An anesthetic drug composition and its application

Abstract

The present invention relates to a pharmaceutical composition of a GABA receptor agonist of different isomers, a preparation method thereof, and an application in the field of central nervous system. The GABA receptor agonist is 2-(1-cyclopropylethyl)-6-isopropyl-phenol (Compound 1) or its stereoisomers, and its pharmaceutically acceptable salts. A A ​

Description

Technical Field

[0001] The present invention relates to a GABA with different isomers A The pharmaceutical composition of GABA receptor agonist and its preparation method and application in the field of central nervous system. A The receptor agonist is 2-(1-cyclopropylethyl)-6-isopropyl-phenol (Compound 1) or its stereoisomers, and pharmaceutically acceptable salts thereof, belonging to the field of biomedicine technology. Background Art

[0002] 2-(1-Cyclopropylethyl)-6-isopropyl-phenol (Compound 1) emulsion injection is an intravenous general anesthetic, clinically intended for digestive endoscopy diagnosis and anesthesia induction. The chemical structure of its main component, Compound 1, has a chiral center and contains two configurations, namely the (R)-configuration isomer 2-[(1R)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound A) and the (S)-configuration isomer 2-[(1S)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound B). Radiolabeled competitive binding assays and patch clamp functional tests found that the two configurations of Compound 1 can compete for GABA binding. A Receptor ion channel sites, directly stimulating or enhancing GABA A The agonist effect of the receptor induces chloride ion influx, thereby inducing inhibitory postsynaptic currents and achieving central inhibition, but the potency of compound A is significantly higher than that of its enantiomer compound B. [1] .

[0003] The loss of righting reflex test (LORR) is a classic test for testing the efficacy of anesthetics. [2][3] In view of the low potency of compound B, it is of great clinical value to find a drug that has a synergistic effect with it and can reduce its side effects, thereby improving its anesthesia effect. Summary of the Invention

[0004] To address the above issues, this study used the mouse LORR test to evaluate the synergistic effect of mixing two enantiomers, (R)-configurational isomer 2-[(1R)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound A) and (S)-configurational isomer 2-[(1S)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound B), in different proportions to improve the anesthetic efficacy.

[0005] The present invention provides a pharmaceutical composition comprising two or more GABA A Optical isomers of receptor agonists.

[0006] In some embodiments, the GABA AThe receptor agonist is an optical isomer of 2-(1-cyclopropylethyl)-6-isopropylphenol (Compound 1) and its pharmaceutically acceptable salts.

[0007] In some embodiments, the optical isomers are 2-[(1R)-1-cyclopropylethyl]-6-isopropylphenol (Compound A) and its pharmaceutically acceptable salts, and 2-[(1S)-1-cyclopropylethyl]-6-isopropylphenol (Compound B) and its pharmaceutically acceptable salts.

[0008] In some embodiments, the weight ratio of Compound A and its pharmaceutically acceptable salts to Compound B and its pharmaceutically acceptable salts is selected from (1-5):(1-5), (1-4):(1-4), (1-3):(1-3), or (1-2):(1-2).

[0009] In some embodiments, the weight ratio of Compound A and its pharmaceutically acceptable salts to Compound B and its pharmaceutically acceptable salts is selected from 5:1, 1:5, 4:1, 1:4, 3:1, 1:3, 2:1, 1:2, or 1:1.

[0010] The present invention also provides a pharmaceutical preparation, comprising the pharmaceutical composition of the above technical solution, and pharmaceutically acceptable excipients or auxiliary components.

[0011] In some embodiments, the preparation is selected from aqueous solution preparations, freeze-dried preparations, or fat emulsions.

[0012] The aqueous solution preparations, freeze-dried preparations, or fat emulsions are prepared by conventional preparation methods.

[0013] The aqueous solution preparations or freeze-dried preparations contain solubilizers, cosolvents, and further may optionally contain fillers, pH regulators, and isotonicity regulators.

[0014] The fat emulsions contain oily components and emulsifiers, and further may optionally contain solubilizers, cosolvents, pH regulators, isotonicity regulators, etc.

[0015] The solubilizers are selected from any one or any mixture in any proportion of Tween-80, Tween-20, polyoxyethylene 35 castor oil, polyoxyethylene 40 hydrogenated castor oil, polyethylene glycol 15 hydroxystearate (i.e., solutol HS15), or poloxamer;

[0016] The cosolvents are selected from any one or any mixture in any proportion of ethanol, glycerol, propylene glycol, or polyethylene glycol.

[0017] The filler described above is selected from any one or any mixture in any proportion of lactose, sucrose, glucose, mannitol, sodium dihydrogen phosphate, sodium phosphate, sodium chloride, disodium hydrogen phosphate, cysteine, glycine, sorbitol, calcium lactate, dextran or polyvinylpyrrolidone.

[0018] The pH regulator described above is selected from any one or any combination of sodium hydroxide, potassium hydroxide, triethanolamine, hydrochloric acid, phosphoric acid, citric acid, acetic acid, malic acid.

[0019] The isotonicity regulator described above is selected from any one or any mixture in any proportion of glycerol, sugars or sugar alcohols.

[0020] The oily component described above is selected from any one or any mixture in any proportion of natural or (and) synthetic oils that meet biocompatibility and can be metabolized in the human body.

[0021] The emulsifier described above is selected from any one or any mixture in any proportion of glyceryl monooleate, Tween-80, Tween-20, poloxamer, polyoxyethylene 35 castor oil, polyoxyethylene 40 hydrogenated castor oil, polyethylene glycol glycerol ester, polyethylene glycol 15 hydroxystearate, egg yolk lecithin, egg yolk phosphatidylcholine, soybean lecithin, soybean phosphatidylcholine, etc.

[0022] The present invention further provides a combined drug, which includes 2-[(1R)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound A) and its pharmaceutically acceptable salts and 2-[(1S)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound B) and its pharmaceutically acceptable salts with the same or different specifications for simultaneous or separate administration, as well as a pharmaceutically acceptable carrier.

[0023] In some embodiments, the weight ratio of Compound A and its pharmaceutically acceptable salts to Compound B and its pharmaceutically acceptable salts is selected from (1-5):(1-5), (1-4):(1-4), (1-3):(1-3) or (1-2):(1-2).

[0024] In some embodiments, the weight ratio of Compound A and its pharmaceutically acceptable salts to Compound B and its pharmaceutically acceptable salts is selected from 5:1, 1:5, 4:1, 1:4, 3:1, 1:3, 2:1, 1:2 or 1:1.

[0025] In addition, the present invention also provides the use of the pharmaceutical composition, pharmaceutical preparation and combined drug described in the above technical solutions in the preparation of drugs for inducing and maintaining anesthesia in animals or humans, promoting sedative hypnosis in animals or humans, treating and / or preventing anxiety, depression, insomnia, nausea, vomiting, migraine, schizophrenia, convulsion and epilepsy.

[0026] Through the mouse LORR test, the research results show that after compounds A and B are mixed in different weight ratios, the LORR maintenance time of the mixtures at all dose ratios is longer than that of compound A alone at the supposed dose ratio. Especially in the groups with dose ratios of 5:1, 4:1, 3:1, 2:1, and 1:2, the LORR maintenance time of the mixtures is longer than that of compound A at the same dose in the mixtures. Particularly in the case of the mixture of 5:1, it exceeds 1.5 times that of compound A alone; when the ratio is 2:1, at the same dose, its sedative effect fully reaches that of compound A alone. Description of the Drawings

[0027] Figure 1 . Hypnotic rate after single subcutaneous injection of subthreshold doses of different ratios of compounds A and B in mice.

[0028] Figure 2 . Duration of disappearance of righting reflex after single subcutaneous injection of anesthetic doses of different ratios of compounds A and B in mice. Detailed Embodiments

[0029] The following details the implementation process and beneficial effects of the present invention through specific embodiments, aiming to help readers better understand the essence and characteristics of the present invention, and shall not be construed as a limitation on the scope of implementation of this case.

[0030] The structure of the compound is determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS). NMR shifts (δ) are given in units of 10-6 (ppm). The NMR measurements are performed using (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic resonance spectrometers, and the solvents for the measurements are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), deuterated acetonitrile (CD3CN), and the internal standard is tetramethylsilane (TMS).

[0031] The MS measurements are performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI)).

[0032] The HPLC measurements are performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C18 100×4.6mm).

[0033] The thin-layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications of the silica gel plates used for thin-layer chromatography (TLC) are 0.15mm - 0.2mm, and the specifications used for thin-layer chromatography separation and purification of products are 0.4mm - 0.5mm.

[0034] Column chromatography generally uses silica gel with a mesh size of 200 - 300 from Yantai Huanghai Silica Gel as the carrier.

[0035] The known starting materials of the present invention can be synthesized by adopting or according to methods known in the art, or can be purchased from companies such as Titan Technology, Aladdin Chemistry, Shanghai Dermachem, Chengdu Kelong Chemical Industry, Shaoyuan Chemical Technology, J&K Scientific, etc.

[0036] Example 1. Preparation of Compound 1

[0037] First step: 2-(2-Isopropylphenoxy)tetrahydropyran (3B)

[0038] Add 2-isopropylphenol (3A) (10.00 g, 73.40 mmol), 3,4-dihydropyran (18.60 g, 220.20 mmol) and dichloromethane (50 mL) to the reaction flask. After stirring evenly, add pyridinium p-toluenesulfonate (1.86 g, 7.40 mmol). Stir at room temperature for 20 hours, add water (30 mL), extract with dichloromethane (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 500:1) to obtain 2-(2-isopropylphenoxy)tetrahydropyran (3B) in the form of a colorless liquid (13.4 g, yield: 82.71%, HPLC: 99.15%).

[0039] 1 HNMR (400 MHz, CDCl3): δ7.25 - 7.20 (m, 1Η), δ7.15 - 7.09 (m, 2H), δ6.97 - 6.93 (m, 1H), δ5.44 - 5.42 (m, 1H), δ3.94 - 3.88 (m, 1H), δ3.65 - 3.62 (m, 1H), δ3.39 - 3.22 (m, 1H), δ1.90 - 1.86 (m, 1H), δ1.73 - 1.67 (m, 2H), δ1.60 - 1.54 (m, 3H), δ1.25 (2d, 6H).

[0040] Second step: Cyclopropyl-(3-isopropyl-2-tetrahydropyran-2-yloxy-phenyl)methanone (3C)

[0041] Add 2-(2-isopropylphenoxy)tetrahydropyran (3B) (10.00 g, 45.40 mmol) and dry tetrahydrofuran (30 mL) to a reaction flask, protect with nitrogen, cool to -20 °C in a dry ice bath, add 2.5 M n-butyllithium (20.00 mL, 50.00 mmol). After addition, warm to room temperature and stir for 1 hour. Then cool to -20 °C in a dry ice bath again, add N-methoxy-N-methylcyclopropanecarboxamide (7.00 g, 54.20 mmol). After addition, warm to room temperature and stir for 2 hours. Add saturated ammonium chloride (30 mL) and stir for several minutes to terminate the reaction. Extract with ethyl acetate (30 mL × 3), wash with saturated brine (30 mL × 3), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain cyclopropyl-(3-isopropyl-2-tetrahydropyran-2-yloxy-phenyl)methanone (3C) as a red liquid (17.4 g, crude product, HPLC: 68.00%), which is directly used in the next step of the reaction.

[0042] Step 3: Cyclopropyl-(2-hydroxy-3-isopropyl-phenyl)methanone (3D)

[0043] Add (3-isopropyl-2-tetrahydropyran-2-yloxy-phenyl)methanone (3C) (17.40 g, crude product) and methanol (50 mL) to a reaction flask, cool to 0 °C in an ice bath, add 2 M hydrochloric acid aqueous solution (35 mL, 70.00 mmol). After addition, warm to room temperature and stir for half an hour. Add saturated sodium bicarbonate aqueous solution to adjust the pH to 6, rotary evaporate the methanol, extract with ethyl acetate (50 mL × 3), wash with saturated brine (50 mL × 3), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The residue is purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 300:1) to obtain cyclopropyl-(2-hydroxy-3-isopropyl-phenyl)methanone (3D) as a colorless liquid (7.23 g, two-step yield: 78.26%, HPLC: 96.29%).

[0044] MS m / z (ESI): 205.1 (M - 1).

[0045] 1 HNMR (400 MHz, DMSO -d6 ): δ 12.98 (s, 1H), δ 8.08 (dd, 1H), δ 7.51 (dd, 1H), δ 6.98 (t, 1H), δ 3.34 - 3.26 (m, 1H), δ 3.04 - 3.01 (m, 1H), δ 1.19 - 1.12 (m, 10H).

[0046] Step 4: 2-(1-cyclopropyl-1-hydroxy-ethyl)-6-isopropyl-phenol (3E)

[0047] Add cyclopropyl-(2-hydroxy-3-isopropyl-phenyl) methyl ketone (3D) (10 g, 48.80 mmol) and dry toluene (50 mL) to a reaction flask. Under nitrogen protection, cool to -30 °C in a dry ice bath, add 3M methylmagnesium bromide n-hexane solution (49.00 mL, 146.30 mmol). After addition, warm to room temperature and stir for 2 hours. Add saturated ammonium chloride (100 mL) to terminate the reaction. Extract with ethyl acetate (100 mL × 3), wash with saturated brine (100 mL × 3), dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 50:1) to obtain 2-(1-cyclopropyl-1-hydroxy-ethyl)-6-isopropyl-phenol (3E) as a light yellow liquid (10.2 g, yield: 95.17%, HPLC: 97.96%).

[0048] MS m / z(ESI): 219.1(M-l).

[0049] 1 HNMR(400MHz, CDCl3): δ7.32(dd, 1H), δ7.24(dd, 1H), δ7.13(t, 1H), δ4.64(s, 1H), δ3.43 - 3.36(m, 1H), δ1.56(s, 3H), δ1.37 - 1.31(m, 1H), δ1.27(d, 6H), δ0.54 - 0.39(m, 4H).

[0050] Step 5: 2-(1-cyclopropylethyl)-6-isopropylphenol (Compound 1)

[0051] Add 2-(1-cyclopropyl-1-hydroxy-ethyl)-6-isopropyl-phenol (3E) (3 g, 13.80 mmol), triethylsilane (6.42 g, 55.21 mmol) and dichloromethane (25 mL) to a reaction flask. Cool to -30 °C in a dry ice bath, slowly add trifluoroacetic acid (12.59 g, 110.40 mmol), control the temperature below 0 °C, stir the reaction for 2 hours and then stop the reaction. Extract with dichloromethane (100 mL × 3), wash with saturated brine (100 mL × 3), dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 100:1) to obtain 2-(1-cyclopropylethyl)-6-isopropylphenol (Compound 1) as a colorless liquid (2.02 g, yield: 71.63%, HPLC: 98.58%).

[0052] MS m / z(ESI): 203.1(M-l).

[0053] 1HNMR (400 MHz, CDCl3): δ 7.13 (dd, 1H), δ 7.08 (dd, 1H), δ 6.90 (t, 1H), δ 4.93 (s, 1H), δ 3.20 - 3.13 (m, 1H), δ 2.53 - 2.46 (m, 1H), δ 1.29 (d, 3H), δ 1.26 (d, 6H), δ 1.07 - 1.05 (m, 1H), δ 0.58 - 0.45 (m, 2H), δ 0.24 - 0.16 (m, 2H).

[0054] Example 2. Preparation of Compound A

[0055] At room temperature, 2-(1-cyclopropylviny)-6-isopropylphenol (2.0 g, 10 mmol) and dichloromethane (8 mL) were added to an autoclave (250 mL). A catalyst, ((4R,5R)-(+)-O-[1-benzyl-1-(5-methyl-2-phenyl-4,5-dihydrooxazol-4-yl)-2-phenylethyl](dicyclohexylphosphineimine)(1,5-COD)iridium(I)tetrakis(3,5-bis(trifluoromethyl)phenyl)borate (17 mg, 0.01 mmol), was added. The mixture was purged with hydrogen three times and then reacted at 40 atm for 6 hours. After concentration, the residue was purified by flash column chromatography (petroleum ether / ethyl acetate (v / v) = 10:1) to obtain 2-[(1R)-1-cyclopropylethyl]-6-isopropylphenol (Compound A) as a pale yellow oil (1.8 g, yield: 90%; chiral HPLC: 91.2%).

[0056] MS m / z (ESI): 203.1 (M - 1).

[0057] 1 HNMR (400 MHz, CDCl3): δ 7.12 (dd, 1H), δ 7.07 (dd, 1H), 6.89 (t, 1H), 6.85 (s, 1H), 3.20 - 3.13 (m, 1H), 2.53 - 2.46 (m, 1H), 1.30 (d, 6H), 1.27 (d, 3H), 1.08 - 1.03 (m, 1H), 0.58 - 0.56 (m, 1H), 0.47 - 0.46 (m, 1H), 0.22 - 0.16 (m, 2H).

[0058] Example 3. Preparation of Compound B

[0059] Add 2-(1-cyclopropylviny)-6-isopropylphenol (102 mg, 0.5 mmol) and dichloromethane (2 mL) to a 250 mL autoclave. Add the catalyst (4S,5S)-(-)-O-[1-benzyl-1-(5-methyl-2-phenyl-4,5-dihydrooxazol-4-yl)-2-phenylethyl](dicyclohexylphosphine)(1,5-COD)iridium(I)tetrakis(3,5-bis(trifluoromethyl)phenyl)borate (8.5 mg, 0.005 mmol). Replace with hydrogen three times and react at 10 atm for 2.5 h. Concentrate and perform flash column chromatography (petroleum ether / ethyl acetate (v / v)=10:1) to obtain 2-[(1S)-1-cyclopropylethyl]-6-isopropylphenol (Compound B) as a pale yellow oil (60 mg, yield: 59%; chiral HPLC: 97.3%).

[0060] MS m / z(ESI): 203.1(M-1).

[0061] 1 HNMR(400 MHz, CDCl3): δ7.12(dd, 1H), δ7.07(dd, 1H), 6.89(t, 1H), 6.85(s, 1H), 3.20 - 3.13(m, 1H), 2.53 - 2.46(m, 1H), 1.30(d, 6H), 1.27(d, 3H), 1.08 - 1.03(m, 1H), 0.58 - 0.56(m, 1H), 0.47 - 0.46(m, 1H), 0.22 - 0.16(m, 2H).

[0062] Biological test examples

[0063] Biological test example 1: Select a subthreshold dose of 1.0 mg / kg of Compound A and explore the effective dose for mouse anesthesia after mixing Compound A and Compound B in different ratios. After single intravenous injection of mixtures of Compound A and Compound B in different ratios, record the sedation level and adverse reactions of the mice.

[0064] The anesthetic and sedative effects of all subthreshold doses of mixtures of Compound A and Compound B in different ratios are weaker than those of 1.2 mg / kg of Compound A( Figure 1 ). However, when Compound A:Compound B = 2:1, the sedative effect is equivalent to that of 1 mg / kg of Compound A (sedation rate: 90%). The specific test result data are shown in Table 1.

[0065]

[0066] Table 1. Anesthetic and sedative effects of compositions with different ratios

[0067] Biological test example 2: Using the anesthetic dose of compound A (3 mg / kg), explore the anesthetic maintenance time and adverse reactions of compound A and compound B after being mixed in different ratios, and test whether the anesthetic maintenance time is prolonged. After a single intravenous injection of the single-configuration compound A at a dose of 3 mg / kg in mice, the LORR maintenance time was 200 s. After a single intravenous injection of the mixture of compound A and compound B with different ratios at the same dose, the LORR maintenance time of all dose ratios of the mixture was longer than that of the single compound A under this dose ratio, especially in the groups with dose ratios of 5:1, 4:1, 3:1, 2:1, and 1:2, the LORR maintenance time was longer than that of compound A in the mixture at the same dose (see Figure 2 ). The specific test result data are shown in Table 2.

[0068]

[0069]

[0070] Table 2: Anesthetic effects of compositions with different ratios of doses

[0071] Conclusion: After compound A and compound B are mixed in different weight ratios, the LORR maintenance time of all dose ratios of the mixture is longer than that of the single compound A under this dose ratio, especially in the groups with dose ratios of 5:1, 4:1, 3:1, 2:1, and 1:2, the LORR maintenance time is longer than that of compound A in the mixture at the same dose. Especially in the case of the mixture of 5:1, it exceeds 1.5 times that of compound A alone; when the ratio is 2:1, at the same dose, its sedative effect completely reaches the sedative effect of compound A alone.

[0072] References

[0073] [1] Linlin Qin.; Lei Ren.; Songlin Wan.; et al. Design, Synthesis, and Evaluation of Novel 2,6-Disubstituted Phenol Derivatives as General Anesthetics. J. Med. Chem. 2017, 60, 3606 - 3617;

[0074] [2]Hill-Venning, C.; Peters, J. A.; Callachan, H.; Lambert, J. J.; Gemmell, D. K.; Anderson, A.; Byford, A.; Hamilton, N.; Hill, D. R.; et al. The anesthetic action and modulation of GABAA receptor activity by the novel water-soluble aminosteroid Org 20599. Neuropharmacology 1996, 35, 1209-1222;

[0075] [3]Lingamaneni, R.; Krasowski, M. D.; Jenkins, A.; Truong, T.; Giunta, A. L.; Blackbeer, J.; MacIver, M. B.; Harrison, N. L.; Hemmings, H. C., Jr. Anesthetic properties of 4-iodopropofol: Implications for mechanism of anesthesia. Anesthesiology 2001; 94:1050–7。

Claims

1. Use of a pharmaceutical composition or its pharmaceutical preparation in the preparation of a drug for inducing and maintaining anesthesia in animals or humans, promoting sedation and hypnosis in animals or humans, treating and / or preventing anxiety, depression, insomnia, nausea, vomiting, migraine, schizophrenia, convulsions and epilepsy, characterized in that: The pharmaceutical composition contains optical isomers of two or more GABAA receptor agonists, The optical isomers are 2-[(1R)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound A) and its pharmaceutically acceptable salts and 2-[(1S)-1-cyclopropylethyl]-6-isopropyl-phenol (Compound B) and its pharmaceutically acceptable salts, The weight ratio of Compound A and its pharmaceutically acceptable salts to Compound B and its pharmaceutically acceptable salts is selected from 5:1, 1:5, 4:1, 1:4, 3:1 or 1:

3.

2. The application according to claim 1, characterized in that The pharmaceutical preparation is selected from aqueous solution preparations, freeze-dried preparations or fat emulsions.

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