JPH09315953A - Physioligically active peptide composition for nasal absorption - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a physiologically active peptide composition for nasal absorption, excellent in the biological absorbability of the physiologically active peptide used as an active ingredient contained in the composition on an intranasal administration (nasal administration). SOLUTION: This physiologically active peptide composition for nasal absorption is prepared by homogeneously dispersing and adhering an active dose of a physiologically active peptide such as a peptide hormone, a physiologically active peptide or an enzymatic protein to a carrier comprising finely ground tourmaline having an average particle diameter of <=250μm. Insulin, calcitonin, a thyroid hormone or a human growth hormone is preferably used as the physiologically active peptide. The active dose of the physiologically active peptide is homogeneously dispersed and adhered to the finely ground tourmaline to prepare the physiological peptide composition for nasal absorption.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a physiologically active peptide composition for nasal absorption, and more particularly, to intranasal administration (nasal administration) of physiologically active peptides such as peptide hormones, physiologically active proteins and enzyme proteins. The present invention relates to a bioactive peptide composition for nasal absorption, which has excellent pharmaceutical stability and excellent absorbability in the body.

[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. Furthermore, as an intranasal administration method, development of an intranasal spray as an intranasal liquid preparation has been examined, and a formulation in which a surfactant as an absorption enhancer is added to calcitonin is also 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 Problems to be Solved by the Inventor
Studies were conducted to develop a nasal formulation that is well absorbed and is not irritating. 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 present inventor can obtain bioavailability equivalent to or higher than that obtained by standard injection administration by the technique of dispersing and adhering bioactive peptides such as calcitonin and insulin in a special carrier proposed by the present invention. After finding out that, the present invention has been completed.

[0004]

Therefore, the present invention provides a nasal absorption physiology in which an effective dose of a physiologically active peptide is evenly dispersed and adhered to a carrier that is tourmaline finely pulverized to an average particle size of 250 μm or less. In providing an active peptide composition.

Tourmaline as a carrier for uniformly dispersing and adhering a physiologically active peptide in the present invention is also called tourmaline, and is a silicate compound having a complicated composition, and its chemical composition is generally as follows. Formula: Na (Mg,
Fe, Li, Al) ₃ Al ₆ Si ₆ O ₁₈ (BO ₃ ) ₃
It is represented by (OH, F) ₄ and is characterized by containing boron. Its production areas are widely distributed all over the world, and Brazil, the former Soviet Union Urals, Yunnan Province of China, Namibia, Zimbabwe, Tanzania, Kenya, Mozambique, Madakaskal, Sri Lanka are the main production areas. In Japan, it is also produced in Ishikawa in Fukushima Prefecture, Kurodaira in Yamanashi Prefecture, Hirose Mine in Tottori Prefecture, etc. Because it has pyroelectricity that is electrically charged when heated, cell activation, corrosion prevention, water quality adjustment, cleaning , Air purification,
It is also used for purposes such as sterilization. As described above, tourmaline has been used for various purposes, but its application as a carrier for nasal preparations has never been examined. 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, tourmaline is finely pulverized to obtain a nasal formulation by encapsulating a physiologically active peptide therein as a carrier by utilizing the porosity of the finely pulverized material having an average particle size of 250 μm or less. It was found that these bioactive peptides were efficiently absorbed from the nasal mucosa when administered to. Carriers in the intranasal preparations conventionally studied are
Although it has been considered that a water-soluble carrier is considered to be good for the bioavailability of the active ingredient, according to the knowledge of the present inventor, the water-soluble carrier is not always recognized as being good, The active ingredient is evenly dispersed in a carrier that does not dissolve in water, such as tourmaline, which is the carrier of the present invention.
It was found that good results were obtained even when they were adhered and administered intranasally.

[0006]

On the other hand, as the physiologically active peptide which is the active ingredient of the present invention, which is uniformly dispersed and adhered to the above carrier,
They are peptide hormones, bioactive proteins, and enzyme proteins. Such usable bioactive peptides include, for example, peptide hormones such as parathyroid hormone (P
TH), calcitonin, insulin, angiotensin, glucagon, gastrin, secretiline, growth hormone (GH), human growth hormone (HGH), prolactin (luteinizing hormone), gonadotropin (gonadotropin), thyrotropin, adrenocorticotropic hormone. , Melanocyte stimulating hormone, vasopressin,
Oxytocin, protilerin, luteinizing hormone-releasing hormone, corticotropin, somatropin, thyrotropin (thyroid stimulating hormone), somatostatin (growth hormone stimulating factor), hypothalamic hormone (GnRH), G-
CSF, erythropoietin, superoxide dismutase (SOD) and the like. Also, interferon,
Interleukins, urokinase, lysozyme, vaccines 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 preferable, and among these peptide hormones, calcitonin, insulin and somatostatin are preferable, and insulin and calcitonin are particularly preferable. 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.

Therefore, the most preferred embodiment of the present invention is a peptide hormone selected from calcitonin, insulin, parathyroid hormone (PTH), human growth hormone (HGH), hypothalamic hormone (GnRH) or its derivatives. Effective dose is average particle size 2
It is an object of the present invention to provide a composition for nasal absorption uniformly dispersed and adhered to a carrier which is tourmaline finely pulverized to 50 μm or less.

The amount of the physiologically active peptide contained in the composition of the present invention to be used as an effective dose depends on the individual active substance to be selected (for example, in the case of calcitonin, its relative activity titer, in the case of insulin). (Insulin unit), target disease to be treated, desired number of administrations, 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 nasally, 4 to 100 units of insulin may be used.

In the case of calcitonin, when the conventional treatment with calcitonin, such as salmon calcitonin, is performed by intramuscular injection, about 50 to 100 MRC (I
U) Unit individual doses are applied at a rate of about once x day to about 3 times 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 is administered at a rate of about once x day to about 3 times x week. The above doses are administered in one application, ie, about 50 to about 400 MRC.
Conveniently, the treatment is effected by the 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 tourmaline having an average particle diameter of 250 μm or less, which is a carrier constituting the composition of the present invention, is, for example, 70 to 99.995%, preferably 80 to 99.99 per 100% by weight of the preparation.
%, More preferably 95 to 99.9%, it was found that good nasal absorption can be obtained.

The physiologically active peptide powder composition of the present invention can be obtained by mixing the carrier finely pulverized tourmaline having an average particle size of 250 μm or less with the 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 tourmaline used as a carrier in the production of the composition of the present invention has an average particle size of 250 μm or less, that is, 20.
˜250 μm, 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.

The bioactive peptide composition for intranasal administration using the composition of the present invention 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 lyophilized powder is kneaded with tourmaline finely pulverized to an average particle size of 250 μm or less, which is the carrier of the present invention, in order of about 2 to 3 times and kneaded at a kneading humidity of about 55% to obtain the purpose. A fine powder (intranasally administered composition) in which the physiologically active peptide is uniformly adhered and bound to the carrier is obtained. The composition for intranasal administration thus obtained is filled in a capsule of gelatin or hydroxypropylmethylcellulose (HPMC) type in order to prevent the loss of the active substance before use (for example, before intranasal administration), and Appropriate packaging, preferably closed packaging. Such closed packaging includes blister packaging-
Good combination with aluminum packaging. In addition, by performing the same treatment for other physiologically active peptides (for example, insulin), the respective target compositions can be obtained.

The specific effects of the composition for nasal absorption of the present invention will be shown below in test examples.

[Test example]

Test Example 1: Insulin was selected as the physiologically active peptide, tourmaline was finely pulverized as a carrier, and the one having a particle size of 40 to 45 μm was used. The insulin content is 2.4 mg / animal (5 insulin units (I
U) / animal) composition of the present invention was prepared. The above composition was nasally administered (single dose) using 6 male New Zealand rabbits per group, and the decrease in mean blood glucose level was measured at 0, 15, 30, 60, 120 and 180 minutes after the administration. 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, 2 IU of insulin / animal dose was subcutaneously injected, and 0, 1, 2, 4, 6 hours after administration.
The decrease in blood glucose level after time was similarly measured. Table 1 shows the results.
Summarized in

[0014]

[Table 1]

From the results in the table, the use of tourmaline as a carrier is excellent in insulin absorbability into blood due to nasal absorption, resulting in a significant decrease in blood glucose (blood glucose level). Prove.

Test Example 2: Salmon calcitonin was selected as the physiologically active peptide, and the composition for nasal absorption of the present invention was used. 200 MRC as salmon calcitonin content
The composition for intranasal administration of the present invention was prepared so as to have a composition of (IU) / 25 mg). In addition, tourmaline as a carrier is pulverized and its average particle size is 40 to 45.
μm was used. The above composition was administered intranasally (single dose) to two normal males, and blood data was 2.5 m.
l, 0 hours, 5, 10, 15, 20, 30, 45,
Blood was collected every 60, 90, 2 hours, 3 hours, 4 hours, 5 hours and 6 hours, and the blood concentration of salmon calcitonin was measured using a standard RIA kit. The results are summarized in Table 2.

[0017]

[Table 2]

As is clear from the results in the table, it is found that the use of tourmaline as a carrier is excellent in the absorption of salmon calcitonin into the blood by nasal absorption.

Test Example 3: PTH (1-34), which is a parathyroid hormone, was selected as the physiologically active peptide and the composition for nasal absorption of the present invention was used. The intranasal composition for administration of the present invention was prepared so that the content of PTH (1-34) was 60 μg / 30 mg. In addition, tourmaline as a carrier is pulverized and its average particle size is 4
The thing of 0-45 micrometers was used. The above composition was administered intranasally (single dose) to two normal males, and 2.5 ml of blood samples were given each for 0 hours, 5, 10, 15, 20, 3
Blood was collected every 0, 45, 60, 90, 2 hours, 3 hours, and 4 hours, and P was collected using the ELISA1-34PTH2 antibody method.
The blood concentration of TH (1-34) was measured. The average blood concentrations obtained as a result are summarized in Table 3.

[0020]

[Table 3]

As is clear from the results in the table, the use of the finely ground product of tourmaline as a carrier proves that the absorption of PTH (1-34) into blood is excellent by nasal absorption. .

The following is an example of the composition of the present invention. 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 product is mixed with 200 mg of tourmaline for 10 minutes using an agate mortar. 300 mg of tourmaline in this mixture
Was added and mixed at a humidity of 55% for 20 minutes, and further 497 mg of tourmaline was added and mixed at a humidity of 55% for 30 minutes to obtain 1,000 mg of the powder composition of the present invention. The tourmaline used was a finely pulverized product having an average particle size of 50 μm.

Powder composition 2: 200 units of insulin, 200 mg of tourmaline and 1 using an agate mortar
Mix for 0 minutes. Add 300m of tourmaline to this mixture
g was added and mixed for 20 minutes at a humidity of 55%, and further 497 mg of tourmaline was added and mixed for 30 minutes at a humidity of 55% to obtain 1,000 mg of the powder composition of the present invention. The tourmaline in this case is also finely pulverized, and its average particle diameter is 50 μm.

[0024]

INDUSTRIAL APPLICABILITY As described above, according to the present invention, regarding the physiologically active peptide which has been difficult to be orally administered until now,
Provided is a composition for a nasal absorption drug which has a good absorbability and has no irritation by nasal administration. Particularly, the powder composition of the present invention in which a physiologically active peptide such as calcitonin and insulin is dispersed in tourmaline which is a specific carrier and has an average particle size of 250 μm or less is administered by the nasal route, that is, nasal mucosa. Therefore, it is suggested that its peptidic absorption in the body is good and effective clinical treatment is possible. Therefore, its medical effects are enormous.

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Claims (4)

[Claims] 1. A bioactive peptide composition for nasal absorption, wherein an effective dose of a bioactive peptide is uniformly dispersed and adhered to tourmaline pulverized to an average particle size of 250 μm or less. 2. The physiologically active peptide is a peptide hormone, a physiologically active protein, or an enzyme protein.
The physiologically active peptide composition for nasal absorption according to the above. 3. The physiologically active peptide for nasal absorption according to claim 1, wherein the peptide hormone is calcitonin, insulin, parathyroid hormone (PTH), human growth hormone (HGH), hypothalamic hormone (GnRH) or a derivative thereof. Composition. 4. An effective dose of a peptide hormone selected from calcitonin, insulin, parathyroid hormone (PTH), human growth hormone (HGH), hypothalamic hormone (GnRH) or a derivative thereof, and an average particle size of 2
A composition for nasal absorption which is uniformly dispersed and adhered to a carrier that is tourmaline finely pulverized to 50 μm or less.