WO2020259440A1 - Nasal spray of dexmedetomidine, preparation method therefor and use thereof - Google Patents

Abstract

Disclosed is a nasal spray of dexmedetomidine. The basic prescription comprises an osmotic pressure regulator, a preservative, a pH regulator, a carrier, and a pharmaceutically effective amount of dexmedetomidine. The nasal spray of dexmedetomidine according to the prescription has a high stability and a broad pH applicability, despite the simple prescription. The nasal spray has a rapid onset, good pharmacokinetic properties, does not cause irritation to the nasal cavity, and also reduces the cost of production.

Description

Dexmedetomidine nasal spray, its preparation method and application

Association application

This application claims the priority of the patent application with the application number CN201910573195.1 filed with the State Intellectual Property Office of China on June 28, 2019. The entire content of the earlier application CN201910573195.1 including the claims and the specification is incorporated by reference. The way to join this article.

Technical field

The invention belongs to the technical field of medicine, and specifically relates to a dexmedetomidine nasal spray, and its preparation method and application.

Background technique

Dexmedetomidine, dexmedetomidine, (+)-4-[(S)-1-(2,3-dimethylphenyl)ethyl]-1H-imidazole, was developed by Orion Pharma and Abbott in cooperation A non-narcotic α-2 adrenergic receptor agonist with anti-sympathetic, sedative and analgesic properties. Its structural formula is as follows:

In terms of the market, dexmedetomidine is currently only approved for marketing in the form of injections. For example, all of the more than ten NMPA approval numbers for dexmedetomidine are injection dosage forms, and the 8 FDA approvals for dexmedetomidine are also injection dosage forms. Contradicting the single marketing authorization injection formulation of dexmedetomidine is the urgent need for clinical application. In view of the fact that dexmedetomidine is more selective in the activation of α-2 adrenergic receptors and has a smaller effective dosage, it has attracted much attention and expectations once it is marketed. However, currently commercially available single injection formulations are used for sedation during surgical tracheal intubation and mechanical ventilation, as well as sedation for non-intubated patients during surgery, before and/or during medical examinations, all require professional medical technicians To proceed. Therefore, its more convenient way of administration has always been a clinical requirement.

In terms of technical research, the current technical research on dexmedetomidine is still focused on the preparation process of dexmedetomidine and injection formulations. In addition, in CN106511273A and CN102639131A, Ruikeo Pharmaceutical Company disclosed the sublingual dexmedetomidine composition, giving a new way of sublingual administration. The dexmedetomidine intranasal drug delivery industry has little research disclosure. Lu Chengxiang et al. (PLoS One.2016; 11(5): e0154192) disclosed the method of using injection nasal drops, but due to the need to use a larger volume of injection, the droplet surface area and the corresponding mucosal absorption area are very limited. It will lead to problems of low bioavailability and greater variation. Xie Zhubin et al. (Am J Emerg Med. 2017Aug; 35(8):1126-1130) disclosed that the injection of MAD Nasal ^TM intranasal nebulization device can be used to administer the injection, which can improve the aforementioned problems to a certain extent, but this administration The medicine method needs to be equipped with injections, syringes and needles, and adapted nozzles, and operations such as aspiration, removal of needles, and installation of nozzles are required, which increases the complexity of the operation and the possibility of errors; repeated use of the syringe makes it susceptible to contamination by microorganisms and other factors; In addition, this method of administration is based on injection, which requires the use of a relatively large volume of liquid medicine. The problem of liquid medicine loss due to the limitation of the nasal cavity volume and further affecting safety and effectiveness is also clearly foreseen.

The characteristics of intranasal administration of dexmedetomidine determine that its requirements for drugs are significantly different from injection administration. The administration of dexmedetomidine injection is usually a one-time administration and is usually used in hospitals. Good storage conditions can ensure its stability by providing a better storage and use environment. In contrast, nasal spray administration usually requires repeated use of a bottle or can for many times, and it is likely to be used in a variety of environments without good storage conditions. On the other hand, dextromethorphan injection is usually completely sealed and packaged in a glass ampoule for one-time use, which is completely isolated from the outside world. In addition to glass containers, the packaging materials of dexmedetomidine nasal spray also include metering pumps, seals, push buttons and other packaging materials that are in contact with the liquid for a long time, and are semi-open structures. The API concentration of nasal spray is also much higher than that of injection. Under the condition of high concentration of dexmedetomidine, its compatibility with packaging materials needs to be considered again. The above factors will affect the storage stability of dexmedetomidine nasal spray. On the other hand, intranasal administration acts on the nasal mucosa, which also puts forward higher requirements for the irritation of dexmedetomidine drugs. In addition, intranasal administration also requires comprehensive consideration of many factors such as bioavailability and absorption stability. For the above reasons, the existing dexmedetomidine injection medicine is not suitable for direct use as a nasal spray. Recoo company provides an intranasal dexmedetomidine composition in CN104470516A, which has carried out a more systematic study on the intranasal administration of dexmedetomidine, but its given intranasal dexmedetomidine The formula composition of the mididine composition is still complicated, and the auxiliary materials are used more, and the stability of the composition is still to be discussed. On the other hand, the specific implementation case of Ricoeur CN104470516A mainly uses phenylethanol as a preservative. Phenylethanol has low antibacterial activity, requires a higher concentration, and has the characteristics of being volatile and easily adsorbed by plastics or rubber. When phenylethanol is stored in a semi-open device of dexmedetomidine nasal spray for a long time, Due to volatilization or adsorption of packaging materials, the content will be insufficient and the antibacterial effect will be weakened; in addition, the presence of light and oxidants will accelerate the degradation of phenylethanol, thereby reducing the antibacterial effect and increasing the impurity components of the nasal spray.

Summary of the invention

Based on the above-mentioned problems existing in the prior art, the present invention has done a series of in-depth research on dexmedetomidine nasal spray, and proposes a new type of dexmedetomidine nasal spray, which has fast onset, excellent and stable effect , Good stability, less irritation and high bioavailability of one or more of the technical advantages.

In CN104470516A, the general concept mentions that many kinds of preservatives can be used, and it reveals the problem of precipitation caused by the use of benzalkonium chloride. It further pointed out that a suitable preservative is phenylethanol, which can keep the preservative not precipitated but in the composition solution. Based on this, the core specific implementation example 1 gives the intranasal formulation prescription and preparation method, using phenylethanol as a preservative and adding EDTA-2Na as a stabilizer.

However, in the practical verification of the present invention, it has been found that the prescription nasal spray formulations using phenylethanol as a preservative have a greater stability problem. Secondly, the various preservatives mentioned in the general concept of CN104470516A have different performances in the formulation stability of dexmedetomidine nasal spray. Furthermore, when potassium sorbate is selected as the preservative, the additional addition of EDTA-2Na has an opposite adverse effect on the stability of the nasal spray formulation. In addition, the formulations specifically provided in CN104470516A are complicated in composition and do not have cost advantages. At the same time, they have more or less certain aspects in the speed of onset, stability, and bioavailability. insufficient.

Based on the above findings, the basic technical idea of the present invention is to optimize the formulation composition of dexmedetomidine nasal spray, and provide a new formulation of dexmedetomidine nasal spray, which overcomes one of the aforementioned or Two or more technical defects, and then achieve the characteristics of fast onset, strong stability and/or high bioavailability, and do not produce mucosal irritation.

The dexmedetomidine nasal spray provided by the present invention comprises a carrier and a therapeutically effective amount of dexmedetomidine or a pharmaceutically acceptable salt, solvate or isomer thereof. Based on dexmedetomidine, the optional concentration of dexmedetomidine in the nasal spray is 0.005% to 1.3%, preferably 0.0125 to 0.7%, more preferably 0.006% to 0.108%. Dexmedetomidine is preferably dexmedetomidine hydrochloride. When it is dexmedetomidine hydrochloride, the preferred concentration of dexmedetomidine hydrochloride in the nasal spray is 0.006% to 1.54%, preferably 0.015 to 0.83%, More preferably, it is 0.007% to 0.13%. Dexmedetomidine nasal spray can release 2.5～650μg of dexmedetomidine per press. The spray particle size Dv90 value of dexmedetomidine nasal spray is preferably 50-152.1 μm, more preferably 52-142 μm, still more preferably 51-66 μm, 66-81 μm, 81-104 μm or 104-152.1 μm. The Dv90 value of the spray particle size can be controlled by the amount of thickener added. Controlling the Dv90 value of the spray particle helps to control and obtain a unique drug-time curve, which is suitable for clinical application for specific purposes.

In the verification study of the example, it was found that when the preservative is selected as benzalkonium chloride, dexmedetomidine nasal spray is compared with "phenylethanol" or "preservative combination of potassium sorbate and EDTA-2Na". The stability of tomidine nasal spray has been significantly enhanced. Further in-depth studies according to specific embodiments of the present invention have found that when the carrier is water, the prescription stability of dexmedetomidine nasal spray is significantly enhanced under the selection of specific preservatives. The specific preservatives are cationic surfactant preservatives such as benzalkonium chloride, benzalkonium bromide or benzethonium bromide, and paraben preservatives such as methyl paraben and paraben Ethyl, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, isopropyl p-hydroxybenzoate, isobutyl p-hydroxybenzoate, sodium propyl p-hydroxybenzoate or sodium methyl p-hydroxybenzoate, non Aromatic alcohol preservatives such as propylene glycol or chlorobutanol. Based on the above findings, the preservatives of the dexmedetomidine nasal spray of the present invention can be selected from benzalkonium chloride, benzalkonium bromide, propylene glycol, chlorobutanol, methyl paraben and paraben One or a combination of two or more propyl esters. When the preservative is selected as benzalkonium chloride or benzalkonium bromide, the concentration can be selected to be 0.002% or more, preferably 0.01% or more, more preferably 0.02% or more, and more preferably 0.01% to 0.2%. When the preservative is selected as methyl p-hydroxybenzoate, its concentration can be selected to be 0.01% or more, preferably 0.01% to 0.25%. When the preservative is selected as propyl p-hydroxybenzoate, the concentration can be selected to be 0.01% or more, preferably 0.02% or more, more preferably 0.02% to 0.075%. When the preservative is selected as propylene glycol, the concentration can be selected to be 0.05% or more, preferably 0.05% to 30%, more preferably 0.1% to 10%. When the preservative is selected as chlorobutanol, its concentration can be selected to be 0.05% or more, preferably 0.05% to 0.75%. When the preservative is selected as a combination of methyl p-hydroxybenzoate and propyl p-hydroxybenzoate, the concentrations of the two can be selected to be more than 0.01% and more than 0.01%, and the concentrations of the two are preferably 0.01% to 0.25% and 0.01% to 0.075%, and the two concentrations are more preferably 0.02% to 0.04% and 0.01% to 0.02% in sequence. When the preservative is selected as a combination of propylene glycol and benzalkonium chloride, the concentrations of the two can be selected to be more than 0.1% and more than 0.01%, and the concentrations of the two are preferably 0.1%-10% and 0.01%-0.2%. The type of carrier water may be a pharmaceutically acceptable type, including but not limited to purified water, water for injection, and the like. The single or compound combination of preservatives selected by the present invention can also be used together with antioxidants. The antioxidants can be EDTA-2Na, tert-butyl hydroxyanisole, 2,6-di-tert-butyl-4 -One or a combination of two or more of methylphenol, sodium metabisulfite, potassium metabisulfite, butylated hydroxyanisole, L-ascorbyl palmitate, sodium thiosulfate and vitamin E. In addition, according to the verification of specific embodiments of the present invention, the preservative of the present invention can also be selected as potassium sorbate, but when the preservative is selected as potassium sorbate, the formulation should not contain EDTA-2Na. The concentration can be selected from 0.01% to 0.2%.

Different thickeners have different effects on the spray performance of nasal spray formulations. According to the specific embodiments of the present invention, in order to obtain good nasal spray spray performance under comprehensive consideration of various factors of viscosity and spray performance, the present invention provides Dexmedetomidine nasal spray also preferably contains a thickener, which is among povidone thickeners, cellulose ether thickeners, mucopolysaccharide thickeners, and polyacrylic thickeners. One or a combination of two or more, povidone thickener can be PVP K30 or PVP K90, cellulose ether thickener can be microcrystalline cellulose-sodium carboxymethyl cellulose composite, Hypromellose, methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose or hydroxyethyl cellulose, mucopolysaccharide thickeners can be sodium hyaluronate or chondroitin sulfate, poly The acrylic thickener can be carbomer. When the thickener is selected as PVP K30, its concentration can be selected from 0.1% to 5.0%. When the thickener is selected as the microcrystalline cellulose-sodium carboxymethyl cellulose composite, the concentration can be selected to be 0.1% to 3.0%. When the thickener is selected as hypromellose, its concentration can be selected from 0.1% to 5.0%, preferably 0.2% to 1.5%. When sodium hyaluronate is selected as the thickener, its concentration can be selected from 0.01% to 1.0%. When the thickener is selected as carbomer, its concentration can be selected from 0.05% to 1.0%.

The dexmedetomidine nasal spray provided by the present invention also preferably contains an osmotic pressure regulator, the osmotic pressure regulator is ionic or non-ionic, and the ionic osmotic pressure regulator includes boric acid, sodium chloride, calcium chloride , Magnesium chloride, zinc chloride or potassium chloride, non-ionic osmotic pressure regulators include anhydrous dextrose, dextrose monohydrate, dextran, glycerol or D-mannitol; the osmotic pressure regulators are preferably boric acid, sodium chloride , Potassium chloride, anhydrous dextrose, dextrose monohydrate, dextran, glycerol and a combination of two or more of D-mannitol, the osmotic pressure regulator is especially preferably sodium chloride or potassium chloride When the osmotic pressure regulator is selected as sodium chloride or potassium chloride, its concentration is preferably 0.7% to 1%.

The pH of the dexmedetomidine nasal spray provided by the present invention is preferably 4-7, more preferably 5-6.5. The pH adjusting agent of the dexmedetomidine nasal spray can be an acidulant and/or an alkalinizer, and the acidulant includes hydrochloric acid, phosphoric acid, tartaric acid, citric acid, malic acid, fumaric acid, lactic acid, ascorbic acid and acetic acid. , Alkalizing agents include meglumine, trimethylolmethane, sodium phosphate, arginine, lysine, glycine, ammonia, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate and borax; The pH adjuster may further preferably be one of hydrochloric acid, phosphoric acid, tartaric acid, citric acid, malic acid, fumaric acid, malic acid, ammonia, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate and borax. One or a combination of two or more; the preferred choice of pH adjusting agent is hydrochloric acid and/or sodium hydroxide.

The general preparation method of the aforementioned dexmedetomidine nasal spray provided by the present invention is as follows, weigh dexmedetomidine or its pharmaceutically acceptable salt, solvate or its isomers and various excipients (excipients include preservatives) , Osmotic pressure regulator, thickener (add if necessary)), add carrier to dissolve, and optionally adjust pH. On this basis, it is further divided into plastic or glass bottle containers, a metering pump is assembled on the container, and a push button is installed.

Based on the performance of dexmedetomidine and the technical solutions and effects of specific embodiments of dexmedetomidine nasal spray of the present invention, the present invention also provides its use in the preparation of sedative, analgesic and/or anti-sympathetic products Further application. The said product may be a further compound preparation used in combination with other types of drugs, or a further complex preparation, etc.

Detailed ways

The present invention will be further explained below in conjunction with specific implementation conditions.

In this article, unless otherwise defined, the technical terms used are in accordance with the industry's common understanding.

In this article, unless otherwise specified, the reagents and instruments mentioned can be obtained through ordinary commercial purchases.

In this article, in accordance with the industry's usual practices, unless there are special additional regulations, the percentages, such as "%", all refer to percentages by weight. In the formulation of the present invention, the density of the formulation solution system is approximately converted at 1 g/mL. For example, if 2.4 mg of dexmedetomidine hydrochloride is added to 10 mL of water, the concentration of the dexmedetomidine hydrochloride system is 0.24 mg/mL, and the density of the prescription solution system is approximately converted to 1 g/mL. The weight percentage concentration of the medetomidine system is 0.024%; in the same way, when dexmedetomidine is used at this time, the converted concentration is 0.2 mg/mL, and the weight percentage concentration is 0.02%.

In this article, the weight conversion coefficient between dexmedetomidine hydrochloride and dexmedetomidine is approximately calculated as 1.1822, and the conversion between them follows the counting retention method of rounding. If there is no special statement or industry customary constraints, the data that needs to be calculated in other similar places in this article also uses the rounding counting retention method.

In this article, the specific data value of each listed component can cover the fluctuation of 10% up and down. For example, "dexmedetomidine concentration 0.02%" can actually cover a concentration range of 0.02% ± 0.002%, that is, 0.018% to 0.022%.

As used herein, "effective amount" generally refers to the amount of an active compound or agent that can cause a biological or medical response. The biological or medical response can be human, animal, system model, tissue, cell in vitro, etc. It is generally obtained by medical researchers, doctors or clinicians in experiments or clinical research. A "biological or medical response" can be the rehabilitation, elimination, alleviation or suppression, or prevention of a disease or symptoms associated with the disease. The "quantity" generally can be determined according to the specific situation of the subject, and a relatively universal range can also be determined for subjects with specific common characteristics. In the present invention, the "effective amount" can also preferably refer to the specific amount described in some specific embodiments or the general range supported by them together.

The terms and abbreviations in the present invention are identified as follows:

DEX: short for dexmedetomidine, that is, dexmedetomidine

EDTA-2Na: Shorthand for disodium edetate

K90: The abbreviation of PVP K90, namely polyvinylpyrrolidone K90, also known as povidone K90, Povidone K90

K30: shorthand for PVP K30, namely polyvinylpyrrolidone K30, also known as povidone K30, Povidone K30

HEC 250M: Short for Hydroxyethylmethyl Cellulose 250M, also abbreviated as HEC-M, that is, hydroxyethyl cellulose 250M

HEC 250HX: Hydroxyethylmethyl Cellulose 250HX abbreviation, also abbreviated as HEC-H, namely hydroxyethyl cellulose 250HX

Sodium Hyaluronate: SODIUM HYALURONATE

CL-611:

Shorthand for

Trade name, produced by FMC BioPolymer company, generic name Microcrystalline Cellulose and Cellulose Gum, Microcrystalline Cellulose and Cellulose Gum

RL-591:

Shorthand for

Trade name, produced by FMC BioPolymer company, generic name Microcrystalline Cellulose and Cellulose Gum, Microcrystalline Cellulose and Cellulose Gum

HPMC E50: short for hydroxypropyl methylcellulose E50, namely hypromellose E50, also written as hydroxypropyl methylcellulose E50, hydroxypropyl methylcellulose E50

TCB: short for trichloro-t-butyl alcohol, also known as 2-Trichloromethyl-2-propanol, Chlorobutanol, namely trichloro-tert-butyl alcohol

PE: short for Phenylethyl alcohol, namely phenylethanol

Methyl ester: short for methyl paraben, also known as methyl paraben, Methylparaben

Propyl ester: short for propyl paraben, also known as propyl paraben, Propylparaben

BKB: Benzalkonium bromide, also known as Benzalkonlum bromide

BKC: Benzalkonium chloride, also known as Benzalkonium chloride

CBP: short for carbopol, namely carbomer

PG: short for Propylene glycol, namely propylene glycol

Examples:

The embodiments described herein specifically provide a method for preparing and obtaining dexmedetomidine nasal spray. The general method steps are: weighing API (referring to dexmedetomidine or its pharmaceutically acceptable salt, solvent Compounds or isomers) and auxiliary materials (the auxiliary materials include osmotic pressure regulators, preservatives, thickeners and antioxidants, etc., and the auxiliary materials are optionally added depending on the formulation), add an appropriate amount of purified water to dissolve, and use a pH regulator to adjust the system The pH is within the predetermined range of 4-7, and then the carrier water is added to make the volume constant, and the dexmedetomidine nasal spray is obtained evenly. It can be further divided into containers, assembled on the metering pump, and installed on the push button.

In some specific embodiments, examples of different active ingredient concentrations are prepared. In the dexmedetomidine nasal spray provided, dexmedetomidine exists in the form of dexmedetomidine hydrochloride, which is By default, the concentrations are 0.006%, 0.02%, 0.0592%, 0.0593%, 0.108%, 0.01%, 0.03%, 0.04%, 0.05%, 0.07%, 0.1%, 0.2%, 0.4%, 0.7%, 1 % Or 1.3%.

In other specific embodiments, several dexmedetomil nasal sprays with different prescription specifications are provided. The nasal spray includes a liquid medicine part and a device part, and the liquid medicine is preloaded in the device. The device includes a metering drug delivery device, which is composed of a metering pump and a container that cooperate with each other. The material of the container is presented in three embodiments: plastic, low borosilicate glass, and medium borosilicate glass. The metering pump is provided with an actuator. Under the control of the actuator, the nasal spray is delivered by a metered drug delivery device. By controlling the concentration of dexmedetomidine in the nasal spray and the total amount of dexmedetomidine per press, the quantification of dexmedetomidine per press is 5μg, 10μg, 15μg, 20μg, 25μg, 30μg, 40μg, 50μg, 60μg , 75μg, 100μg or 125μg administration. For specific use, single or multiple nasal sprays can be administered.

In other specific embodiments, the pH of the nasal spray system is 4-7, which can be adjusted by a pH adjusting agent, acidifying agent and/or alkalizing agent. The acidifying agent includes hydrochloric acid, phosphoric acid, tartaric acid, citric acid, and malic acid. , Fumaric acid and malic acid, alkalizing agents include ammonia, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate and borax; pH adjusters can be further preferably hydrochloric acid, phosphoric acid, tartaric acid, citric acid, One or a combination of two or more of malic acid, fumaric acid, malic acid, ammonia, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate and borax.

In other specific embodiments, the osmotic pressure of the nasal spray can be controlled by the concentration of the osmotic pressure adjusting agent, the osmotic pressure adjusting agent is ionic or non-ionic, and the ionic osmotic pressure adjusting agent includes boric acid and sodium chloride. , Calcium chloride, magnesium chloride, zinc chloride or potassium chloride, non-ionic osmotic pressure regulators include anhydrous glucose, dextrose monohydrate, dextran, glycerol or D-mannitol; the osmotic pressure regulator is preferably boric acid , Sodium chloride, potassium chloride, anhydrous dextrose, dextrose monohydrate, dextran, glycerol and D-mannitol one or a combination of two or more.

The following is a formulation table of several typical examples. The pH in these typical examples is adjusted by 1N sodium hydroxide and 1N hydrochloric acid.

Note [1]: The prescription amount of dexmedetomidine hydrochloride is calculated as dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822

Note [2]: "/" means that this type of substance is not added to the corresponding prescription.

Test example:

The following is a more detailed display of the effect research test examples containing some examples to illustrate the present invention more clearly.

Test Example 1: Stability investigation test of basic prescription

1. Formula and preparation process of dexmedetomidine nasal spray

According to the ratio in Table 1.1, weigh dexmedetomidine hydrochloride and various auxiliary materials, add water to dissolve, adjust the pH to the specified value, divide into glass bottles, and assemble the quantitative pump.

Table 1.1

Note [1]: The prescription amount of dexmedetomidine hydrochloride is calculated based on dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822.

Note [2]: "/" means that the substance is not added in the corresponding prescription.

Note [3]: Use 1N sodium hydroxide or 1N hydrochloric acid as the pH regulator.

2. Development batch of dexmedetomidine nasal spray

Make dexmedetomidine nasal spray according to the formulas in Table 1.1, and produce 80 bottles for each formula, and use them for subsequent stability and other project investigations.

Three, stability investigation

Take formulations 1 to 6 of each development batch, and conduct stability inspections of room temperature, accelerated experiments, and high temperature experiments. The results of each inspection item are shown in the following tables.

1. Investigation results of related substances

The investigation results of related substances under the conditions of room temperature, accelerated experiment and high temperature experiment are shown in Table 1.2 below.

Table 1.2

Note [1]: "/" means that this indicator has not been tested at this point in time.

2. Results of inspection of preservative content

The test results of potassium sorbate under room temperature, accelerated conditions and high temperature conditions are shown in Table 1.3 below.

Table 1.3

Note [1]: Content = actual concentration mg/ml÷theoretical concentration mg/ml×100%, the theoretical concentration of potassium sorbate is 1mg/ml;

Note [2]: Degradation rate = (1-measured content÷prepared content)×100%;

Note [3]: "/" means that this indicator has not been tested at this point in time.

The test results of benzalkonium chloride at room temperature and high temperature are shown in Table 1.4 below.

Table 1.4

Note [1]: Content = actual concentration mg/ml÷theoretical concentration mg/ml×100%, the theoretical concentration of benzalkonium chloride is 0.1mg/ml (this experiment did not perform 8% moisture conversion pure);

Note [2]: Degradation rate = (1-measured content÷prepared content)×100%.

3. Further investigation results of isomers, pH and osmotic pressure of formula 1.

Isomers: The observation experiments at room temperature for 10 days, accelerated for 10 days, and high temperature for 10 days have confirmed that the isomer content of formula 1 is always low, stably maintained in the range of 0.01% to 0.02%.

pH: The observation experiments at room temperature for 10 days, acceleration for 10 days and high temperature for 30 days confirmed that the pH of formula 1 was maintained at about 5.5 during the entire test process, with an upward deviation not exceeding 5.75 and a downward deviation below 5.43.

Osmotic pressure: 10 days at room temperature, 10 days at accelerated (40°C) and 10 days at high temperature (60°C) confirmed that the osmolality of formula 1 is always maintained at 278～286mOsmol/kg.

4. Result analysis

1. According to the results in Table 1.2, no related substances were detected in the dexmedetomidine nasal spray corresponding to formula 1 at room temperature for 10 days and accelerated conditions for 30 days. Even in the 30-day experiment under high temperature conditions, the content of related substances in formula 1 is still in the lower range, and the maximum single impurity and total impurity contents are significantly better than other formulas. Under the three conditions of room temperature, acceleration, and high temperature, the isomers, pH and osmotic pressure of the nasal spray corresponding to formula 1 are all shown to be relatively stable.

2. Selection of preservatives:

According to Tables 1.1 and 1.2, the formulations 1, 4, and 6 using benzalkonium chloride as the preservative, the related substances are significantly lower than the other formulations. Especially for formula 1, no related substances were detected in 10 days at room temperature and 30 days in accelerated experiment. Although related substances were detected during 30 days at high temperature, they were still in a low range. Furthermore, in terms of related substances, compared with the formula 1 that only added benzalkonium chloride, the related substances of the formula 4 that additionally added hypromellose increased. Continuing to add additional antioxidant EDTA-2Na formula 6, its related substances further increased significantly.

Compared with formulas 1, 4 and 6, formulas 2, 3 and 5 with potassium sorbate as preservative have significantly increased related substances. The prior art discloses that the preservative potassium sorbate is usually used in combination with EDTA-2Na. However, when EDTA-2Na was added (formulation 3), and when EDTA-2Na and hypromellose were added (formulation 5), the total impurities exceeded 1% after 10 days of acceleration. In further high temperature experiments It even reached 4.33%, which was significantly beyond the controllable level.

Based on the foregoing analysis, it is found that, compared to the nasal spray preparations with potassium sorbate as the preservative, or the nasal sprays with potassium sorbate as the preservative and the antioxidant EDTA-2Na, the preservative is selected as benzalkonium chloride The related substances in the nasal spray preparations have been significantly reduced, and the prescription stability has been significantly improved.

On the other hand, Table 1.3 shows that the preservative potassium sorbate degrades significantly under room temperature, accelerated, and high temperature conditions, especially under accelerated and high temperature conditions. Table 1.4 shows that under the same conditions of other variables (such as formula 1 and formula 2, formula 5 and formula 6), even if the investigation time is longer, the degradation rate of the preservative benzalkonium chloride in the formula is lower than that of sorbic acid Potassium. This also confirms from another aspect that the choice of benzalkonium chloride as a preservative has improved the stability of nasal sprays.

3. Selection of antioxidant EDTA-2Na:

Under normal circumstances, in order to maintain drug stability and extend the shelf life of the drug, a certain amount of antioxidant/stabilizer is added to the drug. This is a technical method frequently used in pharmaceutical technology. In the research field related to dexmedetomidine nasal spray, the existing technical disclosure also follows this law. For example, the intranasal dexmedetomidine composition disclosed by Recoo Company in CN104470516A. Refer to paragraph [0099] of CN104470516A, and the specific configuration process given in Example 1 "Pure water (USP) is added to the container. Mix anhydrous citric acid, sodium citrate dihydrate, sodium chloride into the water , Phenylethanol and disodium EDTA until dissolved.” This technology discloses the introduction of an antioxidant/stabilizer (disodium EDTA) in the product of dexmedetomidine intranasal preparation.

However, in a series of comparative experiments, it was found that when potassium sorbate is used as the preservative, the introduction of the antioxidant EDTA-2Na will not improve the prescription stability of dexmedetomidine nasal spray, but will lead to dexmedetomidine nasal spray. The prescription stability of medetomidine nasal spray is significantly reduced, which is completely different from the disclosure of the prior art.

For details, refer to Table 1.2 in the investigation items of related substances. Comparing formula 3 with EDTA-2Na added to formula 2, it shows that the total impurity content has increased from 0.1% to 1.19%, an increase of about 11 times; the maximum single impurity has increased from 0.05% to 0.83%, an increase of about 10 days. 16 times. The high temperature for 10 days even showed that the total impurity content rose from 0.03% to 3.32%, an increase of more than 100 times. That is, EDTA-2Na, which is usually used in conjunction with potassium sorbate, has brought about a significant increase in related substances when used in practice with potassium sorbate.

Test Example 2:

Rat pharmacokinetic test 1

1. Formula and preparation process of nasal spray

According to the following table 2.1, weigh the dexmedetomidine hydrochloride and various auxiliary materials, add water to dissolve, adjust the pH to 5.0 with sodium hydroxide, divide into glass bottles, assemble the quantitative pump, and add the push button to obtain dexmedetomidine Tomidine nasal spray.

Table 2.1

Name of raw materials	Dosage
Dexmedetomidine Hydrochloride	35.5mg
Sodium chloride	1350mg
Benzalkonium chloride	30mg
Phenylethanol	534.3mg
water	To 150ml

2. Animal experiment process

Take the above-mentioned dexmedetomidine nasal spray and the commercially available dexmedetomidine hydrochloride injection (2mL: 200μg, Jiangsu Hengrui Pharmaceutical Co., Ltd.) as the test products, and give each of them intranasally and intravenously as shown in Table 2.2. Group Sprague Dawley rats (SD rats) with prescribed doses of drugs. In each group before and after administration, 0.3 mL of blood was collected from the jugular vein at 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 1.5 hours, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours. Concentration detection.

Table 2.2

Note: The dosage is based on dexmedetomidine, and the injection is diluted with saline for use.

3. Experimental results

The measured blood drug concentration was analyzed with Phoenix WinNonlin (v 6.4) as a non-compartmental model, and the results are shown in Table 2.3 below.

Table 2.3

4. Result analysis

After SD rats were given dexmedetomidine hydrochloride nasal spray through the nose, the absolute bioavailability reached 83.8%, indicating that the dexmedetomidine hydrochloride nasal spray provided by the present invention has very good bioavailability; The group peak time (T _max ) is only 7.5 minutes, indicating that the nasal spray can be absorbed quickly and has a rapid onset of action. This feature is very beneficial to clinically wishing to quickly achieve a sedative effect.

Rat pharmacokinetic test 2

1. Formula and preparation process of nasal spray

According to the following table 2.4, weigh the dexmedetomidine hydrochloride and each auxiliary material, add water to dissolve, adjust the pH to 5.0 with sodium hydroxide, divide into glass bottles, assemble the quantitative pump, and add the push button to obtain dexmedetomidine Tomidine nasal spray.

Table 2.4

Name of raw materials	Dosage
Dexmedetomidine Hydrochloride	0.02% (calculated as dexmedetomidine)
Sodium chloride	0.9%
Benzalkonium chloride	0.02%
water	To 100%

2. Animal experiment process

1. Blood sample collection, testing and processing

Take the above-mentioned dexmedetomidine nasal spray and the commercially available dexmedetomidine hydrochloride injection (2mL: 200μg, Jiangsu Hengrui Pharmaceutical Co., Ltd.) as the test products, and give each of them intranasally and intravenously as shown in Table 2.5 Group Sprague Dawley rats (SD rats) with prescribed doses of drugs. Before and after administration, 0.2 mL of blood was collected from the jugular vein at 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours and 24 hours in each group. The blood samples were processed After testing.

Table 2.5

2. Gross anatomy and histopathological examination

Gross anatomy: The 30μg/kg nasal spray group was euthanized one day after the 7th administration (i.e. the eighth day of the test). The main organs (heart, kidney, spleen, liver) and respiratory tract (nasopharynx) of the rats in this group , Bronchus, lung) for general anatomical observation.

Histopathological examination: The nasopharynx, bronchi, and lungs of all rats in the 30μg/kg nasal spray group were examined according to histopathology.

3. Experimental results

1. Pharmacokinetic data results

The concentration of dexmedetomidine in plasma was determined by the validated LC-MS/MS method and the pharmacokinetic parameters were calculated. The main results are shown in the following table 2.6-Dextetomidine in plasma after single nasal/intravenous injection in rats The pharmacokinetic parameters of Midine (mean):

Table 2.6

2. Gross anatomy and histopathological examination

SD rats were infused with dexmedetomidine hydrochloride nasal spray 30μg/kg for 7 consecutive days. After the administration, the gross anatomy showed that none of the rats found naked eyes that may be related to dexmedetomidine hydrochloride nasal spray. Abnormal changes.

In SD rats, nasal drip of dexmedetomidine hydrochloride nasal spray 30μg/kg for 7 consecutive days. After the end of the administration, histopathological examination showed that none of the rats was found to be related to dexmedetomidine hydrochloride nasal spray. Abnormal changes in histopathology.

4. Result analysis

The blood concentration of the dexmedetomidine hydrochloride nasal spray for intranasal administration is slightly lower or is basically equivalent to intravenous administration, the exposure level is also equivalent to that of intravenous administration, the bioavailability is high, and the blood concentration reaches the peak The time is short, and no drug-related toxic reaction is observed within 7 days of administration, which shows the good clinical application prospect of the dexmedetomidine nasal spray of the present invention.

Test example 3: Rhesus monkey pharmacokinetic test

1. The formula and preparation process of dexmedetomidine nasal spray

Weigh dexmedetomidine hydrochloride and various auxiliary materials according to the ratio in Table 3.1, add water to dissolve, adjust the pH to the specified value with sodium hydroxide, put them into glass bottles, assemble the metering pump, and install the push button. Get dexmedetomidine nasal spray.

chart 3.1

Note [1]: The prescription amount of dexmedetomidine hydrochloride is calculated based on dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822.

Note [2]: "/" means that the substance is not added.

2. Spray particle size detection

Take samples of the above formula 1-A, 1-C, 1-D, 1-E, 1-F, and use a laser particle size analyzer to detect Dv90 (particle size value, that is, by volume percentage, 90% of the particle size is smaller than this value ). The detection method is: remove the cap of the sample, discard 1 spray, wait 5 seconds, then discard 1 spray, and repeat this operation until 8 sprays are discarded as a start. According to the particle size and particle size distribution measurement method (Chinese Pharmacopoeia 2015 Edition Four General Rules 0982), place the sample on the trigger of the laser particle size analyzer, measure the spray particle size of 3 consecutive sprays at a distance of 3 cm from the nozzle, and record the Dv90 and average value of each spray. The results are shown in Table 3.2 below.

Table 3.2

It can be seen from the results in the above table that by controlling the concentration of hypromellose to 0-10 mg/ml, the average spray particle size Dv90 is 52-142 μm.

3. Animal experiment process

Take dexmedetomidine nasal spray (formula 1-A, formula 1-B, and formula 1-C above) and commercially available dexmedetomidine hydrochloride injection (2mL: 200μg, Jiangsu Hengrui Pharmaceutical Co., Ltd.) As the test product, each group of healthy male rhesus monkeys were given the prescribed doses of drugs by intravenous and nasal sprays as shown in Table 3.3. Before administration and 5min, 10min, 15min, 20min, 30min, 45min, 1h, 1.5h, 2h, 3h, 4h, 5h, 6h, 8h, and 10h after administration, 1 mL of blood was collected from the vein of the upper extremity for blood concentration detection.

Table 3.3

Note: The dosage is based on dexmedetomidine, and the injection is diluted with saline for use.

4. Experimental results

The measured blood drug concentration was analyzed by Phoenix WinNonlin (v 6.4) as a non-compartmental model, and the results are shown in Table 3.4 below.

Table 3.4

5. Result analysis

1. According to the results in Table 3.4, the absolute bioavailability of dexmedetomidine hydrochloride nasal spray is 48.3% to 68.6%, that is, the dexmedetomidine hydrochloride nasal spray provided by the present invention has stable and good bioavailability Degree; body exposure (AUC) increases with the increase of dose, and the multiple of exposure increase is slightly higher than the multiple of dose increase.

2. Under the same dosage, the bioavailability of the nasal spray formula containing thickener (formulation 1-C) is slightly higher than that of the nasal spray formula without thickener (formula 1-A), which is different from Recoo (CN104470516A, an intranasal dexmedetomidine composition) disclosed that "the added viscosity of a thickener will affect the intranasal delivery effect of the drug,..., the use of dexmedetomidine plasma that can cause a decrease Level and therefore omit "under certain conditions" is very different. That is, the use of thickeners (tackifiers) in dexmedetomidine nasal spray is feasible. Refer to Table 3.1 and Table 3.2, the spray particle size can be controlled by the amount of hypromellose added. If the spray particle size is too small, it will be absorbed quickly after contact with the nasal mucosa, but it also brings high blood concentration after nasal administration and unnecessary drug exposure and medication risks in the lower respiratory tract. On the other hand, the flow of the liquid medicine will also cause an uncontrollable risk of the loss of the medicine, making it difficult to perform accurate quantitative administration. As described in the present invention, the addition of hypromellose is appropriately increased to enhance the adhesion to the nasal mucosa, reduce the occurrence of flow, and thereby avoid unpredictable drug loss, so as to achieve more accurate quantitative dosing control. On the other hand, with the increase of hypromellose, the spray particle size value increases, the dissolution of the drug slows down, and the duration of the dissolution of the drug is prolonged. Avoid multiple nasal sprays or a single high-dose nasal spray, which may be accompanied by rapid drug absorption and rapid increase in blood concentration, which will exceed the treatment window in a short time, which will bring medication risks. Furthermore, through the control of the Dv90 value of the spray particle size, the clinical drug effect of stable drug release is obtained, and the bioavailability is improved to a certain extent.

3. Nasal spray and intravenous injection have a similar terminal half-life, indicating that their in vivo pharmacokinetic properties are similar, that is, it is expected to achieve the same effect as intravenous administration under the same exposure in the body, which is good for clinical dosage The control of effects is of great significance.

Test example 4: irritation test

1. Formula and preparation process of nasal spray

According to the following table 4.1, weigh the dexmedetomidine hydrochloride and various auxiliary materials, add water to dissolve, adjust the pH to 5.0 with sodium hydroxide, divide into glass bottles, assemble the quantitative pump, and add the push button to obtain dexmedetomidine Tomidine nasal spray. Take another 0.9% sodium chloride injection into a glass bottle, assemble a quantitative pump, and add a push button to obtain a negative control nasal spray.

Table 4.1

Note: The prescription amount of dexmedetomidine hydrochloride is calculated based on dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822.

2. Animal experiment process

Take the dexmedetomidine hydrochloride nasal spray and the negative control nasal spray of the above formula 1-D and 1-E, and perform the irritation test of the New Zealand rabbit nasal spray. In the experiment, male New Zealand rabbits were sprayed into the left and right nostrils each time with 200 μl each of the medicinal solution or 0.9% sodium chloride injection, administered once a day for 7 consecutive days. The experimental design is shown in Table 4.2.

Table 4.2

Group	Processing factors	Volume per administration (μl)	Dosing frequency	Dosing cycle	Number of animals
Negative control group	0.9% sodium chloride injection	200+200(left+right)	Once a day	7 days	3
Dexmedetomidine Hydrochloride Nasal Spray 1	Formula 1-D	200+200(left+right)	Once a day	7 days	3
Dexmedetomidine Hydrochloride Nasal Spray 2	Formula 1-E	200+200(left+right)	Once a day	7 days	3

3. Experimental results and conclusions

General situation:

During the experiment, no rabbits died in each group.

During the experiment, the rabbits in each group were in good general condition, normal autonomous activities, clean skin and coat, no abnormal symptoms of breathing and central nervous system, and no local irritation symptoms such as asthma, cough, vomiting, and suffocation.

Gross anatomical observation and histopathological examination:

On the day after the last administration, there were no visible abnormal changes in the parts of the respiratory tract (nose, throat, trachea, lung), oral mucosa tissue and all visible organs of the rabbits in each group.

There were no obvious abnormal histopathological changes in the oral mucosa, tongue, nasal cavity (paranasal sinuses), larynx, trachea, main bronchus, and lung of rabbits in each group.

The above results indicate that the rabbits in each administration group are generally in good condition, and there is no obvious abnormality in the general anatomy of the respiratory tract, oral mucosa, and histopathological examination, indicating that it is not irritating to the respiratory tract.

Test Example 5: Antibacterial efficacy

According to the following table 5.1, weigh the dexmedetomidine hydrochloride and various auxiliary materials, add water to dissolve, adjust the pH to 5.0 with sodium hydroxide, put them into glass bottles, assemble the quantitative pump, and add the push button to obtain dexmedetomidine Tomidine nasal spray.

Table 5.1

Name of raw materials	Dosage (%)
Dexmedetomidine Hydrochloride	0.02
Sodium chloride	0.9
Benzalkonium chloride	0.01
water	Up to 100

The antibacterial efficacy of the above nasal spray was investigated, and the results are shown in Table 5.2 below.

Table 5.2

According to the results of the above table, the nasal spray formulated based on formula 1, its 14-day and 28-day antibacterial efficacy meets the standards A and B of the 2015 edition of the Chinese Pharmacopoeia antibacterial efficacy criteria, and has good antibacterial efficacy.

Test Example 6: pH investigation test

1. Formula and preparation process of dexmedetomidine nasal spray

According to the ratio in Table 6.1, weigh dexmedetomidine hydrochloride and various excipients, add water to dissolve, the resulting solution is divided into 4 parts, and adjust the pH to 4.0 with the pH adjuster hydrochloric acid or sodium hydroxide (formulation 1-I), 5.0 (Formulation 1-J), 6.0 (Formulation 1-K), 7.0 (Formulation 1-L), divided into glass bottles, 30 bottles for each formula, assembled with a quantitative pump.

Table 6.1

Dexmedetomidine hydrochloride (mg)	Sodium chloride (mg)	Benzalkonium chloride (mg)	purified water
240	10800	240	Up to 1200mL

Note: The prescription amount of dexmedetomidine hydrochloride is calculated based on dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822.

2. Investigation results

1. Investigation results of related substances

In the pH investigation, no impurities were detected in formula 1-I to formula 1-L under the conditions of room temperature for 10 days, acceleration for 10 days, and high temperature for 10 days. The test results of related substances at room temperature for 30 days, acceleration for 30 days, and high temperature for 30 days are shown in Table 6.2 below.

Table 6.2

2. Results of investigation on dexmedetomidine content

The test results of dexmedetomidine content under room temperature, accelerated and high temperature conditions are shown in Table 6.3 below.

Table 6.3

3. Results of investigation on the content of benzalkonium chloride

The test results of benzalkonium chloride content under room temperature, accelerated conditions and high temperature conditions are shown in Table 6.4.

Table 6.4

name	30 days at room temperature (%)	30 days accelerated (%)	High temperature 30 days (%)
Formula 1-I (pH=4.0)	92.0	92.2	91.8
Formula 1-J (pH=5.0)	92.2	92.2	92.3
Formula 1-K (pH=6.0)	91.9	92.4	91.9
Formula 1-L (pH=7.0)	92.1	92.6	90.6

4. pH survey results

The pH test results under room temperature, accelerated conditions and high temperature conditions are shown in Table 6.5 below.

Table 6.5

3. Result analysis

1. Related substances and dexmedetomidine content: see Tables 6.2 and 6.3, the formula 1 provided by the present invention corresponds to the dexmedetomidine nasal spray at different pH, at room temperature for 10 to 30 days, accelerated 10 Under the experimental conditions of ～30 days and high temperature of 10～30 days, the content of related substances was extremely low. It further proves the excellent stability of the dexmedetomidine nasal spray of the formula 1 of the present invention, and further proves that the dexmedetomidine nasal spray of the formula 1 of the present invention has a wide range of pH application, at pH 4~ In the range of 7, good stability can be maintained.

2. Benzalkonium chloride: see Table 6.4, based on the overall formula of formula 1 of the present invention, under different preset pH. Under the three experimental conditions of 30 days at room temperature, 30 days at acceleration, and 30 days at high temperature, the content of benzalkonium chloride did not change much and was always stable within the predetermined range. It is further proved that under the environmental conditions of this group, even in a large pH environment deviation, the benzalkonium chloride has good stability.

3. In terms of pH: see Table 6.5, based on the overall formula of formula 1 of the present invention, under room temperature, accelerated and high temperature experimental conditions, the test results of 10 days and 30 days all show that the pH change of the system is deviated from the preset average Not big. That is, under different preset pH conditions of pH 4-7, the pH of the system can still be maintained near a relatively stable initial value.

Preliminary conclusion of the test:

Based on the above discussion, the dexmedetomidine nasal spray provided by the present invention has at least the following technical advantages: First, the present invention is fully aimed at the characteristics of nasal spray administration, and practically provides a suitable dexmedetomidine It is a new nasal spray form with good pharmacokinetic properties. Second, the dexmedetomidine nasal spray of the present invention has a simple formula, but has good stability and antibacterial properties. Third, when the dexmedetomidine nasal spray provided by the present invention uses benzalkonium chloride as the preservative ingredient, the stability of the nasal spray is improved, especially the control of related substances has made significant progress and is unexpected External technical effects. Fourth, different from the technical teachings of the prior art, the present invention finds that when the antioxidant EDTA-2Na is combined with potassium sorbate, it adversely affects the stability of the nasal spray. Fifth, the dexmedetomidine nasal spray formula of the present invention ensures stability and non-irritating nasal administration through the selection of formula types and ratio control. In the pharmacokinetic test of rats and rhesus monkeys, the animals are administered on the same day There were no local or systemic adverse reactions after administration and the acceptance was good. The New Zealand rabbits were given continuous administration for 7 days, and they were generally in good condition. There were no obvious abnormalities in the general anatomical observation of the respiratory tract, oral mucosa, and histopathological examination. Sixth, based on the composition ratio of the present invention, the dexmedetomidine nasal spray of the present invention is applicable to a wide range of pH, and can maintain good stability in the pH range of 4-7.

Test Example 7: Further extended research test of preservatives and thickeners

On the basis of the above examples and test examples 1 to 6, the present invention has carried out further extended tests to in-depth study and consider the various effects of different types of preservatives and thickeners on nasal spray formulations.

1. Prescription design and preparation methods

1. Formulation design to investigate stability

(1) The single-use preservative formulation design is shown in the following table 7.1

Table 7.1

Note [1]: The prescription amount of dexmedetomidine hydrochloride is calculated based on dexmedetomidine free base, and the conversion factor is 1.1822.

Note [2]: "/" means not added.

Note [3]: Adjust the pH to 4.00～6.00 with 1N sodium hydroxide or 1N hydrochloric acid.

(2) The prescription design of compound preservatives is shown in Table 7.2

Table 7.2

Note [1]: The prescription amount of dexmedetomidine hydrochloride is 0.02% based on the free base of dexmedetomidine, and the conversion factor of dexmedetomidine hydrochloride is 1.1822.

Note [2]: "/" means not added.

Note [3]: Adjust the pH to 4.00～6.00 with 1N sodium hydroxide or 1N hydrochloric acid.

2. Prescription design for investigating viscosity and spray performance

(3) The thickener formulation design is shown in Table 7.3 below

Table 7.3

Note [1]: The prescription amount of dexmedetomidine hydrochloride is 0.02% based on the free base of dexmedetomidine, and the conversion factor of dexmedetomidine hydrochloride is 1.1822.

Note [2]: "/" means not added.

Note [3]: Adjust the pH to 4.00～6.00 with 1N sodium hydroxide or 1N hydrochloric acid.

3. Preparation method

(1) Prescription preparation method to investigate stability

According to the prescriptions described in Table 7.1 and Table 7.2, the preparations were prepared in batches of 100ml specifications, and dexmedetomidine hydrochloride and various excipients were weighed out and dissolved in water. Among them, the prescriptions containing chlorobutanol, methyl parahydroxybenzoate and methyl parahydroxybenzoate are dissolved in a water bath, adjusted to pH 4.00～6.00, divided into glass bottles according to 10ml each, and assembled with a quantitative pump The prescription preparations DEX-002～DEX-020 and DEX-022 to be investigated are obtained.

During the preparation process, it was found that the chlorhexidine prescription (prescription DEX-008) was used alone, and the chlorhexidine could not be completely dissolved in the prescription solution, so the prescription DEX-008 was removed from the proposed investigation prescription.

(2) Prescription preparation method to investigate viscosity and spray performance

According to the thickener formulation design in Table 7.3, prepare the prescription in 1000ml batches. Weigh dexmedetomidine hydrochloride and various excipients separately, add water and magnetically stir to dissolve, if necessary, ultrasonic dissolve, that is to get the sample to be tested, of which 900ml of each square sample is used For viscosity measurement, the remaining 100ml is divided into glass bottles according to 10ml each, and a quantitative pump is assembled to obtain a prescription sample to be tested for viscosity and spray performance.

2. Stability investigation

1. Investigation method

The formulations corresponding to the prescriptions in Table 7.1 and Table 7.2 were sampled at high temperature for 10d and 30d respectively, and the related substances and preservative content of the 0d, 10d, and 30d samples were investigated. Among them, the phenylethanol prescription (DEX-004) was compared with the existing technology. Benzalkonium chloride samples (DEX-020) were placed together in a colorless vial and placed under light conditions (4500±500lux) for 10d and 30d to further investigate the content of related substances and preservatives.

2. Investigation results

The test results of related substances in high temperature test samples are shown in the following table 7.4:

Table 7.4

The test results of related substances in the light test samples are shown in the following table 7.5:

Table 7.5

The test results of the preservative content of the high temperature test samples are shown in the following table 7.6:

Table 7.6

The results show that the prior art formulations using phenylethanol as a preservative have more impurities detected in both 10d and 30d high temperature conditions, and at the same time, the content of the formulations is significantly reduced under high temperature conditions. However, the impurity level of the prescription containing chlorobutanol is lower under high temperature conditions, but its chlorobutanol content decreases significantly under high temperature conditions. Since the melting point of chlorobutanol is 61.8℃, it may be considered Caused by volatilization.

The detection of 0.08% single impurities of benzalkonium bromide prescription preparations was lower than the limit requirement of unknown single impurities (0.1%) in 30 days. Adding propylene glycol prescription did not increase the impurity level significantly. No impurities were detected in the formulas of methyl parahydroxybenzoate and propyl parahydroxybenzoate. At the same time, the content of the above-mentioned excipients is relatively stable under high temperature conditions, indicating that these four preservatives can be considered as preservatives for this product, and further investigation is planned. Antibacterial efficacy. Choose to use one preservative such as methyl paraben, propyl paraben, benzalkonium bromide, or two preservatives, such as methyl paraben and propyl paraben, which can be controlled Low impurity levels, while the preservative content remains stable. In this experiment, the formulation of EDTA-2Na and chlorobutanol also had a lower impurity level, indicating that EDTA-2Na and certain preservatives can also achieve better stability. This also reveals that EDTA-2Na has the feasibility of compound use under some preservatives. However, due to the significant decrease in the content of chlorobutanol, considering this reason, such as the use of chlorobutanol as a preservative, the sealing performance of dexmedetomidine nasal spray needs to meet higher standards. , In order to minimize the volatilization loss of the preservative trichlorobutanol. With the support of higher sealing standards and other technical means to reduce the loss of chlorbutamol content, chlorobutanol can also be used as the preservative of this nasal spray.

Under light conditions, the phenylethanol prescription (DEX-004) detected more impurities at 0d and at 10d and 30d, while the benzalkonium chloride prescription (DEX-020) had no significant increase in impurities under light conditions. The level is significantly better than the prescription of phenylethanol.

3. Investigation of viscosity and spray performance

1. Viscosity measurement and results

The viscosity of the prepared samples (DEX-102, 103, 106, 107, 108, 110) was measured with a viscometer 0# rotor, and the viscosity measurement results were recorded as shown in Table 7.7 below.

Table 7.7

The result shows that the order of viscosity of each square is: DEX-107(

)>DEX-110 (HPMC E50)>DEX-103 (carbomer)>DEX-106 (sodium hyaluronate)>DEX-102 (K30)>DEX-108 (blank). Compared with DEX-108 (blank), it has better bioadhesion after viscosity increase, which is beneficial to increase retention in the nasal mucosa, increase bioavailability or prolong the action time.

2. Investigation and results of spray performance

The spray performance of the prepared samples (DEX-102, 103, 106, 107, 108, 110) was investigated, and the spray angle, spray width at 3 cm and 6 cm were measured. The results are shown in Table 7.9 below.

Table 7.9

The results show that from the 3cm and 6cm width and spray angle, the spray width of the selected thickener formulation is slightly smaller than DEX-108 (blank), and the spray angle is approximately equal to or slightly smaller than DEX-108 (blank).

3. Summary of the investigation of viscosity and spray performance

Based on the results of comprehensive viscosity and spray performance, the thickeners selected by the present invention have improved prescription viscosity, better corresponding bioadhesion, reduced loss of nasal drug solution, and improved bioavailability. One theory has been verified by the HPMCE50 prescription (Formulation 1-C) of the pharmacokinetics of rhesus monkeys. At the same time, the addition of thickeners narrowed the spray width, which is not conducive to the increase of the dosing area. Therefore, the spray width of the prescription for adding thickeners was measured. Through the comparison of the results, PVP K30 carbomer, sodium hyaluronate,

Compared with the blank, the spray width is slightly lower, but it is larger than the verified HPMC E50 prescription (formulation 1-C), so PVP K30, carbomer, sodium hyaluronate,

When HPMC E50 is used as a thickening agent, the prescription viscosity increases, which can provide better bioadhesion while the spray width is less affected, and can provide better bioavailability or prolong the action time.

4. Antibacterial efficacy test and results

According to the results of stability, viscosity and spray performance, the antibacterial efficacy test prescription was designed according to the following table 7.10.

Table 7.10

Note [1]: The prescription amount of dexmedetomidine hydrochloride is 0.02% based on dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822.

Note [2]: "/" means not added.

Note [3]: Adjust the pH to 4.00～6.00 with 1N sodium hydroxide or 1N hydrochloric acid.

The bacteria tested in the antibacterial efficacy test were DC, TL, JP, BN and HQ. The results of the antibacterial efficacy test are shown in Table 7.11 below.

Table 7.11

Note: DC refers to Escherichia coli, TL refers to Pseudomonas aeruginosa, JP refers to Staphylococcus aureus, BN refers to Candida albicans, and HQ refers to Aspergillus niger.

The results showed that benzalkonium bromide 0.02% (prescription DEX-116) and propyl p-hydroxybenzoate 0.02% (prescription DEX-115) have good antibacterial effect and can meet the antibacterial B standard of nasal preparations. If the concentration of preservatives is further increased, the antibacterial effect may be further improved; at the same time, the compounding of 0.04% methyl hydroxybenzoate and 0.02% propyl parahydroxybenzoate (prescription DEX-117) will further increase the antibacterial effect, which can reach the nose Using the antibacterial A and B standards of preparations, it is shown that the combination of methylparaben and propylparaben can achieve excellent antibacterial effects. If the concentration of preservatives is further increased, the antibacterial effect may be further improved. For the prescription with 0.01% benzalkonium chloride, according to the results of Test Example 5, the antibacterial efficacy of 14 days and 28 days meets the standards A and B of the antibacterial efficacy judgment standard of the Chinese Pharmacopoeia 2015 edition. In this test, the compound of 0.1% propylene glycol and 0.01% benzalkonium chloride (prescription DEX-118) was added, and the antibacterial effect was further improved, which can reach the antibacterial A and B standards of nasal preparations, indicating that the compound propylene glycol , Benzalkonium chloride can achieve excellent antibacterial effect. The prescription DEX-119 (0.02% benzalkonium chloride, 1% PVP K30) with the thickener PVP K30 has excellent antibacterial effect and meets the antibacterial A and B standards of nasal preparations, indicating that PVP K30 as a thickener, not only It can thicken the prescription viscosity and improve the bioadhesion without adversely affecting the antibacterial efficacy of the prescription.

Therefore, through the antibacterial test, it is preferable to use a preservative, propyl paraben, benzalkonium bromide, or preferably a compound preservative, benzalkonium chloride and propylene glycol, propyl paraben and methyl paraben As the preservative of this product, it can achieve excellent antibacterial effect.

The present invention is not limited to the above-mentioned optional embodiments, and anyone can come to other specific solutions in various forms under the enlightenment of the present invention. The above-mentioned specific embodiments should not be construed as limiting the scope of protection of the present invention, and the scope of protection of the present invention should be defined in the claims, and the description can be used to interpret the claims.

Claims (14)

1. Dexmedetomidine nasal spray is characterized in that it contains a carrier and a therapeutically effective amount of dexmedetomidine or a pharmaceutically acceptable salt, solvate or isomer thereof;

   Based on dexmedetomidine, the optional concentration of dexmedetomidine in the nasal spray is 0.005% to 1.3%, preferably 0.0125% to 0.7%, more preferably 0.006% to 0.108%, more preferably 0.006 %, 0.02%, 0.0592%, 0.0593%, 0.108%, 0.01%, 0.03%, 0.04%, 0.05%, 0.07%, 0.1%, 0.2%, 0.4%, 0.7%, 1% or 1.3%;

   Dexmedetomidine is preferably dexmedetomidine hydrochloride. When it is dexmedetomidine hydrochloride, the preferred concentration of dexmedetomidine hydrochloride in the nasal spray is 0.006% to 1.54%, more preferably 0.015% to 0.015%. 0.83%, more preferably 0.007% to 0.13%.
2. The nasal spray according to claim 1, characterized in that it further comprises a preservative, the carrier is water, and the preservative is a cationic surfactant preservative, a paraben preservative and a non-aromatic alcohol preservative One or a combination of two or more in the agent, the cationic surfactant can be selected as benzalkonium chloride, benzalkonium bromide or benzethonium bromide, and the paraben preservative can be selected as paraben Methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, isopropyl p-hydroxybenzoate, isobutyl p-hydroxybenzoate, sodium propyl p-hydroxybenzoate Or sodium methyl parahydroxybenzoate, the non-aromatic alcohol preservative may be propylene glycol or chlorobutanol.
3. The nasal spray according to claim 2, wherein the preservative is benzalkonium chloride, benzalkonium bromide, propylene glycol, chlorobutanol, methylparaben and propylparaben One or a combination of two or more;

   When the preservative is preferably benzalkonium chloride or benzalkonium bromide, its concentration is preferably 0.002% or more, more preferably 0.01% or more, more preferably 0.02% or more, more preferably 0.01% to 0.2%, more preferably 0.002%, 0.005%, 0.0075%, 0.01%, 0.0125%, 0.015%, 0.02%, 0.05% or 0.1% or 0.2%;

   When the preservative is preferably methyl paraben, its concentration is preferably 0.01% or more, more preferably 0.01% to 0.25%, more preferably 0.01%, 0.02%, 0.04%, 0.05%, 0.1%, 0.15% or 0.25%; when the preservative is preferably propyl paraben, its concentration is preferably 0.01% or more, more preferably 0.02% or more, more preferably 0.02% to 0.075%, more preferably 0.01%, 0.02%, 0.03% , 0.04%, 0.05%, 0.06% or 0.07%;

   When the preservative is preferably propylene glycol, its concentration is preferably 0.05% or more, more preferably 0.05% to 30%, more preferably 0.1% to 10%, more preferably 0.05%, 0.1%, 0.2%, 0.5%, 1% , 5% or 10%;

   When the preservative is preferably chlorobutanol, its concentration is preferably 0.05% or more, more preferably 0.05% to 0.75%, more preferably 0.05%, 0.1%, 0.2%, 0.3%, 0.5% or 0.75%;

   When the preservative is preferably a combination of methyl p-hydroxybenzoate and propyl p-hydroxybenzoate, the concentrations of the two are preferably 0.01% or more and 0.01% or more, and the two concentrations are more preferably 0.01% to 0.25% and 0.01%～0.075%, the two concentrations are more preferably 0.02%～0.04% and 0.01%～0.02%, the two concentrations are more preferably 0.04% and 0.02%, and the two concentrations are more preferably 0.02% and 0.02%. , The two concentrations are more preferably 0.02% and 0.01% in turn;

   When the preservative is preferably a combination of propylene glycol and benzalkonium chloride, the concentrations of the two are preferably 0.1% or more and 0.01% or more, and the two concentrations are more preferably 0.1%-10% and 0.01%-0.2%. The concentration is more preferably 0.1% and 0.01% in this order.
4. The nasal spray according to claim 1 or 2 or 3, characterized in that it further comprises an antioxidant, and the antioxidant may be EDTA-2Na, tert-butyl hydroxyanisole, 2,6-di-tert One or a combination of two or more of butyl-4-methylphenol, sodium metabisulfite, potassium metabisulfite, butylhydroxyanisole, L-ascorbyl palmitate, sodium thiosulfate and vitamin E.
5. The nasal spray according to claim 1, characterized in that it further contains potassium sorbate as a preservative, and does not contain EDTA-2Na; wherein the concentration of potassium sorbate is preferably 0.01% to 0.2%, more preferably 0.1%.
6. The nasal spray according to any one of claims 1 to 5, characterized in that it further comprises a thickener, the carrier is water, and the thickener is a povidone thickener, a cellulose ether thickener, One or a combination of two or more of mucopolysaccharide thickeners and polyacrylic thickeners, povidone thickeners can be PVP K30 or PVP K90, cellulose ether thickeners are optional It is microcrystalline cellulose-sodium carboxymethyl cellulose complex, hypromellose, methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose or hydroxyethyl cellulose, mucopolysaccharides The thickener can be sodium hyaluronate or chondroitin sulfate, and the polyacrylic acid thickener can be carbomer.
7. The nasal spray according to claim 6, wherein the thickening agent is PVP K30, microcrystalline cellulose-sodium carboxymethyl cellulose complex, hypromellose, sodium hyaluronate and carbomer One or a combination of two or more;

   When the thickener is preferably PVP K30, its concentration is preferably 0.1% to 5.0%, more preferably 0.1%, 0.5%, 1%, 3% or 5%;

   When the thickener is preferably a microcrystalline cellulose-sodium carboxymethyl cellulose composite, its concentration is preferably 0.1% to 3.0%, more preferably 0.1%, 0.2%, 0.5%, 1%, 2% or 3% ；

   When the thickener is preferably hypromellose, its concentration is preferably 0.1 to 5.0%, more preferably 0.2% to 1.5%, and more preferably 0.1%, 0.2%, 0.3%, 0.5%, 0.75%, 1.0% , 1.5% or 5.0%;

   When the thickener is preferably sodium hyaluronate, its concentration is preferably 0.01% to 1.0%, more preferably 0.01%, 0.05%, 0.1%, 0.3%, 0.5% or 1.0%;

   When the thickener is preferably carbomer, its concentration is preferably 0.05% to 1.0%, and more preferably 0.05%, 0.1%, 0.3%, 0.5% or 1.0%.
8. The nasal spray according to claim 6, characterized in that: the carrier is water, and further comprises an osmotic pressure regulator, the osmotic pressure regulator is ionic or non-ionic, and the ionic osmotic pressure regulator includes boric acid, Sodium chloride, calcium chloride, magnesium chloride, zinc chloride or potassium chloride, non-ionic osmotic pressure regulators include anhydrous dextrose, dextrose monohydrate, dextran, glycerin or D-mannitol; the osmotic pressure regulator It is preferably one or a combination of two or more of boric acid, sodium chloride, potassium chloride, anhydrous dextrose, dextrose monohydrate, dextran, glycerol and D-mannitol, and the osmotic pressure regulator is particularly preferably When sodium chloride or potassium chloride is selected as the osmotic pressure regulator, the concentration of sodium chloride or potassium chloride is preferably 0.7% to 1%, and more preferably 0.7%, 0.8%, 0.9% or 1.0%.
9. The nasal spray according to claim 6, wherein the pH of the dexmedetomidine nasal spray is 4-7, preferably 5-6.5, more preferably 4, 4.5, 5, 5.5, 6 , 6.5 or 7;

   The pH adjusting agent of the dexmedetomidine nasal spray can be an acidulant and/or an alkalinizer, and the acidulant includes hydrochloric acid, phosphoric acid, tartaric acid, citric acid, malic acid, fumaric acid, lactic acid, ascorbic acid and acetic acid. , Alkalizing agents include meglumine, trimethylolmethane, sodium phosphate, arginine, lysine, glycine, ammonia, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate and borax; The pH adjuster may further preferably be one of hydrochloric acid, phosphoric acid, tartaric acid, citric acid, malic acid, fumaric acid, malic acid, ammonia, sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate and borax. One or a combination of two or more; the preferred choice of pH adjusting agent is hydrochloric acid and/or sodium hydroxide.
10. The nasal spray according to claim 1, wherein the prescription composition of the dexmedetomidine nasal spray is selected from any one of the following table:

    

    

    

    

    

    Note [1]: The prescription amount of dexmedetomidine hydrochloride is calculated based on dexmedetomidine, and the conversion coefficient of dexmedetomidine hydrochloride and dexmedetomidine is 1.1822;

    Note [2]: "/" means that the substance is not added in the corresponding prescription.
11. The nasal spray according to claim 1, characterized in that: dexmedetomidine nasal spray releases 2.5-650 μg of dexmedetomidine per press, preferably 5 μg, 10 μg, 15 μg, 20 μg, 25 μg, 30μg, 40μg, 50μg, 60μg, 75μg, 100μg or 125μg dexmedetomidine.
12. The nasal spray according to claim 1, wherein the spray particle size Dv90 of dexmedetomidine nasal spray is 50-152.1μm, more preferably 52-142μm, and still more preferably 51-66μm, 66 ～81μm, 81～104μm or 104～152.1μm.
13. A preparation method of the nasal spray according to any one of claims 1-12, characterized in that: weigh dexmedetomidine or its pharmaceutically acceptable salt, solvate or isomers thereof and various auxiliary materials, and add Dissolve the carrier, optionally adjust the pH; optionally, further divide it into a plastic or glass bottle container, assemble a metering pump on the container, and install the push button, and it is ready.
14. Use of the nasal spray according to any one of claims 1 to 12 or the nasal spray prepared by the preparation method according to claim 13 in the preparation of sedative, analgesic and/or anti-sympathetic drugs and/or devices.