WO2022078483A1

WO2022078483A1 - Quaternary ammonium steroid compound, preparation method therefor, preparation and use

Info

Publication number: WO2022078483A1
Application number: PCT/CN2021/123999
Authority: WO
Inventors: 李勤耕; 陈捷; 廖建; 申义; 董小虎; 谭波
Original assignee: 江苏恩华络康药物研发有限公司
Priority date: 2020-10-15
Filing date: 2021-10-15
Publication date: 2022-04-21
Also published as: CN116887840A

Abstract

A quaternary ammonium steroid compound represented by formula I, or an addition salt or a solvate of the quaternary ammonium steroid compound: wherein R 1 is H or CH 3CO; R 2 is CH 3 or -CH 2CH＝CH 2, and t is equal to 2 or 3; X is Cl, Br or CH 3COO; Y is Cl, Br or CH 3COO; the values of m and n are both ≥0, and m+n＝1 or 2; and the condition is: when any one of X and Y is Br, m+n＝2. The invention further relates to a preparation method for the compound, a preparation and the use.

Description

Steroidal quaternary ammonium compound and its preparation method, preparation and use

technical field

The present invention relates to steroidal quaternary ammonium compounds, their preparation methods, preparations and their uses in the related medical treatment fields.

Background technique

In the field of medicine, some drugs have a quaternary ammonium salt structure, which can be represented by the following formula:

Wherein R ¹ , R ² , R ³ , R ⁴ are optionally non-hydrogen hydrocarbon groups or substituted hydrocarbon groups, wherein any two or three groups can be linked by chemical bonds, and X ^- is a negative ion.

Existing drugs with quaternary ammonium salt structure such as rocuronium bromide (I), vecuronium bromide (II), pipecuronium bromide (III), pancuronium bromide (IV), hexfluorenium bromide (V), Recuronium bromide (Ⅵ), pinaverium bromide (Ⅶ), dipomonium bromide (Ⅷ), tiotropium bromide (Ⅸ), cetromium bromide (Ⅹ), umeclidinium bromide (ⅩⅠ), oti Bromide (XII), glycopyrrolate (XIII), umeclidinium (XIX), orphenium bromide (XV), hexamethylammonium bromide (XVI), ipratropium bromide (XVII), butorium bromide (XIX), etc. ^, wherein X- are other anions other than chloride anion, such as bromide anion.

It is known that bromide ions are slowly eliminated, and it is easy to accumulate poisoning in the body. Patients with hypertension and edema should not take bromide. Patients with epilepsy should not use ammonium bromide. Patients with severe pulmonary insufficiency, bronchial asthma and respiratory center depression caused by craniocerebral injury It should be avoided, and it should be used with caution in patients with liver and renal insufficiency.

Chemically, it is clear that bromide anion is less stable than chloride anion, and is easily oxidized to bromine or hypobromous acid, and hypobromous acid is further decomposed into bromic acid and elemental bromine or decomposed into bromous acid and hydrobromic acid. Elemental bromine is toxic and irritating. Inhalation of low concentrations of bromine can cause coughing, chest tightness, increased mucosal secretions, headache, dizziness, general discomfort, etc. Some people can cause gastrointestinal symptoms and even some people can develop allergic dermatitis (Cutaneous drug reactions: Pathogenesis and clinical classification. Journal of the American Academy of Dermatology. Bruce U. Wintroub, M.D., and Robert Stem, M.D. San Francisco and Boston. 1985.13(2):167-179). Hydrogen bromide is irritating, toxic, and easily soluble in water. Its aqueous solution is called hydrobromic acid, which is a strong acid.

Comparison of literature data on acute toxicity of sodium bromide and sodium chloride in animals:

[1]Nippon Yakurigaku Zasshi.Japanese Journal of Pharmacology.Vol.56,Pg.377,1960.

[2] Journal of Pharmacology and Experimental Therapeutics.Vol.55,Pg.200,1935.

[3] Endocrinology Vol.24, Pg.523, 1939.

[4] Toxicology and Applied Pharmacology.Vol.20,Pg.57,1971.

[5]"Drug Dosages in Laboratory Animals-A Handbook,"Rev.ed.,Barnes,C.D.,and L.G.Eltherington,Berkeley,Univ.of California Press,1973,Pg.243,1973.

[6]"Abdernalden's Handbuch der Biologischen Arbeitsmethoden."Vol.4,Pg.1289,1935.

Studies have shown that bromine affects the metabolism of iodine in the body and affects thyroid function (Metabolism of Bromide and Its Interference with the Metabolism of Iodine, S. Pavelka, Physiol. Res. 2004, 53 (Suppl. 1): S81-S90). Excessive intake of bromide will affect human health. From the results of toxicity studies, the allowable daily intake of bromine (ADI) is 0.12mg/kg body weight (Toxicity of sodium bromide in rats: effects on endocrine system and reproduction, Van Leeuwen FX, Den Tonkelaar EM, Van Logten MJ, Food Chem Toxicol. 1983, 21(4):383-9)

From the above data, it can be seen that bromide has poorer biocompatibility, higher toxicity and lower safety than chloride.

Rocuronium bromide is a medium-time-acting non-depolarizing muscle relaxant currently used clinically. It has the advantages of rapid onset of action, no release of histamine, and no significant effect on autonomic nerves and cardiovascular disease. Tracheal intubation and intraoperative muscle relaxation were maintained during routine induction of anesthesia.

The 17-position of rocuronium bromide is substituted with an acetoxy group, and its ester bond is chemically unstable and prone to hydrolysis. Both rocuronium bromide and pancuronium bromide currently used clinically are injections. In order to maintain their stability and reduce the generation of impurities and potency due to hydrolysis side reactions, the pH value of the injections must be kept at about 4 or lower. Even so, existing clinical drugs are still unstable and require refrigerated storage at 2-8 degrees Celsius. Others, such as vecuronium bromide, pipecuronium bromide and recuronium bromide, also require a pH of about 4 or lower for clinical use solutions. Due to the low pH and high total concentration of free acids and acid radicals in the above-mentioned steroid muscle relaxant preparations currently used in clinical practice, this product is obviously irritating when it is injected intravenously, which makes the patient feel pain at the injection site, even in the patient In the case of loss of consciousness, there are still reactions such as limb withdrawal caused by pain, which limits the clinical use of such drugs (Pharmacological prevention of rocuronium-induced injection pain or withdrawalal movements: a meta-analysis. Kwak HJ, Kim JY , Kim YB, Min SK, Moon BK, Kim JY., J Anesth. 2013 Oct;27(5):742-9). In addition, in the case of continuous administration of rocuronium bromide, the intake of bromide ions is likely to exceed the safe limit, which poses a safety risk to patients. According to the above-mentioned ADI limit of bromine intake, the safe dose limit of rocuronium bromide is 0.92mg/kg body weight/day. If this dose is exceeded, the intake of bromide ions will exceed the ADI limit, resulting in safety risks. According to the current FDA rocuronium bromide Label: 1) The recommended induction dose of tracheal intubation is 0.6 mg/kg body weight, and its muscle relaxation time is about 30 minutes, followed by 2) intravenous infusion of 10-12 μg/kg body weight at the recommended dose /min to maintain the muscle relaxation effect, the bromine intake safety dose limit is reached when the intravenous infusion is maintained for 27-32 minutes; in the case of rapid sequence intubation (Rapid sequence intubation), the recommended induction dose is 0.6-1.2 mg/kg body weight, the bromide ion introduced by the upper limit dose has exceeded the ADI value, so patients face a very high risk of excess bromine intake in clinical muscle relaxant therapy practice.

The above-mentioned steroid muscle relaxants are all bromides. Studies have shown that bromine can affect the metabolism of iodine in the body and affect thyroid function (Metabolism of Bromide and Its Interference with the Metabolism of Iodine., S. Pavelka, Physiol. Res. 2004, 53 (Suppl. 1): S81-S90). Excessive intake of bromide will affect human health. From the results of toxicity studies, the allowable daily intake of bromine (ADI) is 0.12mg/kg body weight (Toxicity of sodium bromide in rats: effects on endocrine system and reproduction., Van Leeuwen FX , Den Tonkelaar EM, Van Logten MJ, Food Chem Toxicol. 1983, 21(4):383-9). Taking rocuronium bromide as an example, in the case of continuous administration, the intake of bromide ions easily exceeds the safe limit, which poses a safety risk to patients. According to the current routine dosing schedule of rocuronium bromide: 1) intravenous injection of 0.15 mg/kg body weight, then 2) continuous intravenous infusion of 5-10 μg/kg body weight/min, when the dose is 10 μg/kg body weight/min, in Intravenous infusion for 77 minutes has exceeded the safe upper limit of bromine intake, which cannot meet many clinical drug delivery needs.

It can be seen that the development of safer steroidal quaternary ammonium compound drugs with less side effects and more controllable quality has important clinical significance.

SUMMARY OF THE INVENTION

A first aspect of the present invention provides a steroidal quaternary ammonium compound of formula I or an addition salt or a solvate thereof:

in,

A is _CH2 or O;

B is N or

R ¹ is H or CH ₃ CO;

R ² is CH ₃ or -CH ₂ CH=CH ₂ ;

t=2 or 3;

X is Cl, Br or CH ₃ COO;

Y is Cl, Br or CH ₃ COO;

The values of m and n are both ≥0, and m+n=1 or 2;

The condition is: when any one of X and Y is Br, m+n=2.

The steroidal quaternary ammonium compound or its addition salt is preferably selected from the group consisting of:

The solvent in the solvate is water, alcohol, acid, ester, ether, ketone or halogenated hydrocarbon, and the amount of solvent may be present in a stoichiometric or non-stoichiometric ratio. The solvent is water, C _1-6 alcohol, C _1-6 acid, C ₃ -C ₆ ester, C _4-6 ether, C _3-6 ketone or C _1-6 halogenated hydrocarbon; preferably in the following group One or more of: water, methanol, ethanol, propanol, isopropanol, n-butanol, tert-butanol, acetic acid, propionic acid, butyric acid, ethyl acetate, methyl acetate, isopropyl acetate, propyl acetate Methyl acid, ethyl propionate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, acetone, methyl ethyl ketone, dichloromethane, chloroform.

According to the second aspect of the present invention, there is provided a preparation method of a compound comprising the structure of formula I or an addition salt or a solvate thereof.

Preparation method of free base

According to one aspect of the present invention, there is provided a method A1 for preparing the rocuronium chloride of the structure of the formula I-1, which is prepared as follows: the rocuronium bromide is dissolved in water to prepare a solution, and then enters a reversed-phase liquid chromatography , carry out gradient elution with a solution containing hydrochloric acid, collect fractions, the obtained fractions are concentrated under reduced pressure and dried to obtain rocuronium chloride hydrochloride of formula I-2 structure; the obtained rocuronium chloride salt of formula I-2 structure The acid salt is added into organic solvent A and inorganic base, stirred at room temperature, filtered, and the filtrate is concentrated to obtain the rocuronium chloride compound of formula I-1, wherein the organic solvent A includes ethyl acetate, methyl acetate, isopropyl acetate At least one or a combination of ester, methyl propionate, ethyl propionate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, acetone, methyl ethyl ketone, dichloromethane, chloroform and acetonitrile, preferably tetrahydrofuran, acetone, At least one of methylene chloride, chloroform, acetonitrile and methyl tert-butyl ether or a combination thereof, the inorganic base includes sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, potassium carbonate, sodium carbonate, At least one of lithium carbonate, cesium carbonate, calcium carbonate, sodium bicarbonate, potassium bicarbonate, lithium bicarbonate, calcium bicarbonate or its combination, preferably at least one of sodium carbonate, potassium carbonate, lithium carbonate and calcium carbonate species or combinations thereof.

According to one aspect of the present invention, it provides a method A2 for preparing the rocuronium chloride of the structure of the formula I-1, wherein the rocuronium chloride can also be prepared through the following reaction scheme 1:

(Route 1)

Wherein, compound M8 is reacted with allyl chloride at a temperature of 0-80° C., preferably 20-60° C., more preferably 30-50° C., for 12-168 hours, and the reaction solution undergoes a post-treatment procedure to obtain rocuronium chloride.

As an embodiment of the method A2 of the present invention, the post-processing procedure includes the following steps: concentrating the reaction solution to obtain crude rocuronium chloride, preparing by reversed-phase liquid chromatography, using aqueous ammonium acetate and methanol as mobile phases, The acetate fraction was obtained by the first elution and purification, and then entered into reversed-phase liquid chromatography for preparation. Hydrochloric acid solution and methanol were used as mobile phases, and the rocuronium chloride hydrochloride fraction was obtained after the second elution and purification. After concentration under reduced pressure, organic solvent A and inorganic base are added to the residue, stirred at room temperature, filtered, and the filtrate is concentrated to obtain the rocuronium chloride compound of formula I-1, wherein the organic solvent A is selected from the following group: acetic acid Ethyl ester, methyl acetate, isopropyl acetate, methyl propionate, ethyl propionate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, acetone, methyl ethyl ketone, dichloromethane, chloroform and acetonitrile or combinations thereof, preferably is selected from the group consisting of tetrahydrofuran, acetone, dichloromethane, chloroform, acetonitrile, and methyl tert-butyl ether or a combination thereof, and the inorganic base is selected from the group consisting of sodium hydroxide, potassium hydroxide, lithium hydroxide, Calcium hydroxide, potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate, calcium carbonate, sodium bicarbonate, potassium bicarbonate, lithium bicarbonate, calcium bicarbonate or combinations thereof, preferably selected from the group consisting of sodium carbonate, potassium carbonate , lithium carbonate and calcium carbonate or a combination thereof.

As another embodiment of the method A2 of the present invention, the post-processing procedure includes the following steps: concentrating the reaction solution, adding water to dissolve, adding dichloromethane or ethyl acetate for extraction, adjusting the pH of the water phase to 1-6, adding activated carbon for decolorization , filter, neutralize the pH of the filtrate to 7~9, the temperature of the filtrate during the neutralization process is less than or equal to 10 ° C, filter, add sodium chloride to the filtrate until a saturated solution of sodium chloride is formed, add dichloromethane for extraction, and the organic phase is dried. The solvent was dried, filtered, and the filtrate was concentrated, and the concentrate was added to an anhydrous solvent, and the anhydrous solvent included acetonitrile, propionitrile, methanol, ethanol, isopropanol, propanol, butanol, sec-butanol, tert-butanol, dichloromethane At least one of methane, chloroform, N,N-dimethylformamide, dimethyl sulfoxide or a combination thereof, dissolve, filter, drop the filtrate into an organic solvent B, the organic solvent B includes ethers such as diethyl ether , isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone, or esters such as ethyl acetate, methyl acetate, At least one of propyl acetate, butyl acetate, and isopropyl acetate or a combination thereof, crystallize, filter, and dry the filter cake under reduced pressure to obtain rocuronium chloride of the structure of formula I-1.

Preparation method of hydrochloride

The present invention provides a preparation method B1 of the hydrochloride of rocuronium chloride or vecuronium chloride: dissolve the rocuronium chloride or vecuronium chloride compound with an organic solution of hydrogen chloride, wherein the organic solution of hydrogen chloride comprises methanol, ethanol, isomeric Hydrogen chloride solution of at least one of propanol, acetonitrile, propionitrile or a combination thereof, drop an organic solvent B into the solution, the organic solvent B includes ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydrofuran Pyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone or esters such as ethyl acetate, methyl acetate, propyl acetate, butyl acetate, isopropyl acetate At least one or a combination thereof, stirring and crystallizing, filtering, and drying the filter cake under reduced pressure to obtain the rocuronium chloride or vecuronium chloride hydrochloride of the present invention.

The present invention also provides another method B2 for preparing rocuronium chloride hydrochloride with the structure of formula I-2, which comprises preparing rocuronium chloride according to the method of route 1 of the present invention, and then performing post-processing including the following steps on it Procedure: Concentrate the reaction solution, add the organic solvent B including ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexane Ketone, cyclopentanone, or esters such as at least one of ethyl acetate, methyl acetate, propyl acetate, butyl acetate, isopropyl acetate, or a combination thereof, separate out the crude product, filter, dissolve the solid in water, and dissolve the water layer Extract with dichloromethane or ethyl acetate, adjust the pH of the aqueous phase to 1-6, add activated carbon to decolorize, filter, and concentrate the filtrate under reduced pressure at ≤60 °C, preferably at 30-50 °C, add organic solvent B, the organic solvent Solvent B includes ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone, or esters such as acetic acid At least one of ethyl ester, methyl acetate, propyl acetate, butyl acetate, and isopropyl acetate or a combination thereof, dispersed and pulped, filtered, and the filter cake was dried under reduced pressure at ≤ 40°C to obtain the structure of formula I-2. Rocuronium chloride hydrochloride.

Another embodiment of the preparation method B2 of the present invention includes, after preparing rocuronium chloride according to the method of route 1 of the present invention, adding an organic solvent B therein, and the organic solvent B includes ethers such as diethyl ether, isopropyl ether, methyl ether Tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone, or esters such as ethyl acetate, methyl acetate, propyl acetate, butyl acetate At least one of ester, isopropyl acetate or its combination, after dispersing and beating, filtering step, also comprises the following steps: filter cake adds organic solvent C again, and described organic solvent C comprises pentane, hexane, heptane , at least one of petroleum ether, acetone, butanone, cyclohexanone, cyclopentanone or a combination thereof, beating, filtering, and the filter cake ≤ 40 ° C and drying under reduced pressure to obtain the rocuronium chloride salt of formula I-2 structure acid salt.

Yet another embodiment of the preparation method B2 of the present invention includes, after preparing rocuronium chloride according to the method of route 1 of the present invention, performing a post-processing procedure including the following steps: concentrating the reaction solution, adding the organic solvent B, including Ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone, or esters such as ethyl acetate, At least one of methyl acetate, propyl acetate, butyl acetate, and isopropyl acetate or its combination, separate out the crude product, filter, dissolve the solid in water, extract the aqueous layer with dichloromethane or ethyl acetate, and adjust the pH of the aqueous phase To 1-6, add activated carbon to decolorize, filter, the filtrate is concentrated under reduced pressure at ≤60 °C, preferably 30-50 °C under reduced pressure, and dissolved by adding an alcohol solvent, the alcohol solvent includes methanol, ethanol, isopropanol, and propanol. , at least one of butanol, sec-butanol, tert-butanol or a combination thereof, the resulting alcohol solution is added to an organic solvent B, the organic solvent B includes ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran , tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone, or esters such as ethyl acetate, methyl acetate, propyl acetate, butyl acetate, isoacetate At least one of the propyl esters or a combination thereof, crystallize, filter, and dry the filter cake under reduced pressure at a temperature of less than or equal to 40°C to obtain the rocuronium chloride hydrochloride of the structure of formula I-2.

Another embodiment of the preparation method B2 of the present invention includes, after preparing rocuronium chloride according to the method of route 1 of the present invention, performing a post-processing procedure including the following steps: concentrating the reaction solution, adding an organic solvent B, the The organic solvent B includes ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, butanone, cyclohexanone, cyclopentanone, or esters such as At least one of ethyl acetate, methyl acetate, propyl acetate, butyl acetate, and isopropyl acetate or a combination thereof, the crude product was separated out, filtered, the solid was dissolved in water, and the aqueous layer was extracted with dichloromethane or ethyl acetate, The water phase is adjusted to pH 1-6, decolorized by adding activated carbon, filtered, the filtrate is concentrated under reduced pressure at ≤60°C, preferably 30-50°C under reduced pressure, the concentrate is filtered, and the filtrate concentration is 0.1-1500mg/ml, preferably 10-1200mg/ml , freeze-drying to obtain rocuronium chloride hydrochloride of formula I-2 structure.

Preparation method of hydrochloride hydrate

The present invention provides a method for preparing a hydrate of rocuronium chloride hydrochloride having a structure of formula I-2, which comprises the following steps after preparing rocuronium chloride according to the method of route 1 of the present invention Treatment procedure: Concentrate the reaction solution, add organic solvent B, and the organic solvent B includes ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ketones such as acetone, Butanone, cyclohexanone, cyclopentanone or esters such as at least one of ethyl acetate, methyl acetate, propyl acetate, butyl acetate, isopropyl acetate or its combination, separate out the crude product, filter, add water to the solid Dissolve, extract the aqueous layer with dichloromethane or ethyl acetate, adjust the pH of the aqueous phase to 1-6, add activated carbon to decolorize, filter, and concentrate the filtrate under reduced pressure at ≤60 °C, preferably at 30-50 °C, to obtain formula I- The hydrate of rocuronium chloride hydrochloride of the 2 structure.

A third aspect of the present invention provides a lyophilized formulation comprising a steroidal quaternary ammonium compound or an addition salt or a solvate thereof or rocuronium or vecuronium bromide according to the present invention, and one or A variety of pharmaceutically acceptable excipients.

In one embodiment of the present invention, the freeze-dried preparation contains any one or more pharmaceutically acceptable excipients, the adjuvants include freeze-drying support agents and/or protective agents, or powder inhalation supplements, such as mannitol, sorbitan Alcohol, xylitol, sucrose, lactose, glucose, dextran, dextrin, maltose, maltitol, maltodextrin, erythritol, trehalose, calcium gluconate, calcium sulfate, sodium chloride, glycine, hydrolysis At least one of gelatin, human albumin, or a combination thereof.

In one embodiment of the present invention, the freeze-dried preparation contains one or more pharmaceutically acceptable excipients, the excipients include pH adjusters, such as hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, acetic acid, sodium dihydrogen phosphate, diphosphate dibasic Potassium hydrogen phosphate, ammonium dihydrogen phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium phosphate, potassium phosphate, ammonium phosphate, sodium hydrogen sulfate, potassium hydrogen sulfate, ammonium hydrogen sulfate, sodium hydrogen carbonate, hydrogen carbonate Potassium, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, ammonia water, citric acid, sodium dihydrogen citrate, potassium dihydrogen citrate, ammonium dihydrogen citrate, disodium hydrogen citrate, Dipotassium citrate, diammonium citrate, sodium potassium citrate, sodium citrate, potassium citrate, ammonium citrate, lactic acid, sodium lactate, potassium lactate, ammonium lactate, malic acid, apple Sodium, potassium malate, malic acid, sodium hydrogen malate, potassium hydrogen malate, ammonium hydrogen malate, potassium sodium malate, tartaric acid, sodium hydrogen tartrate, potassium hydrogen tartrate, ammonium hydrogen tartrate, potassium sodium tartrate, vitamin C , Sodium Vitamin C, Alginic Acid, Sodium Alginate, Succinic Acid, Sodium Succinate, Potassium Succinate, Ammonium Succinate, Sodium Succinate, Potassium Succinate, Ammonium Succinate, Potassium Sodium Succinate, Acetic Acid, Sodium Acetate, Potassium Acetate , at least one or a combination of ammonium acetate, amino acids and salts thereof.

In one embodiment of the present invention, the freeze-dried preparation contains one or more pharmaceutically acceptable excipients, the excipients include a stabilizer, and the stabilizer includes at least one of an antioxidant or a metal ion complexing agent or a combination thereof , the antioxidant comprises at least one of sodium bisulfite, sodium metabisulfite, sodium sulfite, sodium thiosulfate, vitamin C, sodium thioglycolate, glycine, cysteine or a combination thereof, the metal ion complexing agent Including at least one of disodium EDTA, calcium sodium EDTA, or a combination thereof.

In one embodiment of the present invention, the freeze-dried preparation contains one or more pharmaceutically acceptable excipients, the excipients include analgesics and local anesthetics, such as benzyl alcohol, chlorobutanol, Procaine, Cocaine, Tetracaine, Proparacaine, Orbucaine, Benzocaine, Lidocaine, Cincocaine, Prilocaine, Trimethaine, Bupivacaine, Levobupiva At least one of mepivacaine, ropivacaine, dyclonine, chloroprocaine, articaine, eticaine or a combination thereof.

In one embodiment of the present invention, the freeze-dried preparation contains one or more pharmaceutically acceptable excipients, the excipients include bacteriostatic agents, such as benzyl alcohol, chlorobutanol, benzoic acid and the like. At least one of salts, sorbic acid and its salts, and parabens, or a combination thereof.

According to a fourth aspect of the present invention, there is provided a kit comprising said steroidal quaternary ammonium compound or an addition salt or solvate thereof, or a lyophilized preparation thereof or comprising rocuronium bromide or vecuronium bromide Lyophilized preparation of ammonium.

According to the fifth aspect of the present invention, the steroidal quaternary ammonium compound or its addition salt or its solvate or its freeze-dried preparation or the freeze-dried preparation comprising rocuronium or vecuronium is used for the preparation of bone Muscle relaxants. wherein the drug is administered by gastrointestinal and parenteral routes. For example, the gastrointestinal route is the oral route; the parenteral route is the inhalation, intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, mucosal and deep tissue or ocular, transdermal, topical and other external routes.

According to a sixth aspect of the present invention, there is provided a method of treating a disease or condition in a subject, particularly for endotracheal intubation and maintenance of intraoperative muscle relaxation during routine induction of anesthesia, the method comprising administering to the subject a therapeutically effective amount of a compound according to the invention or an addition salt or a solvate thereof or a lyophilized preparation thereof.

According to the seventh aspect of the present invention, there is provided a preparation method of a freeze-dried preparation, comprising the steps of: wherein a drug, an auxiliary material and a corresponding solvent are mixed to form a solution, and the pH of the solution is adjusted to 1-7, preferably pH 2-6, and more Preferably, the pH is 3.5 to 5.5, and the filtrate is filtered, and the filtrate is freeze-dried to obtain a freeze-dried preparation. Further, in the method for preparing a freeze-dried preparation of a steroidal quaternary ammonium compound, the solvent is water. Further, the preparation method of the steroidal quaternary ammonium compound freeze-dried preparation, wherein the prepared solution, wherein the steroidal quaternary ammonium compound or rocuronium bromide or vecuronium bromide concentration is 100μg/ml-300mg/ml , The concentration of pharmaceutical excipients (lyophilized support agent and/or protective agent, pH adjuster, antioxidant and metal ion complexing agent and other stabilizers, analgesic, bacteriostatic agent) is 0mg/ml-500mg/ml. Further, the preparation method of the described steroidal quaternary ammonium compound freeze-dried preparation adopts freeze-drying, and the process includes:

(1) In the pre-freezing stage, the above solution is cooled to a range of -30°C to -196°C until the solution system is coagulated and frozen;

(2) Optionally, the temperature of the frozen solution system is increased to a temperature in the range of -30°C to -5°C, wherein the temperature in the range of about -30°C to -5°C makes the frozen solution system Keep the frozen state; cool the frozen solution system to below -30°C again, and this process can be repeated;

(3) the primary drying stage, including the sublimation step, by removing the solvent in the frozen solution system in the frozen state described in step (1) or step (2) under reduced pressure, to obtain a partially dried product;

(4) In the secondary drying stage, the residual solvent in the partially dried product in the non-frozen state is removed under reduced pressure to obtain a freeze-dried product, and this stage is usually accompanied by further temperature rise.

The above freeze-dried preparation is used to prepare a muscle relaxant drug.

According to an eighth aspect of the present invention, there is provided a multi-part combination kit comprising a first container and a second container, the first container containing the lyophilized preparation of the steroidal quaternary ammonium compound, the second container The container contains a physiologically acceptable solution for formulating the lyophilized formulation. The acceptable solutions include, but are not limited to, 5% dextrose solution, 0.9% sodium chloride solution, sodium lactated Ringer's solution, 5% dextrose in physiological saline solution.

According to the ninth aspect of the present invention, the multi-part combination kit is used for preparing clinical medicine for muscle relaxation.

A clinical method of skeletal muscle relaxation comprising administering to a subject in need thereof an effective dose of a lyophilized formulation according to the present invention.

The present invention also provides a solution for injection prepared from a freeze-dried preparation of a steroidal quaternary ammonium compound, the pH of the solution for injection is not lower than 4, preferably 4-8, more preferably 5-7, wherein acid-containing solution The concentration is 0 to 0.15M, preferably 0 to 0.1M, and more preferably 0 to 0.03M.

The freeze-dried preparation involved in the present invention has outstanding advantages, and it is formulated into a solution for injection, the pH of the solution for injection is not lower than 4, and the pH is preferably 4-8, more preferably 5-7, wherein acid-containing solution The concentration is 0 to 0.15M, preferably 0 to 0.1M, and more preferably 0 to 0.03M.

In one embodiment of the present invention, the pH of the solution for injection prepared by the freeze-dried preparation is 4.5-6.

In one embodiment of the present invention, a solution for injection prepared from a freeze-dried preparation of rocuronium bromide, the pH of the solution for injection is 4.5-6.

In one embodiment of the present invention, a solution for injection prepared from a freeze-dried preparation of vecuronium bromide, the pH of the solution for injection is 4.5-6.

Compared with bromide, the quaternary ammonium chloride compound provided by the present invention has lower cost.

Compared with bromide, the quaternary ammonium chloride compound provided by the present invention does not form genotoxic impurities, and the chloride ion in the body is better and safer than the bromide ion biocompatibility, so the safety and quality of its pharmaceutical product More controllability.

The existing clinically used rocuronium bromide is a solution preparation with low stability and needs to be transported and stored at a low temperature of 2-8°C. The freeze-dried preparation of rocuronium chloride provided by the present invention is solid, has high stability compared with the above-mentioned rocuronium bromide solution preparation product, does not require low temperature, is more convenient for production, transportation, storage and use of the product, and greatly improves the performance of the drug in each Link quality assurance, thereby improving the safety of medication.

In addition, compared with the rocuronium bromide drug currently used clinically, the freeze-dried preparation of rocuronium bromide, the freeze-dried preparation of vecuronium bromide, the freeze-dried preparation of rocuronium bromide or its hydrochloride and its solvent provided by the present invention The dry product completely eliminates the excess acid such as acetic acid required to maintain the pH of the solution at 4 or lower, and completely eliminates the lower pH and free acid and acid radicals in the current clinical formulations of rocuronium and vecuronium. The disadvantage of high total concentration leads to obvious irritation during intravenous injection, which makes the patient feel pain at the injection site. The advantages of painless injection and clinical practical value.

Vecuronium bromide hydrobromide, vecuronium chloride and vecuronium chloride hydrochloride provided by the present invention also have rocuronium bromide hydrobromide, rocuronium chloride or its hydrochloride provided by the present invention Same advantages.

The present invention overcomes the above-mentioned safety shortcomings of existing bromide-containing anion-containing quaternary ammonium drugs and preparations thereof, and provides a new type of quaternary ammonium compound, a preparation method and preparation thereof, which have higher safety, fewer side effects and better quality. control advantages.

Description of drawings

Figure 1: EMG signals before and after administration of Solution A in Experimental Example 2.

Figure 2: EMG signals before and after administration of solution B in Experimental Example 2.

Figure 3: EMG signals before and after administration of Solution C in Experimental Example 2.

Figure 4: Plot of the ratio of the post-dose response plateau signal integral (8 sec duration) to the pre-dose baseline plateau signal integral (8 sec duration) in each EMG of Solutions A-C.

Fig. 5: EMG signals before and after administration of solution D in Experimental Example 3.

Figure 6: EMG signals before and after administration of solution E in Experimental Example 3.

Fig. 7: EMG signals before and after administration of Solution F in Experimental Example 3.

Figure 8: Plot of the ratio of the post-dose response plateau signal integral (8 seconds duration) to the pre-dose baseline plateau signal integration (8 seconds duration) in each EMG of solutions D-F.

Detailed ways

In order to make the objectives and technical solutions of the present invention clearer, the preferred embodiments of the present invention are described in detail below. It should be noted that the following examples are only used to further illustrate the present invention, and should not be construed as limiting the protection scope of the present invention. Some non-essential improvements and adjustments made by those skilled in the art according to the above content of the present invention belong to the protection scope of the present invention.

Example 1: Preparation of Formula I-2 (Rocuronium Chloride Hydrochloride) (Method A)

Instrument: Hanbang NU3000; Column: Zhongpu Technology, RD-C18 5um, 21.2*250mm

Wavelength: 210nm; Flow Rate: 13ml/min; Column Temperature: Room Temperature

Mobile phase: methanol, 0.15% aqueous HCl

Weigh 200 mg of rocuronium bromide, add an appropriate amount of purified water to dissolve, make the volume about 1 ml, inject the sample to prepare the liquid phase, and collect the fractions by elution as follows, elution gradient:

The obtained fractions were concentrated under reduced pressure not higher than 35° C. and dried in vacuo to obtain 73 mg of rocuronium hydrochloride with a yield of 37%. The products are respectively rocuronium bromide standard (National Institutes for Food and Drug Control, batch number 101361-201902, content 99.6%) and sodium chloride (National Institutes for Food and Drug Control). Food and Drug Control), batch number 100376-201703, content 100%) was used as external standard for content determination, derocurium content was 86.59%, chloride ion content was 11.61%, rocuronium/chlorine=7.458:1 (theoretical value was 7.474 :1). Detected by LC-MS method, ESI-MS m/z[M-HCl-Cl] ⁺ 529.4, ESI-MS m/z[(M-2Cl ^- )/2] ⁺ 265.2, (ESI-) bromide ion signal not detected.

Example 2: Preparation of compound of formula I-1 (rocuronium chloride) (method A)

Add 50 mg of rocuronium chloride hydrochloride prepared according to the method in Example 1, add 10 ml of acetonitrile and 187 mg of sodium carbonate, stir at room temperature for 2 h, filter, concentrate the filtrate, add dichloromethane to the residue and distill it to dryness. Curonium chloride 35mg, purity 99.7%, yield 74.5%.

Example 3: Preparation of compound of formula I-1 (rocuronium chloride)

Add 1.0 g of compound M8, 15 ml of 3-chloropropene, seal the tube, react at an external temperature of about 50-60 ° C for 48 hours, and concentrate to obtain a reddish-brown viscous residue. The obtained residue is subjected to the following reverse phase column chromatography. One purification:

Instrument: Hanbang NU3000; Chromatographic column: Zhongpu Red RD-C18, 5um, 21.2*250mm;

Wavelength: 210nm; Flow Rate: 16ml/min; Column Temperature: Room Temperature

Mobile phase: 30mM ammonium acetate, PH=6.0, methanol;

The above residue was dissolved in an appropriate amount of purified water, and the liquid phase was prepared by injecting the sample. The fractions were eluted and collected according to the following method, and the elution gradient was as follows:

TT	30mM乙酸铵PH＝6.030mM ammonium acetate pH=6.0	甲醇 methanol
00	7575	2525
3030	3535	6565
30.130.1	55	9595
3535	55	9595
35.135.1	7575	2525
4040	7575	2525

The obtained fractions were added with purified water to dilute the methanol ratio to about 20% (v/v), and the second purification was carried out by reversed-phase column chromatography as follows:

Wavelength: 210nm; Flow Rate: 13ml/min; Column Temperature: Room Temperature

Mobile phase: 0.15% HCl, methanol;

The collected fractions were eluted as follows, elution gradient:

TT	0.15％HCl0.15% HCl		甲醇methanol
00	9595	55
55	9595	55

3030	5050	5050
3535	5050	5050

The collected fractions were referred to the operation of Example 1 to obtain rocuronium chloride hydrochloride.

The title compound was obtained by referring to the obtained rocuronium chloride hydrochloride as in Example 2. The obtained sample had a HPLC purity of 99.8%, Mass(ESI+) [M-Cl ⁻ ] ⁺ =529.4, ¹ H-NMR (BRUKER AVANCE III HD 600MHz), D ₂ O solvent, δ(ppm) 0.79(s, 3H), 0.85(s, 3H), 1.03-1.07(m, 1H), 1.09-1.14(m, 1H), 1.16-1.19( t,3H),1.28-1.44(m,3H),1.45-1.59(m,5H),1.61-1.64(m,1H),1.73-1.81(m,3H),1.87-1.92(m,1H), 2.00(s,3H),2.06-2.11(m,4H),3.20-3.22(d,br,1H),3.25-3.27(d,br,1H),3.31-3.37(m,2H),3.49-3.54 (dd,br,1H),3.56-3.59(dd,1H),3.63-3.64(m,3H),3.77-3.81(t,br,1H),3.84-3.88(t,br,1H),4.03- 4.13(m,5H),4.15-4.19(m,1H),5.07-5.08(d,br,1H),5.54-5.57(d,br,1H),5.61-5.62(d,1H),5.97-6.04 (m, 1H).

Example 4: Preparation of formula I-1 (rocuronium chloride)

Put 1.0 g of M8, 5 ml of 3-chloropropene, and 10 ml of dichloromethane into a 75 ml sealed tube, seal, stir, and heat up to 45° C. to stir and react for 5 days. The reaction solution was transferred to a single-necked flask, concentrated, and the external temperature was 30 °C. After the concentration was completed, 10 ml of pure ice water was added, and 10 ml of dichloromethane × 2 was used for extraction. The pH was adjusted to about 1 with 1 mol/L hydrochloric acid, and 150 mg of activated carbon was added to decolorize 2-3 times. Filter, cool the filtrate to 0°C, adjust the pH to 7.88 with 1 mol/L NaOH solution, filter, add sodium chloride to the filtrate to saturation. Extract with dichloromethane 10ml×3, combine the extracts, add anhydrous sodium sulfate and dry overnight. Filter, concentrate, add 3 ml of anhydrous acetonitrile to the concentrate. Dissolve, filter, drop the filtrate into 18 ml of ether for crystallization for 2.0 hours, stir, filter, and dry the filter cake until the residual solvent is qualified, 510 mg of rocuronium chloride, with a purity greater than 99.0% .

Example 5: Preparation of formula I-1 (rocuronium chloride)

Put M8 1.0g, 3-chloropropene 5ml, and dichloromethane 10ml into a 75ml sealed tube, seal, stir, heat up to 30-50°C and stir for 5 days. The reaction solution was transferred to a single-necked flask, concentrated, and the external temperature was 30 ° C. After the concentration was completed, 10 ml of pure ice water was added, and 10 ml of ethyl acetate was used for extraction. Filter, cool the filtrate to 0°C, adjust the pH to 7.88 with 1 mol/L NaOH solution, filter, add sodium chloride to the filtrate to saturation. Extract with dichloromethane 10ml×3, combine the extracts, add anhydrous sodium sulfate and dry overnight. Filter, concentrate, add 3 ml of anhydrous ethanol to the concentrate. Dissolve, filter, drop the filtrate into 18 ml of ethyl acetate for crystallization for 2.0 hours, stir, filter, and dry the filter cake until the residual solvent is qualified. 99.0%.

Example 6: Formula I-2 (rocuronium chloride hydrochloride)

510 mg of rocuronium chloride prepared according to the method of Example 2-5 was dissolved in 3 ml of hydrogen chloride acetonitrile solution, then 18 ml of diethyl ether was added dropwise, and a solid was precipitated, stirred at room temperature for 2-3 hours, filtered, and the filter cake was dried under reduced pressure to The residual solvent is qualified, and the white solid of rocuronium chloride hydrochloride is 458 mg, and the purity is more than 99.0%.

Example 7: Formula I-2 (rocuronium chloride hydrochloride)

Dissolve 510 mg of rocuronium chloride prepared according to the method in Example 2-5, dissolve it in 3 ml of ethanolic hydrogen chloride solution, and then add 18 ml of ether dropwise, a solid is precipitated, stir at room temperature for 2-3 hours, filter, and dry the filter cake under reduced pressure to The residual solvent is qualified, and the white solid of rocuronium chloride hydrochloride is 450 mg, and the purity is more than 99.0%.

Example 8: Preparation of formula I-2 (hydrate of rocuronium chloride hydrochloride)

In a 150ml sealed tube, add 5.0g of M8 and 25ml of allyl chloride, seal, stir, heat up to 30-50℃ for 96.0h, cool down, concentrate to 10ml of the reaction solution at an external temperature of 20±5℃, add ethyl acetate dropwise at room temperature 50ml of ester, solids were precipitated during the dropwise addition, filtered after the dropwise addition, the solid particles were dissolved in 50ml of water, the aqueous layer was extracted 3 times with 50ml of dichloromethane, the aqueous layer was cooled to 5-10°C, and 1N HCl was added to the aqueous phase to adjust pH to 4.0, add activated carbon 1.0g, decolorize for 30min, filter, the filtrate is concentrated to dryness under reduced pressure at not more than 60°C, and after concentration, 4.3g of rocuronium chloride hydrochloride hydrate is obtained, yield 69.8%, purity 99.81%, The clarity is qualified, the dissolved residue is qualified, and the moisture content is 3.0%.

Example 9: Preparation of formula I-2 (hydrate of rocuronium chloride hydrochloride)

In a 150ml sealed tube, add 5.0g of M8 and 25ml of allyl chloride, seal, stir, raise the temperature to 60°C for 12.0h, cool down, concentrate to 10ml of the reaction solution at an external temperature of 20±5°C, add 50ml of acetone dropwise at room temperature, dropwise During the addition, there is solid precipitation. After the dropwise addition is completed, filter the solid particles, dissolve the solid particles with 50 ml of water, extract the aqueous layer twice with 50 ml of dichloromethane, cool the aqueous layer to 5-10 ° C, and add 1N HCl to the aqueous phase to adjust the pH to 6.0, 1.0g of activated carbon was added, decolorized for 30min, filtered, and the filtrate was concentrated to dryness under reduced pressure not exceeding 60°C. After the concentration was completed, 4.2g of hydrate of rocuronium chloride hydrochloride was obtained, with a yield of 68.2%, a purity of 99.8%, and qualified clarity. The dissolved residue is qualified, and the moisture content is 3.1%.

Example 10: Preparation of formula I-2 (hydrate of rocuronium chloride hydrochloride)

In a 150ml sealed tube, add 5.0g of M8, 25ml of allyl chloride, 50ml of dry dichloromethane, seal, stir, heat up to 30-50°C for 90.0h, cool down, and concentrate to 10ml of reaction solution at an external temperature of 20±5°C, 50ml of ether was added dropwise at room temperature, solids were precipitated during the dropwise addition, filtered after the dropwise addition, the solid particles were dissolved in 50ml of water, the aqueous layer was extracted twice with 50ml of ethyl acetate, the aqueous layer was cooled to 5-10°C, and the aqueous phase 1N HCl was added to adjust the pH to 1.0, 1.0 g of activated carbon was added, decolorized for 30 min, filtered, the filtrate was concentrated to dryness under reduced pressure at not more than 60 °C, and 4.2 g of rocuronium chloride hydrochloride hydrate was obtained after the concentration was completed. The purity is 99.8%, the clarity is qualified, the dissolution residue is qualified, and the moisture content is 3.1%.

Example 11: Preparation of formula I-2 (rocuronium chloride hydrochloride)

4.3 g of the hydrate of rocuronium chloride hydrochloride prepared in Example 8 was added to 12 ml of absolute ethanol to dissolve, and the resulting alcohol solution was slowly added dropwise to 72 ml of ether at room temperature, stirred for half an hour, a solid was precipitated, filtered, and the ether 10 ml of the filter cake was washed twice, and the filter cake was vacuum-dried at not higher than 40° C. overnight to obtain 3.2 g of rocuronium chloride hydrochloride with the structure of formula I-2, with a yield of 52.7% and a purity of 99.8%.

Example 12: Preparation of formula I-2 (rocuronium chloride hydrochloride)

Add 4.2 g of the hydrate of rocuronium chloride hydrochloride prepared in Example 9 into 10 ml of anhydrous tert-butanol to dissolve, slowly add the resulting alcohol solution dropwise to 72 ml of methyl tertiary ether at room temperature, stir for half an hour, and precipitate a solid , filtered, the filter cake was washed twice with 10 ml of methyl tertiary ether, and the filter cake was vacuum-dried at not higher than 40°C overnight to obtain 3.3 g of rocuronium chloride hydrochloride with the structure of formula I-2, yield 54.4%, and purity 99.8%.

Example 13: Preparation of formula I-2 (rocuronium chloride hydrochloride)

4.3 g of the hydrate of rocuronium chloride hydrochloride prepared in Example 8 was added to 12 ml of absolute ethanol to dissolve, and the resulting alcohol solution was slowly added dropwise to 72 ml of ether at room temperature, stirred for half an hour, a solid was precipitated, filtered, and the ether The filter cake was washed twice with 10 ml of n-pentane, the filter cake was slurried with 10 ml of n-pentane for 1.0 hours, filtered, washed twice with 5 ml of n-pentane, and the filter cake was vacuum-dried at not higher than 40°C overnight to obtain the rocuronium chloride of formula I-2. 3.6 g of hydrochloride, yield 59.3%, purity 99.8%.

Example 14: Preparation of formula I-2 (rocuronium chloride hydrochloride)

Add 4.2 g of the hydrate of rocuronium chloride hydrochloride prepared in Example 9 into 10 ml of anhydrous tert-butanol to dissolve, slowly add the resulting alcohol solution dropwise to 72 ml of methyl tertiary ether at room temperature, stir for half an hour, and precipitate a solid , filtered, washed the filter cake twice with 10 ml of methyl tertiary ether, slurried the filter cake with 10 ml of hexane for 1.0 hours, filtered, washed twice with 5 ml of hexane, and dried the filter cake under vacuum at not higher than 40 °C overnight to obtain the structure of formula I-2. Rocuronium chloride hydrochloride 3.5g, yield 57.7%, purity 99.7%.

Example 15: Preparation of formula I-2 (rocuronium chloride hydrochloride)

4.3 g of the hydrate of rocuronium chloride hydrochloride prepared in Example 8 was added to 10 ml of acetone to disperse and slurried for 1.5 hours, filtered, washed twice with 5 ml of acetone, and the filter cake was vacuum-dried at not higher than 40 °C overnight to obtain formula I- 3.7 g of rocuronium chloride hydrochloride with structure 2, the yield is 60.9%, and the purity is 99.8%.

Example 16: Preparation of formula I-2 (rocuronium chloride hydrochloride)

4.2 g of the hydrate of rocuronium chloride hydrochloride prepared in Example 9 was added to 10 ml of acetone to disperse and slurried for 1.5 hours, filtered, washed twice with 5 ml of acetone, and the filter cake was then added with 15 ml of heptane to be slurried for 1.0 hours, filtered, and the filter cake It was vacuum-dried at not higher than 40° C. overnight to obtain 3.8 g of rocuronium chloride hydrochloride with the structure of formula I-2, with a yield of 62.6% and a purity of 99.7%.

Example 17: Preparation of formula I-2 (rocuronium chloride hydrochloride)

In a 150ml sealed tube, add 5.0g of M8, 25ml of allyl chloride, 50ml of dry dichloromethane, seal, stir, heat up to 50-80℃ for 96h, cool down, concentrate to 10ml of the reaction solution at an external temperature of 20±5℃, room temperature 50ml of ethyl acetate was added dropwise, solids were precipitated during the dropwise addition, filtered after the addition was completed, the solid particles were dissolved in 50ml of water, the aqueous layer was extracted 3 times with 50ml of dichloromethane, the aqueous layer was cooled to 5-10°C, and the water 1N HCl was added to the phase to adjust the pH to 6, 1.0 g of activated carbon was added, decolorized for 30 min, filtered, the filtrate was concentrated under reduced pressure at no more than 60°C, concentrated and filtered, and the filtrate was freeze-dried to obtain 3.9 g of rocuronium chloride hydrochloride with a yield of 63.3%. , the purity is 99.81%.

Example 18: Preparation of formula I-3 (rocuronium bromide hydrobromide)

Using rocuronium bromide as a raw material, referring to the operation of Example 1, 6 or 7, and using hydrobromic acid instead of hydrochloric acid, the title compound was prepared.

Example 19: Preparation of Formula I-4

Using rocuronium bromide as a raw material, referring to the operation of Example 1, 6 or 7, and using a mixture of hydrobromic acid and hydrochloric acid in different proportions instead of hydrochloric acid, the title compound was prepared.

Example 20: Preparation of Formula I-5

Using rocuronium bromide as a raw material, referring to the operation of Example 1, 6 or 7, and using a mixture of acetic acid and hydrochloric acid in different proportions instead of hydrochloric acid, the title compound was prepared.

Example 21: Preparation of formula I-6 (vecuronium chloride)

Using vecuronium bromide as raw material and referring to the operation of Example 1, the title compound was prepared.

Example 22: Preparation of formula I-7 (vecuronium chloride hydrochloride)

Using vecuronium bromide as a raw material, the title compound was obtained by referring to the operation of Example 1 or 6 or 7.

Example 23: Preparation of Formula I-8 (vecuronium hydrobromide)

Example 24: Preparation of Formula I-9

Using vecuronium bromide as a raw material, referring to the operation of Example 1, 6 or 7, and using a mixture of hydrobromic acid and hydrochloric acid in different proportions instead of hydrochloric acid, the title compound was prepared.

Example 25: Preparation of Formula I-10

Using vecuronium bromide as a raw material, referring to the operation of Example 1, 6 or 7, and using a mixture of acetic acid and hydrochloric acid in different proportions instead of hydrochloric acid, the title compound was prepared.

Example 26: Freeze-drying experiments 1#-3#

Rocuronium chloride and mannitol were dissolved in water for injection into 1# 2mg/mL containing 5% mannitol, 2# 4mg/mL containing 10% mannitol, 3# 6mg/mL containing 15% mannitol solution, adjusted with hydrochloric acid. pH4.0-5.0, filtered, the filtrate was pre-frozen at -70°C for 2.5h, lyophilized by Xinzhi SCIENTZ-12N lyophilizer for 20h, the end point of freeze-drying, the vacuum degree of the system was about 2.7Pa, and the temperature was about 20°C to obtain three All kinds of freeze-dried products are white lumps with smooth structure, and the above three kinds of freeze-dried products are rapidly reconstituted with water for injection.

Reconstitute 2# into a 2mg/mL solution, measure its pH to 5.66, and place it at room temperature (20-25°C). The stability is as follows:

放置时间hput time h	杂质个数Number of impurities	HPLC纯度％ HPLC Purity %
00	44	99.1699.16
44	44	99.3199.31
88	44	99.2999.29
1111	44	99.2999.29
1212	44	99.3199.31
1515	44	99.2899.28
22twenty two	44	99.2399.23
4343	44	99.1899.18
RSD％RSD%	----	0.060.06

It can be seen that the above-mentioned lyophilized preparation complex solution of the present invention has high stability under the experimental conditions.

Example 27: Freeze-drying experiments 4#-7#

Rocuronium chloride and glycine were dissolved in water for injection into 4# 2mg/mL containing 2% glycine, 5# 4mg/mL containing 4% glycine, 6# 6mg/mL containing 6% glycine, 7# 8mg/mL containing 8% glycine Glycine solution, adjusted to pH 4.0-5.0 with hydrochloric acid, filtered, the filtrate was pre-frozen at -70°C for 2.5h, lyophilized by Xinzhi SCIENTZ-12N lyophilizer for 20h, the end point of lyophilization, the vacuum degree of the system was about 2.7Pa, the temperature At about 20° C., the obtained four freeze-dried substances are all white and slightly glossy lumps with smooth structure, and the above-mentioned four freeze-dried substances are rapidly reconstituted with water for injection.

The 5# lyophilisate was reconstituted to 2mg/mL, pH 4.72, and placed at room temperature (20-25°C), the stability is as follows:

放置时间hput time h	杂质个数Number of impurities	HPLC纯度％HPLC Purity %
0.50.5	44	99.2199.21
22	44	99.2199.21
66	44	99.2099.20
1212	44	99.2199.21
1313	44	99.2199.21
1515	44	99.2099.20
23twenty three	44	99.1899.18
4444	44	99.1599.15
RSD％RSD%	----	0.020.02

It can be seen that the above-mentioned lyophilized preparation complex solution of the present invention has high stability under experimental conditions.

Example 28: Freeze-drying experiment 8#-11#

Rocuronium chloride and dextran 40 were dissolved in water for injection into 8# 2mg/mL containing 2% dextran 40, 9# 4mg/mL containing 4% dextran 40, 10# 6mg/mL containing 6% dextran Polysaccharide 40, 11# 8mg/mL solution containing 8% dextran 40, adjusted to pH 4.0-5.0 with hydrochloric acid, filtered, the filtrate was pre-frozen at -70°C for 2.5h, and freeze-dried with Xinzhi SCIENTZ-12N type freeze dryer After drying for 20h, the freeze-drying end point, the vacuum degree of the system is about 2.7Pa, and the temperature is about 20°C, the obtained four kinds of freeze-dried products are all white and smooth in structure, and the above four kinds of freeze-dried products are rapidly reconstituted with water for injection.

Example 29: Freeze-drying experiments 12#-15#

Rocuronium chloride and lactose were dissolved in water for injection into 12#3mg/mL containing 2% lactose, 13#5mg/mL containing 4% lactose, 14#7mg/mL containing 6% lactose, 15#9mg/mL containing 8% lactose, respectively The lactose solution was adjusted to pH 4.0-5.0 with hydrochloric acid, filtered, the filtrate was pre-frozen at -70°C for 2.5 hours, and then lyophilized by Xinzhi SCIENTZ-12N freeze-drying machine for 20 hours. At about 20° C., the obtained four freeze-dried substances are all white and smooth in structure, and the above-mentioned four freeze-dried substances are rapidly reconstituted with water for injection.

Example 30: Freeze-drying experiments 16#-19#

Using rocuronium bromide and mannitol, 16#-19# lyophilisates were prepared with reference to the method in Example 26, all of which were white lumps and smooth in structure, and the above three lyophilisates were rapidly reconstituted with water for injection.

Example 31: Freeze-drying experiments 20#-23#

Using rocuronium bromide and glycine, the 20#-23# lyophilisates were prepared with reference to the method in Example 27, all of which were white lumps and smooth in structure, and the above four lyophilisates were rapidly reconstituted with water for injection.

Example 32: Freeze-drying experiments 23#-26#

Using rocuronium bromide and dextran 40, 23#-26# lyophilisates were prepared with reference to the method in Example 28, all of which were white lumps and smooth in structure, and the above four lyophilisates were rapidly reconstituted with water for injection.

Example 33: Freeze-drying experiments 27#-30#

Using rocuronium bromide and lactose, the 27#-30# lyophilisates were prepared with reference to the method in Example 29, all of which were white lumps and smooth in structure, and the above four lyophilisates were rapidly reconstituted with water for injection.

Corresponding freeze-dried preparations of vecuronium were prepared with reference to Examples 26-33, preferably with a pH of 4.5-6.

With reference to Examples 26-33, corresponding freeze-dried preparations of mixed acid steroid quaternary ammonium compounds were prepared respectively, and the pH was preferably 4.5-6.

With reference to Examples 26-33, a small amount of acid was used to adjust pH≥4, preferably 4.5-6, to prepare corresponding freeze-dried preparations of rocuronium bromide, respectively.

With reference to Examples 26-33, a small amount of acid was used to adjust pH≥4, preferably 4.5-6, to prepare corresponding freeze-dried preparations of vecuronium bromide, respectively.

Experimental Example 1: Experiment of rocuronium chloride and rocuronium bromide in rabbit ear vein injection

1. Test material

1.1. Experimental animals

Japanese white rabbit, ordinary grade, ♂, weight 2-2.5kg.

1.2. Tested product

●Rocuronium chloride lyophilized preparation reconstituted in water for injection (10mg/ml)

●Rocuronium bromide commercial preparation (10mg/ml)

2. Methods and Results

Six Japanese white rabbits were randomly divided into two groups, A and B, with 3 rabbits per group. Neostigmine (0.02 mg/kg) and atropine (0.04 mg/kg) were administered intramuscularly 15 minutes before the injection of muscle relaxants into the ear vein. The animals in group A were given 1ml/mouse of rocuronium chloride freeze-dried reconstituted solution (10mg/ml) via the marginal ear vein, and the animals in group B were given 1ml/mouse of the commercial preparation of rocuronium bromide (10mg/ml) via the marginal ear vein , the liquid is injected at a constant speed in about 15-20 seconds, and the performance of the animals during and after injection is observed. The results are recorded in the following table:

The results show that the compound preparation of the present invention has the effect of eliminating or significantly reducing the injection pain of commercially available rocuronium bromide.

Experimental Example 2: Rat Arterial Injection EMG Detection Experiment (Physiological Saline Solution)

1. Test material

1.1. Experimental animals

SD rats, ♂, body weight 300-350g.

1.2. Test drugs

●Solution A: 5 mg/ml rocuronium chloride in physiological saline solution;

●Solution B: commercial preparation of rocuronium bromide, diluted to 5 mg/ml with normal saline;

• Solution C: physiological saline.

2. Methods and Results

SD rats were given urethane (1.2-1.3g/kg) intraperitoneally. After the animals entered a stable state of anesthesia, the superficial epigastric artery was surgically separated, and electrodes were implanted in the ipsilateral semitendinosus muscle, which was connected to BL-420N biosignal acquisition. With the analysis system, the EMG signal is recorded. 60ul of the above test substances were injected through the superficial epigastric artery respectively, and the administration time was about 3 seconds, and the EMG signals were recorded as shown in Figures 1 to 3 (arrows in the figure).

is the start time of dosing). The ratio of the response stationary signal integral (with a duration of 8 seconds) to the baseline stationary signal integral (with a duration of 8 seconds) before administration in each of the above electromyograms was plotted, as shown in Figure 4 .

The above results show that the electromyographic detection of the vascular injection of the compound of the present invention-physiological saline group shows that its vascular stimulation signal is extremely significantly lower than that of the commercially available rocuronium bromide preparation group, and significantly lower than the same concentration and dose of rocuronium bromide. The ammonium-normal saline group was basically equivalent to the normal saline group. In addition, in the experiment, it was observed that the rats administered with solution B (commercially available formulation of rocuronium bromide) had muscle twitching phenomenon, while the rats administered with solution A and solution C did not have this phenomenon, indicating that rocuronium bromide was administered in the market. The commercially available preparations are irritating to a certain extent and have the side effect of injection pain.

Experimental Example 3: Rocuronium chloride and rocuronium bromide injection pain-myoelectric stimulation test in rats (5% glucose solution)

1. Test material

1.1. Experimental animals

SD rat, ♂, body weight 280g-355g.

1.2. Test drugs

The solution is prepared as follows:

30% urethane solution: Weigh 6.01 g of urethane, add 20 mL of purified water, and vortex to dissolve.

Commercially available rocuronium bromide diluted solution (E): 5 mg/mL, take 2 mL of commercially available rocuronium bromide injection, add 2 mL of 5% glucose injection, and vortex to dissolve.

Rocuronium chloride-5% glucose solution (F): 5 mg/mL, weigh 18.7 mg of rocuronium chloride prepared by the present invention, add 3.74 mL of 5% glucose injection, and dissolve by vortexing.

The rats were intraperitoneally injected with 30% urethane solution. After anesthesia, they were fixed on the operation board. The surface hair of the operation site was cut off. The superficial abdominal artery on one side was found and separated with a thread. A single-use injection needle was inserted into the isolated superficial epigastric artery for drug injection, and the injection needle was replaced with the drug. The blank control group was injected with 5% glucose (ie, solution D) 60uL, and the above test substances, namely solution E and solution F, were injected through the superficial epigastric artery, 40ul each, and the EMG signals were recorded as shown in Figures 5 to 7 (arrows in the figure). is the start time of dosing).

A section of representative data before and after each administration was taken, integrated and averaged, and the stimulation intensity of different solutions was compared by the ratio of the values after administration and before administration, as shown in Figure 8. Specifically, it is a graph of the ratio of the post-dose response plateau signal integral (8 sec duration) to the pre-dose baseline plateau signal integral (8 sec duration) in each EMG of solutions D-F.

It can be seen from the above results that, compared with the commercially available rocuronium bromide, the rocuronium chloride compound of the present invention has less stimulation intensity and less injection pain on rats, and is basically equivalent to the 5% glucose group. In addition, in the experiment, it was observed that the rats administered with solution E (commercially available formulation of rocuronium bromide) had muscle twitching, while the rats administered with solution D and solution F did not have this phenomenon, indicating that rocuronium bromide was administered in the market. The commercially available preparations are irritating to a certain extent and have the side effect of injection pain.

Experimental example 4: Rocuronium chloride and rocuronium bromide mouse rotarod test

experimental method:

(1) Primary screening of mouse rotarod

Three days before the experiment, the rotarod was initially screened, and the rotating speed of the mouse rotarod was set to a constant speed of 20 rpm/min. Five mice were placed on a rotarod with a diameter of 3 cm, the rotarod was activated, and if the mouse fell during the experiment, it was placed on the rod again. Each training is 4 minutes, a total of 5 times, and the interval between two trainings is more than 20 minutes as the fatigue recovery time. The dwell time (time from placing to dropping) of the mouse on the rotarod was recorded on the 3rd, 4th and 5th times. Mice with two dwell times of less than 4 min, immobile axes, poor jumping and physical coordination were excluded. Eligible mice were regrouped after screening. Experiments were carried out under the condition of 20-24°C air-conditioned room temperature.

(2) Rod rod test after administration in mice

After 10 minutes of administration on the day of the experiment, the mice were placed on the rotating rod, and the time (s) from placing the mice to falling down was recorded. The _ED50 was calculated by a linear fit using GraphPad Prism 5 software.

Experimental results:

After 10 minutes of intraperitoneal administration, rocuronium chloride had a dose-dependent effect on skeletal muscle relaxation within the dose range of 0.5-1 mg/kg. With the increase of dose, the residence time of mice on the rotarod was significantly shortened. Compared with the blank group, p<0.05 at doses of 0.7, 1 mg/kg. Its ED ₅₀ is 0.84mg/kg; the positive drug rocuronium bromide has a significant relaxation effect on skeletal muscle in the dose range of 0.39-1mg/kg. With the increase of dose, the residence time of mice on the rotarod is also significantly shortened. Compared with the blank group, p<0.05 at the dose of 0.625, 1 mg/kg. Its ED ₅₀ is 0.83mg/kg;

The results show that the ED ₅₀ of rocuronium chloride exerting muscle relaxation effect is equivalent to that of the positive drug rocuronium bromide, and the rocuronium chloride prepared by the present invention has muscle relaxation effect, and the effect is equivalent to that of rocuronium bromide.

Claims

A steroidal quaternary ammonium compound having the following formula I or an addition salt or a solvate thereof:

in,

A is CH2 or O;

B is N or

R 1 is H or CH 3 CO;

R 2 is CH 3 or -CH 2 CH=CH 2 ;

t=2 or 3;

X is Cl, Br or CH 3 COO;

Y is Cl, Br or CH 3 COO;

The values of m and n are both ≥0, and m+n=1 or 2;

The condition is: when any one of X and Y is Br, m+n=2.
The steroidal quaternary ammonium compound or addition salt or solvate thereof of claim 1, wherein the steroidal quaternary ammonium compound is selected from the group consisting of:
The steroidal quaternary ammonium salt compound or its addition salt or its solvate according to claim 1, wherein the solvent in the solvate is water, alcohol, acid, ester, ether, ketone or halogenated hydrocarbon, Preferred are water, C1-6 alcohols, C1-6 acids, C3 - C6 esters, C4-6 ethers, C3-6 ketones or C1-6 halogenated hydrocarbons.
The steroidal quaternary ammonium salt compound or addition salt or solvate thereof according to claim 3, wherein the solvent is selected from the group consisting of water, methanol, ethanol, propanol, isopropanol, n-butanol , tert-butanol, acetic acid, propionic acid, butyric acid, ethyl acetate, methyl acetate, isopropyl acetate, methyl propionate, ethyl propionate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, acetone, butyl ketones, dichloromethane and chloroform or combinations thereof.
The method for preparing the rocuronium chloride of the structure of formula I-1 according to claim 2, which is prepared as follows: the rocuronium bromide is dissolved in water to prepare a solution, enters a reversed-phase liquid phase, and conducts a gradient with a solution containing hydrochloric acid. Elution, collecting fractions, and the obtained fractions are concentrated under reduced pressure and dried to obtain rocuronium chloride hydrochloride with the following structure of formula I-2;

Adding the rocuronium chloride hydrochloride to an organic solvent A and an inorganic base, stirring at room temperature, filtering, and concentrating the filtrate to obtain the rocuronium chloride of the formula I-1, wherein the organic solvent A is selected from the following In the group: ethyl acetate, methyl acetate, isopropyl acetate, methyl propionate, ethyl propionate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, acetone, methyl ethyl ketone, dichloromethane, chloroform and acetonitrile or Its combination is preferably selected from the following group: tetrahydrofuran, acetone, dichloromethane, chloroform, acetonitrile and methyl tert-butyl ether or a combination thereof, and the inorganic base is selected from the following group: sodium hydroxide, potassium hydroxide, Lithium hydroxide, calcium hydroxide, potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate, calcium carbonate, sodium bicarbonate, potassium bicarbonate, lithium bicarbonate, calcium bicarbonate or a combination thereof, preferably selected from the group consisting of carbonic acid Sodium, potassium carbonate, lithium carbonate and calcium carbonate or combinations thereof.
The method for preparing the rocuronium chloride of the formula I-1 structure according to claim 2, which is prepared by the following reaction scheme 1:

(Route 1)

Wherein, compound M8 is reacted with allyl chloride at a temperature of 0-80° C., preferably 20-60° C., more preferably 30-50° C., for 12-168 hours, and rocuronium chloride is obtained through a post-processing procedure.
The method according to claim 6, wherein the post-processing procedure comprises the following steps: concentrating the reaction solution to obtain crude rocuronium chloride, preparing by reversed-phase liquid chromatography, using aqueous ammonium acetate and methanol as mobile phases, The acetate fraction was obtained by the first elution and purification, and then prepared by reversed-phase liquid chromatography. Hydrochloric acid solution and methanol were used as mobile phases, and the rocuronium chloride hydrochloride fraction was obtained by the second elution and purification. The obtained fraction After concentration under reduced pressure, organic solvent A and inorganic base are added to the residue, stirred at room temperature, filtered, and the filtrate is concentrated to obtain the rocuronium chloride, wherein the organic solvent A is selected from the following group: ethyl acetate, methyl acetate ester, isopropyl acetate, methyl propionate, ethyl propionate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, acetone, methyl ethyl ketone, dichloromethane, chloroform and acetonitrile or combinations thereof, preferably selected from the group : tetrahydrofuran, acetone, dichloromethane, chloroform, acetonitrile and methyl tert-butyl ether or a combination thereof, the inorganic base is selected from the group consisting of sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, carbonic acid Potassium, sodium carbonate, lithium carbonate, cesium carbonate, calcium carbonate, sodium bicarbonate, potassium bicarbonate, lithium bicarbonate, calcium bicarbonate or combinations thereof, preferably selected from the group consisting of sodium carbonate, potassium carbonate, lithium carbonate and carbonic acid calcium or a combination thereof.
The method according to claim 6, wherein the post-processing procedure comprises the following steps: concentrating the reaction solution, adding water to dissolve, adding dichloromethane or ethyl acetate for extraction, adjusting the pH of the aqueous phase to 1-6, adding activated carbon for decolorization, Filter, neutralize the pH of the filtrate to 7-9, the temperature of the filtrate during the neutralization process is less than or equal to 10°C, filter, add sodium chloride to the filtrate until a saturated solution of sodium chloride is formed, add dichloromethane for extraction, and the organic phase is filtered through a desiccant Drying, filtering, and concentrating the filtrate, adding the concentrate to an anhydrous solvent selected from the group consisting of acetonitrile, propionitrile, methanol, ethanol, isopropanol, propanol, butanol, sec-butanol, tert-butyl alcohol alcohol, dichloromethane, chloroform, N,N-dimethylformamide and dimethyl sulfoxide or a combination thereof, dissolve, filter, drop the filtrate into organic solvent B, crystallize, filter, and dry the filter cake under reduced pressure, Rocuronium chloride is obtained, wherein the organic solvent B is selected from the group consisting of ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, or ethers such as acetone, butanone , cyclohexanone, ketones of cyclopentanone, or esters such as ethyl acetate, methyl acetate, propyl acetate, butyl acetate, isopropyl acetate, or combinations thereof.
The method for preparing rocuronium chloride hydrochloride or vecuronium chloride hydrochloride as claimed in claim 2, which is prepared as follows: dissolving rocuronium chloride or vecuronium chloride with an organic solution of hydrogen chloride, said The organic solution of hydrogen chloride is selected from the group consisting of: a hydrogen chloride solution of methanol, ethanol, isopropanol, acetonitrile, propionitrile or a combination thereof, into which is added dropwise an organic solvent B selected from the group such as Diethyl ether, isopropyl ether, tertiary methyl ether, tetrahydrofuran, tetrahydropyran, dioxane ethers, such as acetone, butanone, cyclohexanone, cyclopentanone Ketones, such as ethyl acetate, methyl acetate , propyl acetate, butyl acetate, esters of isopropyl acetate or their combination, stirring and crystallizing, filtering, and drying the filter cake under reduced pressure to obtain rocuronium chloride hydrochloride or vecuronium chloride hydrochloride.
The method for preparing the rocuronium chloride hydrochloride of the formula I-2 structure according to claim 2, which is prepared as follows: preparing rocuronium chloride according to the method according to claim 6, and then performing a post-processing comprising the following steps on it. Processing procedure: Concentrate the reaction solution, add organic solvent B, separate out the crude product, filter, dissolve the solid in water, extract the aqueous layer with dichloromethane or ethyl acetate, adjust the pH of the aqueous phase to 1-6, add activated carbon for decolorization, filter, and the filtrate Concentrating under reduced pressure at ≤60°C, preferably at 30-50°C, adding organic solvent B, dispersing and beating, filtering, and drying the filter cake under reduced pressure at ≤40°C to obtain the rocuronium chloride hydrochloride, wherein The organic solvent B is selected from the group consisting of ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, such as acetone, butanone, cyclohexanone, cyclopentanone ketones, such as ethyl acetate, methyl acetate, propyl acetate, butyl acetate, esters of isopropyl acetate, or combinations thereof.
The method of claim 10, further comprising the steps of: adding an organic solvent C to the filter cake, beating, filtering, and drying the filter cake under reduced pressure at ≤40°C, and the organic solvent C is selected from the group consisting of: alkane, hexane, heptane, petroleum ether, acetone, butanone, cyclohexanone, cyclopentanone, or a combination thereof.
The method for preparing the rocuronium chloride hydrochloride of the formula I-2 structure according to claim 2, which is prepared as follows: preparing rocuronium chloride according to the method according to claim 6, and then performing a post-processing comprising the following steps on it. Processing procedure: Concentrate the reaction solution, add organic solvent B, separate out the crude product, filter, dissolve the solid in water, extract the aqueous layer with dichloromethane or ethyl acetate, adjust the pH of the aqueous phase to 1-6, add activated carbon for decolorization, filter, and the filtrate Concentrate under reduced pressure at ≤60°C, preferably at 30-50°C, add alcohol solvent to dissolve, and the alcohol solvent includes methanol, ethanol, isopropanol, propanol, butanol, sec-butanol, and tert-butanol. at least one or a combination thereof, adding the obtained alcoholic solution to an organic solvent B, crystallizing, filtering, and drying the filter cake under reduced pressure at ≤ 40 °C to obtain the rocuronium chloride hydrochloride, wherein the organic solvent B is selected from In the following group: ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, ketones such as acetone, butanone, cyclohexanone, cyclopentanone, such as ethyl acetate ester, methyl acetate, propyl acetate, butyl acetate, esters of isopropyl acetate, or combinations thereof.
The method for preparing the rocuronium chloride hydrochloride of the formula I-2 structure according to claim 2, which is prepared as follows: preparing rocuronium chloride according to the method according to claim 6, and then performing a post-processing comprising the following steps on it. Processing procedure: Concentrate the reaction solution, add organic solvent B, separate out the crude product, filter, dissolve the solid in water, extract the aqueous layer with dichloromethane or ethyl acetate, adjust the pH of the aqueous phase to 1-6, add activated carbon for decolorization, filter, and the filtrate ≤60°C under reduced pressure concentration, preferably 30～50°C under reduced pressure concentration, the concentrated solution is filtered, the filtrate concentration is 0.1～1500mg/ml, preferably 10～1200mg/ml, freeze-dried to obtain the rocuronium chloride hydrochloride, wherein The organic solvent B is selected from the group consisting of ethers such as diethyl ether, isopropyl ether, methyl tertiary ether, tetrahydrofuran, tetrahydropyran, dioxane, such as acetone, butanone, cyclohexanone, cyclopentanone ketones, such as ethyl acetate, methyl acetate, propyl acetate, butyl acetate, esters of isopropyl acetate, or combinations thereof.
The method for preparing the hydrate of rocuronium chloride hydrochloride having the structure of formula I-2 according to claim 2, which is prepared as follows: preparing rocuronium chloride according to the method of claim 6, and then carrying out the following steps: The post-processing procedure of the step: the reaction solution is concentrated, organic solvent B is added, the crude product is separated out, filtered, the solid is dissolved in water, the aqueous layer is extracted with dichloromethane or ethyl acetate, the pH of the aqueous phase is adjusted to 1-6, activated carbon is added for decolorization, Filtration, the filtrate is concentrated under reduced pressure at ≤60 °C, preferably at 30-50 °C, to obtain the hydrate of rocuronium chloride hydrochloride, wherein the organic solvent B is selected from the following group: such as diethyl ether, isopropyl Ethers, tertiary methyl ether, tetrahydrofuran, tetrahydropyran, ethers of dioxane, such as acetone, butanone, cyclohexanone, cyclopentanone Ketones such as ethyl acetate, methyl acetate, propyl acetate , butyl acetate, esters of isopropyl acetate, or combinations thereof.
A freeze-dried preparation comprising a steroidal quaternary ammonium compound as claimed in any one of claims 1 to 4 or an addition salt or a solvate thereof or rocuronium bromide or vecuronium bromide as active ingredient and One or more pharmaceutically acceptable excipients.
The freeze-dried preparation according to claim 15, wherein the adjuvant comprises a freeze-drying support agent and/or a protective agent, or a powder mist inhalation supplement, preferably selected from the group consisting of: mannitol, sorbitol, xylitol, Sucrose, lactose, glucose, dextran, dextrin, maltose, maltitol, maltodextrin, erythritol, trehalose, calcium gluconate, calcium sulfate, sodium chloride, glycine, hydrolyzed gelatin, human albumin or a combination thereof.
The lyophilized preparation according to claim 15, wherein the auxiliary material comprises a pH adjusting agent, and the pH adjusting agent is preferably selected from the following: hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, acetic acid, sodium dihydrogen phosphate, dihydrogen phosphate Potassium, ammonium dihydrogen phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, diammonium hydrogen phosphate, sodium phosphate, potassium phosphate, ammonium phosphate, sodium hydrogen sulfate, potassium hydrogen sulfate, ammonium hydrogen sulfate, sodium hydrogen carbonate, potassium hydrogen carbonate , sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, ammonia water, citric acid, sodium dihydrogen citrate, potassium dihydrogen citrate, ammonium dihydrogen citrate, disodium hydrogen citrate, citric acid Dipotassium hydrogen citrate, diammonium hydrogen citrate, sodium potassium hydrogen citrate, sodium citrate, potassium citrate, ammonium citrate, lactic acid, sodium lactate, potassium lactate, ammonium lactate, malic acid, malic acid Sodium, potassium malate, malic acid, sodium hydrogen malate, potassium hydrogen malate, ammonium hydrogen malate, potassium sodium malate, tartaric acid, sodium hydrogen tartrate, potassium hydrogen tartrate, ammonium hydrogen tartrate, potassium sodium tartrate, vitamin C, Sodium Vitamin C, Alginic Acid, Sodium Alginate, Succinic Acid, Sodium Succinate, Potassium Succinate, Ammonium Succinate, Sodium Succinate, Potassium Succinate, Ammonium Succinate, Potassium Sodium Succinate, Acetic Acid, Sodium Acetate, Potassium Acetate, Ammonium acetate, amino acids and their salts or combinations thereof.
The lyophilized preparation according to claim 15, wherein the auxiliary material comprises a stabilizer, and the stabilizer comprises at least one of an antioxidant or a metal ion complexing agent or a combination thereof, and the antioxidant is preferably selected from the following In the group: sodium bisulfite, sodium metabisulfite, sodium sulfite, sodium thiosulfate, vitamin C, sodium thioglycolate, glycine, cysteine or a combination thereof, the metal ion complexing agent is preferably selected from the following group: Disodium diaminetetraacetate, calcium sodium EDTA, or a combination thereof.
The lyophilized preparation according to claim 15, wherein the auxiliary materials include analgesics and local anesthetics, and the analgesics and local anesthetics are preferably selected from the group consisting of benzyl alcohol, chlorobutanol, procaine, Cocaine, Tetracaine, Proparacaine, Orbucaine, Benzocaine, Lidocaine, Cincocaine, Prilocaine, Trimethaine, Bupivacaine, Levobupivacaine, Mepivaca Intravenous, ropivacaine, dyclonine, chloroprocaine, articaine, eticaine, or a combination thereof.
The freeze-dried preparation according to claim 15, wherein the auxiliary material comprises a bacteriostatic agent, and the bacteriostatic agent is preferably selected from the group consisting of: benzyl alcohol, chlorobutanol, benzoic acid and its salts, sorbic acid and their salts, parabens, or combinations thereof.
A kit comprising a steroidal quaternary ammonium compound or an addition salt or a solvate thereof according to any one of claims 1-4, or a lyophilized preparation according to any one of claims 15-20.
The steroidal quaternary ammonium compound or its addition salt or its solvate according to any one of claims 1-4 or the freeze-dried preparation according to any one of claims 15-20 is used in the preparation of skeletal muscle Use in relaxation drugs.
23. The use of claim 22, wherein the medicament is administered by the parenteral or parenteral route.
The use according to claim 23, wherein the gastrointestinal route is oral route; the parenteral route is inhalation, intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, mucosal and deep tissue or via eye, via Skin, surface and other external routes.
The preparation method of the freeze-dried preparation according to any one of claims 15-20, comprising the steps of: mixing a compound as an active ingredient, an auxiliary material and a corresponding solvent to prepare a solution, and adjusting the pH of the solution to 1-7, preferably pH 2～6, more preferably pH 3.5～5.5, filter, and freeze-dry the filtrate to obtain a freeze-dried preparation.
The preparation method of claim 25, wherein the solvent is water.
The preparation method according to claim 25, wherein in the prepared solution, the concentration of the compound as the active ingredient is 100 μg/ml-300 mg/ml, and the adjuvant (including lyophilization support agent and/or protection agent) agents, pH adjusters, stabilizers such as antioxidants and metal ion complexing agents, analgesics and/or bacteriostatic agents) at a concentration of 0 mg/ml to 500 mg/ml.
The preparation method of claim 25, wherein the freeze-drying comprises:

(1) in the pre-freezing stage, the solution is cooled to a range of -30°C to -196°C until the solution system is coagulated and frozen;

(2) Optionally, the temperature of the frozen solution system is raised to a temperature in the range of -30°C to -5°C, wherein the frozen solution system is still kept in a frozen state; the frozen solution system is cooled again to - Below 30°C, this process can be repeated;

(3) the primary drying stage, including the sublimation step, by removing the solvent in the frozen solution system in the frozen state described in step (1) or step (2) under reduced pressure, to obtain a partially dried product;

(4) In the secondary drying stage, the residual solvent in the partially dried product in the non-frozen state is removed under reduced pressure to obtain a freeze-dried product, and this stage is usually accompanied by further temperature rise.
A multi-part combination kit comprising a first container containing the lyophilized formulation of any one of claims 15-20 and a second container containing a physiologically acceptable A solution for reconstituting the lyophilized formulation.
A solution for injection, which is prepared by the freeze-dried preparation described in any one of claims 15-20, wherein the pH of the solution for injection is not less than 4, and the pH is preferably 4-8, more preferably 5-7, wherein the concentration of the acid is 0-0.15M, preferably 0-0.1M, more preferably 0-0.03M.
The injectable solution of claim 30, wherein the pH is 4.5-6.
The injectable solution of claim 30, wherein the steroidal quaternary ammonium compound is rocuronium bromide or vecuronium bromide, and the pH is 4.5-6.