JP2001172203A - Medicinal composition for transpulmonary administration - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide medicinal composition for transpulmonary administration that can increase the transpulmonary absorption of macromolecular drugs by administering a composition comprising a biodegradable cationic gelatin and a polymer drug. SOLUTION: This medicinal composition for transpulmonary administration comprises a polymer drug and superfine particles of cationic gelatin.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a pharmaceutical composition for pulmonary administration. More specifically, the present invention relates to a pharmaceutical composition for transpulmonary administration which increases the absorption of a macromolecular drug from the lung epithelium. More specifically, the present invention relates to a pharmaceutical composition for transpulmonary administration in which administration of a polymer drug together with cationized gelatin microparticles increases pulmonary absorption of the polymer drug.

[0002]

2. Description of the Related Art Since the lung epithelium has a large surface area and an abundant vasculature under the mucosa, and has a relatively low activity of degrading enzymes such as proteins, recently, high molecular drugs such as bioactive peptides and proteins have been used. Pulmonary administration of these drugs has been attracting attention as a noninvasive alternative to injection. Other non-invasive methods of administering high molecular drugs include nasal, ocular, transdermal, and rectal administration, but pulmonary administration has a higher molecular weight than these routes. It is advantageous for the absorption of drugs and is also promising for the administration of proteins with a molecular weight of 10,000 or more, such as hematopoietic cytokines.

[0003] Transpulmonary preparations of high molecular drugs can be administered in the form of a liquid by intrabronchial infusion using a bronchoscope, but usually, inhaled steroids intended for local pulmonary action. Similarly, they are delivered to the alveoli by any inhaler in the form of an inhalant. As a dosage form for inhalants,
There are three types: inhalation solutions, chlorofluorocarbon or alternative chlorofluorocarbon formulations, and powdered inhalants. An inhalation solution is usually an aqueous solution of a drug, atomized by a nebulizer into fine droplets, inhaled under spontaneous respiration of the patient, and deposited in the airway in the form of droplets. CFCs or CFC substitutes are preparations in which a drug is dispersed or dissolved in CFCs or CFCs under pressure, and are used after being filled in a pressurized container called a metered dose inhaler (Metered Dose Inhaler; MDI). . At the time of administration, when released from the MDI under pressure, CFCs or CFC substitutes are vaporized, and the dissolved / dispersed drug is usually deposited as fine drug powder in the respiratory tract. The powder inhalant is prepared by filling a fine particle powder mainly containing a drug, for example, as a powdery composition in a container such as a blister together with an excipient, etc., and injecting the fine particle powder in the container from an appropriate dispenser by inhalation of the patient. Is aerosolized into a powder, inhaled, and deposited in the lungs as a drug powder. When these inhalants are actually administered using respective inhalers, usually 10 to 30
Only about 50% by weight of the inhaler is deposited in the respiratory tract. Usually, only the drug deposited in the respiratory tract is absorbed, so that it is necessary to increase the amount of the drug in the preparation to achieve the therapeutically necessary blood concentration. The problem is big.

There have been few attempts to enhance drug absorption in pulmonary formulations. The reason is that it has been considered that the high permeability of the lung mucosa and the large area of the lung epithelium allow the absorption of the high molecular drug without devising the absorption promotion. Meanwhile, Yamamoto et al. (Journal of Pharmacy an
d Pharmacology Vol. 46, 14-18, 1994) report that co-administration of several absorption enhancers and enzyme inhibitors improved the absorption of insulin from the lungs. However, the absorption promoters and enzyme inhibitors used in this report have many problems for practical use in terms of mucosal damage and cost.

Illum et al. (WO 89/03207; JP-T-Hei 3-5)
No. 03160) provides a highly absorbable transmucosal preparation that promotes mucosal absorption of a drug by a drug delivery composition comprising a microsphere such as starch, gelatin, albumin and a peptide drug. In addition, the publication states that microspheres having bioadhesive properties or coated with bioadhesive polymers can increase the bioavailability of polymer drugs by devising release control by any means. I have. However, in the same publication,
There is no description of a method of imparting bioadhesiveness other than coating to microspheres or a specific method of controlling release.

[0006] As described in Japanese Patent Publication No. 3-17014, there is known a technique for maintaining drug efficacy by controlling release based on biodegradation of a synthetic biodegradable polymer microsphere such as polylactic acid. However, this method does not consider the adhesion of the microsphere itself to the lower respiratory tract mucosa, so it cannot prevent clearance into the laryngeal pharynx, and there is no specific information about the effect on the absorption of high molecular drugs. There is no description. WO 96/09814 describes 1 to 10 μm microparticles of a water-soluble material such as albumin by spray drying.
No consideration is given to the clearance of the microparticles themselves due to mucociliary transport as in the case of 17014. WO93 / 25198 describes a technique for maintaining drug efficacy by using hydroxypropylcellulose or hydroxypropylmethylcellulose microspheres having bioadhesive properties. A more preferable form as a base for inhalation from the viewpoint of safety decomposes in the lower respiratory tract,
Although it is a material that can be absorbed, this specification does not consider this point. Further, WO96 / 31198 discloses a technique for sustaining the efficacy of microspheres using a bioadhesive and biodegradable polysaccharide by using a composition for inhaling natural polysaccharide gum. There is a safety problem in that it contains materials suspected to be degraded and absorbed (bioabsorbable), and its optimal use is large, and its effect on the absorbability of high molecular drugs is not specified. There is no typical description.

[0007] However, the pharmaceutical composition for transpulmonary administration having improved absorbability is desirably bioadhesive from the viewpoint of function, but it is necessary that the accumulation is within an allowable range from the viewpoint of safety. . Therefore, when improving the absorption of a drug by using bioadhesive microparticles (microspheres), if the material constituting the microparticles does not have bioabsorbability, it is necessary to design such that the viscosity is reduced by adjusting the molecular weight and the like to prevent accumulation. Are being considered. On the other hand, if it is bioabsorbable, it is considered that its safety is higher because it is biodegraded and absorbed even if it is accumulated, and a pharmaceutical composition for transpulmonary administration comprising a bioabsorbable bioadhesive material is desired. ing.

[0008]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to increase the pulmonary absorption of a polymer drug by administering a composition comprising cationic biodegradable gelatin and a polymer drug. An object of the present invention is to provide a pharmaceutical composition.

The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, have found that a composition of cationized gelatin microparticles (microspheres) and a high molecular drug, preferably a composition in which the high molecular drug is encapsulated in the fine particles. The present inventors have found that pulmonary absorption of the polymer drug is improved, and reached the present invention.

[0010]

That is, the present invention provides a pharmaceutical composition for pulmonary administration comprising a polymer drug and cationized gelatin microparticles.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION The drug of the present invention may be any high molecular drug, but has a molecular weight that acts in the circulating blood.
Examples of high molecular drugs having a molecular weight of 500 to 200,000 include vasopressins, luteinizing hormone-releasing hormones, growth hormone-releasing factor hormones, somatostatin derivatives, oxytocins, hirudin derivatives, enkephalins, corticosteroids, and bradykinin. , Calcitonins, insulins, glucagons, growth hormones, luteinizing hormones, insulin-like growth factors,
Calcitonin gene-related peptides, atrial natriuretic peptide derivatives, interferons, erythropoietin, granulocyte colony formation stimulating factor, macrophage formation stimulating factor, parathyroid hormone, prolactin, thyroid stimulating hormone releasing hormone, angiotensin,
Vaccines, antibodies and the like.

[0012] Among the components of the pharmaceutical composition of the present invention, cationized gelatin microparticles are produced through a cationization step and a microparticle generation step.

The step of cationization may be any method as long as it introduces a functional group (cationic functional group) that can be cationized under physiological conditions. A method of introducing a group or an ammonium group is preferred. For example, there is a method of reacting an alkyldiamine such as ethylenediamine or N, N-dimethyl-1,3-diaminopropane, or a reaction with trimethylammonium acetohydrazide, spermine, spermidine, or diethylamide chloride. Among them, a method of reacting ethylenediamine is preferred because it is simple and versatile. The cationization reaction is usually performed before the fine particle generation step, but may be performed after the fine particle generation step.

[0014] The step of producing fine particles of cationized gelatin is as follows.
For example, there is a method in which a W / O emulsion is prepared in oil and then dropped into water to perform crosslinking (W / O formation method). As a method of crosslinking, there are a method of reacting carbodiimides and diepoxy compounds, a method of heating, and a method of irradiating ultraviolet rays or the like. Further, as long as fine particles are generated, there is no need for a crosslinking reaction, and a method such as a coacervation method using polyethylene glycol or the like, a spray drying method, a spray cooling method, and a supercritical fluid precipitation method can be applied. Among these, the W / O forming method is preferable because spherical fine particles can be easily obtained. An example of production by the W / O forming method is shown below. The reagents used were those manufactured by Wako Pure Chemical Industries, Ltd., except for those specifically described.

Production Example Production of cationized gelatin 1.0 g of gelatin having an isoelectric point of 9.0 (manufactured by Nitta Gelatin Co., Ltd.)
Was dissolved in 50 mL of a 0.1 M aqueous phosphate buffer solution (pH 5.0). 280 mg of ethylenediamine was added to the solution, and 5.4 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was further added. After stirring this mixed solution for 12 hours, it was dialyzed against water for 2 days, freeze-dried and dried.
g of cationized gelatin powder was obtained. Preparation of cationized gelatin microparticles The obtained cationized gelatin powder was adjusted to 100 mg / mL.
The resultant was dissolved in 0.2 mL of purified water, heated to 40 ° C., added to 5 mL of olive oil, and shaken to obtain a W / O emulsion. The obtained W / O emulsion was irradiated with ultrasonic waves by a probe type super sonicator (7 mm diameter, 40 μA, 2 minutes). Immediately thereafter, gelatin was gelatinized by cooling with ice, acetone was added, and the mixture was stirred for 1 hour. The generated gelatin particles were collected by centrifugation, washed with acetone, dispersed in a 37.5 μg / mL aqueous glutaraldehyde solution to a concentration of 10% by weight, and subjected to a crosslinking reaction at 4 ° C. for 40 hours. After the reaction, the mixture was treated with glycine, washed with purified water, and then centrifuged to collect crosslinked cationized gelatin microparticles having an average particle diameter of 3.0 μm (90% by weight of 0.5 to 10 μm).

In the present invention, "cationization" refers to a cationic function which cationizes normally obtainable gelatin (isoelectric point 5 or 9) under physiological conditions (in this case, pH 6.8 corresponding to the lung surface). A group is introduced by a chemical reaction. The inventors have found that the introduction of a cationic functional group imparts bioadhesiveness, and furthermore, an absorption promoting effect that cannot be explained only by bioadhesiveness (see Examples). Although the composition of the present invention is for pulmonary administration, it is considered that the nasal mucosal epithelium, which is a tissue relatively similar to the lung epithelium, has the same absorption promoting effect. The ratio of cationization is preferably higher in order to improve the absorbability of the polymer drug, and when reacting with a carboxyl group, the ratio of introducing a cationic group among all carboxyl groups of gelatin is at least 30% or more, preferably It is at least 50%, more preferably at least 80%.

The particle size of the cationized gelatin fine particles in the present invention is preferably 0.1 to 50 μm, and is preferably 80% by weight.
Is more preferably 0.5 to 10 μm from the viewpoints of improving the deposition of the polymer drug in the lungs and improving the absorption of the drug.

The polymer drug of the present invention may or may not be encapsulated in cationic gelatin microparticles, but the composition is usually produced in an encapsulated state. The encapsulation of the polymer drug in the cationized gelatin can be achieved by, for example, swelling the cationized gelatin in the polymer drug solution.

In the present invention, the composition of the polymer drug and the cationized gelatin can be set in accordance with the required absorption amount. 1.0: 99.0 is preferred.

The composition for transpulmonary administration of the present invention is prepared into a pharmaceutical composition in the form of powder or dispersed in water or the like together with excipients.

The excipients used in the present invention include, for example, lactose, glucose, mannitol, fructose, sucrose, arabinose, xylitol, dextrose, maltose and trehalose and their monohydrates, dextran,
And polysaccharides such as dextrin. Among them, lactose is preferred.

Examples of additives other than the excipient used in the pharmaceutical composition of the present invention include preservatives such as benzalkonium chloride, flavoring agents such as dl-menthol and l-menthol, and fragrances. . Preferably, such components are present in trace amounts, for example, less than 10% by weight of the composition, more preferably less than 5% by weight.

[0023]

Thus, the use of the bioabsorbable composition in a clinical setting according to the present invention is of great significance in systemic therapy by efficiently absorbing a macromolecular drug from the lungs into the blood.

[0024]

The following examples are provided to illustrate the present invention in detail, but are not intended to limit the present invention.

Example 1 Effect of Cationic Gelatin Fine Particles on Pulmonary Absorption of Salmon Calcitonin This example shows the effect of co-administration of cationic cationic microparticles on pulmonary absorption of a high molecular weight drug and the drug efficacy after intrabronchial administration in rats. The results are compared with the case of co-administration of gelatin microparticles in the absence of microparticles and non-cationized gelatin microparticles. The following three compositions were studied. The salmon calcitonin used was from Bachem AG. (1) Salmon calcitonin solution (control composition 1) (2) Salmon calcitonin / gelatin microparticle dispersion (control composition 2) (3) Salmon calcitonin / cationized gelatin microparticle dispersion (composition 1 of the present invention) <control composition Compound 1> was prepared by dissolving salmon calcitonin in a pH 7.0 isotonic phosphate buffer. The gelatin fine particles of <Control composition 2> were produced using gelatin having an isoelectric point of 9 in the same process as in "Production of cationized fine particles" in the production examples (average particle diameter: 3.4 μm). <Inventive composition 1> was produced by the steps shown in Production Examples (average particle diameter: 3.0 μm). <Control composition 2
><Inventive composition 1> both with a pH 7.0 isotonic phosphate buffer solution
10.5 mg of microparticles were added to 0.1 mL of the salmon calcitonin solution adjusted to 25 units / mL, and swelled at 0 ° C. for 3 hours to encapsulate salmon calcitonin in the microparticles. All were prepared by diluting with a pH 7.0 isotonic phosphate buffer solution to give the following dosages.

The pulmonary absorption of the three compositions was evaluated using 8-week-old SD rats (8 rats for each composition). Dosing is Schan
The composition was prepared by bronchial injection known as the ker method, and each composition was prepared so as to have a concentration of 3 units / 0.2 mL / kg rat body weight and administered under pentobarbital anesthesia. The femoral vein is cannulated, blood is collected with time before administration and up to 6 hours after administration, and the calcium concentration in the blood is determined by the ortho-cresol phthalein complexone method (calcium C).
-Test Wako: using Wako Pure Chemical Industries, Ltd.) to measure the blood calcium concentration. FIG. 1 shows the change over time in the rate of decrease in blood calcium. In addition, the time course of blood calcium concentration when a salmon calcitonin pH 7.0 isotonic phosphate buffer solution was administered by intravenous injection of 0.5 unit / kg rat body weight was similarly measured, and the pharmacological utilization of each composition was similarly measured. Table 1 shows the values obtained as the ratios. From Table 1, (1) <(2) <(3),
The composition of the present invention improves pulmonary absorption of salmon calcitonin.

[0027]

[Table 1]

[Example 2] Effect of difference in particle size of cationized gelatin microparticles on salmon calcitonin transpulmonary absorption This example is optimal for expressing the effect of cationized gelatin microparticles to improve the pulmonary absorption of a high molecular drug. It is shown that there are various particle sizes. The following four compositions were studied. (1) Salmon calcitonin solution (control composition 1) (2) Salmon calcitonin / cationized gelatin fine particle dispersion: average particle diameter 3.0 μm (composition 1 of the present invention) (3) Salmon calcitonin / cationized gelatin fine particle dispersion: Average particle diameter 53.1 μm (composition 2 of the present invention) (4) Salmon calcitonin / cationized gelatin fine particle dispersion: average particle diameter 102.1 μm (control composition 3) <Control composition 1> and <composition 1 of the present invention> Is Example 1
Is the same as <Inventive Formulation 2> and <Control Formulation 3>
Was manufactured as follows.

[0029] Cationized gelatin is added to 5 mL of olive oil and shaken to obtain a W / O emulsion.
Immediately cool on ice to gel the gelatin, add acetone and add
Stirred for hours. The generated gelatin particles were collected by centrifugation, washed with acetone, dispersed again in isopropanol, and the particle diameter was adjusted by sieving an appropriate opening. Then 10
The resulting mixture was dispersed in a 37.5 μg / mL aqueous glutaraldehyde solution so as to obtain a weight%, and subjected to a crosslinking reaction at 4 ° C. for 40 hours. After the reaction, the mixture was treated with glycine, washed with purified water, and then collected by centrifugation to obtain crosslinked cationized gelatin microparticles having an average particle size of 53.1 and 102.1 μm.

Each of the compositions was encapsulated in salmon calcitonin in the same manner as in the examples, and administered to rats by intrabronchial injection, and the blood calcium concentration was measured. FIG. 2 shows the change over time in the reduction rate of blood calcium. Table 2 shows the pharmacological utilization rate for intravenous injection. From the results in Table 2, (1) ≒ (4) <
(3) <(2), and the cationized gelatin microparticles are 10
Absorption is improved at 0 μm or less, especially at 10 μm or less. This is considered to be due to an increase in salmon calcitonin release due to an increase in surface area due to a decrease in the particle diameter of the fine particles.

[0031]

[Table 2]

Example 3 Effect of Cationic Gelatin Fine Particles on Permeation of Polymer Drug in Lung Epithelial Cell Monolayer Membrane In this example, cationic gelatin microparticles have a polymer drug absorption promoting effect due to properties other than bioadhesiveness. It is shown that. The following three formulations were evaluated. (1) Salmon calcitonin solution (control composition 1) (2) Salmon calcitonin / gelatin fine particle dispersion (control composition 2) (3) Salmon calcitonin / cationized gelatin fine particle dispersion (composition 1 of the present invention) The same composition as in Example 1.

A lung type II epithelial cell line A-549 (purchased from ATCC)
With 5% fetal bovine serum (Gibco Laboratories) and Dulbecco's Modified Eagles Medium (S
(igmaChemical) and monolayer culture in a 37 ° C.-5% CO 2 incubator. The above three compositions were prepared using Transwell ^™ (Costa
r), the amount of salmon calcitonin in each composition was added to the surface side of the monolayer cells in an amount of 5 μmol / L, and the amount of salmon calcitonin permeated to the basement membrane side after 120 minutes was measured using a high-speed liquid. It was quantified by chromatography. Table 3 shows the ratio of the amount of permeation of the A-549 cell monolayer membrane to the amount added after 120 minutes of each composition. From Table 3, (3)>(2)> (1).
Since there is no mucus layer involved in bioadhesion on the lung epithelial cell monolayer, in this example, the enzymatic degradation resistance and other absorption promoting effects of the microparticles were evaluated. It is considered to have some absorption promoting action other than sex.

[0034]

[Table 3]

[Brief description of the drawings]

FIG. 1. Time course of blood calcium levels after intrabronchial infusion of each composition. The vertical axis shows the blood calcium level when administered.
Displayed as 100. The horizontal axis is the time after administration of each composition.

FIG. 2 shows the time course of blood calcium levels after intrabronchial injection of each composition having a different particle size. The vertical axis shows the blood calcium level as 100 at the time of administration. The horizontal axis is the time after administration of each composition.

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Claims (6)

[Claims] 1. A pharmaceutical composition for pulmonary administration comprising a polymer drug and cationized gelatin microparticles. 2. The pharmaceutical composition for pulmonary administration according to claim 1, wherein the cationized gelatin is obtained by introducing an amino group or an ammonium group into gelatin. 3. The pharmaceutical composition for transpulmonary administration according to claim 1, wherein the polymer drug is selected from the group consisting of a polymer drug having a molecular weight of 500 to 200,000. 4. The method of claim 1, wherein the macromolecular drug is vasopressin, luteinizing hormone-releasing hormone, growth hormone-releasing factor hormone, somatostatin derivative, oxytocin,
Hirudin derivatives, enkephalins, adrenocorticotropic hormones, bradykinins, calcitonins, insulins, glucagons, growth hormones, luteinizing hormones, insulin-like growth factors, calcitonin gene-related peptides, atrial natriuresis One selected from the group consisting of peptide derivatives, interferons, erythropoietin, granulocyte colony stimulating factor, macrophage stimulating factor, parathyroid hormone, prolactin, thyroid stimulating hormone releasing hormone, angiotensin, vaccines, and antibodies The pharmaceutical composition for pulmonary administration according to any one of claims 1 to 3, which is a high molecular drug as described above. 5. The pharmaceutical composition for transpulmonary administration according to claim 1, wherein the cationized gelatin fine particles have a particle size of 0.1 to 50 μm. 6. The cationized gelatin microparticles of claim
The pharmaceutical composition for transpulmonary administration according to any one of claims 1 to 5, wherein the particle size of not less than 0.5% by weight is 0.5 to 10 µm.