Arylalkylamine compound, and preparation method therefor and use thereof

Abstract

The present invention relates to an arylalkylamine compound, and a preparation method therefor and a use thereof. The arylalkylamine compound of the present invention has a structure as shown in formula (I), wherein R1, R2, R3, R4, and R5 are as defined in the description. The compound of the present invention has excellent CaSR agonistic activity, high bioavailability, high in-vivo metabolic stability and high safety.

Description

Arylalkylamine compounds, and preparation method and use thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and in particular relates to an arylalkylamine compound, a preparation method thereof, and a use thereof in allosteric agonizing a calcium-sensing receptor (Cacium-sensing receptor, CaSR). BACKGROUND

[0002] Chronic kidney disease can cause secondary hyperparathyroidism (SHPT). At present, there are many methods for treating SHPT at home and abroad. However, there are disadvantages such as high cost, narrow treatment window, incomplete treatment, easy recurrence, and intolerance. The calcimimetic drug directly acts on the CaSR, can effectively control the synthesis and secretion of PTH, and has more clinical advantages.

[0003] Tanabe Mitsubishi Pharmaceutical Co., Ltd. developed the calcimimetic drug Evocalcet (WO2005115975), which was first approved for marketing in Japan in March 2018. According to the Evocalcet review report, the main adverse reactions include hypocalcemia, vomiting, nausea, chills, QT prolongation, adverse mood, arrhythmia, blood pressure drop, and spasm (Orkedia tablet 1mg, same tablet 2mg review report. March 2018).

[0004] Therefore, it is of extremely important clinical value and social value to develop a calcimimetic drug with higher activity and safety, in particular an arylalkylamine compound, for treating SHPT in patients. SUMMARY

[0005] In order to solve the defects of the prior art, the present application aims to provide an arylalkylamine compound with higher activity and safety. The compound of the present application has excellent CaSR agonizing activity, high bioavailability, high in-vivo metabolic stability, and high safety. The present application is realized by the following technical scheme.

[0006] In one aspect, the present application provides a compound having the structure shown in the following formula (I) or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof:

[0007] wherein R 1 , R 2 , R 3 , R 4 , R 5 are each independently selected from a hydrogen atom; C 2-6 carboxylic acid group; C 1-7alkyl (e.g. methyl, ethyl), C 1-6 carboxylic acid group, amine group, C 3-6 cycloalkyl, aryl C 1-6 alkyloxy (e.g. benzyloxy), hydroxy, C 1-6 alkoxy, halo C 1-6 alkyl and halo C 1-6 alkoxy; halogen; C 1-6 alkyl; halo C 1-6 alkyl;

[0008] provided that: 1) R 1 , R 2 , R 3 , R 4 , R 5 are not simultaneously H; 2) when the substituent is a C 2-6 carboxylic acid group, the substitution site is R 2 -R 4 , and at least 2 of R 1 -R 5 are not H;

[0009] Preferably, R 1 , R 5 are selected from H, halogen (e.g. F, Cl), C 1-6 alkyl (e.g. methyl, ethyl), C 1-6 alkoxy (e.g. methoxy, ethoxy).

[0010] Preferably, at least one of R 2 -R 4 is an unsubstituted or substituted C 2-6 carboxylic acid group, preferably an acetic acid group, and at least 2 of R 1 -R 5 are not H. That is, at least in the para or meta position is an acetic acid group (i.e. -CH2COOH), and at least one other position is substituted.

[0011] Preferably, at least one of R 2 -R 4 is an unsubstituted or substituted acetic acid group, and at least 2 of R 1 -R 5 are not H. That is, at least in the para or meta position is an acetic acid group (i.e. -CH2COOH), and at least one other position is substituted.

[0012] Preferably, R 1 , R 2 , R 4 , R 5 are each independently selected from H, halogen (e.g. F, Cl), C 1-6alkyl (e.g., methyl, ethyl), C 1-6 alkoxy (e.g., methoxy, ethoxy, etc.) substituted with a substituent selected from the group consisting of hydroxy, C 1-6 alkoxy, and R 1 , R 2 , R 4 , R 5 are not simultaneously H; R 3 is a C 2-6 carboxylic acid group.

[0013] Preferably, R 1 , R 2 , R 4 , R 5 are each independently selected from the group consisting of H, halogen (e.g., F, Cl), C 1-6 alkyl (e.g., methyl, ethyl), C 1-6 alkoxy (e.g., methoxy, ethoxy, etc.) substituted with a substituent selected from the group consisting of hydroxy, C 1-6 alkoxy, and R 1 , R 2 , R 4 , R 5 are not simultaneously H; R 3 is a C

[0014] In one embodiment, the compound of formula (I) is selected from the group consisting of:

[0015] The compound of general formula (I) of the present application can be in a free form or a pharmaceutically acceptable salt form. A pharmaceutically acceptable inorganic acid salt such as a hydrochloride, a sulfate, a phosphate, a hydrobromide, or the like. A pharmaceutically acceptable organic acid salt such as an acetate, a fumarate, an oxalate, a citrate, a methanesulfonate, a benzenesulfonate, a p-toluenesulfonate, or a maleate, or the like. In addition, when the compound has an acidic group such as a carboxyl group, a salt with a metal ion such as a sodium salt, a potassium salt, a calcium salt, or the like, an alkali metal salt or an alkaline earth metal salt can be formed.

[0016] In another aspect, the present application provides a method for preparing the aforementioned compound of formula (I), the method comprising the step of reacting a compound of formula A with a compound of formula (II) to form a compound of formula (I):

[0017] wherein X represents a leaving group selected from the group consisting of halogen, hydroxy, lower alkylsulfonyloxy (e.g., C 1-6 alkylsulfonyloxy), sulfonyloxy, or the like, preferably Br, Cl, trifluoromethylsulfonyloxy.

[0018] In still another aspect, the present application provides a pharmaceutical composition comprising the aforementioned compound or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient.

[0019] In particular, the compound of formula (I) or a pharmaceutically acceptable salt thereof of the present application, as an effective ingredient for medical use, can be used together with an inert carrier suitable for each administration method, and can be formulated into a conventional pharmaceutical preparation such as a tablet, granule, capsule, powder, solution, suspension, emulsion, injection, etc. When used as a solid preparation, the inert carrier includes a binder (acacia, gelatin, sorbitol, polyvinylpyrrolidone, etc.), an excipient (lactose, galactose, corn starch, sorbitol, etc.), a lubricant (magnesium stearate, talc, polyethylene glycol, etc.), a disintegrant (potato starch, etc.), etc. When used as an injection, it can be formulated with distilled water for injection, physiological saline, an aqueous glucose solution, etc.

[0020] The compound of formula (I) or a pharmaceutically acceptable salt thereof for medical use of the present application can be administered orally, intravenously, intramuscularly, subcutaneously, transdermally, etc., and the administration dose and volume can be determined according to the characteristics of the drug, the administration route, the age, weight or disease state of the patient, etc.

[0021] The present application provides the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof, or a pharmaceutical composition comprising the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer or tautomer thereof and a pharmaceutically acceptable carrier or excipient. It produces various pharmacological effects by activating CaSR, and is used for preventing and treating diseases related to CaSR, such as inhibiting the production of PTH, regulating the calcium and phosphorus levels in blood by activating CaSR, and is used for treating primary hyperparathyroidism, secondary hyperparathyroidism, tertiary hyperparathyroidism, chronic renal failure (with or without dialysis), chronic kidney disease (with or without dialysis), parathyroid adenoma, parathyroid hyperplasia, parathyroid cancer, vascular calcification and valve calcification, abnormal calcium homeostasis (such as hypercalcemia), abnormal phosphorus homeostasis (such as hypophosphatemia), bone-related diseases or complications caused by hyperparathyroidism, chronic kidney disease or parathyroid cancer, bone loss after kidney transplantation, osteogenesis imperfecta, adynamic bone disease, renal osteodystrophy, cardiovascular complications caused by hyperparathyroidism or chronic kidney disease, Ca 2+ certain malignancies of abnormally high.

[0022] The following terms as used in the specification and claims have the meanings given unless otherwise stated.

[0023] The present application includes all stereoisomeric forms of the compounds described. Unless specified to the contrary, the application is intended to include all such isomeric forms of the compounds. Each asymmetric center in a compound of the present application can independently have the (R) or (S) configuration. When a bond is drawn in a structural formula of the present application as a straight line, it is understood that both the (R) and (S) configurations of the chiral carbon, as well as both enantiomers and mixtures thereof, are included in the formula. When a particular configuration is depicted, either the enantiomer (R) or (S) is intended. Similarly, when a compound name is recited without a chiral designation for a chiral carbon, it is understood that both the (R) and (S) configurations of the chiral carbon, as well as the individual enantiomers and mixtures thereof, are included by the name. The preparation of particular stereoisomers or mixtures thereof can be identified in embodiments where such stereoisomers or mixtures are obtained, but this in no way limits the scope of the disclosure to include all stereoisomers and mixtures thereof.

[0024] The term "carboxyl" is a fundamental functional group in organic chemistry, consisting of one carbon atom, two oxygen atoms, and one hydrogen atom, with the chemical formula -COOH.C 1-6 carboxyl, C 2-6 Carboxyl is a lower alkyl further containing a carboxyl group.C 1-6 Carboxyl can be formyl (e.g., -COOH), acetyl, propionyl, butyryl, valeryl, hexanoyl.C 2-6 Carboxyl can be acetyl, propionyl, butyryl, valeryl, hexanoyl.

[0025] The compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof as an active ingredient, by allosteric activation of CaSR on parathyroid cells, inhibits the synthesis and secretion of parathyroid hormone, thereby reducing the level of parathyroid hormone in the blood, and further stabilizing the blood calcium and blood phosphorus concentrations, and can be used for the prevention or treatment of hyperparathyroidism, having excellent CaSR allosteric agonistic effect and reducing the effect of PTH in vivo. By CaSR allosteric activation effect experiment, SD rat adenine model or SD rat 5 / 6 nephrectomy model, it is confirmed that the compound has CaSR allosteric activation effect and reduces the effect of PTH in vivo.

[0026] The compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof as an active ingredient, not only has excellent activity, but also has certain in vivo stability. The present application carries out rat liver microsomal incubation experiment to verify that the compound indeed has metabolic stability. The compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof as an active ingredient, not only has excellent CaSR activation effect, but also has high safety. Compared with the prior art, the present application not only has excellent activity and good safety, but also has high in vivo metabolic stability and high in vivo relative bioavailability. DETAILED DESCRIPTION

[0027] The present disclosure is further described in conjunction with the following examples, which do not limit the scope of the present disclosure.

[0028] The known starting materials of the present disclosure can be synthesized by using or following the methods known in the art, or purchased from chemical companies such as Angene, Bailingwei Technology, Macklin, Araldin, Bide Pharmaceutical, etc.

[0029] Unless otherwise specified in the examples, the solution refers to an aqueous solution, the room temperature refers to 20-30°C, the inert gas refers to argon or nitrogen, the developing solvent for thin layer chromatography (TLC) monitoring reaction and column chromatography purification is dichloromethane / methanol system, n-hexane / ethyl acetate system, petroleum ether / ethyl acetate system, petroleum ether / ethyl acetate / methanol system, etc., the proportion of the system components is adjusted according to the polarity of the compound, and a small amount of triethylamine and basic or acidic reagents such as acetic acid can also be added for adjustment.

[0030] Unless otherwise specified in the examples, the mass spectrometry (Mass spectrum, MS), nuclear magnetic resonance (Nuclear Magnetic Resonance, NMR) and high performance liquid chromatography (High performance liquid chromatography, HPLC) for structure confirmation are Agilent InfinityLab LC / MSD iQ G6160A, Bruker AVANCE 400 or Bruke AVANCE NEO 600 and Shimadzu LC-2010AHT, respectively. The determination solvent of NMR is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) or deuterated heavy water (D2O), and the internal standard is tetramethylsilane (TMS).

[0031] Example 1: Synthesis of (3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}pyrrolidine dihydrochloride (A)

[0032] The synthesis route is as follows:

[0033] Step 1: 2-nitrobenzenesulfonic acid-(3R)-1-{[(2-methylpropan-2-yl)oxy]carbonyl}pyrrolidin-3-yl ester (A-2)

[0034] 500 mL three-necked flask, compound A-1 (25.00 g, 0.13 mol), triethylamine (17.57 g, 0.17 mol), trimethylamine hydrochloride (2.55 g, 0.03 mol) and dichloromethane 125 mL were added and stirred; cooled to 0-10 °C, dropwise added 2-nitrobenzenesulfonyl chloride (32.55 g, 0.15 mol) in dichloromethane 125 mL; after the reaction was completed, 150 mL of purified water was added, and the pH was adjusted to 2.0-4.0 with 6N hydrochloric acid; allowed to stand until the layers were clearly separated, the phases were separated, the organic phase was retained, the aqueous phase was extracted once with 50 mL of dichloromethane, and the combined organic phases were dried over anhydrous sodium sulfate and filtered; the filtrate was concentrated under reduced pressure to constant weight to obtain compound A-2 (47.40 g, y = 95%). LCMS (ESI) m / z: 373.1 [M+H] + .

[0035] Step 2: (3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydropyrrole-1-carboxylic acid-2-methylprop-2-yl ester (A-3)

[0036] 100 mL three-necked flask, compound A-2 (45.00 g, 0.12 mol), (1R)-1-(naphthalen-1-yl)ethan-1-amine (17.60 g, 0.10 mol), potassium phosphate (21.23 g, 0.10 mol) and acetonitrile 150 mL were added and warmed to 75 °C, and stirred for 24 h; cooled to room temperature, filtered, the filter cake was washed with 60 mL of acetonitrile, and concentrated under reduced pressure to distill off the solvent; 90 mL of ethyl acetate and 90 mL of saturated brine were added and stirred for 0.5 h, allowed to stand until the layers were clearly separated, the phases were separated, the organic phase was retained, the aqueous phase was extracted once with 45 mL of ethyl acetate, the combined organic phases were dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to constant weight to obtain compound A-3 (32.62 g, y = 96%). LCMS (ESI) m / z: 341.2 [M+H] + .

[0037] Step 3: (3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydropyrrole dihydrochloride (A)

[0038] 500 mL three-necked flask, compound A-3 (30.00 g, 0.09 mol) and ethyl acetate 150 mL were added and stirred, cooled to 0-10 °C, dropwise added concentrated hydrochloric acid 22.5 mL, after the dropwise addition was completed, stirred for 0.5 h, warmed to 55 °C and reacted for 2.0 h; cooled to room temperature, filtered, the filter cake was washed with 50 mL of ethyl acetate, and dried at 45 °C under a blast of air to constant weight to obtain compound A (19.87 g, y = 92%). LCMS (ESI) m / z: 241.2 [M+H] + . 1H NMR (400 MHz, MeOH-d4): δ 8.29 (d, J = 8.8 Hz, 1H), 8.04-7.97 (m, 2H), 7.94 (d, J = 7.2 Hz, 1H), 7.72-7.58 (m, 3H), 5.60-5.51 (m, 1H), 3.98-3.87 (m, 1H), 3.65-3.54 (m, 1H), 3.51-3.38 (m, 2H), 3.29-3.20 (m, 1H), 2.55-2.44 (m, 1H), 2.43-2.32 (m, 1H), 1.89 (d, J = 6.8 Hz, 3H).

[0039] Example 2: Synthesis of {2-fluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1- yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]phenyl}acetic acid (1)

[0040] The synthetic route is as follows:

[0041] Step 1: (3-bromo-2-fluorophenyl) ethyl acetate (1-1)

[0042] 250 mL single neck flask, compound B (2.00 g, 8.58 mmol) and 60 mL of ethanol were stirred and cooled to 0 °C, 1 mL of concentrated sulfuric acid was added dropwise, and the reaction was warmed to reflux for 3.0 h; cooled to room temperature, concentrated under reduced pressure to remove solvent, 50 mL of ethyl acetate and 50 mL of purified water were added and stirred, the pH was adjusted to 8.0 by adding saturated sodium carbonate solution dropwise, and the mixture was allowed to stand until it was clearly separated into two layers, the organic phase was separated, the aqueous phase was extracted with 50 mL of ethyl acetate once, the combined organic phase was dried over anhydrous sodium sulfate, and filtered; the filtrate was concentrated under reduced pressure to constant weight to give compound 1-1 (2.00 g, y = 89%). LCMS (ESI) m / z: 261.0 [M+H] + .

[0043] Step 2: {2-fluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H- pyrrol-1-yl]phenyl} ethyl acetate (1-2)

[0044] 250 mL single necked flask, was charged with free compound A (1.00 g, 4.16 mmol), compound 1-1 (1.31 g, 4.99 mmol), Pd(OAc)2(50 mg, 0.20 mmol), X-Phos (100 mg, 0.20 mmol), Cs2CO3(4.10 g, 12.48 mmol) and 50 mL of toluene, replaced with nitrogen three times, warmed to 100 °C and stirred for 12.0 h, cooled to room temperature, stirred with 30 mL of saturated brine, allowed to settle to distinct layers, separated, retained the organic phase, extracted the aqueous phase with 20 mL of ethyl acetate once, combined the organic phases, dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to no solvent was distilled off, purified the residue by column chromatography (200-300 mesh silica gel, petroleum ether / ethyl acetate: 80 / 20→ 50 / 50), concentrated under reduced pressure to give compound 1-2 (0.70 g, y = 40%). LCMS (ESI) m / z: 421.2 [M+H] + .

[0045] Step 3: {2-Fluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1- yl]phenyl}acetic acid (1)

[0046] 100 mL single necked flask, was charged with 1-2 (0.60 g, 1.43 mmol), sodium hydroxide (572 mg, 14.3 mmol), purified water 10 mL and ethanol 10 mL, stirred at 60 °C for 3.0 h, cooled to room temperature, adjusted the pH to solid precipitated obviously with 1 N hydrochloric acid, filtered, washed the filter cake with 10 mL of purified water, air dried at 45 °C to constant weight to give compound 1 (0.30 g, y = 53%). LCMS (ESI) m / z: 393.3 [M+H] + . 1 H NMR (600 MHz, DMSO-d6) δ 8.34 (d, J = 8.4 Hz, 1H), 7.96 (d, J = 7.9 Hz, 1H), 7.87 (d, J = 8.6 Hz, 2H), 7.56 (dd, J = 14.2, 5.9 Hz, 3H), 7.05 (t, J = 8.6 Hz, 1H), 6.24 (d, J = 8.9 Hz, 2H), 4.96 (s, 1H), 3.43 (s, 3H), 3.36 (q, J = 7.7 Hz, 2H), 3.13 (q, J = 7.8 Hz, 2H), 2.07 (m, 2H), 1.52 (s, 3H).

[0047] Example 3: Synthesis of {3-Fluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H- pyrrol-1-yl]phenyl}acetic acid (2)

[0048] The synthetic route is as follows:

[0049] Step 1: (4-bromo-3-fluorophenyl) ethyl acetate (2-1)

[0050] 250 mL single neck flask, add compound C (2.00 g, 8.58 mmol) and 60 mL of ethanol, stirring, cooling to 0 °C, drop 1 mL of concentrated sulfuric acid, synthesis and post-processing operation reference compound 1-1, compound 2-1 (1.80 g, y = 81%) was obtained. LCMS (ESI) m / z: 261.0 [M+H] + .

[0051] Step 2: {3-fluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl] phenyl} ethyl acetate (2-2)

[0052] 250 mL single neck flask, add free compound A (1.00 g, 4.16 mmol), compound 2-1 (1.31 g, 4.99 mmol), Pd(OAc)2(50 mg, 0.20 mmol), X-Phos (100 mg, 0.20 mmol), Cs2CO3(4.10 g, 12.48 mmol) and 50 mL of toluene, synthesis and post-processing operation reference compound 1-2, compound 2-2 (0.89 g, y = 51%) was obtained. LCMS (ESI) m / z: 421.2 [M+H] + .

[0053] Step 3: {2-fluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl] phenyl} acetic acid (2)

[0054] 100 mL single neck flask, add 2-2 (0.80 g, 1.90 mmol), sodium hydroxide (760 mg, 19.0 mmol), purified water 10 mL and ethanol 10 mL, synthesis and post-processing operation reference compound 1, compound 2 (0.39 g, y = 52%) was obtained. LCMS (ESI) m / z: 393.3 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.33 - 8.26 (m, 1H), 7.93 (dd, J = 7.0, 2.3 Hz, 1H), 7.79 (d, J = 8.1 Hz, 1H), 7.72 (d, J = 7.2 Hz, 1H), 7.51 (dtd, J = 10.2, 7.2, 4.8 Hz, 3H), 6.91 (dd, J = 14.9, 2.0 Hz, 1H), 6.85 (dd, J = 8.2, 2.0 Hz, 1H), 6.56 (t, J = 9.0 Hz, 1H), 4.72 (q, J = 6.6 Hz, 1H), 3.42 - 3.28 (m, 4H), 3.19 (p, J = 6.9, 5.9 Hz, 2H), 3.09 (ddd, J = 8.9, 5.7, 2.7 Hz, 1H), 1.96 (dq, J = 12.3, 6.1 Hz, 1H), 1.83 (dq, J = 13.6, 7.2 Hz, 1H), 1.40 (d, J = 6.5 Hz, 3H).

[0055] Example 4: Synthesis of {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-3-methylphenyl}acetic acid (3)

[0056] The synthetic route is as follows:

[0057] Step 1: (4-bromo-3-methylphenyl) ethanoate (3-1)

[0058] 250 mL single necked flask, compound D (1.00 g, 4.37 mmol) and 60 mL ethanol were stirred, cooled to 0 °C, dropwise added 1 mL of concentrated sulfuric acid, the synthesis and post-processing operation reference compound 1-1, compound 3-1 (0.87 g, y = 77%) was obtained. LCMS (ESI) m / z: 257.0 [M+H] + .

[0059] Step 2: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-3- methylphenyl} ethanoate (3-2)

[0060] 250 mL single neck flask, add free compound A (1.00 g, 4.16 mmol), compound 3-1 (1.28 g, 4.99 mmol), Pd(OAc)2(50 mg, 0.20 mmol), X-Phos (100 mg, 0.20 mmol), Cs2CO3(4.10 g, 12.48 mmol) and 50 mL of toluene, synthesis and post-processing operations refer to compound 1-2, to obtain compound 3-2 (0.73 g, y = 42%). LCMS (ESI) m / z: 417.2 [M+H] + .

[0061] Step 3: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-3- methylphenyl}acetic acid (3)

[0062] 100 mL single neck flask, add 3-2 (0.70 g, 1.68 mmol), sodium hydroxide (0.34 g, 8.40 mmol), purified water 10 mL and ethanol 10 mL, synthesis and post-processing operations refer to compound 1, to obtain compound 3 (0.44 g, y = 67%). LCMS (ESI) m / z: 389.2 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.32 (d, J = 8.0 Hz, 1H), 7.93 (dd, J = 7.3, 2.1 Hz, 1H), 7.80 (d, J = 8.1 Hz, 1H), 7.73 (d, J = 7.1 Hz, 1H), 7.51 (qd, J = 8.1, 7.3, 2.3 Hz, 3H), 6.97 - 6.86 (m, 2H), 6.71 - 6.64 (m, 1H), 4.71 (q, J = 6.5 Hz, 1H), 3.38 (s, 2H), 3.18 (s, 1H), 3.13 - 3.06 (m, 2H), 3.02 (m, 1H), 2.92 (dd, J = 9.2, 6.3 Hz, 1H), 2.11 (s, 3H), 1.99 (dq, J = 12.6, 6.5 Hz, 1H), 1.86 - 1.73 (m, 1H), 1.41 (d, J = 6.5 Hz, 3H).

[0063] Example 5: Synthesis of {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H- pyrrol-1-yl]-2-methylphenyl}acetic acid (4):

[0064] The synthesis route is as follows:

[0065] Step 1: (4-bromo-2-methylphenyl) ethyl acetate (4-1)

[0066] 250 mL single neck flask, add compound E (1.00 g, 4.37 mmol) and 60 mL ethanol, stirring, cooling to 0 °C, dropwise addition of concentrated sulfuric acid 1 mL, synthesis and post-processing operation for reference compound 1-1, compound 4-1 (0.93 g, y = 83%) was obtained. LCMS (ESI) m / z: 257.0 [M+H] + .

[0067] Step 2: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-3- methylphenyl}acetic acid ethyl ester (4-2)

[0068] 250 mL single neck flask, add free compound A (1.00 g, 4.16 mmol), compound 4-1 (1.28 g, 4.99 mmol), Pd(OAc)2(50 mg, 0.20 mmol), X-Phos (100 mg, 0.20 mmol), Cs2CO3(4.10 g, 12.48 mmol) and 50 mL toluene, synthesis and post-processing operation for reference compound 1-2, compound 4-2 (0.73 g, y = 42%) was obtained. LCMS (ESI) m / z: 417.2 [M+H] + .

[0069] Step 3: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-2- methylphenyl}acetic acid (4)

[0070] 100 mL single neck flask, add 4-2 (0.50 g, 1.20 mmol), sodium hydroxide (0.24 g, 6.00 mmol), purified water 5 mL and ethanol 5 mL, synthesis and post-processing operation for reference compound 1, compound 4 (0.32 g, y = 69%) was obtained. LCMS (ESI) m / z: 389.2 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.30 (d, J = 8.0 Hz, 1H), 7.92 (dd, J = 7.3, 2.1 Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.73 (d, J = 7.1 Hz, 1H), 7.53 (qd, J = 8.1, 7.3, 2.3 Hz, 3H), 6.97 - 6.86 (m, 2H), 6.71 - 6.64 (m, 1H), 4.71 (q, J = 6.5 Hz, 1H), 3.38 (s, 2H), 3.18 (s, 1H), 3.13 - 3.06 (m, 2H), 3.02 (m, 1H), 2.91 (dd, J = 9.2, 6.3 Hz, 1H), 2.10 (s, 3H), 1.99 (dq, J = 12.6, 6.5 Hz, 1H), 1.86 - 1.72 (m, 1H), 1.43 (d, J = 6.5 Hz, 3H).

[0071] Example 6: Synthesis of {2,6-difluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1- yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]phenyl}acetic acid (5)

[0072] The synthetic route is as follows:

[0073] Step 1: {2,6-difluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro- 1H-pyrrol-1-yl]phenyl} ethyl acetate (5-1)

[0074] 250 mL single necked flask, free compound A (861 mg, 3.58 mmol), compound F (1.00 g, 3.58 mmol), Pd(OAc)2(80 mg, 0.36 mmol), X-Phos (171 mg, 0.36 mmol), Cs2CO3(3.50 g, 10.74 mmol) and 50 mL of toluene, synthesis and workup procedure refer to compound 1-2, to give compound 5-1 (1.44 g, y = 74%). LCMS (ESI) m / z: 439.2 [M+H] + .

[0075] Step 2: {2,6-difluoro-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro- 1H-pyrrol-1-yl]phenyl} acetic acid (5)

[0076] 100 mL single necked flask, was added 5-1 (1.44 g, 3.28 mmol), sodium hydroxide (0.66 g, 16.43 mmol), purified water 5 mL and ethanol 5 mL, synthesis and post-treatment operation reference compound 1, compound 5 (1.25 g, y = 93%) was obtained. LCMS (ESI) m / z: 411.2 [M+H] + . 1 H NMR (600 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.94 (s, 1H), 7.85-7.78 (m, 1H), 7.73 (dd, J = 10.9, 6.0 Hz, 1H), 7.57-7.47 (m, J = 6.5, 5.5 Hz, 3H), 6.08 (dq, J = 10.2, 5.2 Hz, 2H), 4.75 (dd, J = 11.7, 6.0 Hz, 1H), 3.42 (q, J = 5.5, 5.0 Hz, 2H), 3.37-3.23 (m, 3H), 3.11 (s, 1H), 2.99 (s, 1H), 2.05-1.98 (m, 1H), 1.94-1.88 (m, 1H), 1.42 (p, J = 5.7 Hz, 3H).

[0077] Example 7: Synthesis of {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-3-methoxyphenyl}acetic acid (6)

[0078] The synthesis route is as follows:

[0079] Step 1: methyl 4-bromo-3-methoxybenzoate (6-1)

[0080] 100 mL single necked flask, was added 5-1 (1.44 g, 3.28 mmol), sodium hydroxide (0.66 g, 16.43 mmol), purified water 5 mL and ethanol 5 mL, synthesis and post-treatment operation reference compound 1, compound 5 (1.25 g, y = 93%) was obtained. LCMS (ESI) m / z: 411.2 [M+H] + .

[0081] Step 2: (4-bromo-3-methoxyphenyl)methanol (6-2)

[0082] 500 mL three-necked flask, compound 6-1 (10.50 g, 23.8 mmol) and 100 mL of anhydrous tetrahydrofuran were added and stirred, cooled to 0 °C, lithium aluminum hydride (1.95 g, 51.4 mmol) was added in portions, incubated for 4.0 h, 150 mL of 1 N hydrochloric acid was added slowly and stirred, left to settle until clear separation, the organic phase was retained, the aqueous phase was extracted twice with 50 mL of ethyl acetate, the combined organic phases were dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure until solvent evaporation, the residue was purified by column chromatography (200-300 mesh silica gel, petroleum ether / ethyl acetate: 95 / 5→80 / 20), concentrated under reduced pressure to obtain compound 6-2 (5.80 g, y = 62%). LCMS (ESI) m / z: 217.0 [M+H] + .

[0083] Step 3: 1-bromo-4-(bromomethyl)-2-methoxybenzene (6-3)

[0084] 500 mL three-necked flask, compound 6-2 (5.00 g, 23.00 mmol), triphenylphosphine (7.30 g, 27.60 mmol), carbon tetrabromide (8.40 g, 25.30 mmol) and 100 mL of dichloromethane were added and stirred overnight, the filtrate was concentrated under reduced pressure until solvent evaporation, the residue was purified by column chromatography (200-300 mesh silica gel, petroleum ether / ethyl acetate: 95 / 5→85 / 15), concentrated under reduced pressure to obtain compound 6-3 (5.80 g, y = 90%). LCMS (ESI) m / z: 279.0 [M+H] + .

[0085] Step 4: (4-bromo-3-methoxyphenyl)acetonitrile (6-4)

[0086] 500 mL three-necked flask, compound 6-3 (5.00 g, 17.9 mmol), TBAF (5.60 g, 21.4 mmol) and 50 mL of acetonitrile were added and stirred, cooled to 0 °C, TMSCN (2.13 g, 21.4 mmol) was added in portions, stirred overnight, the filtrate was concentrated under reduced pressure until solvent evaporation, the residue was purified by column chromatography (200-300 mesh silica gel, petroleum ether / ethyl acetate: 95 / 5→85 / 15), concentrated under reduced pressure to obtain compound 6-4 (3.60 g, y = 89%). LCMS (ESI) m / z: 226.0 [M+H] + .

[0087] Step 5: (4-bromo-3-methoxyphenyl) ethyl acetate (6-5)

[0088] 500 mL three-necked flask, compound 6-4 (3.00 g, 13.27 mmol) and 50 mL of ethanol were stirred, cooled to 0 °C, and 7.5 mL of concentrated sulfuric acid was added dropwise. The synthesis and post-processing operations were referred to compound 1-1 to obtain compound 6-5 (3.20 g, y = 88%). LCMS (ESI) m / z: 273.0 [M+H] + .

[0089] Step 6: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-3- methoxyphenyl}acetic acid ethyl ester (6-6)

[0090] 100 mL single-necked flask, free compound A (1.00 g, 4.61 mmol), compound 6-5 (1.10 g, 4.99 mmol), Pd(OAc)2(19 mg, 0.08 mmol), X-Phos (40 mg, 0.08 mmol), Cs2CO3(4.00 g, 12.50 mmol) and 20 mL of toluene were added. The synthesis and post-processing operations were referred to compound 1-2 to obtain compound 6-6 (1.20 g, y = 67%). LCMS (ESI) m / z: 433.2 [M+H] + .

[0091] Step 7: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-3- methoxyphenyl}acetic acid (6)

[0092] 100 mL single-necked flask, 6-6 (1.00 g, 2.31 mmol), sodium hydroxide (0.28 g, 6.93 mmol), 5 mL of purified water and 5 mL of ethanol were added. The synthesis and post-processing operations were referred to compound 1 to obtain compound 6 (0.31 g, y = 33%). LCMS (ESI) m / z: 405.2 [M+H] + . 1H NMR (600 MHz, DMSO-d6) δ 8.32 (d, J = 8.2 Hz, 1H), 7.92 (dd, J = 7.7, 1.8 Hz, 1H), 7.79 (d, J = 8.2 Hz, 1H), 7.72 (dd, J = 7.3, 1.2 Hz, 1H), 7.55 - 7.46 (m, 3H), 6.73 (d, J = 2.0 Hz, 1H), 6.65 (dd, J = 8.1, 1.9 Hz, 1H), 6.48 (d, J = 8.1 Hz, 1H), 4.71 (s, 1H), 3.64 (s, 3H), 3.40 (s, 2H), 3.22 (dt, J = 7.9, 6.0 Hz, 2H), 3.14 (dt, J = 9.4, 6.8 Hz, 2H), 3.01 (t, J = 7.9 Hz, 1H), 1.93 (dt, J = 13.2, 6.6 Hz, 1H), 1.77 (dd, J = 12.5, 6.7 Hz, 1H), 1.40 (d, J = 6.6 Hz, 3H).

[0093] Example 8: Synthesis of {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-2-methoxyphenyl}acetic acid (7)

[0094] The synthetic route is as follows:

[0095] Step 1: 4-bromo-2-methoxyphenyl acetate (7-1)

[0096] A 500 mL three-necked flask was charged with compound H (10.00 g, 40.80 mmol) and 80 mL of ethanol, stirred, cooled to 0 °C, and 1.5 mL of concentrated sulfuric acid was added dropwise. The synthesis and post-processing operations were referred to compound 1-1 to obtain compound 7-1 (10.50 g, y = 94%). LCMS (ESI) m / z: 245.0 [M+H] + .

[0097] Step 2: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-2- methoxyphenyl} acetic acid ethyl ester (7-2)

[0098] 100 mL single necked flask, add free compound A (1.47 g, 6.11 mmol), compound 7-1 (2.00 g, 7.32 mmol), Pd(OAc)2(70 mg, 0.31 mmol), X-Phos (150 mg, 0.31 mmol), Cs2CO3(5.96 g, 12.50 mmol) and 20 mL of toluene, synthesis and work-up procedure refer to compound 1-2, get compound 7-2 (1.60 g, y = 61%). LCMS (ESI) m / z: 433.2 [M+H] + .

[0099] Step 3: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-2- methoxyphenyl}acetic acid ethyl ester (7)

[0100] 100 mL single necked flask, add 7-2 (1.60 g, 3.70 mmol), sodium hydroxide (0.59 g, 14.75 mmol), purified water 5 mL and ethanol 5 mL, synthesis and work-up procedure refer to compound 1, get compound 7 (0.50 g, y = 33%). LCMS (ESI) m / z: 405.2 [M+H] + . 1 H NMR (600 MHz, DMSO-d6) δ 8.33 (d, J = 8.3 Hz, 1H), 7.93 (d, J = 7.8 Hz, 1H), 7.81 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 7.2 Hz, 1H), 7.51 (q, J = 7.9, 6.8 Hz, 3H), 6.88 (d, J = 8.2 Hz, 1H), 5.98 (d, J = 2.3 Hz, 1H), 5.96 (dd, J = 8.4, 2.2 Hz, 1H), 4.76 (s, 1H), 3.70 (s, 3H), 3.32 (s, 4H), 3.12 (dt, J = 9.2, 7.3 Hz, 1H), 2.99 (s, 1H), 2.04 (s, 1H), 1.91 (s, 1H), 1.42 (s, 3H).

[0101] Example 9: Synthesis of {2-ethoxy-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]phenyl}acetic acid (8)

[0102] The synthesis route is as follows:

[0103] Step 1: 4-bromo-2-ethoxybenzoic acid ethyl ester (8-1)

[0104] 250 mL three-necked flask, compound I (2.00 g, 7.72 mmol), potassium carbonate (3.20 g, 23.00 mmol), ethyl bromide (0.93 g, 8.40 mmol) and 50 mL of acetonitrile were added, and the reaction was stirred at 60 °C for 24.0 h; cooled to room temperature, concentrated under reduced pressure to remove solvent, 50 mL of ethyl acetate and 50 mL of purified water were added and stirred, and allowed to stand until the layers were clearly separated, the organic phase was separated, the aqueous phase was extracted with 50 mL of ethyl acetate once, the combined organic phase was dried over anhydrous sodium sulfate, and filtered; the filtrate was concentrated under reduced pressure to constant weight to obtain compound 8-1 (2.02 g, y = 91%). LCMS (ESI) m / z: 287.0 [M+H] + .

[0105] Step 2: {2-Ethoxy-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1- yl]phenyl}acetic acid ethyl ester (8-2)

[0106] 100 mL single-necked flask, free compound A (1.00 g, 4.16 mmol), compound 8-1 (2.00 g, 7.32 mmol), Pd(OAc)2 (70 mg, 0.31 mmol), X-Phos (150 mg, 0.31 mmol), Cs2CO3 (4.51 g, 13.90 mmol) and 20 mL of toluene were added, and the synthesis and post-treatment operations were performed with reference to compound 1-2 to obtain compound 8-2 (1.60 g, y = 86%). LCMS (ESI) m / z: 447.3 [M+H] + .

[0107] Step 3: {2-Ethoxy-4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1- yl]phenyl}acetic acid (8)

[0108] 100 mL single-necked flask, 8-2 (1.60 g, 3.53 mmol), sodium hydroxide (0.60 g, 15.00 mmol), 10 mL of purified water and 10 mL of ethanol were added, and the synthesis and post-treatment operations were performed with reference to compound 1 to obtain compound 8 (0.52 g, y = 35%). LCMS (ESI) m / z: 419.2 [M+H] + . 1H NMR (600 MHz, DMSO-d6) δ 11.70 (s, 1H), 10.23 (s, 1H), 9.78 (s, 1H), 8.39 (d, J = 8.5 Hz, 1H), 8.12 (s, 1H), 8.02 (t, J = 8.9 Hz, 2H), 7.64 (d, J = 7.5 Hz, 2H), 7.60 (t, J = 7.5 Hz, 1H), 6.93 (d, J = 8.0 Hz, 1H), 6.02 (d, J = 9.1 Hz, 2H), 5.42 (s, 1H), 3.96 (q, J = 7.0 Hz, 2H), 3.82 (s, 1H), 3.40 (t, J = 7.1 Hz, 3H), 3.15 (q, J = 8.1 Hz, 1H), 2.34 (s, 1H), 2.26 (s, 1H), 1.76 (s, 3H), 1.29 (t, J = 7.0 Hz, 3H).

[0109] Example 10: Synthesis of {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-2-(propan-2-yloxy)phenyl}acetic acid (9)

[0110] The synthetic route is as follows:

[0111] Step 1: [4-bromo-2-(propan-2-yloxy)phenyl]acetic acid ethyl ester (9-1)

[0112] Into a 250 mL three-necked flask, was placed compound I (2.00 g, 7.72 mmol), potassium carbonate (3.20 g, 23.15 mmol), bromoisopropane (1.04 g, 8.49 mmol) and 50 mL of acetonitrile. The synthesis and work-up procedure was referenced to compound 8-1 to give compound 9-1 (1.95 g, y = 84%). LCMS (ESI) m / z: 301.0 [M+H] + .

[0113] Step 2: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino} tetrahydro-1H-pyrrol-1-yl]-2- (propan-2-yloxy)phenyl}acetic acid ethyl ester (9-2)

[0114] 100 mL single necked flask, add free compound A (0.80 g, 3.32 mmol), compound 9-1 (1.20 g, 3.32 mmol), Pd(OAc)2(40 mg, 0.16 mmol), X-Phos (80 mg, 0.16 mmol), Cs2CO3(3.25 g, 9.96 mmol) and 20 mL of toluene, synthesis and work-up procedure refer to compound 1-2, get compound 9-2 (0.80 g, y = 52%). LCMS (ESI) m / z: 461.3 [M+H] + .

[0115] Step 3: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-2- (prop-2-yloxy)phenyl}acetic acid (9)

[0116] 100 mL single necked flask, add 9-2 (0.80 g, 1.70 mmol), sodium hydroxide (0.20 g, 5.20 mmol), purified water 10 mL and ethanol 10 mL, synthesis and work-up procedure refer to compound 1, get compound 9 (0.50 g, y = 68%). LCMS (ESI) m / z: 433.2 [M+H] + . 1 H NMR (600 MHz, DMSO-d6) δ 8.32 (d, J = 8.0 Hz, 1H), 7.92 (dd, J = 7.2, 2.3 Hz, 1H), 7.79 (d, J = 8.1 Hz, 1H), 7.74 (d, J = 7.2 Hz, 1H), 7.51 (qd, J = 7.2, 5.1 Hz, 3H), 6.87 (d, J = 8.7 Hz, 1H), 5.94 (dq, J = 4.4, 2.3 Hz, 2H), 4.73 (d, J = 6.9 Hz, 1H), 4.50 - 4.42 (m, 1H), 3.29 (d, J = 4.1 Hz, 3H), 3.24 (t, J = 6.3 Hz, 1H), 3.20 (dd, J = 9.0, 6.6 Hz, 1H), 3.09 (dt, J = 9.1, 7.2 Hz, 1H), 2.92 (dd, J = 9.1, 5.5 Hz, 1H), 2.02 (dd, J = 12.2, 6.3 Hz, 1H), 1.87 (s, 1H), 1.40 (d, J = 6.6 Hz, 3H), 1.21 (d, J = 6.0 Hz, 6H).

[0117] Example 11: Synthesis of {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H- pyrrol-1-yl]-2-(propyloxy)phenyl}acetic acid (10)

[0118] The synthetic route is as follows:

[0119] Step 1: [4-bromo-2-(propyloxy)phenyl]acetic acid ethyl ester (10-1)

[0120] Into a 250 mL three-necked flask, was placed compound I (2.00 g, 7.72 mmol), potassium carbonate (3.20 g, 23.15 mmol), n-propyl bromide (1.04 g, 8.49 mmol) and 50 mL of acetonitrile. The synthesis and work-up procedure was referenced to compound 8-1 to give compound 10-1 (1.57 g, y = 68%). LCMS (ESI) m / z: 301.0 [M+H] + .

[0121] Step 2: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-2- (propyloxy)phenyl}acetic acid ethyl ester (10-2)

[0122] Into a 100 mL single-necked flask, was placed free compound A (1.04 g, 4.31 mmol), compound 10-1 (1.57 g, 5.20 mmol), Pd(OAc)2(50 mg, 0.22 mmol), X-Phos (100 mg, 0.22 mmol), Cs2CO3(4.25 g, 13.04 mmol) and 20 mL of toluene. The synthesis and work-up procedure was referenced to compound 1-2 to give compound 10-2 (0.45 g, y = 23%). LCMS (ESI) m / z: 461.3 [M+H] + .

[0123] Step 3: {4-[(3S)-3-{[(1R)-1-(naphthalen-1-yl)ethyl]amino}tetrahydro-1H-pyrrol-1-yl]-2- (propyloxy)phenyl}acetic acid (10)

[0124] Into a 100 mL single-necked flask, was placed 10-2 (0.45 g, 0.98 mmol), sodium hydroxide (0.12 g, 3.00 mmol), purified water 10 mL and ethanol 10 mL. The synthesis and work-up procedure was referenced to compound 1 to give compound 10 (0.38 g, y = 90%). LCMS (ESI) m / z: 433.2 [M+H] +1H NMR (600 MHz, DMSO-d6) δ 8.33 (d, J = 8.3 Hz, 1H), 7.94 (d, J = 7.8 Hz, 1H), 7.82 (s, 1H), 7.76 (s, 1H), 7.52 (dt, J = 15.4, 8.8 Hz, 3H), 6.88 (d, J = 8.3 Hz, 1H), 5.95 (s, 2H), 4.78 (s, 1H), 3.84 (t, J = 6.3 Hz, 2H), 3.32 (s, 4H), 3.27 (s, 1H), 3.11 (q, J = 7.8 Hz, 1H), 2.99 (s, 1H), 2.06 (s, 1H), 1.94 (s, 1H), 1.68 (h, J = 6.7 Hz, 2H), 1.44 (s, 3H), 0.97 (t, J = 7.4 Hz, 3H).

[0125] Experimental Example 1: CaSR Allosteric Activation Effect in Vitro Experiment

[0126] CaSR belongs to the C family of G protein-coupled receptors (GPCRs). When extracellular Ca 2+ concentration increases. Therefore, the hCaSR-CHO cell line is used, and the intracellular Ca 2+ concentration is used as an index for detection. By detecting the change in intracellular Ca 2+ concentration, the allosteric activation effect of the compound on CaSR is evaluated. 2+

[0127] I. Experimental Steps:

[0128] 1. Preparation before plating: 12 h before plating, the 384-well cell culture plate is coated with 0.1 mg / mL polylysine.

[0129] 2. Cell plating: The digested CaSR-CHO cells are resuspended with complete medium, counted with a cell counter, and the cell density is adjusted to 5 x 10 5 cells / mL. The cell suspension is inoculated in each well at a volume of 25 μL. After cell inoculation, the cells are incubated in a 37°C, 5% CO2 incubator for 18 h.

[0130] ​3. Preparation of test compound solution: Dilute the test compound to the required concentration (10 μM, 2 μM, 400 nM, 80 nM, 16 nM, 3.2 nM and 0.64 nM) and add to the 384-well plate, ready for use. Here, since there are 25 μL of Fluo-4 NW detection reagent in the cell culture plate to be tested, the instrument sets the dosing system to 20 μL, so the concentration of the compound in the sample plate at this time is 2.25 times the actual test concentration, i.e. the concentration of 10 μM compound in the sample well is 22.5 μM, and so on.

[0131] 4. Preparation of detection reagent: According to the requirements of the reagent kit Fluo-4 NW Calcium Assay Kits (Thermo, F36206), add 1 mL of assay buffer to Component B to prepare a 250 mM probenecid solution; then take 100 μL of the probenecid solution and add it to 10 mL of assay buffer, mix well and then add it to Component A to prepare a 1X dye loading solution ready for use.

[0132] 5. Add fluorescent dye: Discard the complete medium, use the multidrop automatic dispenser to add 25 μL of 1X dye loading solution to the 384-well plate, place it in the dark at 37°C for 30 min, transfer it to room temperature and place it for another 30 min, and then transfer it to the Fliper Tetra (Molecular Devices) instrument.

[0133] 6. Read the raw experimental data: Add 20 μL of test compound solution, set the excitation wavelength to 470-495 nm, detect the emission light in the range of 515-575 nm, and after the instrument quality control plate is read and determined to be correct, start the detection, with a continuous detection time length of 660 seconds, and after reading is completed, export the values.

[0134] 7. Data processing: Subtract the minimum fluorescence value (MIN, taken as the average of 14-21 seconds) from the maximum fluorescence value (MAX, taken as the average of 40-80 seconds) after the addition of the drug concentration to obtain the fluorescence intensity change value, which represents the intracellular calcium signal change. Using GraphPad Prism 8.0, plot the drug concentration on the horizontal axis and the fluorescence intensity change on the vertical axis, and calculate the EC 50 value of each compound.

[0135] II. Experimental results:

[0136] The in vitro efficacy results of some of the compounds of the present application are shown in Table 1.

[0137] Table 1. Cell efficacy experiment data of compounds

[0138] Experimental Example 2: In vivo experiment of inhibiting PTH concentration in model rats

[0139] A SD male rat hyperparathyroidism model was constructed by resection of 5 / 6 kidney + special feed induction, and parathyroid hormone (PTH) in blood was used as a detection index. The change in PTH concentration in rat plasma after administration of the compound was detected by ELISA enzyme-linked immunoassay, and was used to evaluate the in vivo activity of the compound.

[0140] I. Experimental steps:

[0141] 1. Model construction: SPF level 7-8 week old male SD rats were used. Normal feed was adaptively fed for 1 week; 2 / 3 of the left kidney was resected and normal feed was recovered for 1 week; the right kidney was resected and normal feed was recovered for 1 week; special feed (calcium content 0.5% and total phosphorus content 1.2%) was fed for 2 weeks.

[0142] 2. Detection and grouping: about 300 μL of blood was collected from the tail vein into a coagulation tube, which was placed at room temperature for 30 minutes, centrifuged at 12000 rpm for 3 minutes, and serum samples were obtained, and the concentration of full-length PTH (1-84) in the serum was detected by MicroVue Intact PTH EIA-96Test (Quidel, catalog No. 60-2500); according to the PTH concentration level of the rats, the rats in each group had substantially the same mean PTH concentration;

[0143] 3. Dosing and activity testing: a certain dose of compound was administered by single gavage, and blood samples before and after administration were collected, and serum was obtained by the above method, and PTH content was detected by ELISA. According to the method, it was confirmed that the compound had the effect of reducing the PTH level in the blood of the model rats.

[0144] II. Experimental results:

[0145] The effect of the compounds of the present application on reducing the PTH of the model rats is shown in Table 2.

[0146] Table 2. In vivo efficacy experiment data of compounds

[0147] Note: compared with the PTH concentration at 0h, the efficiency of reducing PTH at 1h after administration.

[0148] Experimental Example 3: Rat liver microsome stability test

[0149] Incubate rat liver microsomes, testosterone (positive control), tolbutamide (internal standard), test compound and PBS (pH 7.4) at a certain concentration at 37℃, quench with methanol at 0, 5, 10, 20, 30 and 60 minutes, pretreat the samples, and detect by LC-MS method, plot the data (test compound peak area / internal standard peak area as ordinate and time as abscissa), and calculate t 1 / 2 The test results of some compounds of the present application are shown in Table 3.

[0150] Table 3. Rat liver microsomal stability experiment data of compounds

[0151] Note: The liver microsomal stability test is affected by the activity of liver microsomes, the longest incubation time in vitro is 60 min, and t 1 / 2 > 60 min is estimated to have high metabolic stability in vivo.

[0152] Experimental Example 4: SD rat pharmacokinetic experiment

[0153] I. Experimental steps:

[0154] 1) Animal information: SD normal rats, 7-8 weeks old, male, 3-5 rats / group

[0155] 2) Dosing information: fast for more than 12 hours before dosing, single gavage.

[0156] 3) Blood sampling points: tail vein blood sampling before dosing and at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after dosing, 100 μL blood sample was collected at each sampling point.

[0157] 4) Sample processing and storage: after blood sampling, transfer to 1.5 mL EP tube, 50 μL per tube, 2 tubes per sample, complete centrifugation within 2 hours, whole blood samples are stored in ice bath before centrifugation, centrifugation conditions: 4000 r / min, 2-8℃, 10 min. After aspirating the supernatant, store at -80℃ if not detected immediately.

[0158] 5) Detection

[0159] a) Column: Infinitylab poroshell 120SB C18 2.1*100mm, 2.7μm; column temperature: 30℃; flow rate: 0.3ml / min; run time: 9.5min; post-run time: 2min; injection volume: 2μL; mobile phase: methanol-0.1% formic acid. Gradient elution is performed according to the following table:

[0160] b) Mass spectrometry conditions:

[0161] c) Preparation of stock solution, working solution sample:

[0162] All prepared stock solutions were stored in -80 °C refrigerator, working solution was placed in 4 °C refrigerator. Evocalcet stock solution (0.2 mg / mL): precisely weighed 10 mg of the product into a 50 ml volumetric flask, dissolved with methanol and made up to the mark, shaken well, and obtained. (Compound stock solution preparation is the same as Evocalcet)

[0163] Preparation of standard solution: accurately measured Evocalcet, compound 2 and compound 3 stock solution, diluted with 50% methanol, prepared standard solution with concentration of 80, 200, 1000, 5000, 10000, 20000 and 40000 ng / mL.

[0164] Preparation of internal standard clenbuterol hydrochloride solution: precisely weighed 10 mg of clenbuterol hydrochloride into a 10 ml volumetric flask, dissolved with methanol and made up to the mark, shaken well, and obtained clenbuterol hydrochloride stock solution (1.0 mg / mL). Accurately measured the stock solution, diluted with methanol to 2000 ng / mL of internal standard working solution.

[0165] d) Sample processing process:

[0166] Standard curve sample processing: take 90 μL of blank matrix, add 10 uL of each standard series solution, 50 μL of internal standard solution, vortex mix for 30 s, add 350 μL of methanol to precipitate the protein solution, vortex mix, centrifuge the sample at 12000 rpm for 10 min, take 200 μL of supernatant, add 600 μL of water to mix, take the supernatant for sample analysis.

[0167] Blood sample processing: take 50 μL of plasma sample, add 25 μL of internal standard solution, vortex mix for 30 s, add 175 μL of methanol to precipitate the protein solution, vortex mix, centrifuge the sample at 12000 rpm for 10 min, take 100 μL of supernatant, add 300 μL of water to mix, take the supernatant for sample analysis.

[0168] II. Experimental results

[0169] The results are shown in Table 4.

[0170] Table 4

Claims

1. A compound having a structure according to Formula (I): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof. wherein R 1 、R 2 、R 3 、R 4 、R 5 are each independently selected from a hydrogen atom; C 2-6 Carboxylic acid group; C 1-7 Alkoxy, which is substituted by a substituent selected from the group consisting of halogen, cyano, nitro, oxygen, C 1-6 Carboxylic acid group, amino group, C 3-6 Cycloalkyl, aryl C 1-6 Alkyloxy, hydroxyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl and halogenated C 1-6 Alkoxy; Halogen; C 1-6 Alkyl; Halogenated C 1-6 alkyl; provided that: 1) R 1 , R 2 , R 3 , R 4 , R 5 are not simultaneously H; 2) when the substituent is a C 2-6 carboxylic acid group, the substitution site is R 2 -R 4 , and at least 2 of R 1 -R 5 are not H.

2. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein, R 1 , R 5 is selected from H, halogen, C 1-6 1-6alkyl, C 1-6 1-6alkoxy.

3. The compound or pharmaceutically acceptable salt, stereoisomer, or tautomer thereof of claim 1 or 2, wherein, R 2 -R 4 at least one of R 2-6 is unsubstituted or substituted C 1 -R 5 at least two of R 2 -R 4 at least one of R 1 -R 5 at least two of R 4. The compound or pharmaceutically acceptable salt, stereoisomer, or tautomer thereof of any one of claims 1 to 3, wherein, R 1 , R 2 , R 4 , R 5 are each independently selected from the group consisting of H, halogen, C 1-6 1-6alkyl, C 1-6 1-6alkoxy, which is substituted by a substituent selected from the group consisting of hydroxy, C 1-6 1-6alkoxy, and R 1 , R 2 , R 4 , R 5 are not simultaneously H; R 3 is C 2-6 carboxylic acid group; R 1 , R 2 , R 4 , R 5 are each independently selected from the group consisting of H, halogen, C 1-6 1-6alkyl, C 1-6 1-6alkoxy, which is substituted by a substituent selected from the group consisting of hydroxy, C 1-6 1-6alkoxy, and R 1 , R 2 , R 4 , R 5 are not simultaneously H; R 3 is acetoxy.

5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein, The compound of formula (I) is selected from 6. A method of making a compound of formula (I) as claimed in any one of claims 1 to 5, the method comprising the step of reacting a compound of formula A: ###0002### with a compound of formula (II): ###0003### to form a compound of formula (I): ###0004### (I) ###0005### (II) A wherein X represents a leaving group selected from halogen, hydroxy, lower alkylsulfonyloxy (e.g. C 1-6 alkylsulfonyloxy), sulfonyloxy and the like, preferably Br, Cl, trifluoromethylsulfonyloxy.

7. A pharmaceutical composition comprising a compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient.

8. Use of a compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, or a pharmaceutical composition of claim 6, for the manufacture of a medicament for the prevention and / or treatment of a disease associated with CaSR.

9. Use according to claim 8, wherein, The disease is selected from the group consisting of primary hyperparathyroidism, secondary hyperparathyroidism, tertiary hyperparathyroidism, chronic renal failure, chronic kidney disease, parathyroid adenoma, parathyroid hyperplasia, parathyroid carcinoma, vascular calcification and valve calcification, abnormal calcium homeostasis, abnormal phosphorous homeostasis, bone-related diseases or complications caused by hyperparathyroidism, chronic kidney disease or parathyroid carcinoma, bone loss after kidney transplantation, osteitis fibrosa cystica, adynamic bone disease, renal osteodystrophy, cardiovascular complications caused by hyperparathyroidism or chronic kidney disease, Ca 2+ abnormally high levels of certain malignancies.

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