JPH0480008B2 - - Google Patents

Description

[Detailed description of the invention]

<Industrial Application Field> The present invention relates to a powdered polypeptide composition useful for nasal administration. More specifically, the present invention is directed to the use of bioactive polypeptides such as calcitonin and insulin in combination with ascorbic acids such as ascorbic acid and basic amino acid salts such as arginine hydrochloride as absorption enhancers. A polypeptide composition useful for nasal administration, which is a powdered composition consisting of an absorbable solid base, in which the polypeptide is absorbed extremely efficiently through the nasal mucosa when sprayed into the nasal cavity. relating to things. <Problems to be solved by the prior art and the invention> Peptide hormones such as insulin and calcitonin have large molecular weights and are easily degraded by proteolytic enzymes such as pepsin, trypsin, or chymotrypsin, so they are difficult to absorb when administered orally. Currently, it is unable to exert effective pharmacological effects and is therefore administered as an injection. However, since administration by injection is painful, various other administration methods have been attempted. For example, an intrarectal administration method using a suppository using salicylic acid derivatives such as sodium salicylate, sodium 3-methoxysalicylate, and 5-methoxysalicylic acid as an absorption enhancer [Journal of
Pharmacy and Pharmacology (J.
Pharm. Pharmacol.), 33, 334 (1981)].
Other methods under consideration include intratracheal administration [Diabetes, 20, 552, (1971)] and eye drop administration (Diabetes Society Abstracts, 237, (1974)). However, since all of these methods require a higher dose than injection and have the disadvantage that absorption tends to fluctuate, very few methods have reached practical use yet. On the other hand, as an attempt at intranasal administration, a method of intranasal administration of an aqueous solution of insulin using a surfactant such as sodium glycodeoxycholate as an absorption enhancer is known [Proceedings of the National Academy of
Science (Proc.Natl.Acad.Sci.USA) 82,
pp7419-7423 (1985)]. Alternatively, nasal administration of an aqueous solution of insulin, glucagon, etc. using a fusidic acid derivative such as sodium-tauro-24,25-dihydroxyfusidate as an absorption enhancer is known (Japanese Patent Laid-Open Publication No. 33126/1982). There is. However, these methods are highly irritating to the nasal mucosa during administration, and since all dosage forms are liquid, when administered nasally, the liquid drug tends to flow out of the nasal cavity, making it difficult for the drug to be administered efficiently than the nasal mucosa. It is difficult to say that it is well absorbed. Furthermore, a liquid preparation for nasal administration of insulin combined with cyclodextrin (Japanese Patent Application Laid-open No. 189118/1983);
Calcitonin nasal preparations containing basic and/or neutral amino acids (Japanese Patent Application Laid-open No. 118325/1982) and intranasal calcitonin containing aldose (Japanese Patent Application Laid-open No. 126034/1989) are known. There is. In these methods, the irritation to the nasal mucosa during administration is considerably reduced, and in this respect, the methods are satisfactory to some extent. However, even with these methods, it is still difficult to say that the drug is absorbed efficiently through the nasal mucosa. On the other hand, as a powdered formulation for nasal administration,
No. 59-163313 and Japanese Patent Application Laid-open No. 60-224616 disclose powdered preparations for nasal administration consisting of a water-absorbing base and peptide hormones;
In the publication, the present inventors have disclosed a powder preparation for nasal administration comprising a quaternary ammonium compound and a cellulose lower alkyl ether. These preparations cause almost no irritation to the nasal mucosa upon administration, and can absorb high molecular weight peptide hormones through the nasal mucosa relatively efficiently. Furthermore, as a powdered preparation for nasal administration, US Pat. No. 4,294,829 discloses a preparation comprising a cellulose lower alkyl ether drug. This formulation is characterized in that cellulose lower alkyl ether absorbs water on the nasal mucosa, becomes a viscous liquid, flows on the nasal mucosa, and gradually releases the drug. However, even with these methods, it is difficult to say that the drug is absorbed through the nasal mucosa with sufficient efficiency, and there is still room for improvement. Therefore, there is a need for a formulation for nasal administration in which the drug is efficiently absorbed through the nasal mucosa and is less irritating to the nasal mucosa. <Means for solving the problem> The present inventors have developed a nasal polypeptide that can efficiently absorb physiologically active polypeptides such as calcitonin through the nasal mucosa and is suitable for intranasal administration. As a result of intensive research, we have developed an absorption enhancer consisting of a combination of polypeptides, two specific absorption enhancers, and a water-absorbing solid with the aim of obtaining a formulation for nasal administration that causes almost no irritation to the mucous membranes. The present invention was achieved by discovering that a powder composition comprising a base material can surprisingly achieve the above-mentioned objects. That is, the present invention provides (a) physiologically active polypeptides, (b) as an absorption enhancer () ascorbic acids and () basic amino acids and/or their pharmaceutically acceptable salts, and (c) ) A powder composition for nasal administration, comprising a water-absorbing solid base suitable for application to the nasal mucosa. Specific examples of preferred ascorbic acids of the present invention include ascorbic acid and erythorbic acid. Further, specific examples of the basic amino acids preferred in the present invention include arginine, histidine, lysine, orthinine, ergothioneine, and the like. Among these basic amino acids, arginine, histidine, and lysine are particularly preferred. These basic amino acids may be in the L-form, D-form, or racemic form. Pharmaceutically acceptable salts of these basic amino acids are any salts that maintain enhanced absorption of physiologically active polypeptides through the nasal mucosa and are not harmful; Specific examples include salts of organic acids such as citric acid, tartaric acid, malic acid, and fumaric acid, and mineral acids such as hydrochloric acid. Among these salts, hydrochloride is particularly preferred. The preferred concentration of ascorbic acids in powdered compositions useful for nasal administration of the present invention is about
The content is 0.1 to 20% by weight, more preferably 1 to 15% by weight, and even more preferably 5 to 10% by weight. Further, the preferred concentration of the basic amino acid and/or its pharmaceutically acceptable salt in the powdered composition useful for nasal administration of the present invention is about 0.1% of the total composition.
-20% by weight, more preferably 1-15% by weight
and more preferably 5 to 10% by weight. In the present invention, the target drug is polypeptides having physiological activity. Polypeptides have molecular weights in the range of 300,000 to 300,000.
It is preferable because it is more easily absorbed than the nasal mucosa.
The molecular weight is particularly preferably in the range of 300 to 150,000. Preferred specific examples of physiologically active polypeptides include the following. For example, insulin, proinsulin, angiotensin,
Vasopressin, desmopressin, feripressin, protirelin, luteinizing hormone-releasing hormone, corticotropin, prolactin, somatotropin, thyrotropin, luteinizing hormone, calcitonin, kallikrein, glucagon, oxytocin, gastrin, secretin, serum gonadotropin, lipomodulin, atrial sodium Diuretic peptide [alpha human laboratory polypeptide (α-hANP)]
etc.], growth hormone releasing hormone, growth hormone,
Pupetide hormones such as erythropoietin, urogastrone, renin, parathylin (PTH) and its antagonists, corticotropin release factor,
Its precursors, its regulatory factors, or its derivatives; bioactive proteins such as interferon, interleukin, tumor necrosis factor (TNF), transferrin, histaglobulin, macrocortin, and blood coagulation factor; enzymes such as lysozyme and urokinase Proteins include vaccines or vaccine components such as pertussis vaccine, diphtheria vaccine, tetanus vaccine, influenza vaccine, lymphocyte pleocytosis factor, and fibrillar hemagglutination factor. Among these, peptide hormones are particularly preferred, and among peptide hormones, calcitonin, insulin, α-hANP,
Preferred are luteinizing hormone releasing hormone, corticotropin, desmopressin, vasopressin, glucagon, oxytocin, PTH or growth hormone. Furthermore, calcitonin, insulin, α
-hANP is preferred. In the composition for nasal administration of the present invention, physiologically active polypeptides in powder form are preferably used. Polypeptides that are not in powder form are preferably lyophilized before use. The amount of the above-mentioned polypeptides to be used is appropriately determined depending on the strength of the medicinal efficacy of each polypeptide. The above-mentioned polypeptides may be used in combination with human serum albumin, mannitol, sorbitol, aminoacetic acid, non-basic amino acids, sodium chloride, phospholipids, etc., in order to stabilize them or as bulking agents in addition to stabilization. Powdered compositions useful for nasal administration of the present invention use a water-absorbing solid base as the base. Preferred examples of the water-absorbing solid base include a base that is water-absorbent and poorly soluble in water, and a base that is water-absorbent and easily soluble in water. Such bases may be used alone or in combination of two or more depending on the purpose. Here, "water-absorbent and poorly water-soluble" means "water-absorbent and poorly water-soluble" on the human nasal mucosa or in an environment similar to this, i.e., in water at a pH of about 7.4 and a temperature of about 36°C to about 37°C. This means that it has the property of being poorly soluble in water. Furthermore, water-absorbing and water-soluble means that on the human nasal mucosa or in an environment similar to this, that is, at a pH of about 7.4 and a temperature of about 36°C to about 37°C.
This means that it has the property of being water-absorbing and easily soluble in water at ℃. Preferred specific examples of the water-absorbing and poorly water-soluble base of the present invention include the following. For example, water-absorbing and poorly water-soluble celluloses such as crystalline cellulose, α-cellulose, and cross-linked carboxymethylcellulose sodium; water-absorbing and water-resistant celluloses such as hydroxypropyl starch, carboxymethyl starch, cross-linked starch, amylose, amylopectin, and pectin. Soluble starches; Water-absorbing and poorly water-soluble proteins such as gelatin, casein, and sodium caseinate; Water-absorbing and poorly water-soluble gums such as gum arabic, gum tragacanth, and glucomannan; Polyvinyl polypyrrolidone, cross-linked polyester Examples include water-absorbing and poorly water-soluble crosslinked vinyl polymers such as acrylic acid and its salts, crosslinked polyvinyl alcohol, and polyhydroxyethyl methacrylate. Among these, water-absorbing and poorly water-soluble celluloses are preferred, with crystalline cellulose, α-cellulose, or crosslinked sodium carboxymethylcellulose being particularly preferred, and crystalline cellulose being even more preferred. The amount of the water-absorbing and poorly water-soluble base to be used varies depending on the type of polypeptide used and cannot be generalized, but it is usually in the range of at least 1 times the weight of the polypeptide, especially 15 more than double the weight, and even
A range of 20 times or more by weight is preferred. Preferred specific examples of the water-absorbing and easily water-soluble base of the present invention include the following. For example, water-absorbing and easily water-soluble cellulose lower alkyl ethers such as hydroxypropyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and sodium carboxymethyl cellulose; dextrin, cyclodextrin (α-, β-, γ
- or dimethyl α- or dimethyl β-), water-absorbing and easily water-soluble starches such as pullulan; water-absorbing and easily water-soluble starches such as polyvinyl alcohol, polyvinylpyrrolidone, and sodium salts of carboxyvinyl polymers. Polymers: Water-absorbing and easily water-soluble polyacrylates such as sodium polyacrylate, potassium polyacrylate, and ammonium polyacrylate; Water-absorbing and easily water-soluble proteins such as chitin and chitosan Examples include water-absorbing and easily water-soluble saccharides such as lactose, glucose, maltose, and sucrose. Among these water-absorbing and easily water-soluble bases, hydroxypropylcellulose, methylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, dextrin, cyclodextrin (β-, dimethylβ- or dimethylα -), chitin, chitosan, lactose, sodium polyacrylate, polyethylene glycol, and polyvinylpyrrolidone are preferred, and hydroxyprolcellulose, β-cyclodextrin, dimethyl β-cyclodextrin, dimethyl α-cyclodextrin, and lactose are particularly preferred, More preferred are hydroxypropylcellulose, β-cyclodextrin, and lactose. The amount of the water-absorbing and easily water-soluble base to be used varies depending on the type of polypeptide used and cannot be generalized, but it is usually in the range of at least 1 times the weight of the polypeptide, especially More than 15 times the weight, and even
A range of 20 times or more by weight is preferred. In the powder composition of the present invention, it is preferred that 90% by weight or more of the particles have an effective particle size between 10 and 250 microns. By making the particles have a particle size within this range, when administered into the nasal cavity, they are widely distributed on the nasal mucosa and remain well at the adhesion site, and can also be sprayed into the nasal cavity through the nostrils as a powder. When administered, it can be efficiently administered intranasally. If particles with an effective particle diameter of less than 10 microns account for more than 10% by weight, when administered by methods such as spraying, many particles reach the lungs or dissipate outside the nasal cavity when sprayed. .
Furthermore, it is difficult to maintain a high drug concentration at the attachment site. On the other hand, when particles with an effective particle diameter exceeding 250 microns account for more than 10% by weight, when administered into the nasal cavity, even if they adhere to the nasal mucosa, they tend to separate from the mucous membrane, resulting in low local retention of the drug. Therefore, it is undesirable. Particularly preferred is one in which 90% by weight or more of the particles have an effective particle diameter of between 20 and 150 microns. The powder composition of the present invention can be produced, for example, as follows. That is, physiologically active polypeptides, ascorbic acids, basic amino acids and/or their pharmaceutically acceptable salts, and a water-absorbing solid base are mechanically mixed, and then sieved. ,Preferably
It can be produced by obtaining a composition in which 90% by weight or more of the particles have an effective particle size of 10 to 250 microns. Alternatively, physiologically active polypeptides, ascorbic acids, basic amino acids, and/or pharmaceutically acceptable salts thereof are uniformly dissolved or suspended in an appropriate amount of purified water, and then added to the nasal mucosa. A water-absorbing solid base suitable for application to the base, that is, a water-absorbing and slightly water-soluble base and/or a water-absorbing and easily water-soluble base, is added to the base and/or the water-absorbing solid base.
Or, after uniformly containing and/or adsorbing polypeptides and ascorbic acids, basic amino acids and/or pharmaceutically acceptable salts thereof on the surface of the base, or polypeptides and ascorbic acids, and A water-absorbing solid base is uniformly dissolved, suspended or kneaded together with a basic amino acid and/or a pharmaceutically acceptable salt thereof in an appropriate amount of purified water, and then frozen, and then the frozen composition is prepared. By freeze-drying the product and then grinding it by conventional methods and then sieving, or by grinding it and then mixing it homogeneously with the desired water-absorbing solid base. , preferably 90% by weight or more of the particles have an effective particle size of 10 to 250 microns. Alternatively, polypeptides and ascorbic acids, basic amino acids and/or pharmaceutically acceptable salts thereof may be mixed into a water-absorbing solid base, that is, a water-absorbing and poorly water-soluble base, or a water-absorbing solid base. The resulting mixture is then compressed under pressure, and the resulting compressed product is crushed and sieved to obtain a powder containing preferably at least 90% by weight of particles. It is manufactured by obtaining a composition consisting of an effective particle size of 10 to 25 microns. Or polypeptides, ascorbic acids, basic amino acids and/or pharmaceutically acceptable salts thereof, and water-absorbing solid bases, i.e., water-absorbing and poorly water-soluble bases, or water-absorbing and poorly water-soluble bases; It can also be obtained by adding an easily water-soluble base to an appropriate amount of purified water and uniformly dissolving or suspending it, or by kneading it well, drying it by a conventional method, and then sieving. . The powder composition of the present invention has physical properties as a formulation,
In order to improve the appearance or odor, known lubricants, colorants, preservatives, preservatives, flavoring agents, etc. may be added as necessary. Examples of lubricants include talc, stearic acid and its salts; colorants include copper chlorophyll, β-carotene, Red No. 2, Blue No. 1, etc.; preservatives include stearic acid, ascorbic acid stearate, etc. ; Examples of preservatives include quaternary ammonium compounds such as benzalkonium chloride, paraoxybenzoic acid esters, phenol, chlorobutanol, etc.;
Examples of the flavoring agent include menthol, citrus flavor, and the like. The composition of the present invention can be made into a unit dosage form of a powder. Such powders can be filled into capsules, such as hard gelatin capsules, as a preferred form for administration. A method for administering a powder by spraying into the nasal cavity is, for example, by setting a capsule filled with the powder in a special spray device equipped with a needle, passing the needle through it, and making minute holes at the top and bottom of the capsule. Next, there is a method of blowing out the powder by blowing air in with a rubber ball or the like. <Examples> Hereinafter, the present invention will be explained in detail with reference to Examples, but the present invention is not limited thereto. Example 1 () A powdered composition useful for nasal administration of the present invention was obtained as follows. (a) Salmon calcitonin (4000 MRC units/mg)
0.1mg and L-ascorbic acid 29.8mg and L-
Take 35.6 mg of arginine hydrochloride in a test tube,
Add 250μ of purified water and dissolve uniformly, then
A homogeneous composition was obtained by adding 500 mg of microcrystalline cellulose to the mixture, sprinkling it well, and freeze-drying it. Next, by sieving this composition, 90% by weight or more of the particles are removed.
A homogeneous powder composition with a particle size of 46-149 microns was obtained. The powdered composition thus obtained contains 0.0527 mg of L-ascorbic acid, 0.0630 mg of L-arginine hydrochloride and 0.707 MRC units of salmon calcitonin in 1 mg. (b) Hydroxypropylcellulose 500mg and L
- Take 29.8 mg of ascorbic acid and 35.6 mg of L-arginine hydrochloride in a mortar, and add salmon calcitonin (4000 MRC units/mg) to the mortar.
By adding 0.1 mg and mixing well, a uniform powder composition was obtained in which more than 90% by weight of the particles had a particle size of 46 to 149 microns. The powdered composition obtained in this way is
0.0527mg of L-ascorbic acid in 1mg
0.0630mg L-arginine hydrochloride and
Contains 0.707 MRC units of salmon calcitonin. (c) Salmon calcitonin (4000MRC units/mg)
0.06mg and L-ascorbic acid 17.8mg and L-
After uniformly dissolving 21.3 mg of arginine hydrochloride in 150μ of purified water, 300 mg of lactose was added and sprinkled well (at this time, some of the lactose was dissolved). This was then freeze-dried to obtain a uniform composition. By sieving this composition, a uniform powder composition was obtained in which 90% by weight or more of the particles had a particle size of 46 to 149 microns. The powder composition thus obtained was 1 mg
Contains 0.0525mg of L-ascorbic acid and 0.0628mg of L-ascorbic acid.
Contains mg L-arginine hydrochloride and 0.708 MRC units of salmon calcitonin. (d) Powdered compositions containing salmon calcitonin shown in (a), (b), and (c) are placed in the prescribed capsules for 10 to 10 minutes.
A salmon calcitonin preparation for nasal administration to humans was obtained by filling 50 mg. () In order to compare with the composition of the present invention, the following comparative composition containing no absorption enhancer was obtained. (a′) Salmon calcitonin (4000 MRC units/mg)
0.1 mg was taken into a test tube, 250μ of purified water was added to dissolve it uniformly, and then 500mg of microcrystalline cellulose was added thereto, the mixture was thoroughly sprinkled, and freeze-dried to obtain a uniform composition.
Then, by sieving this composition
A homogeneous powder composition was obtained in which more than 90% by weight of the particles had a particle size of 46 to 149 microns. The powdered composition thus obtained contains 1mg of
Contains 0.800 MRC units of salmon calcitonin. (b′) 500 mg of hydroxypropyl cellulose and 0.1 mg of salmon calcitonin (4000 MRC units/mg)
A uniform powdery composition was obtained by placing the ingredients in a mortar and mixing them thoroughly. The powdered composition thus obtained contains 1 mg of
Contains 0.800 MRC units of salmon calcitonin. (c′) Salmon calcitonin (4000 MRC units/mg)
After uniformly dissolving 0.06mg in 150μ of purified water, 300mg of lactose was added and sprinkled well (at this time, some of the lactose was dissolved). This was then freeze-dried to obtain a uniform composition. By sieving this composition, a uniform powder composition in which 90% by weight or more of the particles had a particle size of 46 to 149 microns was obtained. The powder composition thus obtained was 1 mg.
Contains 0.800 MRC units of salmon calcitonin. Example 2 (Nasal administration experiment of powdered salmon calcitonin preparation in domestic rabbits) The (a), (b), ( c), (a′), (b′),
(c′)
Salmon calcitonin preparations prepared in
1.4 MRC units/Kg administered, 30 before and after administration
Blood was collected from the ear vein of the rabbit at minutes, 1 hour, 2 hours, 4 hours, and 6 hours. After blood collection, the blood was centrifuged at 2800 rpm for 10 minutes using a centrifuge to obtain plasma. The powder was administered without anesthesia using a sprayer modified for animals. Specifically, this was done as follows. That is, first, Example 1
The six types of salmon calcitonin preparations prepared above were filled into prescribed capsules at a dosage of 1.4 MRC units/Kg. The capsule is then placed in a nebulizer and fitted with a modified animal nozzle. Further, by placing the cap on the capsule, a hole was made in the capsule using the needle attached to the inside of the cap, and the nozzle was immediately inserted into the nasal cavity of the rabbit to perform intranasal administration by spraying with air pressure. Plasma calcium concentrations were measured before and after administration, and the absorbability of salmon calcitonin through the nasal mucosa was investigated. Plasma calcium was measured using a calcium measurement kit manufactured by Yatron. Figures 1, 2, and 3 show changes in plasma calcium in terms of plasma calcium lowering rate (%). The values shown in each figure are the average value ± standard error of 4 to 5 domestic rabbits.
For comparison, 1.4 MRC units/50μ/Kg of a salmon calcitonin aqueous solution dissolved in 17.5mM citrate buffer (PH approximately 6) containing 0.16% gelatin and 0.7% sodium chloride was injected intravenously. Changes in plasma calcium at different times are also shown by broken lines in each figure. Figure 1 shows the case where microcrystalline cellulose is used as the base, and (1) is a preparation containing a combination of L-ascorbic acid and L-arginine hydrochloride of the present invention ((a) of Example 1). ), and (2) shows the case where a formulation ((a') of Example 1) that did not use any of these absorption enhancers was administered. Figure 2 shows the case where hydroxypropylcellulose was used as the base, and (1) shows a formulation containing L-ascorbic acid and L-arginine hydrochloride in combination ((b) of Example 1). (2) is a formulation that does not use any of those absorption enhancers ((b') of Example 1)
The figure shows the case where the drug was administered. Figure 3 shows the case using lactose as the base;
(1) is a formulation containing L-ascorbic acid and L-arginine hydrochloride ((c) of Example 1),
(2) shows the case where a formulation ((c') in Example 1) without any of these absorption enhancers was administered. As is clear from Figures 1, 2 and 3,
The powdered composition of the present invention containing L-ascorbic acid and L-arginine hydrochloride in combination has excellent absorption of salmon calcitonin, and both can provide good physiological activity almost equivalent to intravenous injection. Indicated. Example 3 () 100mg of porcine insulin to 2.5mg of 0.1N hydrochloric acid
After dissolving in water, add 37.5mg of water, then 0.1N
- Add approximately 3.2ml of sodium hydroxide aqueous solution to pH
By adjusting to 7.4 and then freeze-drying,
Water-soluble insulin powder (23.5 units/mg)
I got it. Using this insulin powder, the following composition of the present invention was obtained. (a) Water-soluble insulin powder (23.5 units/
mg), 20 mg of erythorbic acid, and 20 mg of L-histidine hydrochloride were placed in a mortar, and 300 mg of microcrystalline cellulose was added thereto, and the three components were thoroughly mixed to obtain a uniform powder composition. . The powdered composition thus obtained contained 0.057 mg of erythorbic acid and L-histidine hydrochloride and 0.67 mg of L-histidine hydrochloride in 1 mg, respectively.
Contains units of insulin. () Using the above insulin powder, the following comparative compositions were obtained for comparison with the compositions of the present invention. (b) Water-soluble insulin powder (23.5 units/
10 mg) and 340 mg of microcrystalline cellulose were placed in a mortar and mixed well to obtain a uniform powder composition. The powdered composition thus obtained contains 0.67 units of insulin in 1 mg. Example 4 (Intranasal administration experiment of insulin preparation in domestic rabbits) The powder prepared in (a) and (b) of Example 3 was administered into the nasal cavity of a white native male domestic rabbit (weight 2.5-3.5 kg). Each insulin preparation was administered at 1.21 units/kg, and blood was collected before administration and at 5 minutes, 10 minutes, 20 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and 6 hours after administration. The powder was administered in the same manner as in Example 2. Plasma glucose and insulin concentrations were measured before and after administration, and the absorbability of insulin from the nasal mucosa was investigated. The concentration of insulin in plasma was measured by radioimmunoassay. The results are shown in Figure 4. Plasma glucose concentration was measured by a method using orthotoluidine (Clinical Chemistry 8,
215 (1962)). The results are shown in Figure 5 as blood sugar lowering rate (%). The values shown in FIGS. 4 and 5 are the average values of five domestic rabbits. As shown in Figure 4a, the plasma insulin level after administration of the powder containing the combination of erythorbic acid and L-histidine hydrochloride of the present invention prepared in Example 3 (a) was The highest level was reached 10 minutes after administration, increasing more than 50 times the basal value. On the other hand b
As shown in (b) of Example 3, when the powder was administered as a comparative example, the highest level was reached 30 minutes after administration; It was at a slightly lower level. The above facts indicate that administration of the powder of the present invention allows insulin to be absorbed particularly efficiently from the nasal mucosa, and that absorption is efficiently sustained. In FIG. 5, a represents the case where the powder containing erythorbic acid and L-histidine hydrochloride of the present invention prepared in Example 3 (a) was administered as an absorption enhancer.
b shows the blood sugar lowering rate when the powder containing it, prepared as a comparative example in (b) of Example 3, was administered. As shown in Figure 5, the fluctuation pattern of the blood sugar lowering rate correlates well with the fluctuation pattern of the insulin level after administration of each preparation, and the powder of the present invention is particularly superior in terms of the blood sugar lowering rate. I understand that. Example 5 A powdered composition useful for nasal administration of the present invention was obtained as follows. (1) α-hANP 0.5mg and L-ascorbic acid 29.8mg
and 29.8mg of L-arginine were uniformly dissolved in 250μ of purified water, and then microcrystalline cellulose was added to this.
Add 500 mg and sprinkle well. This was then freeze-dried to obtain a uniform composition. By sieving this composition, 90
A uniform powder composition was obtained in which more than % by weight of the particles had a particle size of 46 to 149 microns. The composition thus obtained contained 0.893 μg of α in 1 mg.
-hANP and 0.0532 mg each of L-ascorbic acid and L-arginine. (2) Hydroxypropylcellulose 500mg and L-
Ascorbic acid 29.8 mg and L-lysine hydrochloride 29.8
mg and into a mortar, and then add α-
By adding 0.5 mg of hANP and mixing thoroughly, a uniform powder composition in which 90% by weight or more of the particles had a particle size of 46 to 149 microns was obtained.
The powder composition thus obtained was 1 mg
0.893μg α-hANP and 0.0532mg each
It contains L-ascorbic acid and L-lysine hydrochloride. (3) In exactly the same manner as in (2) except that 500 mg of lactose was used instead of 500 mg of hydroxypropyl cellulose, 0.893 μg of α-hANP and 0.0532 mg of L-ascorbic acid and L-lysine hydrochloride were added in 1 mg. A homogeneous powder composition was obtained. (4) L-ascorbic acid and L-arginine or L-lysine hydrochloride and α-
A preparation for human nasal administration was obtained by filling a powdered composition containing hANP into a predetermined capsule. Example 6 470 mg of β-cyclodextrin was placed in a mortar,
This includes 30 mg of erythorbic acid and 30 mg of histidine hydrochloride.
mg and 10 mg of lyophilized vasopressin (70-100 units/mg) were added and mixed well to obtain a homogeneous powder composition. The powdered composition thus obtained contained 0.056
mg erythorbic acid and histidine hydrochloride and 1.30
Contains ~1.85 units of vasopressin. A preparation for human nasal administration was obtained by filling the obtained powder composition into a predetermined capsule. Example 7 Take 870 mg of dimethyl-β-cyclodextrin in a mortar, add 60 mg of erythorbic acid, 60 mg of histidine hydrochloride, and 10 mg of freeze-dried luteinizing hormone-releasing hormone, and mix well to form a uniform powder. A composition was obtained. The powdered composition thus obtained contains 1 mg of each
0.06mg of erythorbic acid and histidine hydrochloride and
A formulation for human nasal administration containing 0.01 mg of luteinizing hormone-releasing hormone was obtained by filling it into a predetermined capsule. Example 8 Take 879 mg of dimethyl-α-cyclodextrin in a mortar, add 60 mg of L-ascorbic acid, 60 mg of histidine hydrochloride, and 1 mg of freeze-dried desmopressin acetate, and mix well to obtain a uniform powder composition. I got something. The powdered composition thus obtained contained 0.06 mg of L-ascorbic acid, histidine hydrochloride and
A formulation for human nasal administration containing 0.001 mg of desmopressin acetate was obtained by filling it into a predetermined capsule. Example 9 A powdered composition useful for nasal administration of the present invention was obtained as follows. (1) Take 850 mg of microcrystalline cellulose in a mortar and add 50 mg of L-ascorbic acid and arginine hydrochloride to it.
50 mg and 50 mg of interferon (100,000 units/mg) freeze-dried with human serum albumin were added and mixed well to obtain a uniform powder composition. The powdered composition thus obtained contains 0.05 mg of L-ascorbic acid, arginine hydrochloride, and 5000 units of interferon in 1 mg each. (2) Except using 850 mg of hydroxypropyl cellulose instead of 850 mg of microcrystalline cellulose (1)
0.05 of each in 1mg
mg of L-ascorbic acid and arginine hydrochloride and
A homogeneous powder composition containing 5000 units of interferon was obtained. (3) 850mg of lactose instead of 850mg of microcrystalline cellulose
Exactly as in (1) except for using 1mg
A homogeneous powder composition containing 0.05 mg of L-ascorbic acid, arginine hydrochloride, and 5000 units of interferon, respectively, was obtained. (4) By filling a powdered composition containing L-ascorbic acid, arginine hydrochloride, and interferon shown in (1), (2), and (3) into a prescribed capsule, human A formulation for nasal administration was obtained. Example 10 879 mg of hydroxypropylcellulose was placed in a mortar, and 60 mg of L-ascorbic acid, 60 mg of arginine hydrochloride, and 1 mg of PTH were added thereto and mixed well to obtain a uniform powder composition. The powdered composition thus obtained contains 0.05 mg of L-ascorbic acid, arginine hydrochloride, and 0.001 mg of PTH in 1 mg, and by filling it into a predetermined capsule, it can be used to A formulation for nasal administration was obtained. Example 11 1 mg of hemagglutinin, which is a component of Bordetella pertussis, 1 mg of detoxified pertussis toxin, 38 mg of erythorbic acid, 38 mg of histidine hydrochloride, and 922 mg of hydroxypropyl cellulose were placed in a mortar and mixed well to form a uniform powder. A composition was obtained. The powdered composition thus obtained is 1
Each mg contained 0.038 mg of erythorbic acid, arginine hydrochloride, and 0.002 mg of Bordetella pertussis components, and was filled into a predetermined capsule to obtain a formulation for human nasal administration. Examples 12-17, Comparative Examples 1-2 Examples 12-17 For comparison with the powdered composition useful for nasal administration of the present invention ((b) of Example 1), ), 0.707 MRC units of salmon calcitonin, 0.0527 mg of L-ascorbic acid, and various basic amino acids shown in Table 1 were added to 1 mg of a powdered composition based on hydroxypropylcellulose.
Powder compositions containing 0.0630 mg were prepared (Examples 12 to 17).

【table】

[Table] Comparative Examples 1-2 For further comparison with the formulation of the present invention, Example 1
In (b), instead of L-ascorbic acid and arginine hydrochloride as absorption enhancers, a powder composition containing L-ascorbic acid alone (Comparative Example 1) and a powder composition containing arginine hydrochloride alone ( Comparative Example 2) was created. In these cases, the amount of hydroxypropyl cellulose as a base was adjusted so that the doses of salmon calcitonin and each absorption enhancer were the same as those mentioned above. Control Example As a control example, a powdered composition containing 0.707 MRC units of salmon calcitonin per mg and having hydroxypropylcellulose as a base was prepared. Comparative administration experiments were conducted on white native male domestic rabbits (body weight 3~
The experiment was conducted in the same manner as in Example 2 using 3.8 kg (4 to 5 birds per group). The results when the dose was 1.4 MRC units/Kg are shown in Table 2 as plasma calcium lowering rate. As is clear from the comparison of the plasma calcium lowering rate (%) 2 hours after administration and the total plasma calcium lowering rate (%・hr) up to 4 hours after administration shown in Table 2, L-ascorbic acid and base The powders of the present invention (Examples 12 to 17) in which a sexual amino acid and/or a pharmaceutically acceptable salt thereof were used in combination were better than L-ascorbic acid alone (Comparative Example 1) prepared for comparison. Alternatively, it can be seen that the nasal mucosal absorption of salmon calcitonin is superior to the powder used with arginine hydrochloride alone (Comparative Example 2), or to the control powder containing neither of these.

【table】 [Brief explanation of the drawing]

Figures 1, 2, and 3 show the absorption of salmon calcitonin in terms of plasma calcium lowering rate (%) when the composition of the present invention using salmon calcitonin as a polypeptide was administered nasally. It is.
Figures 4 and 5 show the absorption of insulin in terms of insulin concentration (Figure 4) and blood glucose lowering rate (Figure 5), respectively, when the composition of the present invention using insulin was administered nasally. be. In FIGS. 1, 2, and 3, 1 indicates the case where an absorption enhancer is used in combination, and 2 indicates the case where no absorption enhancer is included. The dotted line indicates the case where the same dose of calcitonin as the intranasal administration was administered intravenously. In FIGS. 4 and 5, a shows the case where an absorption enhancer is used in combination, and b shows the case where they are not included.

Claims (1)

[Scope of Claims] 1(a) Polypeptides having physiological activity (b) As an absorption enhancer () Ascorbic acids and () Basic amino acids and/or their pharmaceutically acceptable salts and (c ) A powder composition for nasal administration, comprising a water-absorbing solid base suitable for application to the nasal mucosa. 2. The composition according to claim 1, wherein the ascorbic acid is ascorbic acid or erythorbic acid. 3. The composition according to claim 1, wherein the basic amino acid is arginine, histidine or lysine. 4. Claim 1 or 3, wherein the pharmaceutically acceptable salt of the basic amino acid is a hydrochloride.
Compositions as described in Section. 5 Ascorbic acids 0.1 to 20% by weight of the total weight
Claim 1 or 2 present in a concentration of
Compositions as described in Section. 6. Claims 1, 3, or 4, wherein the basic amino acid and/or its pharmaceutically acceptable salt is present in a concentration of 0.1 to 20% by weight of the total weight.
Compositions as described in Section. 7. The composition according to claim 1, wherein the water-absorbing solid base is a water-absorbing and poorly water-soluble cellulose, starch, protein, crosslinked vinyl polymer, or gum. 8 Water-absorbing and poorly water-soluble cellulose is
The composition according to claim 7, which is crystalline cellulose, α-cellulose, or crosslinked sodium carboxymethylcellulose. 9. The composition according to claim 7, wherein the water-absorbing and poorly water-soluble crosslinked vinyl polymer is a crosslinked polyvinylpyrrolidone or a crosslinked carboxyvinyl polymer. 10 Claims in which the water-absorbing solid base is water-absorbing and easily water-soluble cellulose lower alkyl ethers, sugars, starches, vinyl polymers, polyacrylates, or proteins. The composition according to item 1. 11. The composition according to claim 10, wherein the water-absorbing and easily water-soluble cellulose lower alkyl ether is hydroxypropylcellulose, methylcellulose, carboxymethylcellulose, hydroxyethylcellulose, or hydroxyperopylmethylcellulose. 12. The composition according to claim 10, wherein the water-absorbing and easily water-soluble saccharide is lactose. 13 Water-absorbing and water-soluble starches are β-
The composition according to claim 10, which is cyclodextrin, dimethyl-β-cyclodextrin and/or dimethyl-α-cyclodextrin. 14. The composition according to any one of claims 1 and 7 to 13, which uses one type of water-absorbing solid base or an appropriate combination of two or more types. 15. The composition according to claim 1, wherein the physiologically active polypeptide has a molecular weight of 300 to 300,000. 16. The biologically active polypeptide according to claim 1 or 15, wherein the physiologically active polypeptide is a peptide hormone, a precursor thereof, a regulator thereof, a derivative thereof, a physiologically active protein, an enzyme protein, or a vaccine or vaccine component. Composition. 17 Peptide hormones include calcitonin, insulin, alpha human laboratory polypeptide (α-hANP), luteinizing hormone-releasing hormone, desmopressin,
Claim 16 which is growth hormone, parathylin (PTH), vasopressin or oxytocin
Compositions as described in Section. 18. The composition according to claim 16, wherein the physiologically active protein is interferon. 19. The composition according to claim 16, wherein the enzyme protein is lysozyme, urokinase, or a precursor or regulator thereof. 20. The composition according to claim 16, wherein the vaccine is an influenza vaccine or a pertussis vaccine. 21. The composition according to claim 1, wherein 90% by weight or more of the particles in the powder composition have an effective particle size of between 10 and 250 microns.