A class of environment-sensitive α1-adrenergic receptor near-infrared fluorescent ligands and their applications

Abstract

The present invention relates to a class of environment-sensitive α1-adrenergic receptor near-infrared fluorescent ligands and applications thereof, the general formula of which is (I), wherein: R1 is the pharmacophore quinazoline or phenylpiperazine; R2 is a different substituent; X1 is a different type of linker (carbon chain or triazole ring); n1 means that the carbon number is 1-6, and X2 is iodine or bromide ion. The fluorescent ligand molecule shows a high affinity to the α1-adrenoceptor, and the fluorescence will be significantly enhanced due to the decrease of the polarity or the increase of the viscosity of the surrounding environment. Upon binding to α1‑adrenergic receptors, the molecule enters a hydrophobic environment and releases a strong fluorescent signal, allowing for easier visualization and a better signal-to-noise ratio. The fluorescent ligand can be used as a tool drug for the research on the pharmacology and physiological characteristics of α1-adrenergic receptors. In addition, the preparation method of this type of compound is mature, the reaction condition is mild, the raw material is cheap and easy to obtain, and the operation and post-treatment are convenient.

Description

technical field

[0001] The invention belongs to the technical field of medicines, in particular to a class of environment-sensitive alpha 1 - Adrenergic receptor near-infrared fluorescent probe and its 1 Applications in pharmacology and physiology studies such as adrenergic receptor cell imaging, subcellular localization and expression level detection. Background technique

[0002] G-protein-coupled receptors with seven transmembrane structures are the largest family of cell surface receptors, and their coding genes account for about 4% of the human protein-coding genome [Zhang, R.; Xie, X. Tools for GPCR drug discovery. ActaPharmacol. Sin. 2012, 33, 372-84.]. As multifunctional molecular machines, G-protein-coupled receptors regulate most physiological responses mediated by hormones and neurotransmitters and are the target of nearly 26% of marketed prescription drugs. Although gratifying progress has been made in the chemical and structural biology of G-protein coupled r...

Examples

example 1

[0030] Example 1: Synthesis of PTC-1~3

[0031]

[0032] Reagents and conditions: (a) 3-iodopropionic acid, acetonitrile, reflux, 48h, 81%; (b) methyl iodide, acetonitrile, reflux, 12h, 86%; (c) hydrochloric acid-N-(3-benzene Amino-2-propenylidene)aniline, AcOH / AC 2 O, 120℃, 4h; Pyridine / AcOH, 120℃, 4h; 30% two steps; (d) propargylamine, EDCI, DMAP, dichloromethane, r.t., 6h, 53%; (e) piperazine, H 2 O, 100°C, 81%; (f) acid chloride, TEA, acetonitrile, 1h, 0°C, 62-75%; (g) NaN 3 , DMSO, 100℃, 6h, 35-88%; (h) CuSO4, sodium ascorbate, tBuOH / H 2 O=2:1, 50°C, 1h, 48.9-67.3%.

[0033] Intermediate C3: 1-(2-carboxyethyl)-2,3,3-trimethyl-3H-indole-1-iodonium

[0034] 2,3,3-Trimethyl-3H-indole (1.0g, 6.23mmol) and 3-iodopropionic acid (1.86g, 9.3mmol) were dissolved in 15mL of acetonitrile, refluxed for 48 hours, and cooled to room temperature. After the solvent was spin-dried, 100 mL of ethyl acetate was slowly added under ultrasonic vibration, and a large amount of precipita...

example 2

[0061] Example 2: Synthesis of PTC-4~6

[0062]

[0063] Reagents and conditions: (a) H 2 O / acetonitrile,NaN 3 , 80℃, 20h, 43-74%; (b) TsCl, triethylamine, chloroform, r.t., 24h, 65-88%; (c) K 2 CO 3 , acetonitrile, 90°C, 64-69%; (d) CuSO4, sodium ascorbate, tBuOH / H 2 O=2:1, 50°C, 1h, 55.2-72.5%.

[0064] Intermediate 11a: 2-Azidoethanol

[0065] NaN 3 (3.12g, 48mmol) was carefully added to a mixed solution of 2-bromoethanol (2g, 16mmol) in acetonitrile (10mL) and water (40mL), stirred vigorously at 80°C for 20h, then cooled to room temperature. The reaction solution was extracted with dichloromethane (3×50 mL), dried over anhydrous magnesium sulfate, filtered, and the solvent was distilled off to obtain 0.7 g of a transparent oil with a yield of 50%. 1 HNMR (300MHz, CDCl3): δ3.80 (td, 2H, J 1 =4.8Hz,J 2 = 3.0Hz), 3.27(t, 2H, J = 5.4Hz), 1.96(s, 1H).

[0066] Intermediate 11b: 3-Azido-1-propanol

[0067] Synthesis of intermediate 11b Using 3-bromo-1-propanol as r...

example 3

[0088] Example three: the synthesis of PHC-1~3

[0089]

[0090] Reagents and conditions: (a) K 2 CO 3 , acetonitrile, 90°C, 73-96%; (b) CuSO4, sodium ascorbate, tBuOH / H 2 O=2:1, 50°C, 1h, 49.4-57.7%.

[0091] Intermediate PH1: 1-(2-azidoethyl)-4-(2-methoxyphenyl)piperazine

[0092] To the acetonitrile solution (30 mL) of 1-(2-methoxyphenyl)piperazine (665 mg, 3.46 mmol) and 11a (918 mg, 3.8 mmol) was added 1.43 g of potassium carbonate, and the mixture was refluxed for 24 h, then filtered immediately to obtain the filtrate. The solvent was evaporated and purified by silica gel column chromatography to obtain 690 mg of yellow oil with a yield of 76%. 1 HNMR (300MHz, CDCl 3 ): δ7.02-6.84(m, 4H), 3.80(s, 3H), 3.41(t, 2H, J=6.0Hz), 3.11(m, 4H), 2.73-2.65(m, 6H); ESI -MS:m / z[M+H] + calcd for C 13 h 20 N 5 o + 262.2, found 262.4.

[0093] Intermediate PH2: 1-(2-azidopropyl)-4-(2-methoxyphenyl)piperazine

[0094] Synthesis of intermediate P7 Using intermediate 10b (...

technical field

[0001] The invention belongs to the technical field of medicines, in particular to a class of environment-sensitive alpha 1 - Adrenergic receptor near-infrared fluorescent probe and its 1 Applications in pharmacology and physiology studies such as adrenergic receptor cell imaging, subcellular localization and expression level detection. Background technique

[0002] G-protein-coupled receptors with seven transmembrane structures are the largest family of cell surface receptors, and their coding genes account for about 4% of the human protein-coding genome [Zhang, R.; Xie, X. Tools for GPCR drug discovery. ActaPharmacol. Sin. 2012, 33, 372-84.]. As multifunctional molecular machines, G-protein-coupled receptors regulate most physiological responses mediated by hormones and neurotransmitters and are the target of nearly 26% of marketed prescription drugs. Although gratifying progress has been made in the chemical and structural biology of G-protein coupled r...

Examples

example 1

[0030] Example 1: Synthesis of PTC-1~3

[0031]

[0032] Reagents and conditions: (a) 3-iodopropionic acid, acetonitrile, reflux, 48h, 81%; (b) methyl iodide, acetonitrile, reflux, 12h, 86%; (c) hydrochloric acid-N-(3-benzene Amino-2-propenylidene)aniline, AcOH / AC 2 O, 120℃, 4h; Pyridine / AcOH, 120℃, 4h; 30% two steps; (d) propargylamine, EDCI, DMAP, dichloromethane, r.t., 6h, 53%; (e) piperazine, H 2 O, 100°C, 81%; (f) acid chloride, TEA, acetonitrile, 1h, 0°C, 62-75%; (g) NaN 3 , DMSO, 100℃, 6h, 35-88%; (h) CuSO4, sodium ascorbate, tBuOH / H 2 O=2:1, 50°C, 1h, 48.9-67.3%.

[0033] Intermediate C3: 1-(2-carboxyethyl)-2,3,3-trimethyl-3H-indole-1-iodonium

[0034] 2,3,3-Trimethyl-3H-indole (1.0g, 6.23mmol) and 3-iodopropionic acid (1.86g, 9.3mmol) were dissolved in 15mL of acetonitrile, refluxed for 48 hours, and cooled to room temperature. After the solvent was spin-dried, 100 mL of ethyl acetate was slowly added under ultrasonic vibration, and a large amount of precipita...

example 2

[0061] Example 2: Synthesis of PTC-4~6

[0062]

[0063] Reagents and conditions: (a) H 2 O / acetonitrile,NaN 3 , 80℃, 20h, 43-74%; (b) TsCl, triethylamine, chloroform, r.t., 24h, 65-88%; (c) K 2 CO 3 , acetonitrile, 90°C, 64-69%; (d) CuSO4, sodium ascorbate, tBuOH / H 2 O=2:1, 50°C, 1h, 55.2-72.5%.

[0064] Intermediate 11a: 2-Azidoethanol

[0065] NaN 3 (3.12g, 48mmol) was carefully added to a mixed solution of 2-bromoethanol (2g, 16mmol) in acetonitrile (10mL) and water (40mL), stirred vigorously at 80°C for 20h, then cooled to room temperature. The reaction solution was extracted with dichloromethane (3×50 mL), dried over anhydrous magnesium sulfate, filtered, and the solvent was distilled off to obtain 0.7 g of a transparent oil with a yield of 50%. 1 HNMR (300MHz, CDCl3): δ3.80 (td, 2H, J 1 =4.8Hz,J 2 = 3.0Hz), 3.27(t, 2H, J = 5.4Hz), 1.96(s, 1H).

[0066] Intermediate 11b: 3-Azido-1-propanol

[0067] Synthesis of intermediate 11b Using 3-bromo-1-propanol as r...

example 3

[0088] Example three: the synthesis of PHC-1~3

[0089]

[0090] Reagents and conditions: (a) K 2 CO 3 , acetonitrile, 90°C, 73-96%; (b) CuSO4, sodium ascorbate, tBuOH / H 2 O=2:1, 50°C, 1h, 49.4-57.7%.

[0091] Intermediate PH1: 1-(2-azidoethyl)-4-(2-methoxyphenyl)piperazine

[0092] To the acetonitrile solution (30 mL) of 1-(2-methoxyphenyl)piperazine (665 mg, 3.46 mmol) and 11a (918 mg, 3.8 mmol) was added 1.43 g of potassium carbonate, and the mixture was refluxed for 24 h, then filtered immediately to obtain the filtrate. The solvent was evaporated and purified by silica gel column chromatography to obtain 690 mg of yellow oil with a yield of 76%. 1 HNMR (300MHz, CDCl 3 ): δ7.02-6.84(m, 4H), 3.80(s, 3H), 3.41(t, 2H, J=6.0Hz), 3.11(m, 4H), 2.73-2.65(m, 6H); ESI -MS:m / z[M+H] + calcd for C 13 h 20 N 5 o + 262.2, found 262.4.

[0093] Intermediate PH2: 1-(2-azidopropyl)-4-(2-methoxyphenyl)piperazine

[0094] Synthesis of intermediate P7 Using intermediate 10b (...

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