JP2802488B2 - Bioactive peptide composition for nasal absorption - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a physiologically active peptide composition for nasal absorption, and more particularly to a peptide hormone,
The present invention relates to a physiologically active peptide composition for nasal absorption, which is excellent in pharmaceutical stability and in vivo absorption when a physiologically active peptide such as a physiologically active protein or an enzyme protein is administered intranasally (intranasal administration).

[0002]

BACKGROUND OF THE INVENTION Physiologically active peptides such as calcitonin, insulin and the like are specific high-molecular compounds which are used in various medical applications in the field of treatment because of their specific physiological activities. However, these physiologically active peptides are degraded by proteases in the digestive tract, or have high molecular weight and high polarity, so that they are hardly absorbed as they are from the intestinal mucosa, and are difficult to administer orally. Is limited to injection administration. By the way, such an administration method gives pain at the injection site to the patient, and is not preferable.
When administered at regular intervals, the patient suffers significantly. Therefore, the development of a non-injectable administration method that is simple and easy to administer safely and frequently is desired. As one such administration method,
For example, as for calcitonin, an aerosol for nasal inhalation of a powder suspension using a fluorocarbon as a propellant has been developed. Further, the development of a nasal spray as an intranasal solution as a nasal administration method has been studied, and a formulation in which a surfactant as an absorption enhancer is added to calcitonin has been proposed. Recently, some nasal administration type powders having improved absorbability by adsorbing calcitonin to polysaccharide cellulose have been proposed. Considering the nasal administration method that has been actively studied recently, a venous plexus has developed in the nasal mucosa lamina propria of the nasal cavity, which is the administration site, and the drug passes through this nasal mucosa. It is excellent as a method of administering a physiologically active peptide, which is a drug that is difficult to be orally administered because it is absorbed and enters the systemic circulation,
Although various application examples of the preparations have been proposed, the preparation examples so far are not sufficient in terms of drug absorption or local irritation, and none of them have yet been put to practical use.

[0003]

DISCLOSURE OF THE INVENTION The present inventors have studied the development of a physiologically active peptide which had been difficult to administer orally until now, to develop a nasal preparation having good absorbability and no irritation. . That is, an object of the present invention is to provide a composition for intranasal administration which is superior in bioavailability (bioavailability) as compared with the intranasal preparations proposed so far. The present inventors have conducted intensive studies to solve such problems and found that bioactive peptides such as calcitonin and insulin were uniformly dispersed and adhered to a specific carrier which had not been previously studied as a carrier for nasal administration preparations. It has been found that an effective clinical treatment is possible by administering the composition to the nasal route, that is, by applying it to the nasal mucosa. That is,
The inventor of the present invention has proposed that a bioactive peptide such as calcitonin and insulin is dispersed and adhered to a special carrier proposed by the present invention, thereby achieving bioavailability equivalent to or higher than that obtained by standard injection administration. The inventors have completed the present invention.

[0004]

Means for Solving the Problems The present invention provides an effective dose of a physiologically active peptide in an average particle size of 5 μm to 20 μm.
An object of the present invention is to provide a bioactive peptide composition for nasal absorption, which is uniformly dispersed and adhered to a carrier which is a 00 μm porous crystallized glass.

In the present invention, the porous crystallized glass as a carrier for uniformly dispersing, adhering, and binding a physiologically active peptide is an artificial bone filling material used for filling a defective portion of a bone or a root, Easy to integrate with the organization,
Although it is widely used in the field of orthopedic surgery because of its high mechanical strength, application of a nasal administration preparation as a carrier has not been studied at all. It has now been found by the present inventors that it can be a carrier for the administration of preparations by the nasal route for the first time. That is, utilizing the porosity of the porous crystallized glass, those having an average particle size of 5 μm to 2000 μm are:
It has been found that these physiologically active peptides are efficiently absorbed through the nasal mucosa when administered into the nasal cavity as a transnasal formulation by enclosing a bioactive peptide therein as a carrier. Carriers in intranasal administration preparations that have been conventionally studied are considered to be water-soluble carriers for the absorption of the active ingredient in vivo, but according to the findings of the present inventors, it is not always necessary. A water-soluble carrier is not considered to be good, and the active ingredient is uniformly dispersed, adhered and bound to a carrier that does not dissolve in water such as the porous crystallized glass which is the carrier of the present invention. It has been found that good results are obtained.

[0006] Therefore, the carrier which does not dissolve in water is not limited to the porous crystallized glass provided in the present invention, but may be any carrier which does not adversely affect the human body in intranasal administration. Besides porous crystallized glass, for example, poorly water-soluble celluloses such as crystalline cellulose, α-cellulose, crosslinked sodium carboxymethylcellulose; hydroxypropyl starch, carboxymethyl starch, crosslinked starch, amylose, amylopectin Poorly water-soluble starches such as pectin, pectin; poorly water-soluble proteins such as gelatin, casein and sodium caseinate; poorly water-soluble gums such as gum arabic, tragacanth gum and glucomannan; polyvinylpyrrolidone, cross-linked polyacrylic acid and salts thereof; Crosslinked polyvinyl alcohol, poly Crosslinked vinyl polymer hardly soluble in water, such as mud carboxyethyl methacrylate; Amberlite (Am
berlite (registered trademark), Dowex (Dowe)
x: registered trademark), Sephadex:
(Registered trademark) and the like; fine particles of various synthetic fiber compounds such as nylon and vinylon; general plastic fine particles; activated carbon and the like are also effective as carriers for intranasal administration of physiologically active peptides. That is, these carriers also have an average particle size of 5 μm.
When the composition had a size of ~ 2000 µm, a composition for nasal absorption in which a physiologically active peptide was uniformly dispersed and adhered was found to provide good bioavailability.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The porous crystallized glass as a carrier in the composition provided by the present invention is, among others, SiO ₂ : 22 to 50% by weight and P ₂ O ₅ : 8 to 30 by weight%.
%, CaO: 20 to 53%, MgO: 1 to 16%, F
_{2: 0.1~2%, Al 2 O} 3: 0~9%, B 2 O 3:
It has a composition of 0 to 5%, and is made of apatite (Ca ₁₀ (PO ₄ )
₆ O) and at least one of wollastonite (CaO.SiO ₂ ) and diopside (CaO.MgO.2SiO ₂ ) having an average pore diameter of 20 to 2000 μm.
m, the porosity was 66 to 95% by volume, and it was found that a porous crystallized glass having a three-dimensional network structure was good. Such porous crystallized glass, for example,
Artificial bone substitutes on the market under the trade name Seraborne (registered trademark) may be mentioned. Such a porous crystallized glass is finely pulverized, and the average particle diameter is 5 μm to 2000 μm.
m was found to be particularly excellent as a carrier for nasal absorption of a bioactive peptide.

On the other hand, the physiologically active peptide which is an active ingredient of the present invention which is uniformly dispersed and adhered to the above-mentioned carrier is a peptide hormone, a physiologically active protein, and an enzyme protein. Such usable bioactive peptides include, for example, peptide hormones such as paratolmon (parathyroid hormone), calcitonin, insulin,
Angiotensin, glucagon, gastrin, secretillin, growth hormone, prolactin (luteinizing hormone), gonadotropin (gonadotropin), thyotropic hormone, adrenocorticotropic hormone, melanocyte stimulating hormone, vasopressin, oxytocin, protirelin, luteinizing hormone, corticotropin , Somatropin, thyrotropin (thyroid stimulating hormone), somatostatin (growth hormone stimulating factor), hypothalamic hormone (GnRH) and its derivatives, G-CSF, erythropoietin, superoxide dismutase (SOD)
And so on. In addition, interferon, interleukin, urokinase, lysozyme, vaccine and the like can also be mentioned. These physiologically active peptides are not limited to those described above, and it goes without saying that the composition of the present invention can be used as long as it can be administered intranasally.

Among the above-mentioned physiologically active peptides, peptide hormones are particularly preferred. Among these peptide hormones, calcitonin, insulin and somatostatin are preferred, and insulin and calcitonin are particularly preferred. As calcitonin, salmon calcitonin, human calcitonin, salmon human chimeric calcitonin,
Pig calcitonin, chicken calcitonin, bovine calcitonin, ell (eel) calcitonin and the like can be mentioned, and all of these calcitonins are naturally occurring, extractable and commercially available. In these calcitonins, the stability is said to be in the order of el-calcitonin-human calcitonin-salmon calcitonin, but even salmon calcitonin, which is said to be relatively inferior in stability, is special in the present invention. By dispersing and adhering to a carrier to form the composition of the present invention, it has been found that, for example, bioavailability and effective concentration in blood are extremely good. Therefore, so-called commercially available calcitonin is the most suitable as the bioactive peptide used in the present invention.

[0010]

Accordingly, in a most preferred embodiment of the present invention, the effective dose of insulin is determined by the following carrier: wt% SiO ₂ : 22.
_{~50%, P 2 O 5:} 8~30%, CaO: 20~53
%, MgO: 1~16%, F 2: 0.1~2%, Al 2
It has a composition of O ₃ : 0 to 9% and B ₂ O ₃ : 0 to 5%, and precipitates at least one of apatite, wollastonite and diopside, and has an average pore diameter of 20 to 2000 μm.
m, a carrier which is a porous crystallized glass having a porosity of 66 to 95% by volume, a three-dimensional network structure, and an average particle size of 5 μm to 2000 μm; An object of the present invention is to provide a physiologically active peptide composition for nasal absorption.

Further, in another most preferred embodiment of the present invention, the effective dose of calcitonin is as follows: Carrier: 22-50% of SiO _{2 by} weight%, P ₂ O ₅ :
8-30%, CaO: 20-53%, MgO: 1-16
_{%, F 2: 0.1~2%,} Al 2 O 3: 0~9%, B 2
O ₃ : having a composition of 0 to 5%, and precipitating at least one of apatite, wollastonite and diopside, having an average pore diameter of 20 to 2000 μm and a porosity of 66 to 9;
5% by volume, a carrier which is a porous crystallized glass having a three-dimensional network structure and an average particle diameter of 5 μm to 2000 μm; a physiologically active peptide composition for nasal absorption, which is uniformly dispersed and adhered to a carrier. To provide things.

The amount of the physiologically active peptide contained in the composition of the present invention used as an effective dose depends on the individual active substance to be selected (for example, calcitonin, its relative activity titer, and insulin, its insulin). Insulin unit), the target disease to be treated, the desired number of doses, the individual therapeutic effect required, and the like. When the compositions of the invention are used for intranasal administration, for example, the therapeutic effect of a formulation containing the active substance can be determined in comparison to bioavailability with other known formulations. For example, taking insulin as an example, when used for diabetes, 4 to 20 insulin units are injected subcutaneously at the initial stage, and the maintenance amount is usually 4 to 100 per day.
The unit is used, and the maximum amount is 800 units per day. Therefore, when the composition according to the present invention is administered by nasal administration, 4 to 100 units of insulin may be used.

In the case of calcitonin, when the treatment with a conventional calcitonin such as salmon calcitonin is performed by intramuscular injection, about 50 to 100 MRC (I
U) Individual doses of units are applied at a rate of about 1 x day to about 3 x week. Thus, when the composition according to the invention is administered by the nasal route, from about 50 to about 400 MR.
C units, more preferably about 100 to about 200 MRC
Suitably, the unit dose will be administered from about 1 x day to about 3 x weekly for treatment. The dose is administered in a single application, ie, from about 50 to about 400 MRC.
Conveniently, the treatment is effected by administration of a single intranasal dose consisting of units, preferably about 100 to about 200 MRC units of calcitonin. Therefore, when producing the composition of the present invention, the content of the bioactive peptide,
For example, 0.005 to 30% per 100% of the formulation weight,
Preferably 0.01-20%, more preferably 0.1-
It is better to mix 5.0%. The amount of the porous crystallized glass which is a carrier constituting the composition of the present invention is, for example, 70 to 99.99 per 100% of the weight of the preparation.
It has been found that good nasal absorption can be obtained by adding 5%, preferably 80 to 99.99%, more preferably 95 to 99.9%.

The powder of the physiologically active peptide of the present invention can be obtained by mixing a porous crystallized glass as a carrier with a physiologically active peptide. For example, this mixing is performed while applying pressure or shearing force as in the case of mixing using a mortar. The average particle size of the porous crystallized glass as a carrier used in the production of the composition of the present invention is 5 μm to 2000 μm, that is, preferably 20 to 2 μm.
The thickness is preferably 50 μm, more preferably 30 to 60 μm. On the other hand, the physiologically active peptide is preferably as fine powder as possible, and its average particle size is usually 20 μm or less, preferably 10 μm or less.

In order to prepare a physiologically active peptide composition for intranasal administration using the composition of the present invention, it can be produced, for example, as follows. That is, when salmon calcitonin or el calcitonin is used as a physiologically active peptide, the effective amount of calcitonin is 1% for gelatin as a stabilizer for calcitonin, and 0.1 to 0.5% for aspartic acid, for example, preferably 0.
It is mixed with an aqueous solution having a pH of 4.5 to 5.5 containing about 38%, and this mixture is freeze-dried. Next, this freeze-dried powder was added to the carrier of the present invention having an average particle size of 5 μm to 20 μm.
A fine powder having a bioactive peptide uniformly adhered and bonded to a target carrier (intranasal) is obtained by dividing a porous crystallized glass of 00 μm into two or three times sequentially and kneading at a kneading humidity of about 55%. Administration composition). The composition for intranasal administration obtained in this way is intended to prevent the loss of the active substance before use (for example, before administration into the nasal cavity), so as to prevent low-gre.
After filling into ase type capsule, suitable packaging,
Preferably, the package is hermetically sealed. As such hermetically sealed packaging,
A good combination is blister packaging-aluminum packaging. In addition, by performing the same treatment for other physiologically active peptides (for example, insulin), the respective target compositions can be obtained.

[0016]

EXAMPLES The specific effects of the composition for nasal absorption of the present invention are shown below by test examples.

[0017]

Test Example Test Example 1: Insulin was selected as a physiologically active peptide, and finely pulverized Ceraborn (registered trademark), which is a porous crystallized glass, was used as a carrier. An intranasal composition of the present invention was prepared which had a composition of insulin of 2.4 mg / animal (5 insulin units (IU) / animal). The carrier having a particle diameter of 40 to 45 μm was used as the carrier.
The above composition was intranasally administered (single) using 6 male New Zealand rabbits per group.
The decrease in mean blood glucose at 0, 60, 120 and 180 minutes was measured. The blood sugar level at 0 hour after administration was defined as 100%, and the decrease in blood sugar level at each measurement was determined as a percentage.
As a control group, subcutaneous injection of 2 IU of insulin / animal was performed, and the decrease in blood glucose level at 0 hours, 1, 2, 4, and 6 hours after administration was measured in the same manner. Table 1 summarizes the results.

[0018]

[Table 1]

From the results in the table, it can be seen that the use of porous crystallized glass as a carrier results in excellent absorption of insulin into the blood by nasal absorption, and as a result significantly lowers blood glucose (blood sugar level). Turns out to be.

Test Example 2 Salmon calcitonin was selected as a physiologically active peptide, and Ceraborn (registered trademark), which is a porous crystallized glass, was selected as a carrier, and the composition for nasal absorption of the present invention was used. The composition for intranasal administration of the present invention was prepared so that the composition of salmon calcitonin was 200 MRC (IU) / 25 mg). The cerabone as a carrier was finely pulverized and used having an average particle diameter of 40 to 45 μm. The above composition was intranasally administered (single dose) to three normal male subjects, and blood samples were 2.5 ml each for 0 hours, 5, 10, 15, and 2
Blood was collected every 0, 30, 45, 60, 90 minutes, 2 hours, and 3 hours, and the concentration of salmon calcitonin in blood was measured using a standard RIA kit. Table 2 summarizes the results.

[0021]

[Table 2]

As is clear from the results shown in the table, the use of porous crystallized glass (Seraborn: registered trademark) as a carrier is superior in the absorption of salmon calcitonin into blood by nasal absorption. Prove.

Examples of the composition of the present invention are shown below. Powder composition 1: salmon calcitonin (L-calcitonin is produced similarly) 3 mg / 3,000-3,500 M
The RC is dissolved in an aqueous solution containing 1% gelatin and 0.38% aspartic acid as a stabilizer at pH 4.5-5.5 and freeze-dried. Then, the obtained freeze-dried powder was poured into a porous crystallized glass 20 using an agate mortar.
Mix with 0 mg for 10 minutes. To this mixture was added 300 mg of porous crystallized glass, mixed at 55% humidity for 20 minutes, and further added 497 mg of porous crystallized glass.
The mixture was mixed at 55% humidity for 30 minutes to obtain 1,000 mg of the powder composition of the present invention. As the porous crystallized glass, a finely pulverized product of cerabone having an average particle diameter of 50 μm was used.

Powder composition 2: Insulin is 200 units, and aporous crystallized glass 200 is prepared using an agate mortar.
Mix with mg for 10 minutes. To this mixture was added 300 mg of porous crystallized glass, mixed at 55% humidity for 20 minutes, and further added with 497 mg of porous crystallized glass and mixed at 55% humidity for 30 minutes to obtain a powder composition of the present invention. 1,
000 mg were obtained. The porous crystallized glass in this case is also
Cerabone is finely pulverized and has an average particle size of 50.
μm.

[0025]

As described above, according to the present invention, a physiologically active peptide which has been difficult to administer orally up to now,
Provided is a composition for a nasal absorption drug which has a good absorbability and has no irritation by nasal administration. In particular, a physiologically active peptide such as calcitonin or insulin is converted to a specific carrier having an average particle diameter of 5 μm to 2000 μm, preferably 20 μm to 250 μm, more preferably 30 μm.
The powder composition of the present invention dispersed in 結晶 60 μm porous crystallized glass can be administered by the nasal route, that is, applied to the nasal mucosa, so that its peptide-based body absorbability is good and effective clinical It suggests that treatment is possible. That is, as is clear from the results of the above-described test examples, the composition of the present invention such as calcitonin and insulin shows a bioabsorbability equal to or higher than that of conventional intravenous administration by intranasal administration, It is found that nasal absorption is effective. Therefore, its medical effects are enormous.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 38/43 A61K 37/26 47/02 37/30 37/48

Claims (8)

(57) [Claims] 1. A physiologically active peptide composition for nasal absorption wherein an effective dose of a physiologically active peptide is uniformly dispersed and adhered to a carrier which is a porous crystallized glass having an average particle diameter of 5 μm to 2000 μm. 2. The carrier, which is a porous crystallized glass,
SiO ₂ : 22 to 50% by weight, P ₂ O ₅ : 8 to 30 by weight%
%, CaO: 20 to 53%, MgO: 1 to 16%, F
_{2: 0.1~2%, Al 2 O} 3: 0~9%, B 2 O 3:
It has a composition of 0 to 5%, precipitates at least one of apatite, wollastonite and diopside, has an average pore size of 20 to 2000 μm, a porosity of 66 to 95% by volume, and has a three-dimensional network structure. The physiologically active peptide composition for nasal absorption according to claim 1, which is a pharmaceutically acceptable carrier. 3. The effective dose of the physiologically active peptide is as follows: Carrier: weight%, SiO ₂ : 22 to 50%, P ₂ O
₅ : 8 to 30%, CaO: 20 to 53%, MgO: 1 to 1
_{16%, F 2: 0.1~2%} , Al 2 O 3: 0~9%,
B ₂ O _3: have a 0-5% composition, precipitating least one apatite and wollastonite and Jiopusaido, average pore size 20～2000Myuemu, porosity 66
９５95% by volume, a carrier which is a porous crystallized glass having a three-dimensional network structure and an average particle size of 5 μm to 2,000 μm; Stuff. 4. The bioactive peptide is a peptide hormone, a bioactive protein, or an enzyme protein.
The physiologically active peptide composition for nasal absorption according to the above. 5. The physiologically active peptide composition for nasal absorption according to claim 1, wherein the peptide hormone is calcitonin or insulin. 6. The calcitonin is composed of the following carriers: SiO ₂ : 22 to 50% by weight, P ₂ O ₅ : 8 to 30% by weight,
CaO: 20~53%, MgO: 1~16 %, F 2:
_{0.1~2%, Al 2 O 3:} 0~9%, B 2 O 3: 0~
Having a composition of 5%, precipitating at least one or more of apatite, wollastonite and diopside, having an average pore diameter of 20 to 2000 μm, a porosity of 66 to 95% by volume, and a three-dimensional network structure; The average particle size is 5 μm
A bioactive peptide composition for nasal absorption, which is uniformly dispersed and adhered to a carrier which is a porous crystallized glass having a size of ２０００2000 μm. 7. The method according to claim 1, wherein the insulating carrier is SiO ₂ : 22 to 50% by weight, P ₂ O ₅ : 8 to 30% by weight,
CaO: 20~53%, MgO: 1~16 %, F 2:
_{0.1~2%, Al 2 O 3:} 0~9%, B 2 O 3: 0~
Having a composition of 5%, precipitating at least one or more of apatite, wollastonite and diopside, having an average pore diameter of 20 to 2000 μm, a porosity of 66 to 95% by volume, and a three-dimensional network structure; The average particle size is 5 μm
A bioactive peptide composition for nasal absorption, which is uniformly dispersed and adhered to a carrier which is a porous crystallized glass having a size of ２０００2000 μm. 8. A physiologically active peptide composition for nasal absorption in which an effective dose of a physiologically active peptide is uniformly dispersed and adhered to a carrier which is an ion exchange resin having an average particle diameter of 5 μm to 2000 μm.