Opioid receptor agonists and methods of making and using the same

Abstract

The application provides a kind of oxygen heterospirocyclic small molecule compound, and the pharmaceutical composition containing the compound and its purposes as therapeutic agent, especially as MOR receptor agonist and in the preparation of treating and / or preventing pain and other related diseases.The MOR receptor agonist provided by the application has novel structure, shows high activity, and has higher selectivity to MOR, and its maximum efficiency E max Also has obvious improvement.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a class of oxaspirocyclic small molecule compounds, their preparation methods, pharmaceutical compositions containing the compounds, and their use as therapeutic agents, particularly as MOR receptor agonists, and in the preparation of remedies for treating and preventing pain and related diseases. Technical Background

[0002] Opioid receptors are G protein-coupled receptors (GPCRs) and are targets for the binding of endogenous opioid peptides and opioid drugs. Multiple opioid receptors exist in the human body, mainly including muopioid receptors (MOR), delta opioid receptors (DOR), and kappaopioid receptors (KOR), which are widely distributed in peripheral tissues such as the central nervous system, heart, digestive tract, blood vessels, and kidneys (Nature, 2016, 537(7619):185). MOR has the strongest binding affinity for morphine peptides and is the primary receptor protein site for the action of analgesics such as morphine and fentanyl. Zadina et al. found that the binding affinity of the MOR receptor to morphine 1 (360 pM) was 4,000 times and 15,000 times greater than that of the DOR receptor and KOR receptor to morphine 1, respectively (Science 2001 Vol.293 No:311-315; Biochem Biophys Res Commun 235:567-570; Life Sci 61:409-415).

[0003] Studies have found that GPCRs mediate and regulate physiological functions mainly through the activation of G protein pathways and β-arrestin pathways. G protein signaling pathways mainly include second messenger systems such as calcium ions, adenylate cyclase, and mitogen-activated protein kinases. The β-arrestin pathway has three main aspects: (1) acting as a negative regulator and interacting with GPCR kinases to induce receptor desensitization of GPCRs, thereby terminating G protein signal transduction; (2) acting as a scaffold protein to recruit endocytic proteins and induce GPCR endocytosis; (3) acting as an adaptor protein to form a complex with downstream signaling molecules of GPCRs, thereby activating signal transduction molecules in a G protein-independent manner. Early studies showed that endogenous enkephalins and the opioid drug etorphine can stimulate G proteins and induce receptor endocytosis, while morphine does not induce receptor endocytosis. This is because morphine exerts its physiological functions through the G protein signaling pathway rather than the β-arrestin pathway (Zhang et al., Proc Natl Acad Sci USA, 1998, 95(12):7157-7162). Studies have found that morphine injection into β-arrestin2 gene knockout mice resulted in stronger and longer-lasting analgesic effects mediated by G protein signaling (Bohn et al., Science, 1999). This suggests that the difference in ligand-stimulated G protein and / or β-arrestin signaling determines the ligand-specific cellular biological effects of GPCRs. If such ligands exhibit a stronger negative β-arrestin preference, they may even escape β-arrestin-mediated receptor desensitization, leading to prolonged G protein signaling transmission and a stronger analgesic effect. Recent studies have also found that the β-arrestin pathway is associated with several side effects of MOR agonists, such as constipation, respiratory depression, and analgesic tolerance (Science 1999 Vol. 286:2495-2498: J. Pharmacol. Exp. Ther. 2005, 314:1195-1201). Therefore, developing a "biased" MOR agonist drug that can selectively activate the G protein signaling pathway, namely a drug designed with a negative β-arrestin-biased ligand for MOR, would reduce β-arrestin-mediated side effects and have significant clinical value and social significance in the field of analgesia.

[0004] In August 2020, the FDA approved Trevena Inc.'s drug Olinvyk (WO2012129495A1) for marketing. Currently, patents reported on the research and development of G protein-biased MOR agonists include WO2017063509A1, WO2019205983A1, CN109206417A, WO2019072235A1, CN111662284A, WO2019052557A1, CN111836807A, and CN112789276A. Although these patents have disclosed a series of G protein-biased MOR agonists, their molecular structures are quite different from the structure provided in this invention, and their efficacy and safety have not yet been confirmed. Clinically, it is still necessary to develop new molecular structures to obtain MOR agonists with better efficacy, selectivity, drug safety, and drug metabolism results. Summary of the Invention

[0005] In response to the needs of the prior art, the present invention provides a novel compound that can be used as a MOR receptor agonist. This type of compound exhibits high activity, significantly improved Emax, and high selectivity for MOR.

[0006] This invention provides a compound of formula (I), its solvate, stereoisomer, deuterated compound, or a pharmaceutically acceptable salt thereof.

[0007]

[0008] in,

[0009] Ring A is selected from substituted or unsubstituted 4- to 6-membered cycloalkyl or heterocycloalkyl groups;

[0010] R1 and R2 together with the attached carbon atom form a substituted or unsubstituted 5- or 6-membered monocyclic heteroaromatic ring, which forms a fused bicyclic ring with the attached A ring.

[0011] The compound is not selected from:

[0012]

[0013] In some preferred embodiments provided by the present invention, R1 and R2 together with the connected carbon atom form a substituted or unsubstituted 5-membered monocyclic heteroaromatic ring containing an S heteroatom.

[0014] In some embodiments provided by the present invention, R1 and R2 together with the attached carbon atom form a substituted or unsubstituted 5- or 6-membered monocyclic heteroaromatic ring selected from the following structures:

[0015]

[0016] in The two ring atoms connected represent adjacent atom pairs shared when fused with ring A.

[0017] In some preferred embodiments provided by the present invention, R1 and R2 together with the attached carbon atom form a substituted or unsubstituted 5-membered monocyclic heteroaromatic ring containing an S heteroatom selected from:

[0018] In some preferred embodiments provided by the present invention, the ring A is selected from 5-membered cycloalkyl groups.

[0019] In some embodiments provided by this invention, the compound is selected from:

[0020]

[0021] Another aspect of the present invention relates to a pharmaceutical composition comprising compounds represented by the above general formulas, their solvates, stereoisomers, deuterated compounds or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

[0022] Another aspect of the invention provides the use of compounds represented by the above general formulas, their solvates, stereoisomers, deuterated compounds or pharmaceutically acceptable salts thereof, or the above pharmaceutical compositions in the preparation and / or in the treatment of medicaments for treating MOR receptor agonist-mediated diseases.

[0023] Among them, the diseases mediated by MOR receptor agonists are selected from pain, immune dysfunction, inflammation, esophageal reflux, neurological and psychiatric diseases, urinary and reproductive diseases, cardiovascular diseases and respiratory diseases; the pain mentioned above is selected from postoperative pain, cancer pain, neuropathic pain, traumatic pain and pain caused by inflammation.

[0024] This invention mainly modifies the structure of the substituent Z in the general formula (II) disclosed in the prior art, and designs and synthesizes a class of compounds that can be used as MOR receptor agonists. Surprisingly, the compounds provided by this invention have better in vivo or in vitro activity than compounds with similar structures in the prior art, exhibiting high activity.

[0025]

[0026] The present invention uses compounds with similar structures to those in the prior art as control groups as follows: TRV130 (racemic), control group 1 and control group 2 have the following structural formulas.

[0027]

[0028] Terminology Explanation

[0029] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon group, "C 3-8 "Cycloalkyl" refers to a cyclic hydrocarbon group containing 3 to 8 carbon atoms, preferably C12. 4-6 Cycloalkyl groups. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptanetrienyl, cyclooctyl, etc., with cyclopropyl, cyclopentyl, and cyclohexenyl being preferred.

[0030] The term "5- to 6-membered monocyclic heteroaryl ring" refers to a monocyclic heteroaryl ring containing 5 to 6 ring atoms, including (but not limited to): thiophene ring, N-alkyl ring, pyrrole ring, furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, 1,2,3-triazole ring, 1,2,4-triazole ring, 1,2,5-triazole ring, 1,3,4-triazole ring, tetraazole ring, isoxazole ring, oxadiazole ring, 1,2,3-oxadiazole ring, 1,2,4-oxadiazole ring, 1,2,5-oxadiazole ring, 1,3,4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring, etc.

[0031] The term "heterocyclic alkyl" (or "heterocyclic aliphatic") refers to a monovalent, non-aromatic ring system whose ring atoms consist of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus, and are connected to a parent nucleus or other groups by a single bond; common heterocyclic alkyl groups include (but are not limited to) ethylene oxide, oxetane-3-yl, azirone-3-yl, tetrahydrofuran-2-yl, pyrrolidine-1-yl, pyrrolidine-2-yl, tetrahydro-2-hydropyran-2-yl, tetrahydro-2-hydropyran-4-yl, piperidin-2-yl, piperidin-4-yl, etc.

[0032] The substituents in the present invention, namely "substituted or unsubstituted 4- or 6-membered cycloalkyl or heterocycloalkyl" and "substituted or unsubstituted 5- or 6-membered monocyclic heteroaromatic ring", may include, for example, groups selected from the group consisting of: halogen, cyano, hydroxyl, carboxyl, nitro, C 1-6 Alkyl, C 1-6 Alkoxy, alkynyl, alkenyl, amino, and the substituents may be further substituted, such as by halogen, hydroxyl, alkoxy, alkylamino, 3-6 membered cycloalkyl and heterocycloalkyl, 5-6 membered aryl and heteroaryl, etc.

[0033] The term "stereoisomer" in the present invention refers to the enantiomer produced when the compound contains asymmetric carbon atoms; the cis-trans isomer produced when the compound contains carbon-carbon double bonds or a cyclic structure; and the tautomer produced when the compound contains ketones or oximes. All enantiomers, diastereomers, racemic isomers, cis-trans isomers, tautomers, geometric isomers, epimers, and mixtures thereof are included within the scope of this invention. Specific Implementation

[0034] The following embodiments illustrate the technical solution of the present invention in detail, but the scope of protection of the present invention includes, but is not limited to, these embodiments.

[0035] Synthesis of intermediates:

[0036] Synthesis of intermediate A

[0037] Synthetic route

[0038]

[0039] Step 1: Synthesis of intermediate A-3

[0040] At room temperature, 375 mL of 75% H₂SO₄ was slowly added to a 2 L single-necked flask containing intermediate A-1 (150 g, 1.79 mol) and intermediate A-2 (139 g, 1.88 mol) at -10 °C. After the addition was complete, the reaction mixture was reacted at 5 °C for 16 h. The reaction solution was then slowly poured into 2 L of ice water, and the pH was adjusted to 9 by slowly adding (1 N) NaOH solution. The mixture was extracted with ethyl acetate (2 L × 5), dried over Na₂SO₄, filtered, and concentrated to obtain an orange oily liquid intermediate A-3 (215 g, 77% yield). (TLCV) PE V EA =1:1, R f =0.2, PMA color development).

[0041] Step 2: Synthesis of intermediate A-4

[0042] At room temperature, Dess-Matin reagent (585 g, 1.38 mol) was slowly added to dichloromethane (1 L). Under ice bath conditions, a dichloromethane solution (500 mL) of intermediate A-3 (205 g, 1.31 mol dissolved in 500 mL LDCM) was slowly added dropwise. After the addition was complete, the mixture was slowly heated to room temperature and stirred overnight. After the reaction was complete, 2 L of n-hexane was added, and the mixture was stirred for 10 min. The mixture was filtered, and the filtrate was concentrated to 500 mL. Another 2 L of n-hexane was added, and the mixture was filtered again. The solution was evaporated to dryness, and 200 mL of tert-methyl ether was added. The mixture was stirred for 10 min, filtered, and the filtrate was washed successively with saturated sodium sulfite solution (200 mL) and saturated sodium chloride solution (200 mL). The solution was dried over anhydrous sodium sulfate, filtered, and the filtrate (which passed the test with starch-potassium iodide paper) was concentrated to obtain an orange oily liquid intermediate A-4 (160 g, 79% yield). (TLCV) PE V EA =1:1, R f =0.6, PMA color development).

[0043] Step 3: Synthesis of intermediate A-6

[0044] At room temperature, intermediates A-4 (168 g, 1.09 mol), A-5 (139 g, 1.15 mol), ammonium acetate (22 g, 0.28 mol), and acetic acid (13 g, 0.22 mol) were dissolved in 1.5 L of toluene. A water separator was added, and the reaction was carried out at 130 °C for 4 h until completion. The solution was concentrated to 500 mL, EA (1 L) was added, and the mixture was washed with water (1 L × 2), then with 0.5 N NaOH (1 L × 2). The organic phase was dried and concentrated to give an orange oily liquid intermediate A-6 (222 g, 87% yield). (TLCV) PE V EA =1:1).

[0045] Step 4: Synthesis of intermediate A-7

[0046] Under nitrogen protection, at 25°C, a solution of 2-bromopyridine (178.89 g, 1.13 mmol, 1.2 eq) in tetrahydrofuran (1500 mL) was slowly added dropwise to a solution of magnesium isopropyl chloride in tetrahydrofuran (2 M, 615 mL). After the addition was complete, the reaction was carried out at 30°C for 3 h, and then cuprous iodide (17.97 g, 94.35 mmol, 0.1 eq) was added, and the reaction was continued for another 0.5 h. The reaction solution was cooled to 0°C, and a THF solution of intermediate A-6 (222.0 g, 943.54 mmol, 1 eq) in 500 mL was slowly added dropwise. The mixture was stirred overnight at room temperature. After the reaction was complete, a saturated ammonium chloride solution (1 L) was added to quench the reaction. The mixture was extracted with ethyl acetate (2000 mL × 3). The combined organic phases were washed with 1 L of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, a black oily intermediate A-7 (310 g, purity 81%, yield 85%), [M+H]. + :315.12.

[0047] Step 5: Synthesis of intermediate A-8

[0048] Intermediate A-7 (310 g, 986.05 mmol, 1 eq) was dissolved in ethylene glycol (2 L), and potassium hydroxide (110.65 g, 1.97 mol, 2 eq) was added. The mixture was heated to 120 °C and reacted for 3 h. After cooling to room temperature, water (3000 mL) was added to the reaction solution, and the aqueous phase was extracted with ethyl acetate (2 L × 3). The combined organic phases were washed with brine (2 L × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product, a black solid intermediate A-8 (230 g, purity 96.6%, yield 90.0%), [M+H]. + :257.15.

[0049] Step 6: Synthesis of Intermediate A

[0050] Intermediate A-8 (20 g, 78.02 mmol) was dissolved in tetrahydrofuran (200 mL). Lithium aluminum hydride (5.92 g, 156.04 mmol) was slowly added under ice-water bath conditions. After the addition was complete, the reaction was carried out at 60 °C for 5 h. The mixture was then cooled to room temperature, and water (5.92 mL), sodium hydroxide solution (5.92 mL, 10%), and water (5.92 mL) were added slowly in sequence under ice-water bath conditions. 5 g of anhydrous sodium sulfate was added, and the mixture was stirred at room temperature for 0.5 h. The mixture was filtered, and the filtrate was evaporated to dryness to obtain a pale yellow oily intermediate A (16 g, yield 8.76%).

[0051] Synthesis of intermediate 6-1

[0052] Synthetic route

[0053]

[0054] Step 1: Synthesis of intermediate 6-3

[0055] At room temperature, intermediate 6-2 (2.5 g, 19.50 mmol), triphenylphosphine (5.03 g, 21.45 mmol), DCM (25 mL), imidazole (1.59 g, 23.40 mmol), and elemental iodine (5.54 g, 21.45 mmol) were added sequentially to a single-necked flask and stirred at room temperature for 1 h. The mixture was then quenched with saturated ammonium chloride solution (20 mL). Extraction was performed with dichloromethane (20 mL × 3). The organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (V). PE V EA =3:1) to obtain colorless oily intermediate 6-3 (2.5 g, yield 58.63%).

[0056] Step 2: Synthesis of intermediate 6-4

[0057] Under ice-water bath conditions, diethyl malonate (3.40 g, 21.23 mmol), THF (40 mL), and sodium hydroxide (0.83 g, 20.80 mmol, 60%) were added to a tetrahydrofuran solution (40 mL). The reaction was carried out at room temperature for 1 h. Intermediate 6-3 (2.5 g, 10.60 mmol) was slowly added to the reaction solution, and the reaction was carried out at room temperature for 12 h. Saturated ammonium chloride solution (100 mL) was slowly added to the reaction solution. Ethyl acetate (20 mL × 3) was added for extraction. The organic phases were combined, washed with saturated brine (35 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (V). PE V EA =3:1) to obtain colorless oily intermediate 6-4 (2.0 g, yield 70.54%).

[0058] Step 3: Synthesis of intermediate 6-5

[0059] Intermediate 6-4 (2.0 g, 7.40 mmol), potassium hydroxide (830 mg, 14.80 mmol), ethanol (40 mL), and water (40 mL) were added to a 250 mL single-necked flask. The mixture was reacted at room temperature for 12 h. The ethanol was removed by concentration. The pH of the solution was adjusted to 4 with 1 N hydrochloric acid solution. The mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined and washed with saturated brine (35 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a colorless oily intermediate 6-5 (1.5 g, yield 92.64%).

[0060] Step 4: Synthesis of intermediate 6-6

[0061] Intermediate 6-5 (1.5 g, 7.0 mmol) and diethyl ether diethanolate (150 mL) were added to a 250 mL single-necked flask. The mixture was reacted at 160 °C for 2 h. The diethyl ether diethanolate was then concentrated to remove the ether. A saturated ammonium chloride solution (15 mL) was slowly added to the reaction mixture. Ethyl acetate (20 mL × 3) was added for extraction. The organic phases were combined, washed with saturated brine (35 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (V). PE V EA =3:1) to obtain colorless oily intermediate 6-6 (0.8 g, yield 67.2%).

[0062] Step 5: Synthesis of intermediate 6-1

[0063] Under nitrogen protection, methanesulfonic acid (30 mL) and phosphorus pentoxide (1.2 g) were added to a 150 mL single-necked flask and stirred at room temperature for 0.5 h. Intermediate 6-6 (0.8 g, 4.70 mmol) was added to the reaction mixture, and the reaction was allowed to proceed at room temperature for 2 h. Water (100 mL) was then slowly added to the reaction mixture. Extraction was performed with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (35 mL), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (V...). PE V EA =3:1) to obtain colorless oily intermediate 6-1 (0.56 g, yield 72.28%).

[0064] Synthesis of intermediate 7-1

[0065] Synthetic route

[0066]

[0067] Step 1: Synthesis of intermediate 7-3

[0068] Intermediate 7-2 (2.05 g, 13.0 mmol) was dissolved in DMF (20 mL), followed by the addition of potassium carbonate (3.59 g, 26.0 mmol) and ethyl bromoacetate (2.16 g, 13.0 mmol). The reaction mixture was stirred at room temperature for 3 h. TLC monitoring showed that the reaction proceeded completely. After the reaction was complete, ethyl acetate (20 mL) and water (20 mL) were added to the reaction mixture. The mixture was extracted with ethyl acetate (20 mL × 3), and the organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was then concentrated, mixed with 100-200 mesh silica gel, and the mixture was subjected to column chromatography (V) PE V EA =2:1) ​​Purification yielded 3.16 g of white solid intermediate 7-3.

[0069] Step 2: Synthesis of intermediate 7-4

[0070] Potassium tert-butoxide (1.12 g, 10.0 mmol) dissolved in THF (7.5 mL) was added to the reaction flask. The reaction mixture was then placed under an inert gas atmosphere in an ice-water bath and stirred for 30 min. Intermediate 7-3 (1.22 g, 5.0 mmol) was then dissolved in THF (3.0 mL) and added dropwise to the reaction mixture. The reaction was carried out at 0 °C for 30 min. TLC monitoring showed that the starting material reacted completely. After the reaction was complete, the reaction mixture was quenched with saturated ammonium chloride solution, followed by acidification. The mixture was then extracted with ethyl acetate (20 × 3 mL), and the organic phase was collected and dried over anhydrous sodium sulfate. The concentrated organic phase was used directly in the next reaction step.

[0071] Step 3: Synthesis of intermediate 7-1

[0072] Intermediate 7-4 (1.06 g, 5.0 mmol) was dissolved in dioxane (7.5 mL) and dilute hydrochloric acid (3 M, 16.5 mL). The reaction mixture was stirred at room temperature for 12 h. TLC monitoring showed that the reaction proceeded completely. After the reaction was complete, water (30 mL) and methyl tert-butyl ether (30 mL) were added to the reaction mixture, followed by extraction with methyl tert-butyl ether (20 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, concentrated, and mixed with 100-200 mesh silica gel. The mixture was then subjected to column chromatography (V) PE V EA Elution was performed at a ratio of 10:1, and the purified product was 155 mg.

[0073] 1H NMR (400MHz, Chloroform-d) δ7.98 (d, J = 28.6Hz, 1H), 6.83 (q, 1H), 5.04 (s, 2H).

[0074] Example 1

[0075] Synthesis of nitrogen-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-5,6-dihydro-4H-cyclopentane[b]thiophene-4-ethylamine (compound 1)

[0076]

[0077] Synthetic route

[0078]

[0079] Under nitrogen protection, intermediate A (100 mg, 0.37 mmol), intermediate 1-1 (80 mg, 0.58 mmol), DCE (10 mL), and tetraisopropyl titanate (1.5 mL) were sequentially added to a single-necked flask and stirred at 60 °C for 16 h. Sodium borohydride (44 mg, 1.2 mmol) was added, and the mixture was stirred at 60 °C for 2 h. The mixture was then quenched with water (5 mL). Dichloromethane (10 mL) was added, and the mixture was filtered. The filtrate was extracted with dichloromethane (5 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and evaporated to dryness. The crude product was purified by column chromatography (V... DCM V MeOH =10:1) to give yellow viscous compound 1 (30 mg, yield 20%), [M+H) + :383.3.

[0080] 1 H NMR (400MHz, CDCl3) δ8.57(d,J=4.4Hz,1H),7.63(d,J=7.6Hz,1H),7.31(d,J=8.0Hz 1H),7.14-7.08(m,2H),6.72-6.60(m,1H),4.01-3.98(m,1H),3.78-3.73(m,2H),2.9 4-2.88(m,1H),2.77-2.52(m,3H),2.48-2.45(m,1H),2.36-2.32(m,1H),2.19-1.93 (m,4H),1.75-1.62(m,5H),1,49-1.38(m,3H),1.12-1.09(m,1H),0.73-0.64(m,1H).

[0081] Example 2

[0082] Synthesis of nitrogen-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-5,6-dihydro-4H-cyclopentathiophene-6-ethylamine (compound 2)

[0083]

[0084] Synthesis route:

[0085]

[0086] At room temperature, intermediate 2-1 (100 mg, 0.724 mmol) was dissolved in dichloromethane (10 mL), and then intermediate A (188 mg, 0.724 mmol) was added. The reaction was carried out at 50 °C under nitrogen protection. After 16 h of reaction, sodium borohydride (138 mg, 3.620 mmol) was added and the reaction was carried out for 2 h. After the reaction was complete, 15 mL of water was added for quenching, followed by extraction with ethyl acetate (20 mL × 2), washing with saturated sodium chloride solution (10 mL × 2), drying the organic phase with anhydrous sodium sulfate, filtering, and purifying the crude product by column chromatography (V). DCM V MeOH =20:1-10:1) yielded a pale yellow viscous solid compound 2 (13 mg, yield 4.69%, purity 96.00%), [M+H] + :383.21.

[0087] 1 H NMR (400MHz, Chloroform-d) δ8.57 (q, J=3.4, 2.9Hz, 1H), 7.64 (ddt, J=7.7, 4.8, 2.4Hz, 1H), 7.32 (dd, J=8.1, 3.9Hz,1H),7.21–7.07(m,2H),6.75(t,J=4.8Hz,1H),4.19–4.08(m,1H),3.83–3.68(m,2H),2.87–2.52(m,4H) ,2.50–2.43(m,1H),2.35(dd,J=13.8,2.0Hz,1H),2.21(dd,J=47.2,4.8Hz,1H),2.11–1.97(m,2H),1.95(d,J= 2.8Hz,1H),1.71(ddd,J=33.3,19.2,8.4Hz,4H),1.53–1.33(m,4H),1.12(s,1H),0.70(dt,J=8.3,4.0Hz,1H).

[0088] Example 3

[0089] Synthesis of N-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-6,7-dihydro-5H-thiophene[3,2-b]pyran-7-ethylamine (compound 3)

[0090]

[0091] Synthetic route

[0092]

[0093] Under nitrogen protection, intermediate A (100 mg, 0.38 mmol), intermediate 3-1 (80 mg, 0.51 mmol), DCM (10 mL), and tetraisopropyl titanate (1.5 mL) were sequentially added to a single-necked flask and stirred at room temperature for 16 h. Sodium borohydride (44 mg, 1.2 mmol) was added, and the mixture was stirred for 2 h. The mixture was then quenched with water (2 mL). Dichloromethane (10 mL) was added, and the mixture was filtered. The filtrate was extracted with dichloromethane (5 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and evaporated to dryness. The crude product was purified by column chromatography and concentrated at room temperature (V). DCM V MeOH =10:1) to give yellow viscous compound 3 (10 mg, yield 6.4%), [M+H] + :399.2.

[0094] 1 HNMR(400MHz, CDCl3)δ8.62-8.60(m,1H),7.72-7.70(m,1H),7.54-7.53(m,1H),7.40-7.38(m,1H ),7.28-7.19(m,1H),6.52-6.50(m,1H),3.81-3.78(m,2H),3.26-3.25(m,1H),2.95-2.88(m,1H) ,2.54-2.51(m,1H),2.43-2.42(m,1H),2.39-2.35(m,2H),2.15-2.13(m,1H),1.97-1.91(m,2H), 1.89-1.78(m,2H),1,68-1.61(m,1H),1,53-1.43(m,5H),1,27-1.17(m,3H),0.74-0.70(m,1H).

[0095] Example 4

[0096] Synthesis of N-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-5,6,7,8-tetrahydroquinoline-8-ethylamine (compound 4)

[0097]

[0098] Synthetic route

[0099]

[0100] Intermediate 4-1 (100 mg, 660 μmol), intermediate A (176.8 mg, 660 μmol), and tetraisopropyl titanate (386 mg, 1.36 mmol) were added sequentially to THF (5 mL) and dissolved. The gas was purged three times with nitrogen, and the mixture was heated to 60 °C and stirred overnight (12 h). After the reaction was complete, the mixture was cooled to 0 °C, and NaBH4 (50.3 mg, 1.36 mmol) was added to the reaction flask. The mixture was stirred at low temperature for 30 min, then 1 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 2 h. After the reaction was stopped, the mixture was cooled to room temperature, and the reaction solution was filtered through diatomaceous earth. The filtrate was extracted with dichloromethane (5 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, evaporated to dryness, and the crude product was purified by column chromatography (eluent: V). DCM V MeOH =50:1-10:1) yielded a pale yellow viscous oil compound 4 (20 mg, yield 23%, purity 96%), [M+H) + :392.4.

[0101] 1 HNMR(400MHz,Chloroform-d)δ8.60(s,1H),8.35(s,1H),7.65(s,1H),7.35( d,J=8.0Hz,2H),7.13(d,J=6.5Hz,1H),7.08–7.02(m,1H),3.80(d,J=7.8Hz,2 H),3.64(d,J=25.7Hz,2H),2.72(s,2H),2.51(s,2H),2.37(d,J=13.8Hz,1H), 2.16–1.91(m,5H),1.59–1.45(m,5H),1.28(s,4H),1.15(s,2H),0.90(s,1H).

[0102] Example 5

[0103] Synthesis of N-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-5,6,7,8-tetrahydroquinoline-5-ethylamine (compound 5)

[0104]

[0105] Synthetic route

[0106]

[0107] Intermediate 5-1 (100 mg, 660 μmol), intermediate A (176.8 mg, 660 μmol), and tetraisopropyl titanate (386 mg, 1.36 mmol) were added sequentially to THF (5 mL) for dissolution. The gas was purged three times with nitrogen, and the mixture was heated to 60 °C and stirred overnight. After the reaction was complete, the mixture was cooled to 0 °C, and NaBH4 (50.3 mg, 1.36 mmol) was added to the reaction flask for reduction. The mixture was stirred at low temperature for 30 min, then 1 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 2 h. After the reaction was stopped, water (5 mL) was added, and the reaction solution was filtered through diatomaceous earth. The filtrate was extracted with dichloromethane (5 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, evaporated to dryness, and the crude product was purified by column chromatography (V). DCM V MeOH =50:1-20:1) yielded a pale yellow viscous oil compound 5 (25 mg, yield 23%, purity 95%), [M+H). + :392.3.

[0108] 1 HNMR(400MHz,Chloroform-d)δ8.58–8.49(m,1H),8.38(d,J=4.3Hz,1H),7.69–

[0109] 7.54(m,2H),7.30(d,J=8.1Hz,1H),7.15–7.09(m,1H),7.02(dd,J=7.5,4.8Hz,1H),3.81–

[0110] 3.68(m,3H),2.95–2.88(m,1H),2.86–2.80(m,1H),2.64(td,J=10.9,5. 4Hz,1H),2.53(dd,J=11.0,5.6Hz,1H),2.47–2.29(m,4H),2.06(s,2H), 1.92(dd,J=13.7,3.8Hz,2H),1.85–1.60(m,7H),1.42–1.36(m,1H),1.1 1(s,1H),0.88(dd,J=15.7,9.4Hz,1H),0.68(dq,J=8.8,4.4,3.9Hz,1H).

[0111] Example 6

[0112] Synthesis of N-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-4,5,6,7-tetrahydrobenzo[b]thiophene-7-ethylamine (compound 6)

[0113]

[0114] Synthetic route

[0115]

[0116] Under nitrogen protection, intermediate A (120 mg, 0.46 mmol), intermediate 6-1 (70 mg, 0.46 mmol), DCM (10 mL), and tetraisopropyl titanate (1.5 mL) were sequentially added to a single-necked flask and stirred at room temperature for 16 h. Sodium borohydride (44 mg, 1.2 mmol) was added, and the mixture was stirred for 2 h. The mixture was then quenched with water (5 mL). Dichloromethane (10 mL) was added, and the mixture was filtered. The filtrate was extracted with dichloromethane (5 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and evaporated to dryness. The crude product was purified by column chromatography (V... DCM V MeOH =10:1) to give a yellow viscous compound 6 (30 mg, yield 16%), [M+H) + :397.2.

[0117] 1 HNMR(400MHz, CDCl3)δ8.60-8.58(m,1H),7.67-7.63(m,1H),7.36-7.28( m,1H),7.16-7.10(m,2H),7.16-7.10(m,1H),3.81-3.80(m,2H),3.79-3. 70(m,1H),2.58-2.52(m,4H),2.49-2.40(m,1H),2.37-2.36(m,1H),1.98 -1.91(m,2H),1.85-1.41(m,13H),1.30-1.27(m,1H),0.75-0.74(m,1H).

[0118] Example 7

[0119] Synthesis of nitrogen-(2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)-2H,3H-thienyl[3,2-b]furan-3-ethylamine (compound 7)

[0120]

[0121] Synthetic route

[0122]

[0123] Under nitrogen protection, intermediate A (120 mg, 0.46 mmol), intermediate 7-1 (65 mg, 0.46 mmol), DCM (10 mL), and tetraisopropyl titanate (1.5 mL) were sequentially added to a single-necked flask and stirred at room temperature for 16 h. Sodium borohydride (44 mg, 1.2 mmol) was added, and the mixture was stirred for 2 h. The mixture was then quenched with water (5 mL). Dichloromethane (10 mL) was added, and the mixture was filtered. The filtrate was extracted with dichloromethane (5 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and evaporated to dryness. The crude product was purified by column chromatography (V... DCM V MeOH =10:1) to give a yellow viscous compound 7 (30 mg, yield 17%), [M+H) + :385.2.

[0124] 1 HNMR(400MHz, CDCl3)δ8.63-8.61(m,1H),7.74-7.71(m,1H),7.56-7.54(m,1H),7.40-7.38(m,1 H),7.30-7.21(m,1H),6.55-6.52(m,1H),3.84-3.79(m,2H),3.27-3.25(m,1H),2.93-2.88(m,1H ),2.52-2.50(m,1H),2.43-2.41(m,1H),2.41-2.36(m,2H),2.18-2.14(m,1H),1.96-1.90(m,2H) ,1.89-1.78(m,2H),1,69-1.62(m,1H),1,52-1.41(m,3H),1,25-1.14(m,3H),0.74-0.70(m,1H).

[0125] Biological evaluation

[0126] Op-Mu agonist cAMP assay

[0127] The compounds of this invention can activate the μ-opioid receptor (MOR). Activated MOR can regulate intracellular cAMP levels, which then enter the cell nucleus and bind to the CRE region of the reporter gene luciferase, initiating reporter gene expression. Luciferase reacts with its substrate to emit fluorescence, and the agonistic activity of the compound can be reflected by measuring the fluorescence signal.

[0128] Experimental methods

[0129] The activity of the compound in inducing MOR and affecting downstream cAMP levels was tested using the following methods.

[0130] 1. Materials and Reagents

[0131]

[0132]

[0133] 2. Experimental Procedure

[0134] Detection buffer: 1×stimulation buffer, 500uM IBMX, ddH2O.

[0135] Compound preparation: The compound was dissolved in DMSO to prepare a stock solution with a final concentration of 10 mM. This stock solution was then diluted to a working concentration of 0.08 mM. The compound was serially diluted 4-fold using an Echo analyzer, with an initial concentration of 0.08 mM and 10 concentration gradients. 50 nmol of each gradient was added to 384 cell culture plates in duplicate, with a final concentration of 0.4 μM. The cell culture plates were then centrifuged at 1000 rpm for 1 min. 50 nmol of Forskolin (final concentration 1 μM) was then transferred to the 384 cell culture plates using an Echo analyzer.

[0136] Cell plating: Thaw the frozen cells, centrifuge at 1000 rpm for 5 min, discard the supernatant, wash the cells twice with HBSS buffer, resuspend the cells in assay buffer, and adjust the cell density to 5.0 × 10⁶ cells / min. 5 Add 10 μL of the desired cell count / mL to each well of a 384-well plate (5000 cells per well). Shake for 20 seconds, centrifuge at 1000 rpm for 1 minute, and incubate the plate at 23°C for 60 minutes.

[0137] Preparation of standard curve: The cAMP standard was serially diluted 4-fold with detection buffer to a total of 8 concentration points, with the highest concentration being 800 nM. 10 μL was added to each well according to the microplate layout diagram.

[0138] Preparation of detection reagents: Dilute AnticAMP-Cryptate and AMP-d2 to 1× with lysis buffer. Add 10 μL of detection reagent to each well according to the microplate layout diagram, shake for 20 s, centrifuge at 1000 rpm for 1 min, and incubate the cell plate in a 23℃ incubator for 60 min. Read the plate on an Envision microplate.

[0139] 3. Results Analysis

[0140] The percentage of activity was calculated using Microsoft Excel. For agonists, the formula %Effect = 100 × (Sample Raw Value - Low Control Average) / (High Control Average - Low Control Average) was used. GraphPad Prism 5 data analysis software was then used, with the Dose-response-Stimulation—log[agonist] vs. response-variable slope mode selected for fitting analysis to obtain the EC5 values ​​for each tested sample. 50 value.

[0141] The effects of the compounds of this invention on downstream cAMP levels by activating MOR were determined through the above experiments. The experimental results show that this series of compounds exhibits a strong Op-Mu activating effect, among which the EC50 of the typical representative compounds was measured. 50 The values ​​are shown in Table 1. TRV130 (racemic) has the following structural formula, and its preparation method is described in patent CN103702561A; E max This represents the maximum efficacy of the compound in inducing changes in cAMP levels.

[0142]

[0143] Table 1: Effects of test compounds on cAMP levels of MOR receptors (EC5) 50 and E max

[0144] Compound numbering <![CDATA[EC 50 (nM)]]> <![CDATA[E max ]]> Compound 1 3.850 96.1％ Compound 2 0.7795 88.8％ TRV130 (Rash) 5.74 81.8％ Control group 1 4.538 72.5％ Control group 2 4.162 69.7％

Claims

1. The compound represented by formula (Ⅰ) or a pharmaceutically acceptable salt thereof, (Ⅰ) in, Ring A is selected from a 5-membered cycloalkyl group; The R1 and R2, together with the attached carbon atom, form a substituted or unsubstituted 5-membered monocyclic heteroaromatic ring selected from the following structures: , , , , , ,in" The two ring atoms connected by the symbol "" are adjacent atom pairs shared when fused with ring A.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, R1 and R2, together with the attached carbon atom, form a substituted or unsubstituted 5-membered monocyclic heteroaromatic ring containing an S heteroatom, selected from: .

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The compound is selected from: 、 。 4. A pharmaceutical composition comprising the compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, or the pharmaceutical composition according to claim 4, in the preparation of a medicament for treating MOR receptor agonist-mediated diseases.

6. The use according to claim 5, wherein the MOR receptor agonist-mediated related diseases are selected from pain, immune dysfunction, inflammation, esophageal reflux, neurological and psychiatric diseases, urinary and reproductive diseases, cardiovascular diseases, and respiratory diseases.

7. The use according to claim 6, characterized in that, The pain is selected from postoperative pain, cancer-related pain, neuropathic pain, traumatic pain, and inflammation-related pain.

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