -C 2 haloalkoxy, C 6 -C 10 aryl, or monocyclic or polycyclic heteroaryl;

R 3 is independently selected from C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 acyl;

R 4 is independently selected from C 1 -C 8 acyl, aryl acyl or heteroaryl acyl;

Or salts, stereoisomers, or tautomers of the above compounds.
The application of a long-acting compound in the preparation of antidepressant, anesthesia, analgesic, cognitive function improvement, lung protection, amyotrophic lateral sclerosis or complex regional pain syndrome drugs, characterized in that: the compound structural formula is as follows Shown:

among them

R 3 is independently selected from C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 acyl, C 6 -C 10 aryl or monocyclic or polycyclic Heteroaryl

R 4 is independently selected from C 1 -C 8 acyl, aryl acyl or heteroaryl acyl;

Or salts, stereoisomers, or tautomers of the above compounds.
The application of a long-acting compound in the preparation of antidepressant, anesthesia, analgesic, cognitive function improvement, lung protection, amyotrophic lateral sclerosis or complex regional pain syndrome drugs, characterized in that: the compound structural formula is as follows Shown:

Where m is an integer of 0-3, and n is an integer of 0-4;

R 1 and R 2 are independently selected from H, halogen, hydroxyl, amino, cyano, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, mono- and di-C 1 -C 4 alkylamino , 1 or more of C 1 -C 2 haloalkyl, C 1 -C 2 haloalkoxy, C 6 -C 10 aryl, or monocyclic or polycyclic heteroaryl;

R 3 is independently selected from C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 acyl;

R 4 is independently selected from C 1 -C 8 acyl, aryl acyl or heteroaryl acyl;

Or salts, stereoisomers, or tautomers of the above compounds.
The application of a long-acting compound in the preparation of antidepressant, anesthesia, analgesic, cognitive function improvement, lung protection, amyotrophic lateral sclerosis or complex regional pain syndrome drugs, characterized in that: the compound structural formula is as follows Shown:

among them

R 3 is independently selected from C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 acyl, C 6 -C 10 aryl or monocyclic or polycyclic Heteroaryl

R 4 is independently selected from C 1 -C 8 acyl, aryl acyl or heteroaryl acyl;

Or salts, stereoisomers, or tautomers of the above compounds.
The application according to any one of claims 1-4, characterized in that: the structural formula of the compound is as follows:
The application according to any one of claims 1-4, characterized in that: the structural formula of the compound is as follows:
The application of a long-acting compound in the preparation of antidepressant, anesthesia, analgesic, cognitive function improvement, lung protection, amyotrophic lateral sclerosis or complex regional pain syndrome drugs, characterized in that: the compound structural formula is as follows Shown:

Where m is an integer of 0-3, and n is an integer of 0-4;

R 1 and R 2 are independently selected from H, halogen, hydroxyl, amino, cyano, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, mono- and di-C 1 -C 4 alkylamino , 1 or more of C 1 -C 2 haloalkyl, C 1 -C 2 haloalkoxy, C 6 -C 10 aryl, or monocyclic or polycyclic heteroaryl;

R5 and R6 are protecting groups;

Or salts, stereoisomers, or tautomers of the above compounds.
The application according to claim 7, wherein the structural formula of the compound is as follows:

Wherein R 5 and R 6 are H or a protecting group;

Or salts, stereoisomers, or tautomers of the above compounds.
The application according to claim 8, wherein the structural formula of the compound is as follows:
The application according to claim 9, characterized in that: the structural formula of the compound is as follows:
The application according to any one of claims 1-10, characterized in that the pain comprises: chronic pain or neuropathic pain.
The application according to any one of claims 1-10, wherein the depression includes: bipolar depression, major depression and/or depression accompanied by neurodegenerative diseases.
The application according to any one of claims 1-10, wherein the improvement of cognitive function includes: improvement of cognitive function includes prevention or treatment of Alzheimer's dementia and/or Parkinson's.
The use according to any one of claims 1-10, characterized in that the medicine is formulated as a pill, capsule, tablet or syrup.