JPH0741428A - Peptide or protein medicine transnasal-transpulmonary preparation - Google Patents

Abstract

PURPOSE:To provide a peptide or protein medicine transnasal.transpulmonary preparation improved in absorbability from nasal mucosa membranes or alveoli. CONSTITUTION:The peptide or protein medicine transnasal.transpulmonary preparation containing SLPI (secretory leukocyte protease-inhibiting protein) and a peptide or protein medicine such as a calcitonin compound, an insulin compound, LHRH or GHRH in the form of an aqueous solution agent or powder agent. The SLPI prevents the decomposition of the medicines with the protease in a nasal cavity or lung.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a nasal / pulmonary preparation of a peptide or protein drug having improved absorbability. More specifically, it relates to a peptide having improved absorbability comprising SLPI (secretory leukocyte proteinase inhibitor protein), a protein drug nasal / pulmonary preparation, and more specifically, administered to the nasal cavity or the lung. Degradation of peptide and protein drugs
The present invention relates to a nasal / pulmonary preparation of a peptide or protein drug, which has improved absorption from the nasal cavity or lung mucosa into the systemic bloodstream by being suppressed by LPI.

[0002]

2. Description of the Related Art With the recent advances in biotechnology, the discovery of peptides and proteinaceous compounds having physiological activity has been one after another, and their production has become easy. Most of these peptides and protein drugs are used only as injections because of their low stability and low absorbability. However, administration of these drugs by injection
The current situation is that the patient is frequently administered therapeutically, and as a result, the patient suffers a heavy burden due to pain, necessity of going to the hospital, and the like.

Therefore, as a non-invasive method of administering peptide or protein drugs instead of injection, for example, rectal administration [J. Pharma. Pharmcol. 33 334 (1981)], eye drop administration [Diabetes Society Abstracts 237] (1964)], transdermal administration by iontophoresis and the like are being studied. However, any of these methods are difficult to put into practical use because they require high doses as compared with injections, their absorption is likely to fluctuate, and their safety is problematic.

On the other hand, the nasal cavity has attracted attention because of its advantages such as well-developed vascular system and rapid and less variable absorption of drug. Intranasal administration, that is, nasal administration has already been put to practical use. There are some that have reached. However, under the present circumstances, the absorption rate is not always satisfactory, and various studies have been made to increase the absorption rate.

For example, Nolte et al. (Hormone Metabolic
Research 22, 170-174, 1990), Moses (Pharmaceutisch
Week-blad-Scientific Edition 10, 45-46, 1988), Br
uce et al. (Diabetic Medicine 8, 366-370, 1991) reported intranasal administration of insulin containing sodium glycocholate or sodium taurofusidate as an absorption enhancer. However, these absorption-promoting agent-containing preparations have not been put to practical use due to the problem of irritation to the nasal mucosa.

Further, since one layer of epithelial cells is in contact with capillaries in the alveoli and its thickness is only 0.5 to 1 μm, it has recently attracted attention as an absorption site. But,
Its absorbability is not well understood (Latest Biopharmaceuticals, published by Nankodo, 1991, 66-67).
page).

[0007]

The main cause of poor absorption of peptides and protein drugs from the nasal mucosa is Illum.
(Trends Biotechnol 9, 284-289, 1991), WALee (Bio
Pharm. Manuf. 1, 30-37, 1988), Edman (Advanced Drug
Delivery Reviews 8, 165-177, 1992) and the like mention low drug permeability of mucous membranes, elimination of drugs by ciliary movement, and degradation of drugs by proteolytic enzymes in the nasal cavity.

[0008] Of these, the main findings regarding the proteolytic enzyme in the nasal cavity include O'Hagan et al. [Pharm. Res.
772 (1990)], Hussain et al. (Pharm. Res. 6 186).
(1989)], that nasal absorption of insulin, growth hormone, peptides such as enkephalin and protein drugs is improved by administration with aminopeptidase inhibitors such as amastatin, bestatin and α-aminoboronic acid in rats. , Animal studies with sheep.

Also, Illum et al.
15) discloses a proteolytic enzyme inhibitor effective for nasal preparations based on studies in rats, rabbits and sheep.
Actinonin, amastatin, bestatin, chloroacetyl -HO-Leu-Ala-Gly- NH 2, diprotin A and B,
Everactone A and B, E-64, H- (tBu) -Phe-Pr
o-OH, kallikrein inhibitor I, chymotrypsin inhibitor I, trypsin inhibitor III-0, leupeptin, pepstatin, phosphoramidon, aprotinin, chymostatin, benzamidine are mentioned.

In addition, Morimoto et al.
1 and 112), TAM, which has a nasal absorption rate of salmon calcitonin, is a synthetic substrate for aprotinin and trypsin.
It has been shown by animal experiments using rats that the administration is improved with a trypsin inhibitor such as E.

However, these findings are all based on animal experiments using rats, sheep and rabbits, and it is unknown in the case of humans. Therefore, in the present circumstances, it is not clear whether the method of improving the nasal absorption rate of peptide or protein drugs in animals such as rats, sheep and rabbits is effective for humans.

Therefore, it is desired to provide a nasal preparation of a peptide or protein drug which suppresses the degradation of the peptide or protein drug in the human nasal cavity, has an improved nasal absorption rate, and is safe for the living body.

On the other hand, the absorbability of peptides and protein drugs from the lung mucosa (alveoli) has not been sufficiently studied, but the drug inhaled from the oral cavity reaches the alveoli. It is recognized that the efficiency of doing so is not sufficient. However, regarding this point, in recent years, the administration device has been improved and a nebulizer and a powder inhaler having a high alveolar reach are being developed. However, even if it reaches the alveoli, it is known that various enzyme groups exist in the alveoli,
It is fully expected that they will degrade the drug.

Therefore, it is desired to provide a peptide / protein drug transpulmonary preparation which suppresses the degradation of the peptide / protein drug in the lung, has an improved transpulmonary absorption rate and is safe for the living body.

[0015]

[Means for Solving the Problems] The present inventors have diligently studied a method for improving the absorption rate of a peptide or protein drug from the nasal or pulmonary mucosa with respect to a peptide or protein drug transnasal / pulmonary preparation. As a result, S existing in the respiratory tract
It was found that the addition of LPI (secretory leukocyte protease inhibitor protein) improves the absorption rate of peptides and protein drugs from human nasal mucosa or lung mucosa and is safe for living organisms. Arrived

That is, the present invention is a nasal / pulmonary preparation of a peptide or proteinaceous drug having improved absorbability, which comprises SLPI (secretory leukocyte protease inhibitor protein).

SLPI is the Secretary Leukocyte Protea
Abbreviation of se Inhibitor, a 12 kDa molecular weight serine protease inhibitor, which is considered to be a protease inhibitor that protects against neutrophil elastase mainly in the upper respiratory tract because it does not contain sugar chains and is produced by airway secretory gland cells. ing. SLPI consists of 107 amino acids, contains 8 disulfide bonds in the molecule, and there are domains with high homology in the first half (1-54AA) and the second half (55-107AA), and the tertiary structure is The two domains have a boomerang-like structure.

The protease inhibitor found in the lung is not limited to SLPI but has a molecular weight of 52 KD.
α1 antitrypsin of a, α1 antichymotrypsin of 68 kDa molecular weight, TIMP of 29 kDa molecular weight, and α2 macroglobulin of 720 kDa molecular weight are known.

When the airways are normal, protease and protease inhibitors are balanced in the airways, but chronic bronchitis, diffuse panbronchiolitis, emphysema, and other chronic obstructive pulmonary diseases and pulmonary fibrosis. In chronic respiratory diseases such as, it is thought that the matrix is damaged due to imbalance and excessive protease. Therefore, in such diseases, administration of SLPI, which is a protease inhibitor, neutralizes excess protease, and is expected to be effective in the treatment of these diseases (Table 62-502162). Publication, special table Sho 62-501291 publication, WO89 / 06239
Issue).

However, a protease inhibitor present in the respiratory tract is administered to the respiratory tract, more specifically, intranasally or alveolarly at the same time as a peptide or proteinaceous drug to suppress the degradation of the peptide or proteinaceous drug by protease. It has not been reported until now that the absorption of the peptide or protein drug from the nasal cavity or alveoli into the systemic bloodstream is improved. Specifically, O'Hagon et al. Or Hussain et al., Il.
The use of the protease inhibitor as an absorption enhancer is not mentioned at all in the patent of lum et al. or in the report of Morimoto et al. Furthermore, even among the above-mentioned protease inhibitors naturally present in the respiratory tract, it is not possible to know from the above description that SLPI has a remarkably large absorption promoting effect.

SLPI can be extracted from parotid secretions, but can be easily produced from microorganisms such as Escherichia coli by a gene recombination method. As the method, for example, the method described in JP-A-62-501262 can be cited.

In the present invention, the peptides and proteinaceous drugs include calcitonins, insulins, glucagon, luteinizing hormone stimulating hormone (LHRH) and its derivatives, growth hormone, growth hormone promoting hormone (GHRH) and its derivatives, growth. Hormone-releasing factor (GRF), enkephalin and its derivatives, insulin-like growth factor (IGF) s, calcitonin gene-related peptide (CGRP), vasopressin and its derivatives, atrial natriuretic peptide (ANF), interferons, erythropoietin, granulocytes A colony formation stimulating factor (G-CFS) can be mentioned. In the nasal / pulmonary preparation of the present invention, one or more peptides or proteinaceous drugs selected from the group consisting of these drugs are used.

Among these drugs, calcitonin is a peptide hormone that regulates calcium metabolism in the body, has a function of inhibiting bone resorption of calcium and reabsorbing calcium excreted from the kidney. However, salmon calcitonin, eel calcitonin, human calcitonin, pig calcitonin, chicken calcitonin, bovine calcitonin, sheep calcitonin, rat calcitonin are known, and in the present invention, any of these calcitonins and their derivatives can be used. You can

In the present invention, the amount of the peptide or proteinaceous drug is a therapeutically effective amount, and is an amount specific to each peptide or proteinaceous drug. The therapeutically effective amount is usually preferably the same amount to 20 times as much as the amount of each peptide or proteinaceous drug used for injection administration, and particularly preferably 2 times to 10 times the amount.

On the other hand, SLPI is preferably in a molar ratio of about 0.1 to 10 times, and particularly preferably about 1 to 5 times the therapeutically effective amount of the peptide or protein drug.

The nasal preparation of the peptide or proteinaceous drug of the present invention is usually administered in the form of liquid or powder. The liquid is usually an aqueous liquid, and is produced by dissolving and mixing one or more kinds of fine powdery peptides or proteinaceous drugs and SLPI in an aqueous base. As the aqueous base, water, physiological saline,
Buffer agents may be mentioned.

Such an aqueous liquid preparation contains, in addition to one or more of peptides and proteinaceous drugs and SLPI, known colorants, preservatives, preservatives, thickeners, stabilizers, if necessary, You may add a flavoring agent, a tonicity agent, etc.

Examples of colorants include copper chlorophyll, β-carotene, red No. 2 and blue No. 1; preservatives such as ascorbic acid, erythorbic acid and salts thereof; preservatives such as For example, paraoxybenzoic acid ester, phenol, benzethonium chloride, benzalkonium chloride and the like; thickeners such as hydroxypropyl cellulose, carboxymethyl cellulose and salts thereof; stabilizers,
For example, human serum albumin, mannitol, sorbitol, and the like; examples of the flavoring agent include menthol and citrus flavors, and examples of the tonicity agent include glucose.

On the other hand, the powder is prepared by mixing at least one of finely powdered peptides and proteinaceous drugs, SLPI, and a water-absorbing and poorly water-soluble base. The mixing is carried out using an ordinary mixer such as a mortar and a high speed mixer. As used herein, a water-absorbent and sparingly water-soluble base means on a human nasal mucosa or in an environment close to this, that is, with respect to water having a pH of about 7.4 and a temperature of about 36 to about 37 ° C. It means that it has absorbency and poorly soluble property.

Preferred specific examples include water-absorbing and sparingly water-soluble celluloses such as crystalline cellulose, α-cellulose, cross-linked sodium carboxymethyl cellulose and derivatives thereof; hydroxypropyl starch, carboxymethyl starch, cross-linked starch, amylose, Water-absorbing and poorly water-soluble polysaccharides such as amylopectin, pectin and their derivatives; Water-absorbing and poorly water-soluble gums such as gum arabic, tragacanth gum, glucomannan and their derivatives; polyvinyl polypyrrolidone, cross-linked poly Mention may be made of crosslinked vinyl polymers such as acrylic acid and its salts, crosslinked polyvinyl alcohol, polyhydroxyethylmethacrylate and their derivatives. Among these, water-absorbent and sparingly water-soluble celluloses are preferable, and crystalline cellulose and its derivatives are particularly preferable.

Such powders include, in addition to one or more of peptides, protein drugs, SLPI and the above-mentioned bases,
If necessary, known lubricants, coloring agents, preservatives, preservatives,
You may add a flavoring agent.

Lubricants such as talc, stearic acid and salts thereof; colorants such as copper chlorophyll, β-carotene, red No. 2 and blue No. 1;
As preservatives, for example, stearic acid, ascorbic acid stearate, etc .; as preservatives, for example, quaternary ammonium compounds such as benzalkonium chloride, paraoxybenzoic acid esters, phenol, chlorobutanol, etc .; flavoring agents Examples thereof include menthol and citrus flavors.

The nasal preparation of the peptide or protein drug of the present invention is usually administered intranasally by the following administration method. That is, in the case of an aqueous solution, for example, the solution is placed in a nasal drop container, a spray container or a similar container suitable for applying such a solution into the nasal cavity, and a method of dripping or spraying into the nasal cavity, or There is a method of using an inserter that can be easily and quantitatively administered into the nasal cavity, and then administering a prescribed amount of the liquid agent into the nasal cavity.

In the case of powder, for example, a capsule filled with powder is set in a dedicated spray device equipped with a needle, and the needle is penetrated, thereby forming a microscopic hole at the top and bottom of the capsule, and then air. There is a method of sending powder with a rubber ball and ejecting powder.

On the other hand, the peptide or protein drug transpulmonary preparation of the present invention is usually in the form of an aqueous liquid inhaled by a nebulizer, a metered dose inhaler filled in a pressure vessel together with a propellant, or a powdered inhalant. Administered into human lungs.

Aqueous solutions include peptides and protein drugs.
It is manufactured by dissolving and mixing seeds and SLPI with an aqueous base. It is provided in the solid state and may be dissolved in an aqueous base when used. As the aqueous base used, water, physiological saline,
Examples thereof include buffer solutions. In addition to peptides, proteinaceous drugs and SLPI, such aqueous solutions include
Known colorants, preservatives, preservatives, thickeners, stabilizers, flavoring agents, isotonic agents and the like may be added as required. Examples of these include the same compounds as those used in the nasal formulation.

These aqueous solutions are atomized by a nebulizer and administered into the lungs. As the nebulizer, a normal one such as a jet nebulizer or an ultrasonic nebulizer is used.

On the other hand, in the case of a metered dose inhaler, a peptide, a proteinaceous drug and SLPI are dispersed in a propellant such as Freon together with appropriate additives such as a dispersion aid and filled in a pressure vessel. Manufactured. After injection, the drug flying in the respiratory tract and SLPI are dispersed in the propellant by adjusting the particle size such that the drug has a particle size sufficient to reach the alveoli, that is, about 0.5 to 5 μm. It is necessary.

Further, a powdered inhalant is produced by preparing a peptide, a proteinaceous drug and SLPI together with an appropriate additive such as an excipient in lactose to a particle size of about 0.5 to 5 μm. To be done. The manufactured powder is filled in a capsule or the like and inhaled into the lung by a unit dose administration device, or administered into the lung by a multidose administration device.

The present invention provides a nasal / pulmonary preparation of a peptide or protein drug having improved absorbability, which is of great medical value.

[0041]

EXAMPLES The present invention will be described in detail below with reference to Examples and Reference Experimental Examples, but these are only for explaining the present invention and do not limit the present invention.

Example 1 A homogenate solution was prepared from human nasal mucosa so that the amount of protein was 1 mg / ml.
5 ml of salmon calcitonin (SCT) solution (0.1
mg / ml) 0.5 ml was added, and the aqueous solutions of the three human-derived protease inhibitors shown in Table 1 were added to 5.0
ml was added and incubated at 37 ° C., and the amount of undegraded salmon calcitonin after a certain period of time was determined by HPLC.
The SCT decomposition inhibition rate of these protease inhibitors was calculated by comparing with the amount of undecomposed salmon calcitonin when distilled water was added instead of the aqueous solution of protease inhibitors. The results are shown in Table 1.

[0043]

[Table 1] Inhibitory effect of human-derived protease inhibitors on SCT degradation by homogenates As SLPI, one manufactured according to the method described in WO89 / 06239 was used. As α1-antitrypsin and α2-macroglobulin, those manufactured by Sigma were used.

As shown in Table 1, it was found that the coexistence of SLPI markedly inhibited the decomposition of salmon calcitonin in human nasal mucosa homogenate,
This inhibitory effect is significantly higher than any of the other four human-derived protease inhibitors.

Example 2 Salmon calcitonin 40 μg, SLPI 15 μg, microcrystalline cellulose 30 mg, magnesium stearate 15 μm
g was mixed in a mortar to prepare SLPI-added calcitonin nasal powder. Calcitonin amount 100 IU (20μ
g) and filled in No. 2 capsule, and then administered into the nasal cavity on the right side of 3 normal volunteers with a pubizer (registered trademark, Teijin Ltd.). After a certain time from the administration, 5 ml of blood was collected from the forearm vein, and the salmon calcitonin concentration in plasma was measured by the RIA method. The results are shown in FIG. 1 as a time curve of plasma salmon calcitonin concentration.

Control Example 1 A calcitonin nasal dust containing no SLPI was prepared in the same manner as in Example 1 except that 15 μg of SLPI was not added, and the calcitonin amount was 100 IU (20 μg). After administration to 3 normal volunteers in the same manner as in Example 1, the salmon calcitonin concentration in plasma was measured after a certain period of time. The results are also shown in FIG.

Example 3 300 μg of salmon calcitonin and 100 μg of SLPI were dissolved in distilled water to make a total volume of 5 ml. This was made into a mist by a compressor type nebulizer, and under anesthesia, the amount of calcitonin in the tracheotomized rat was 100 IU (20 μm).
g). Plasma salmon calcitonin concentrations were measured by the RIA method after a certain time from the administration. The results are shown in the plasma salmon calcitonin concentration time curve in FIG.
Shown in.

Control Example 2 A SLPI-free pulmonary liquid preparation was prepared in the same manner as in Example 2 except that 100 μg of SLPI was not added, and was administered to rats in the same manner as in Example 2, and the plasma salmon calcitonin concentration was measured. . The results are also shown in FIG.

[Brief description of drawings]

FIG. 1 shows the time course of plasma salmon calcitonin concentration after administration of a salmon calcitonin nasal powder in normal volunteers obtained in Example 2 and Control 1.

FIG. 2 shows the time-dependent changes in the plasma salmon calcitonin concentration after administration of a transpulmonary salmon calcitonin solution in rats obtained in Example 3 and Control Example 2.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Office reference number FI Technical indication location A61K 38/28 38/26 38/23 47/42 E 8314-4C A61K 37/26 8314-4C 37 / 28 8314-4C 37/30 (72) Inventor Yoshiki Suzuki 4-3-2 Asahigaoka, Hino-shi, Tokyo Inside Teijin Limited Tokyo Research Center

Claims (6)

[Claims] 1. A nasal / pulmonary preparation of a peptide or protein drug having improved absorption, comprising SLPI (secretory leukocyte proteinase inhibitor protein). 2. Peptides and protein drugs are calcitonin, insulins, glucagon, luteinizing hormone stimulating hormone (LHRH) and its derivatives, growth hormone, growth hormone stimulating hormone (GHRH) and its derivatives, growth hormone releasing factor. (GRF), enkephalin and its derivatives, insulin-like growth factor (IG
F), calcitonin gene-related peptide (CGR
P), vasopressin and its derivatives, atrial natriuretic peptide (ANF), interferons, erythropoietin, granulocyte colony stimulating factor (GC)
The nasal / pulmonary preparation of a peptide or proteinaceous drug according to claim 1, which is a peptide or a proteinaceous drug selected from the group consisting of FS). 3. The peptide / protein drug according to claim 1, wherein the peptide / protein drug is calcitonin.
Transpulmonary formulation. 4. Calcitonins are salmon calcitonin,
The peptide or proteinaceous drug according to claim 2 or 3, which is one or more kinds of calcitonin selected from the group consisting of eel calcitonin, human calcitonin, porcine calcitonin, chicken calcitonin, bovine calcitonin, sheep calcitonin, rat calcitonin and derivatives thereof. Nasal / lung formulation. 5. The nasal / pulmonary preparation of the peptide or protein drug according to claim 1, wherein the nasal / pulmonary preparation is an aqueous liquid preparation. 6. The nasal / pulmonary preparation of peptide or protein drug according to claim 1, wherein the nasal / pulmonary preparation is a powder.