Medicinal salt of cucurbituril derivative, crystal form and application thereof

Abstract

The invention relates to a medicinal salt of a cucurbituril derivative, and a crystal form and application thereof. Specifically, the invention provides pharmaceutically acceptable salts, crystal forms and preparation methods of the compound shown in the formula 1, and the corresponding salts have good stability and can be better used for clinical treatment.

Description

Technical Field

[0001] This disclosure belongs to the field of pharmaceutical technology and relates to a pharmaceutically acceptable salt, crystalline form, and uses of a cucurbituril derivative. Background Technology

[0002] Muscle relaxation is one of the three essential elements of general anesthesia. While muscle relaxants meet the needs of endotracheal intubation and surgery, they also bring safety risks—residual muscle relaxation. This can lead to subjective discomfort in patients and a series of pulmonary complications such as hypoxemia, regurgitation, and aspiration. To reduce the incidence of residual muscle relaxation, measures are often taken, including the use of intermediate- or short-acting muscle relaxants, optimized intraoperative muscle relaxation management, postoperative antagonism of muscle relaxant effects, and objective perioperative muscle relaxation monitoring.

[0003] Postoperative muscle relaxant antagonism refers to the use of muscle relaxant antagonists to reverse the residual effects of non-depolarizing muscle relaxants. Currently, commonly used muscle relaxant antagonists can be broadly classified into two categories: competitive muscle relaxant antagonists, including neostigmine as an acetylcholine inhibitor; and selective muscle relaxant antagonists, including sugambogia sodium as a steroid muscle relaxant antagonist and cysteine ​​as a benzylisoquinoline muscle relaxant antagonist.

[0004] PCT / CN2023 / 131007 discloses a non-closed-ring CB[n] type molecular container with a cucurbituril structure, having the following structure. It can bind efficiently to benzyl isoquinoline and steroid muscle relaxants, and by covering the quaternary ammonium sites of benzyl isoquinoline and steroid muscle relaxants, it prevents the muscle relaxants from binding to neuromuscular cholinergic receptors, thereby rapidly reversing the muscle relaxant effect.

[0005] Salt formation can improve certain undesirable physicochemical or biological properties of drugs. Developing salts with superior physicochemical or pharmaceutical properties compared to compounds of Formula 1 is of great significance. Given the importance of the crystal form and stability of solid drugs in clinical treatment, in-depth research into the polymorphisms of pharmaceutically viable salts of Formula 1 compounds is also crucial for developing drugs suitable for industrial production and possessing good biological activity. Summary of the Invention

[0006] This disclosure provides a pharmaceutically acceptable salt of a compound of formula 1, wherein the pharmaceutically acceptable salt is selected from sodium salts, potassium salts, calcium salts, choline salts, ethanolamine salts, diethanolamine salts, diethylamine salts, tromethamine salts, arginine salts, lysine salts, meglumine salts, and ammonium salts.

[0007]

[0008] This disclosure also provides a method for preparing a pharmaceutically acceptable salt of a compound of formula 1, comprising the step of reacting the compound of formula 1 with a base, wherein the base is selected from sodium hydroxide, potassium hydroxide, calcium hydroxide, choline hydroxide, tromethamine, arginine, lysine, ethanolamine, diethylamine, meglumine, diethanolamine, and ammonia.

[0009] The solvents used in the salt formation of this disclosure are selected from one or more of tetrahydrofuran, methanol, ethanol, isopropanol, acetone, ethyl acetate, acetonitrile, isopropyl acetate, methyl tert-butyl ether, 2-butanone, 2-methyltetrahydrofuran, methyl isobutyl ketone, dichloromethane, cyclohexane, 1,4-dioxane, n-heptane, water, isopropyl ether, and dimethyl sulfoxide.

[0010] Furthermore, in an optional embodiment, the method for preparing the aforementioned pharmaceutically usable salt also includes steps such as crystallization, filtration, washing, or drying.

[0011] In an optional embodiment, the chemical ratio of the compound of Formula 1 to the base is 4:1 to 1:4, including but not limited to 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, and 1:4.

[0012] In another embodiment, the chemical ratio of the compound of formula 1 to the base is 2:1 to 1:4.

[0013] In an optional embodiment, the chemical ratio of the compound of Formula 1 to sodium ions is 1:2 or 1:4.

[0014] In some embodiments, the sodium salt of Formula 1 is amorphous, and its X-ray powder diffraction pattern has no obvious characteristic peaks in the diffraction angle 2θ range of 3-50°.

[0015] The sodium salt crystal form I of the compound of formula 1 provided in this disclosure has characteristic peaks at 8.586, 11.983, 15.016, 17.701, 19.507, and 21.120 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0016] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form I of the compound of formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 8.586, 10.524, 11.983, 13.711, 15.016, 15.976, 17.701, 19.507, and 21.120.

[0017] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form I of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 2 As shown.

[0018] This disclosure also provides a method for preparing sodium salt of Formula 1, crystal form I, the method comprising adding sodium salt of Formula 1 to 10% water / isopropanol and stirring.

[0019] The sodium salt crystal form II of the compound of formula 1 disclosed herein has characteristic peaks at 12.098, 18.586, 20.928, 24.268, and 27.646 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0020] In some embodiments, the sodium salt crystal form II of the compound of formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles of 2θ, has characteristic peaks at 8.528, 12.098, 13.135, 14.824, 18.586, 20.928, 22.464, 24.268, and 27.646.

[0021] In some embodiments, the sodium salt crystal form II of the compound of formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 8.528, 12.098, 13.135, 14.824, 17.243, 18.586, 20.928, 22.464, 23.654, 24.268, and 27.646.

[0022] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form II of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 3 As shown.

[0023] This disclosure also provides a method for preparing sodium salt of Formula 1, crystal form II, the method comprising adding sodium salt of Formula 1 to 10% water / acetone and stirring.

[0024] The sodium salt crystal form III of the compound of formula 1 disclosed herein has characteristic peaks at 6.027, 16.935, 18.087, 20.391, 23.001, and 24.232 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0025] In some embodiments, the sodium salt crystal form III of the compound of formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles of 2θ, has characteristic peaks at 6.027, 12.905, 13.979, 16.014, 16.935, 18.087, 20.391, 23.001, and 24.232.

[0026] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form III of the compound of formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 6.027, 12.905, 13.979, 16.014, 16.935, 18.087, 20.391, 21.619, 21.926, 23.001, 24.232, and 26.072.

[0027] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form III of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 4 As shown.

[0028] This disclosure also provides a method for preparing sodium salt of Formula 1, crystal form III, the method comprising the steps of adding sodium salt of Formula 1 to water / methanol (v / v = 1:3) and stirring.

[0029] The sodium salt crystal form IV of the compound of formula 1 provided in this disclosure has characteristic peaks at 5.940, 10.678, 12.290, 14.139, 20.122, and 21.926 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0030] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form IV of the compound of Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.940, 7.914, 10.678, 12.290, 14.139, 15.745, 20.122, 21.926, and 25.458.

[0031] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form IV of the compound of Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.940, 7.914, 10.678, 12.290, 14.139, 15.745, 17.243, 17.818, 20.122, 21.926, 23.922, and 25.458.

[0032] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form IV of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 5 As shown.

[0033] This disclosure also provides a method for preparing sodium salt of Formula 1 crystal form IV, the method comprising dissolving sodium salt of Formula 1 in water and adding ethanol or isopropanol while stirring.

[0034] The sodium salt crystal form V of the compound of formula 1 provided in this disclosure has characteristic peaks at 5.905, 10.333, 13.309, 16.436, 21.981, and 25.496 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0035] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form V of the compound of Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.905, 6.444, 10.333, 12.222, 13.309, 14.901, 16.436, 18.855, 20.045, 21.981, and 25.496.

[0036] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form V of the compound of formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.646, 5.905, 6.444, 8.292, 10.333, 12.222, 13.309, 14.325, 14.901, 16.436, 17.934, 18.855, 20.045, 21.981, 23.577, 25.496, 25.995, and 30.295.

[0037] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form V of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 6 As shown.

[0038] This disclosure also provides a method for preparing sodium salt crystal form V of compound of formula 1, the method comprising the step of heating sodium salt crystal form IV of compound of formula 1 to 170°C.

[0039] The sodium salt crystal form VI of the compound of formula 1 provided in this disclosure has characteristic peaks at 5.013, 11.062, 13.645, 18.855, 21.427, and 24.921 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0040] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form VI of the compound of formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.013, 6.724, 10.332, 11.062, 13.645, 16.475, 17.473, 18.855, 19.546, 20.544, 21.427, 24.038, 24.921, and 27.915.

[0041] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form VI of the compound of formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.013, 6.724, 9.104, 10.332, 11.062, 12.751, 13.645, 15.093, 16.475, 17.473, 18.855, 19.546, 20.544, 21.427, 21.965, 24.038, 24.921, 25.573, 26.610, and 27.915.

[0042] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form VI of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 7 As shown.

[0043] This disclosure also provides a method for preparing sodium salt crystal form VI of compound 1, the method comprising the steps of subjecting sodium salt crystal form IV of compound 1 to DVS, 25°C, humidity variation of 50%-95%-0-95%-50% RH, in 10% increments (5% in the final step), dm / dt≤0.002%, and repeating the process.

[0044] The sodium salt crystal form VII of the compound of formula 1 provided in this disclosure has characteristic peaks at 5.824, 10.625, 12.332, 13.919, 19.889, and 21.543 in its X-ray powder diffraction pattern expressed as a diffraction angle of 2θ.

[0045] In some embodiments, the sodium salt crystal form VII of the compound of formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles of 2θ, has characteristic peaks at 5.824, 7.797, 9.409, 10.625, 12.332, 13.919, 15.495, 17.632, 19.889, and 21.543.

[0046] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form VII of the compound of formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.824, 7.797, 9.409, 10.625, 12.332, 13.919, 15.495, 17.632, 19.889, 21.543, 22.468, and 26.741.

[0047] In some embodiments, the X-ray powder diffraction pattern of the sodium salt crystal form VII of compound 1, expressed as a diffraction angle 2θ, is as follows: Figure 8 As shown.

[0048] This disclosure also provides a method for preparing sodium salt crystal form VII of compound 1, wherein the method is selected from any of the following methods:

[0049] Method 1: Place the sodium salt crystal form IV of compound 1 under 92.5% RH conditions for 1 month.

[0050] Method 2: Disperse a small amount of sodium salt crystal type VII seed crystals of Formula 1 in ethanol, add an aqueous solution of sodium salt of Formula 1, and stir.

[0051] In some embodiments, this disclosure also provides a method for preparing an amorphous sodium salt of compound formula 1, said method being selected from any of the following methods:

[0052] Method 1: Add the sodium salt of compound 1 to solvent I and stir. Solvent I is selected from one of the following: tetrahydrofuran, methanol, ethanol, isopropanol, acetone, ethyl acetate, acetonitrile, isopropyl acetate, methyl tert-butyl ether, 2-butanone, tetrahydrofuran, 2-methyltetrahydrofuran, methyl isobutyl ketone, dichloromethane, cyclohexane, 1,4-dioxane, n-heptane, 10% water / methanol, 7% water / ethanol, EA / ethanol (1:1), and isopropyl ether.

[0053] Method 2: Dissolve the sodium salt of compound 1 in water, add solvent II, and stir. Solvent II is selected from methanol, acetone, acetonitrile, and tetrahydrofuran.

[0054] Method 3: Dissolve the sodium salt of compound 1 in solvent III and evaporate the solvent, wherein solvent III is selected from water and dimethyl sulfoxide.

[0055] This disclosure also provides a pharmaceutical composition comprising the aforementioned sodium salt crystal form I, sodium salt crystal form II, sodium salt crystal form III, sodium salt crystal form IV, sodium salt crystal form V, sodium salt crystal form VI, sodium salt crystal form VII, or amorphous sodium salt or any of the aforementioned pharmaceutically acceptable salts, and a pharmaceutical excipient optionally selected from pharmaceutically acceptable excipients.

[0056] This disclosure also provides a pharmaceutical composition prepared from the aforementioned sodium salt crystal form I, sodium salt crystal form II, sodium salt crystal form III, sodium salt crystal form IV, sodium salt crystal form V, sodium salt crystal form VI, sodium salt crystal form VII, or amorphous sodium salt or any of the aforementioned pharmaceutically acceptable salts, and optionally a pharmaceutically acceptable excipient.

[0057] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned sodium salt crystal form I, sodium salt crystal form II, sodium salt crystal form III, sodium salt crystal form IV, sodium salt crystal form V, sodium salt crystal form VI, sodium salt crystal form VII, or sodium salt amorphous or any of the aforementioned pharmaceutically acceptable salts, with a pharmaceutically acceptable excipient.

[0058] This disclosure also provides the use of the aforementioned sodium salt crystal form I, sodium salt crystal form II, sodium salt crystal form III, sodium salt crystal form IV, sodium salt crystal form V, sodium salt crystal form VI, sodium salt crystal form VII, or amorphous sodium salt, or any of the aforementioned pharmaceutically acceptable salts, or the aforementioned compositions, in the preparation of medicaments for reversing drug-induced neuromuscular blockade and / or anesthesia.

[0059] The "2θ or 2θ angle" mentioned in this disclosure refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degree; the error range of 2θ for each characteristic peak is ±0.20 (including the case where the number has more than one decimal place after rounding), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0060] The numerical values ​​in this disclosure, such as those relating to the content of certain substances, are calculated data and inevitably contain a certain degree of error. Generally, ±10% is within the reasonable error range. The error may vary to some extent depending on the context in which it is used, but this variation shall not exceed ±10%, and may be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%, preferably ±5%.

[0061] The starting material used in the crystal form preparation method disclosed herein can be any form of compound, including but not limited to: amorphous, arbitrary crystal form, hydrate, solvate, etc.

[0062] The drying temperature described in this disclosure is generally 25℃-100℃, preferably 40℃-70℃, and can be dried under normal pressure or reduced pressure.

[0063] The crystallization methods described in this disclosure include room temperature crystallization, cooling crystallization, solvent evaporation crystallization, and seed crystallization induction. The cooling temperature is selected from below 65°C, preferably from -10°C to 60°C. Stirring can also be performed during the crystallization process.

[0064] The “differential scanning calorimetry or DSC” described in this disclosure refers to measuring the temperature difference and heat flow difference between the sample and the reference material during the sample heating or isothermal process, in order to characterize all physical and chemical changes related to thermal effects and obtain phase transition information of the sample.

[0065] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "Guiding Principles on Hygroscopicity of Drugs" in Part IV of the 2015 edition of the Chinese Pharmacopoeia,

[0066] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0067] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0068] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0069] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0070] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0071] The “excipients” described in this disclosure include, but are not limited to, any adjuvants, carriers, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock. Attached Figure Description

[0072] Figure 1 The TOF 0.9 time for cisatracurium administration of muscle relaxant antagonists was as follows: *p<0.05, **p<0.01, ***p<0.001.

[0073] Figure 2 XRPD spectrum of sodium salt crystal form I of compound 1.

[0074] Figure 3 XRPD spectrum of sodium salt form II of compound 1.

[0075] Figure 4 XRPD spectrum of sodium salt of compound 1, crystal form III.

[0076] Figure 5 XRPD spectrum of sodium salt of compound 1, crystal form IV.

[0077] Figure 6 XRPD spectrum of sodium salt crystal form V of compound 1.

[0078] Figure 7 XRPD spectrum of sodium salt crystal form VI of compound 1.

[0079] Figure 8 XRPD spectrum of sodium salt of compound VII of formula 1.

[0080] Figure 9 XRPD spectrum of the amorphous sodium salt of compound 1. Detailed Implementation

[0081] The present disclosure will be explained in more detail below with reference to embodiments or experimental examples. The embodiments or experimental examples in the present disclosure are only used to illustrate the technical solutions in the present disclosure and are not intended to limit the substance and scope of the present disclosure.

[0082] Test conditions of the instruments used in the experiment:

[0083] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ ohms. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE NEO 500M NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0084] MS measurements were performed using an Agilent 1200 / 1290DAD-6110 / 6120 Quadrupole MS LC-MS system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD system (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), or a THERMO Ultimate3000-Q Exactive system (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0085] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1200DAD, an Agilent HPLC 1200VWD, and a Waters HPLCe2695-2489 HPLC system.

[0086] High performance liquid chromatography (HPLC) was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson-281 preparative chromatographs.

[0087] Silica gel column chromatography generally uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0088] XRPD (X-ray Powder Diffraction) was used for analysis: measurements were performed using a BRUKER D8 X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα1 rays. Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2θ range): 5°~45°.

[0089] DSC stands for Differential Scanning Calorimetry: Measurements were performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10℃ / min. The specific temperature range was referenced from the corresponding spectra (mostly 25-350℃), and the nitrogen purging rate was 50mL / min.

[0090] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer, with a heating rate of 10℃ / min, and the specific temperature range was referenced from the corresponding spectrum (mostly 25-350℃). The nitrogen purging rate was 50mL / min.

[0091] DVS stands for Dynamic Moisture Adsorption: The detection method is SMSDVS Advantage. At 25℃, the humidity changes from 50% to 95% to 0% to 95% to 50%, with a step of 10% (the last step is 5%). (The specific humidity range is subject to the corresponding spectrum. The methods listed here are the most commonly used methods.) The judgment standard is that dm / dt is not greater than 0.002%.

[0092] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0093] Unless otherwise specified in the examples, all reactions can be carried out under an argon or nitrogen atmosphere.

[0094] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0095] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system used for column chromatography to purify the compounds, and the developing solvent system for TLC included: A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system. The volume ratio of the solvent was adjusted according to the polarity of the compounds, and small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0096] Example 1

[0097]

[0098]

[0099] Step 1: Urea (38.89 g, 647.46 mmol), 0.3 M dilute hydrochloric acid (80 mL), and 1,2-cyclohexanedione 1a (22.0 g, 196.20 mmol) were added to a 500 mL three-necked flask. The mixture was heated to 50 °C and stirred for 16 hours. The reaction mixture was cooled to room temperature, filtered, and the filter cake was washed with 100 mL of water and 100 mL of anhydrous ethanol. After drying, cyclohexylglycerol, compound 1b (pale yellow body, 27.4 g, yield: 71%) was obtained.

[0100] MSm / z(ESI):197.1[M+1]+.

[0101] 1H NMR (400MHz, DMSO-d6): δ7.02 (s, 4H), 1.72-1.68 (m, 4H), 1.42-1.35 (m, 4H).

[0102] Step 2: Add compound 1b (27.4 g, 139.65 mmol), 140 mL of 9 M hydrochloric acid, and paraformaldehyde (20.9 g, 698.23 mmol) to a 1 L three-necked flask. Stir the reaction mixture at room temperature for 24 hours. Add 500 mL of water to the reaction system and continue stirring at room temperature for 16 hours. Filter the reaction mixture, wash, and dry to obtain cyclohexylglycerol diether and compound 1c (white solid, 20.2 g, yield: 52%).

[0103] MS(ESI):281.1[M+1]+.

[0104] 1H NMR (400MHz, DMSO-d6): δ5.20(d,4H,J=11.6), 4.91(d,4H,J=11.2), 2.25-2.18(m,4H), 1.56-1.50(m,4H).

[0105] Step 3: Compound 1c (2.73 g, 9.73 mmol) was weighed into a dry three-necked flask, purged with argon, dissolved in methanesulfonic acid (10 mL), and then glycourea dimer 1d (1 g, 3.24 mmol, prepared using the known method "WO2012051407A2") was added. The mixture was stirred at room temperature for 24 hours. The reaction solution was slowly added to 100 mL of water (cooled in an ice-water bath), and then brought to room temperature. The mixture was filtered. 1.77 g of crude product was obtained by drying. The crude product was heated and dissolved in TFA (4 mL), then 16 mL of water was added, stirred, filtered, and dried under vacuum to obtain compound 1e (1.21 g, yield: 44.9%).

[0106] MS(ESI):837.3[M+1]+.

[0107] 1H NMR (400MHz, CDCl3): δ5.72~5.37(m,10H),5.15(d,4H),4.75(d,4H),4.15~4.11(m,6H),2.28(br,4H),2.05(br,4H),1.45(br,8H).

[0108] Step 4: Weigh compound 1e (1.06 g, 1.27 mmol) into a dry three-necked flask, replace with argon gas, add TFA (10 mL) to dissolve, then add sodium 3,3'-(naphthalene-1,4-dioxy)bis(propane-1-sulfonic acid) and compound 1f (1.42 g, 3.17 mmol, prepared by the known method "WO2012051407A2"). After the addition is complete, heat the reaction to 60 °C and stir for 3 hours. Evaporate the TFA under reduced pressure, add 20 mL of ethanol to the resulting solid, heat under reflux for 2 hours, cool to room temperature, and filter. The filter cake was washed with ethanol and dried. The resulting solid was dissolved in 10 mL of water by heating, and then 30 mL of ethanol was added. The mixture was filtered, and the filter cake was then purified by high performance liquid chromatography (column: SharpSil-T, 30*150 mm, 5 μm; mobile phase: aqueous phase and acetonitrile, gradient ratio: aqueous phase 25%-42%) to obtain compound 1 (0.27 g, yield: 12.6%).

[0109] MSm / z (negative ion ESI): 801.0 [M / 2-1] -

[0110] 1 H NMR (400MHz, D2O): δ7.85-7.61(m,4H),7.37-6.99(m,4H),5.58-5.42(m,6H),5.32-5.10(m,8H), 4.42-4.25(m,4H),4.17-3.81(m,14H),3.20-3.05(m,8H),2.28-1.92(m,16H),1.56-1.19(m,8H).

[0111] Test Example 1. Test of the in vitro binding activity of the disclosed compounds against muscle relaxants.

[0112] I. Experimental Objective:

[0113] The in vitro binding activity of the disclosed compound with the muscle relaxant was tested by isothermal titration calorimetry (ITC), and the Kd value of the binding was evaluated.

[0114] II. Experimental Materials:

[0115] Isothermal titration calorimeter (including computer host and supporting software), CB2, compound 1, cisatracurium ammonium, deionized water.

[0116] III. Experimental Methods and Procedures:

[0117] Using a preset washing program, wash the tubing, sample cell, and titration needle of the isothermal titration calorimetry system with deionized water. Prepare an aqueous solution of cisatracurium ammonium and an aqueous solution of the analyte, with a cisatracurium ammonium:analyte ratio of 10:1 to 20:1. Fill the sample cell with CB2 and the analyte using the dispensing needle, and then pipette cisatracurium ammonium into the titration needle using the dispensing program. Place the titration needle into the sample cell, start stirring, and allow the system to equilibrate for 5–10 minutes. Set the titration parameters: 2.5 μL of sample per drop, 20 drops in total, and a titration interval of 150 s. Begin the titration and record the thermal curve.

[0118] After titration, the Kd value of the combination of the test compound and cisatracurium is calculated by thermal curve. The tubing, sample cell and titration needle are then washed using a washing program before testing the next compound.

[0119] CB2 (prepared using the known method "WO2012051407A2") has the following structure:

[0120]

[0121] IV. Test Results:

[0122] The binding capacity of CB2 and compound 1 to cisatracurium in vitro was tested using the ITC method. The dissociation constant Kd values ​​are shown in Table 1. The results show that compound 1 has a stronger ability to bind cisatracurium in vitro than CB2.

[0123] Table 1: In vitro binding affinity of the test compounds to cisatracurium

[0124] Test compound <![CDATA[K d (M)]]> CB2 2.506E-5 Compound 1 7.475E-6

[0125] Test Example 2. Test on the antagonistic effect of the disclosed compound on muscle relaxants.

[0126] I. Experimental Objective

[0127] The effects of the compounds disclosed herein on the reversal of muscle relaxant effects in the gastrocnemius muscle were tested in a rat neuromuscular model. Onset time, TOF and other indicators were evaluated and compared with CB2 and neostigmine.

[0128] II. Experimental Materials

[0129] SPF grade male SD rats, BL-420A biological function experimental system (main unit, stimulator, tension transducer), ventilator, electronic scale, surgical instruments, syringe, clippers, electronic scale, iron stand, foam board, urethane, sodium chloride, CB2, compound 1, cisatracurium, succinylcholine, neostigmine, sterile water, 95% alcohol.

[0130] III. Experimental Methods

[0131] SPF-grade male SD rats that have passed quarantine were used and housed in an environment with a room temperature of 22±0.5℃, an air exchange rate of 20-50 / h, an airflow velocity of 0.05-0.18m / s, and a 12 / 12-hour day / night light / dark cycle. The animals were allowed to acclimatize in the facility for at least 3 days, with 6 rats per cage, until their weight reached 220g-250g before the start of the experiment.

[0132] The test sample was reconstituted using 0.9% sodium chloride. After calculating the content, the required amount of test sample was weighed, dissolved in 0.9% sodium chloride injection (within 30 minutes), and then mixed thoroughly using a vortex mixer. CB2 was reconstituted with purified water.

[0133] Actual drug weight (mg) = Dosage concentration A (mg / mL) × Solvent volume (mL) / Content (%)

[0134] Rats were randomly divided into CB2, compound 1, and neostigmine groups, with 5 rats in each group (based on the actual number of rats enrolled). The drug volume was 2 mL / kg for each group.

[0135] CB2 (prepared using the known method "WO2012051407A2") has the following structure:

[0136]

[0137] After anesthetizing rats, the sciatic nerve and gastrocnemius muscle were isolated. The sciatic nerve was stimulated, and muscle tone signals were recorded using a tension transducer. The rats were then intubated and mechanically ventilated using a small animal ventilator. After a period of stable muscle tone recording, the drug was administered: the initial dose was twice the ED. 90 Administer a dose (0.8 mg / kg) of muscle relaxant (cistracurium). The muscle tone curve should decrease at this point. Inject the antagonist (test sample and neostigmine) 30-60 seconds after administration. Once the muscle tone curve recovers to above 95%, administer the ED. 90 When administering a muscle relaxant (succinylcholine) at a dose of 0.9 mg / kg, a decrease in muscle tone should be observed. The experiment can be stopped once muscle tone naturally recovers to above 95%. Muscle tone signals should be continuously recorded during the experiment, and indicators such as onset time and clinical efficacy should be statistically analyzed. By comparing muscle tone signals after administration, the reversal effect of the antagonist can be determined and compared.

[0138] IV. Experimental Procedure

[0139] 4.1 Weighing and anesthetizing rats

[0140] The rats were weighed and, once they were in a stable mood, 25% urethane was prepared from ethyl carbamate and anesthetized by intraperitoneal injection at a rate of 1 mL / 100 g. After the pain reflex disappeared, the rats were fixed in a prone position on a foam board, and hair was removed from the buttocks and the outer side of the right thigh.

[0141] 4.2 Separation of the sciatic nerve

[0142] Behind the hip joint, make an incision along the outer edge of the femur in the middle of the thigh, lift the skin and superficial fascia, bluntly dissect the muscles to expose the sciatic nerve. Be careful to use a glass dissecting needle during dissection to prevent damage to the nerve with metal instruments.

[0143] 4.3 Separation of the gastrocnemius muscle

[0144] Cut open the skin of the lower leg at the ankle joint, cut the ligaments at the front of the ankle joint, separate the gastrocnemius muscle, tie a suture at the gastrocnemius tendon at the ankle, and cut the tendon at the distal end of the suture.

[0145] 4.4 Signal Collection

[0146] The gastrocnemius muscle ligation suture was connected to the tension transducer, and the stimulator was connected to the sciatic nerve. The input signal was set to tension, and the parameters were set as square wave, fine voltage, series stimulation, delay 0.05ms, pulse width 0.2ms, frequency 2Hz, intensity 0.225±0.025V, intensity increment 0, series length 4, main period 12s, and number of stops 30000. The muscle contraction curve was recorded. During the measurement, the muscle nerve was kept moist with physiological saline every 3-5 minutes.

[0147] 4.5 Connect to ventilator and administer muscle relaxants and antagonists.

[0148] Wipe the ventilator tubing with alcohol, make an incision in the neck skin to locate the jugular vein and trachea, cut the trachea, connect the ventilator, and set the parameters as follows: tidal volume 6 mL, respiratory rate to exhalation ratio 5:4, and respiratory rate 80 breaths / min. After stabilizing for approximately 5 minutes, administer the drug via the jugular vein. Based on the reference settings, administer 2 times the initial dose. 90 After administering the appropriate dose of muscle relaxant (cistracurium), the muscle tone curve should decrease. Inject the antagonist (test sample or neostigmine) 30-60 seconds later. Once the muscle tone curve recovers to over 95%, administer the ED. 90 After administering the appropriate dose of muscle relaxant (succinylcholine), wait until the muscle tone curve recovers to above 95% before stopping the experiment. Continue recording the muscle tone curve throughout the experiment.

[0149] Table 2

[0150]

[0151] 4.6 Statistics on Timeliness Indicators

[0152] The onset time and secondary muscle relaxation onset time were statistically analyzed using a biological function experimental system, and the statistical standards are as follows.

[0153] 1) Time to 90% recovery of TOF (TOF 0.9): The time for the T4 / T1 value of TOF string stimulation to recover to approximately 90% - the time of antagonist administration.

[0154] 2) Duration of muscle relaxation by succinylcholine: Time to reduce muscle tone to its lowest value - Time to administer succinylcholine

[0155] V. Experimental Conclusions

[0156] Depend on Figure 1 It can be seen that compound 1 can achieve the efficacy level of neostigmine at 20 mg / kg, which is superior to CB2.

[0157] Example 2: Preparation of sodium salt of compound 1

[0158] Compound 1 (2.2 g, 1.0 eq) was added to pure water (11 ml, 5 V) and stirred until dissolved. Then, 5.2 mL of 2 M sodium hydroxide solution was added dropwise, and the mixture was stirred for 1 hour after the addition was complete. Next, DMF (32.6 ml, 14.8 V) was added dropwise, during which a solid precipitated. After the addition was complete, the mixture was stirred for 18 hours. The filter cake was collected and dried under reduced pressure at 40 °C to constant weight to obtain 1.0 g of product. X-ray powder diffraction analysis showed that the product was amorphous, and the XRPD spectrum is shown below. Figure 9 Ion analysis results showed that the sodium ion content was 5.4%.

[0159] Example 3: Amorphous Preparation of Sodium Salt

[0160] 125 mg of the sodium salt prepared in Example 2 was added to 2 mL of tetrahydrofuran, stirred at room temperature, centrifuged, and dried under vacuum to obtain the title product. X-ray powder diffraction analysis showed that the product was amorphous.

[0161] Example 4: Amorphous Preparation of Sodium Salt

[0162] Add 5 mg of the sodium salt prepared in Example 2 to 0.5 mL of the solvent in Table 3, stir at room temperature for 2 days, centrifuge, and dry the solid under vacuum to obtain the title product.

[0163] Table 3

[0164]

[0165]

[0166] Example 5: Amorphous Preparation of Sodium Salt

[0167] 5 mg of the sodium salt prepared in Example 2 was dissolved in 0.05 mL of water, and then added to solvent 1 in Table 4. The mixture was stirred at room temperature to crystallize, centrifuged, and dried under vacuum to obtain the title product.

[0168] Table 4

[0169]

[0170] Example 6: Preparation of amorphous sodium salts

[0171] 5 mg of the sodium salt prepared in Example 2 was dissolved in the solvent in Table 5, and the solvent was evaporated at room temperature to obtain the title product.

[0172] Table 5

[0173] solvent Solvent volume result water 0.05mL Sodium salts are amorphous Dimethyl sulfoxide 0.1mL Sodium salts are amorphous

[0174] Example 7 Preparation of Sodium Salt Crystal Form I

[0175] Five mg of the sodium salt prepared in Example 2 was added to 0.1 mL of 10% water / isopropanol, stirred at room temperature for 2 days, centrifuged, and vacuum dried to obtain a solid. X-ray powder diffraction analysis identified the product as sodium salt crystal form I, and the XRPD spectrum is shown below. Figure 2 The positions of its characteristic peaks are shown in Table 6. The DSC spectrum shows that the endothermic peak has a peak value of 80.16℃. The TGA spectrum shows that the weight loss is 12.40% from 30℃ to 220℃.

[0176] Table 6

[0177]

[0178] Example 8 Preparation of Sodium Salt Crystal Form II

[0179] Five mg of the sodium salt prepared in Example 2 was added to 0.1 mL of 10% water / acetone, stirred at room temperature for 2 days, centrifuged, and vacuum dried to obtain a solid. X-ray powder diffraction analysis identified the product as sodium salt crystal form II. The XRPD spectrum is shown below. Figure 3 The positions of its characteristic peaks are shown in Table 7. The TGA spectrum shows a weight loss of 10.51% between 30℃ and 222℃.

[0180] Table 7

[0181]

[0182] Example 9 Preparation of Sodium Salt Crystal Form III

[0183] 5 mg of the sodium salt prepared in Example 2 was added to 1 mL of water / methanol (1:3), stirred, filtered, and the supernatant was allowed to evaporate at room temperature, resulting in crystal precipitation. The crystals were then centrifuged and dried under vacuum to obtain a solid. X-ray powder diffraction analysis identified the product as sodium salt crystal form III. The XRPD spectrum is shown below. Figure 4 The positions of its characteristic peaks are shown in Table 8.

[0184] Table 8

[0185]

[0186]

[0187] Example 10 Preparation of Sodium Salt Crystal Form IV

[0188] 150 mg of the sodium salt prepared in Example 2 was dissolved in 1.5 mL of water, and 4.5 mL of ethanol was added. The mixture was stirred at room temperature to induce crystallization, centrifuged, and vacuum dried to obtain a solid. X-ray powder diffraction analysis identified the product as sodium salt crystal form IV. The XRPD spectrum is shown below. Figure 5 The positions of its characteristic peaks are shown in Table 9. The TGA spectrum shows a weight loss of 7.64% between 30℃ and 168℃.

[0189] Table 9

[0190]

[0191] Example 11 Preparation of Sodium Salt Crystal Form IV

[0192] 5 mg of the sodium salt prepared in Example 2 was dissolved in 0.05 mL of water, and solvent 1 from Table 10 was added. The mixture was stirred at room temperature to crystallize, centrifuged, and dried under vacuum to obtain the title product.

[0193] Table 10

[0194]

[0195] Example 12 Preparation of sodium salt crystal form V

[0196] The sodium salt crystal form IV from Example 10 was heated to 170°C to obtain the title product. X-ray powder diffraction analysis identified this product as sodium salt crystal form V, and the XRPD spectrum is shown below. Figure 6 The positions of its characteristic peaks are shown in Table 11. The TGA spectrum shows a weight loss of 5.16% at 30℃-264℃.

[0197] Table 11

[0198]

[0199] Example 13 Preparation of Sodium Salt Crystal Form VI

[0200] The title product was obtained by testing the sodium salt crystal form IV under DVS at 50%-95%-0-95%-50% RH.

[0201] X-ray powder diffraction analysis determined the product to be sodium salt crystal form VI. The XRPD spectrum is shown below. Figure 7 The positions of its characteristic peaks are shown in Table 12. The TGA spectrum shows a weight loss of 14.72% between 31℃ and 264℃.

[0202] Table 12

[0203]

[0204]

[0205] Example 14 Preparation of sodium salt crystal form VII

[0206] The sodium salt crystal form IV was placed under 92.5% RH conditions for 1 month to obtain the title product.

[0207] X-ray powder diffraction analysis determined the product to be sodium salt crystal form VII. The XRPD spectrum is shown below. Figure 8 The positions of its characteristic peaks are shown in Table 13. The TGA spectrum shows a weight loss of 8.85% between 31℃ and 164℃.

[0208] Table 13

[0209]

[0210] Example 15 Preparation of sodium salt crystal form VII

[0211] A small amount of sodium salt crystal form VII from Example 14 was added to 0.3 ml of ethanol; 10 mg of the sodium salt of the compound shown in Formula 1 was dissolved in 0.1 ml of water, and the aqueous solution was added to the ethanol solution. The mixture was stirred overnight at room temperature, centrifuged, and the solid was dried under vacuum to obtain the title product.

[0212] Example 16: Study on Crystal Form Stability

[0213] Sodium salt crystal form IV and amorphous samples were laid out in open containers to investigate their stability under high temperature (60℃), high humidity (RH 92.5%), and light conditions, respectively. The sampling period was 14 or 30 days.

[0214] Table 14

[0215]

[0216] Conclusion: The influencing factor experiment showed that sodium salt crystal form IV exhibited good physical and chemical stability after 14 days under high temperature (60℃) and high humidity (92.5% RH) conditions; and good physical stability but slightly poor chemical stability after 14 days under light conditions. Sodium salt amorphous form exhibited good physical stability but poor chemical stability after one month under high temperature (60℃) and light conditions; and poor physical stability but good chemical stability after one month under high humidity (92.5% RH) conditions.

[0217] Example 17: Long-term / Accelerated Stability

[0218] The stability of sodium salt crystal form IV and amorphous form was investigated under conditions of 25℃ / 60%RH and 40℃ / 75%RH, respectively.

[0219] Table 15

[0220]

[0221]

[0222] Conclusion: Long-term accelerated experiments show that sodium salt crystal form IV and amorphous form have good physical and chemical stability under long-term (25℃ / 60%RH) and accelerated (40℃ / 75%RH) conditions for 14 days.

Claims

1. A pharmaceutically acceptable salt of the compound shown in Formula 1, said pharmaceutically acceptable salt being selected from sodium salts, potassium salts, calcium salts, choline salts, ethanolamine salts, diethanolamine salts, diethylamine salts, tromethamine salts, arginine salts, lysine salts, meglumine salts, and ammonium salts.

2. The medicinal salt according to claim 1, characterized in that, The chemical ratio of the compound of Formula 1 to the base is 4:1-1:8, preferably 4:1-1:4, and more preferably 1:2 or 1:

4.

3. The method for preparing the pharmaceutically acceptable salt according to any one of claims 1-2, comprising the step of reacting the compound of formula 1 with a base, wherein the base is selected from sodium hydroxide, potassium hydroxide, calcium hydroxide, choline hydroxide, tromethamine, arginine, lysine, ethanolamine, diethylamine, meglumine, diethanolamine, and ammonia.

4. A sodium salt crystal form IV of the compound shown in Formula 1, characterized in that, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 5.940, 10.678, 12.290, 14.139, 20.122, and 21.926, preferably at 5.940, 7.914, 10.678, 12.290, 14.139, 15.745, 20.122, 21.926, and 25.458, and more preferably at 5.940, 7.914, 10.678, 12.290, 14.139, 15.745, 17.243, 17.818, 20.122, 21.926, 23.922, and 25.

458.

5. The sodium salt crystal form IV according to claim 4, characterized in that, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 5.

6. A method for preparing sodium salt crystal form IV as described in claim 4 or 5, the method comprising the steps of dissolving the compound of formula 1 in water and adding ethanol or isopropanol while stirring.

7. A sodium salt crystal form VII of the compound shown in Formula 1, characterized in that, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 5.824, 10.625, 12.332, 13.919, 19.889, and 21.543, preferably at 5.824, 7.797, 9.409, 10.625, 12.332, 13.919, 15.495, 17.632, 19.889, and 21.543, and more preferably at 5.824, 7.797, 9.409, 10.625, 12.332, 13.919, 15.495, 17.632, 19.889, 21.543, 22.468, and 26.

741.

8. The sodium salt crystal form VII according to claim 7, characterized in that, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 8.

9. A method for preparing sodium salt crystal form VII as described in claim 7 or 8, wherein the method is selected from any of the following methods: Method 1: Place the sodium salt crystal form IV of compound 1 under 92.5% RH conditions for 1 month. Method 2: Disperse a small amount of sodium salt crystals of Formula 1 (type VII) in ethanol and add an aqueous solution of sodium salt of Formula 1.

10. The crystal form according to any one of claims 4-5 and 7-8, wherein the 2θ angle error range is ±0.

20.

11. A pharmaceutical composition comprising the salt of any one of claims 1-2, or the crystal form of any one of claims 4-5, 7-8, and optionally a pharmaceutically acceptable excipient.

12. A method for preparing a pharmaceutical composition, comprising the following steps: The step of mixing the salt according to any one of claims 1-2, or the crystal form according to any one of claims 4-5, 7-8, and a pharmaceutically acceptable excipient.

13. Use of the salt of any one of claims 1-2, or the crystal form of any one of claims 4-5, 7-8, or the pharmaceutical composition of claim 11 in the preparation of a medicament for reversing drug-induced neuromuscular blockade and / or anesthesia.

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