CN115192554A - Propellant-free peptide-containing inhalation solution and preparation method thereof - Google Patents

Abstract

The invention discloses a propellant-free peptide-containing inhalation solution and a preparation method thereof, wherein the propellant-free peptide-containing inhalation solution comprises an active substance, a solvent and a surfactant, wherein the active substance is GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide, the concentration of the active substance dissolved in the solvent is 0.1-100 mg/ml, and the concentration of the surfactant dissolved in the solvent is 0.01-0.1 mg/ml. The peptide-containing inhalation solution without the propellant is used as a new drug formulation, is convenient to use, has high effective part deposition rate, is convenient to produce and store because the feed liquid is in a solution state, does not contain the propellant in the formula, reduces the influence of the product on the environment, is safe and effective, has good adaptability compared with an injection preparation, has higher bioavailability than an oral preparation, and has good application prospect.

Description

Propellant-free peptide-containing inhalation solution and preparation method thereof

Technical Field

The invention relates to the technical field of pharmaceutical preparations. More specifically, the invention relates to a propellant-free peptide-containing inhalation solution and a preparation method thereof.

Background

In recent years, diabetes and its complications have become health concerns of global concern as chronic non-infectious diseases that seriously affect human health and quality of life, so that governments have all over the world paid great attention to the development of diabetes therapeutic drugs. For many pharmaceutical manufacturing enterprises, overcoming diabetes as early as possible is not only the line of social responsibility but also the trend of huge economic benefits. The diabetes is mainly divided into type I diabetes, type II diabetes and other special types of diabetes, wherein the disease proportion of the type II diabetes is the highest and reaches more than 90 percent. Obesity is currently considered to be a major risk factor for diabetes. Clinically, obese type II diabetic patients have three high characteristics, including hyperglycemia, hyperlipidemia, hypertension, etc., obesity is the most dangerous signal among various complex factors inducing diabetes, and the control of body weight is necessary to prevent and treat diabetes.

At present, the sugar-reducing medicines for type II diabetes mellitus are various, and the somaglutide is used as a new long-acting glucagon-like peptide-1 (GLP-1) analogue, insulin secretion is promoted and glucagon secretion is inhibited by a glucose concentration dependent mechanism, so that the blood sugar level of a type II diabetes mellitus patient can be greatly improved, and the risk of hypoglycemia symptoms after the patient takes the somaglutide is low. Meanwhile, the somalutide can reduce appetite and food intake, and has obvious weight-losing effect. The medicine is developed by Novonide, and injection preparation and oral preparation are approved to be on the market at present.

However, there is a high probability that the injectable and oral formulations of somaglutide will cause gastrointestinal adverse reactions such as nausea, vomiting, etc. In addition, the injection preparation has high requirements on transportation and storage conditions, must be carried out under cold chain conditions, and is administrated once per week by subcutaneous injection, which is very painful for diabetic patients requiring long-term treatment and even lifelong treatment, not only has poor compliance, but also is easy to cause infection, and brings physical and psychological burdens to the patients. The requirements for oral preparations are very demanding: must be swallowed with <100ml of white water on an empty stomach, neither food or drink nor other medicines can be taken within half an hour after administration, the convenience is greatly compromised compared to other oral medicines, patient compliance may be affected, and the bioavailability of oral formulations is low, only about 1%.

Disclosure of Invention

An object of the present invention is to solve at least the above problems and to provide at least the advantages described later.

The invention also aims to provide a propellent-free peptide-containing inhalation solution and a preparation method thereof, compared with an injection preparation, the propellent-free peptide-containing inhalation solution has good adaptability, higher bioavailability than an oral preparation, good medicine stability, convenient administration, safety and effectiveness.

To achieve these objects and other advantages in accordance with the present invention, there is provided a propellant-free peptide-containing inhalation solution formulation comprising an active substance, a solvent, a surfactant, wherein the active substance is a GLP-1 peptide or a pharmaceutically acceptable salt of a GLP-1 peptide, and the active substance is dissolved in the solvent at a concentration of 0.1 to 100mg/ml.

The present invention relates to GLP-1 analogs and derivatives having Trp at a position corresponding to position 8 of GLP-1 (7-37) and pharmaceutical uses thereof, e.g. in the treatment of type 2 diabetes. Such Trp8 compounds are extremely stable to DPP-IV degradation while retaining the ability to bind to and activate the GLP-1 receptor. Such derivatives have one or two substituents (P-L) attached via an optional branching group to a Lys residue of a GLP-1 analog, where P is a protracting moiety, such as a fatty diacid, and L is a linker consisting of one or more linker elements, such as 8-amino-3, 6-dioxaoctanoic acid.

In some embodiments, the compositions of the invention comprise a peptide. In some embodiments, the peptide comprises a lipophilic side chain, such as a peptide comprising an alkyl moiety having at least 14 carbon atoms. In some embodiments, the peptide is an acylated peptide. In some embodiments, the peptide comprises a substituent comprising a fatty acid or fatty diacid, such as formula (X)

Wherein n is at least 13.

In some embodiments, the peptide comprises one or more 8-amino-3, 6-dioxaoctanoic acids (OEG).

Preferably, the solvent is water.

Preferably, the surfactant is dissolved in the solvent at a concentration of 0.01 to 0.1mg/ml.

Preferably, the surfactant is any one or more of polysorbate 80, poloxamer, polyoxyethylene castor oil, polyethylene glycol 15-hydroxystearate and polyvinylpyrrolidone.

Preferably, the propellant-free peptide-containing inhalation solution produces an aerosol having a mass median aerodynamic particle size of less than 10 μm upon aerosolization.

Preferably, the propellant-free peptide-containing inhalation solution produces an aerosol having a median particle diameter D50 of less than 10 μm after nebulization.

Preferably, the propellant-free peptide-containing inhalation solution is nebulized by a metered dose, spring-pressurized inhalation aerosol inhaler.

The invention also provides a preparation method of the propellant-free peptide-containing inhalation solution, which mainly comprises the following steps:

firstly, precisely weighing a surfactant, putting the surfactant into a beaker, adding a solvent, and fully stirring to dissolve the surfactant;

step two, precisely weighing GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide, adding the GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide into a beaker, and fully stirring to ensure that the concentration of the GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide is 0.1-100 mg/ml, thereby obtaining a mixed solution;

and step three, filtering the mixed solution by a filter membrane to obtain the propellant-free peptide-containing inhalation solution.

The invention at least comprises the following beneficial effects:

the invention relates to a propellent-free peptide-containing inhalation solution, wherein GLP-1 peptide or pharmaceutically acceptable salts of the GLP-1 peptide are used as active substances and used in combination with a quantitative spring pressure type inhalation spray inhaler, and the active substances are directly inhaled through an oral cavity and enter a blood system from the lung.

Secondly, the propellent-free peptide-containing inhalation solution of the invention is used as a new drug formulation, and has no problem of uniformity of inhalation suspension/aerosol or layering of active drug ingredients and carriers in dry powder inhalation formulations (DPI) due to the solution state; the formula does not contain a propellant, so that the influence of the product on the environment is reduced to the maximum extent; meanwhile, the feed liquid is in a solution state, so that the humidity in the production and storage processes is not strictly required.

Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.

Drawings

Fig. 1 is the ACI measurement of the somaglutide inhalation solution of example 1;

fig. 2 is the ACI measurement of the somaglutide inhalation solution of example 2;

FIG. 3 is a concentration-time curve of a subcutaneously injected cynomolgus monkey administered sogluteptide subcutaneous injection;

fig. 4 is a concentration-time curve of the cynomolgus monkey after administration of the somaglutide inhalation solution by airway administration;

fig. 5 is a schematic view showing the construction of a metered dose, spring-pressurized inhaler according to another embodiment of the present invention.

Detailed Description

The present invention is further described in detail below with reference to the drawings and the detailed description so that those skilled in the art can practice the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

It is to be noted that the experimental methods described in the following embodiments are all conventional methods unless otherwise specified, and the reagents and materials, if not otherwise specified, are commercially available; in the description of the present invention, it should be noted that unless otherwise explicitly stated or limited, the terms "mounted," "connected," and "disposed" are to be construed broadly and can, for example, be fixedly connected, disposed, detachably connected, disposed, or integrally connected and disposed. The specific meanings of the above terms in the present invention can be understood in a specific case to those of ordinary skill in the art. The terms "lateral," "longitudinal," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in the orientation or positional relationship indicated in the drawings for convenience in describing the invention and to simplify the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the invention.

The invention provides a propellent-free peptide-containing inhalation solution and a preparation method thereof, and the propellent-free peptide-containing inhalation solution comprises an active substance, a solvent and a surfactant, wherein the active substance is GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide, and the concentration of the active substance dissolved in the solvent is 0.1-100 mg/ml. The present solution provides a formulation of an inhalation solution comprising somaglutide or a pharmaceutically acceptable salt of somaglutide that meets the required high standards in order to enable optimal nebulization of the solution.

In another technical scheme, the solvent is water.

In another technical scheme, the concentration of the surfactant dissolved in the solvent is 0.01-0.1 mg/ml.

In another technical scheme, the surfactant is any one or more of polysorbate 80, poloxamer, polyoxyethylene castor oil, polyethylene glycol 15-hydroxystearate and polyvinylpyrrolidone.

In another technical scheme, the mass median aerodynamic particle size of aerosol generated by atomizing the propellant-free peptide-containing inhalation solution is less than 10 mu m.

In another technical scheme, the median particle diameter D50 of aerosol generated by atomizing the propellant-free peptide-containing inhalation solution is less than 10 μm.

In another technical solution, the propellant-free peptide-containing inhalation solution is atomized by a metered dose, spring-pressurized inhalation aerosol inhaler.

A liquid formulation without a gaseous propellant and administered using a suitable inhaler is desired to obtain a better distribution of the active substance in the lungs. In addition, it is also desirable to increase the deposition of drugs administered by inhalation in the lungs. Currently, conventional pressurized metered dose inhalation (pMDI) or Dry Powder Inhalation (DPI) devices can only deliver about 20% of the drug in the formulation to the lungs, resulting in large amounts of drug being deposited in the mouth and throat, eventually into the stomach, and possibly causing adverse side effects and/or secondary absorption by the digestive system. There is therefore a need to improve the delivery of inhaled drugs by increasing pulmonary deposition. The metered dose, spring-loaded inhaler disclosed in the prior art (as shown in figure 5) can significantly increase pulmonary deposition of inhalable drug. Such inhalers can aerosolize small quantities of the liquid formulation in seconds into an inhalable aerosol suitable for treating local or systemic diseases. Such inhalers are particularly suitable for use with the liquid formulations of the present invention. A metered dose, spring-loaded inhalation aerosol inhaler suitable for administration of a solution-type pharmaceutical formulation of the present invention can aerosolize less than about 70 microliters, e.g., less than about 30 microliters, more specifically less than about 15 microliters of the pharmaceutical solution in a single spray such that the respirable portion of the aerosol corresponds to a therapeutically effective amount. The aerosol formed by one spray has an average particle size of less than 15 microns, or less than 10 microns. Such devices for the metered administration of propellant-free liquid pharmaceutical inhalable formulations have been described in detail, for example in US 20190030268.

The invention also provides a preparation method of the propellant-free peptide-containing inhalation solution, which mainly comprises the following steps:

firstly, precisely weighing a surfactant, putting the surfactant into a beaker, adding a solvent, and fully stirring to dissolve the surfactant;

secondly, precisely weighing GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide, adding the GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide into a beaker, and fully stirring to ensure that the concentration of the GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide solution is 0.1-100 mg/ml, thereby obtaining a mixed solution;

and step three, filtering the mixed solution by a filter membrane to obtain the propellant-free peptide-containing inhalation solution.

< example 1>

The peptide-containing inhalation solution without the propellant is prepared by the specific steps of precisely weighing 0.001g of polysorbate 80 in a glass beaker, adding 10g of purified water, stirring to dissolve the purified water, precisely weighing 0.5mg of common commercially available Somalide, adding the Somalide into the beaker, stirring to dissolve, filtering the solution by using a 0.22 mu m filter membrane to prepare the Somalide solution with the concentration of 0.5mg/ml, namely the peptide-containing inhalation solution without the propellant.

< example 2>

The peptide-containing inhalation solution without the propellant is prepared by the specific steps of precisely weighing 0.001g of poloxamer into a glass beaker, adding 10g of purified water, stirring to dissolve the poloxamer, precisely weighing 0.5mg of ordinary commercially available thaumatin sodium salt, adding the thaumatin sodium salt into the beaker, stirring to dissolve the thaumatin sodium salt, and filtering the solution by using a filter membrane of 0.22 mu m to prepare the thaumatin sodium salt solution with the concentration of 0.5mg/ml, namely the peptide-containing inhalation solution without the propellant.

< comparative example 1>

The commercially available someropeptin subcutaneous injection, ozampic.

< spray particle size distribution test >

The droplet size distribution of the sprays of examples 1 and 2 was measured using the INHALER module of the New Partach laser particle sizer with test environmental conditions of 98% RH. + -. 2% RH, 20-25 deg.C. The results are shown in Table 1. Wherein, D10 refers to the corresponding particle diameter when the cumulative particle size distribution number of a sample reaches 10 percent, and the unit is micrometer. The D50 is the particle size in microns corresponding to the cumulative particle size distribution of 50% of a sample. D90 is the particle size in microns corresponding to 90% of the cumulative particle size distribution for a sample. Test results show that the median particle diameter D50 of the generated aerosol is less than 10 mu m by using the propellant-free peptide-containing inhalation solution provided by the invention in combination with a quantitative spring pressure type inhalation aerosol inhaler, and the aerosol meets the standard of an atomized preparation.

TABLE 1

	D10/μm	D50/μm	D90/μm
				Example 1	2.12	4.38	7.99
Example 2	2.07	4.26	8.05

< aerodynamic particle size distribution test >

"aerodynamic diameter (Da)" also called aerodynamic equivalent diameter (particle diameter) is an imaginary particle diameter (particle diameter) that expresses particle motion. Stober (w.stober) defines it as: unit density (ρ) ₀ ＝1g/cm ³ ) The diameter of the sphere (2) when the sphere (2) has reached the same final settling velocity (Vs) as that of an actual particle when the sphere is moved at a low reynolds number in a still air atmosphere. I.e. the actual particle size is exchanged for an equivalent diameter (or equivalent diameter) having the same aerodynamic properties. Since the actual particle size and density cannot usually be determined, aerodynamic particle size can be determined directly from the kinetic measurements, which allows a uniform measure of particle size with different shapes, densities, optical and electrical properties. The aerodynamic particle size can be calculated with reference to the following method: the particle diameter (volume particle diameter) Dv of the powder sample was measured by a laser granulometer according to Da = (ρ/ρ) ₁ ) ^1/2 Dv gives the aerodynamic particle size Da. Where ρ is the density of the particles, ρ ₁ ＝1g/cm ³ And Dv is the average particle diameter of the particles. The value of p can be estimated by tap density, which is about 1.26 times the tap density. The term "mass median aerodynamic particle size" or "MMAD (mass median aerodynamic diameter)" refers to: when the total mass of particles of various sizes in the particulate matter that are smaller than a certain aerodynamic size accounts for 50% of the total particulate matter mass (i.e., the sum of all different particle sizes), then this size is referred to as the mass median aerodynamic size. The term "effective site deposition rate" or "FPF (fine particle fraction)" refers to the percentage of particle dose equal to or less than 5 μm of the total delivered dose, calculated as follows:

wherein:

FPD is the fine particle dose, namely the particle dose with the mass median aerodynamic particle diameter less than or equal to 5 μm, and is calculated according to the drug mass of each level of an Anderson Cascade Impactor (ACI) or a New Generation Impactor (NGI) and the corresponding cut-off particle diameter of each level under the test flow rate;

emitted Dose is the total delivered Dose, which refers to the sum of the drug mass at each level of the Anderson Cascade Impactor (ACI) or the Next Generation Impactor (NGI) excluding capsule and device residues.

ACI device environmental humidity was controlled within the range of 92% rh ± 2% rh, the test results of which are shown in table 2, the ACI measurement results of example 1 and example 2 are shown in fig. 1 and fig. 2, respectively, and the resulting somaglutide inhalation solution had effective site deposition rates of 27.27% and 27.67% and mass median aerodynamic particle diameters of 6.01 μm and 6.07 μm, by calculation.

TABLE 2

Detecting items	FPF/％	MMAD/μm
			Example 1	27.27	6.01
Example 2	27.67	6.07

< non-clinical pharmacokinetic Studies >

a test drugs

Agents in example 1 and comparative example 1

b test animals

The weight of the cynomolgus monkey is 3-5kg, and the weight is half of that of the male and female; 3-5 years old.

c route of administration and dosage

Example 1 airway inhalation group (0.1 mg/kg): after isoflurane is inhaled for anesthesia, the tested sorangin inhalation solution with the concentration of 0.1mg/kg is sprayed into the airway through a laryngoscope.

Comparative example 1 subcutaneous injection group (0.1 mg/kg): the calculated weight of the cynomolgus monkey was followed by subcutaneous injection of the corresponding volume of the subcutaneous injection of somalutide.

d dosing regimen

The cynomolgus monkeys 4, divide into 2 groups, male and female half. The soxhlet inhalation solution and the soxhlet subcutaneous injection prepared in example 1 and comparative example 1 were administered separately, fasting was not less than 12 hours before administration, water was freely drunk, and 4 hours after administration, and unified food was taken.

The specific arrangement is shown in table 3 below:

TABLE 3

e sample Collection and processing

In the subcutaneous injection group, 0.5mL of blood is taken from forelimb veins before and after the administration for 0.5h, 1h, 2h, 4h, 8h, 24h and 48 h. The airway inhalation group was bled 0.5mL via the forelimb vein at 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h, 48h and 72h before and after administration. Placing in EDTA-K2 anticoagulation tube, centrifuging at 3000rpm for 10min (4 deg.C), separating plasma within 2 hr, and storing at-70 deg.C for testing. Blood sampling to centrifugation process was performed under ice bath conditions.

Analysis of biological samples

And determining the concentration of the somaglutide in the plasma of the cynomolgus monkey by adopting an LC-MS/MS method.

g test results

The concentration-time curve of the subcutaneous injection of 0.1mg/kg of the somatid is shown in figure 3, and the concentration-time curve of the inhalant of the somatid is shown in figure 4.

TABLE 4

After the cynomolgus monkey is injected with 0.1mg/kg of the somaglutide subcutaneous injection, the AUC (0-t) is 64023.13 mu g/L per hour, the tmax is 16.00h, and the Cmax is 1295 mu g/L.

After 0.1mg/kg of the Somalide inhalation solution is administrated to the cynomolgus monkey airway, the AUC (0-t) is 10151.10 mu g/L h, the tmax is 16.00h, and the Cmax is 219 mu g/L.

After the cynomolgus monkey is administrated with 0.1mg/kg of the somaglutide by different administration modes, the proportion of the plasma exposure amount of the different administration modes is subcutaneous injection: airway administration =1:0.16, ratio of cmax for subcutaneous injection: airway administration =1:0.17, relative bioavailability of the somaglutide inhalation solution relative to subcutaneous injection after the administration of the somaglutide inhalation solution in the cynomolgus monkey airway administration mode can reach 16%. Whereas the relative bioavailability of oral tablets of somaglutide given by oral administration was only 1.0% according to the description in the norshanode published patent CN 104487056A. Therefore, compared with the oral tablet administration mode, the peptide-containing inhalation solution without the propellant is matched with a quantitative spring pressure type inhalation spray inhaler for use, and the bioavailability is greatly improved.

Therefore, although the bioavailability of subcutaneous injection administration is higher, the administration frequency is once per week, the adaptability is poor, and physical and psychological burdens are easily brought to patients.

The number of apparatuses and the scale of the process described herein are intended to simplify the description of the present invention. Applications, modifications and variations of the present invention will be apparent to those skilled in the art.

While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in various fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the general concept defined by the appended claims and their equivalents.

Claims (9)

1. A propellent-free peptide-containing inhalation solution is characterized by comprising an active substance, a solvent and a surfactant, wherein the active substance is GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide, and the concentration of the active substance dissolved in the solvent is 0.1-100 mg/ml.

2. A propellant-free peptide-containing inhalation solution of claim 1, wherein the peptide comprises a substituent comprising a fatty acid or a fatty diacid of formula (X):

wherein n is at least 13;

and wherein the peptide comprises one or more 8-amino-3, 6-dioxaoctanoic acids (OEG).

3. The propellant-free peptide-containing inhalation solution of claim 2, wherein the GLP-1 peptide or a pharmaceutically acceptable salt of a GLP-1 peptide is somatid or a pharmaceutically acceptable salt of somatid.

4. The propellant-free peptide-containing inhalation solution of claim 1, wherein the concentration of said surfactant dissolved in the solvent is 0.01 to 0.1mg/ml.

5. The propellant-free peptide-containing inhalation solution of claim 4, wherein the surfactant is any one or more of polysorbate 80, poloxamer, polyoxyethylene castor oil, polyethylene glycol 15-hydroxystearate, and polyvinylpyrrolidone.

6. The propellant-free peptide-containing inhalation solution of claim 1, wherein the mass median aerodynamic particle size of the aerosol generated by nebulization of the propellant-free peptide-containing inhalation solution is less than 10 μm.

7. The propellant-free peptide-containing inhalation solution of claim 6, wherein said propellant-free peptide-containing inhalation solution upon aerosolization produces an aerosol having a median particle size D50 of less than 10 μm.

8. Propellant-free peptide-containing inhalation solution according to claim 7, characterized in that it is nebulized by a metered dose, spring-pressure inhaler.

9. The process for preparing a propellant-free peptide-containing inhalation solution of claim 1, comprising essentially the steps of:

step one, precisely weighing a surfactant, adding the surfactant into a beaker, and fully stirring to dissolve the surfactant;

secondly, precisely weighing GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide, adding the GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide into a beaker, and fully stirring to ensure that the concentration of the GLP-1 peptide or pharmaceutically acceptable salt of the GLP-1 peptide solution is 0.1-100 mg/ml, thereby obtaining a mixed solution;

and step three, filtering the mixed solution by a filter membrane to obtain the propellant-free peptide-containing inhalation solution.

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