JPH07206699A - Nasal preparation of peptide and proteinaceous agent - Google Patents

Abstract

PURPOSE:To obtain a nasal preparation of peptide and proteinaceous agent having improved absorbability. CONSTITUTION:This nasal preparation of peptide and proteinaceous agent having improved absorbability contains one or more kinds of enzyme inhibitors selected from gabexate mesilate, nafamostat mesilate and camostat mesilate in powdery form.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a nasal preparation of a peptide or protein drug having improved absorbability. Specifically, it relates to a nasal preparation of a peptide or proteinaceous drug, which comprises an enzyme inhibitor such as gabexate mesylate and has improved absorbability.

[0002]

2. Description of the Related Art With recent advances in biotechnology, peptides and proteinaceous compounds having physiological activity have been discovered 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, it is desirable that these drugs be administered frequently because of their therapeutic use, and in that case, administration by injection causes a burden on the patient due to pain to the patient, necessity of going to the hospital, etc. .

Therefore, various other administration methods have been studied as non-invasive administration methods of peptide and protein drugs instead of injection. Nasal administration has recently attracted attention as such an administration method, but at present, a sufficient absorption rate has not been achieved, and various studies have been conducted to improve the absorption rate. Degradation by enzymes in the nasal cavity is considered to be one of the main causes of such low absorption rate, and animal studies have shown that absorption rate is improved by adding various proteolytic enzyme inhibitors. There is.

For example, Morimoto et al. Of Osaka Pharmaceutical University (Japan Society of Pharmaceutical Sciences 111 and 112) have shown that the absorption rate of salmon calcitonin in nasal solutions is improved by administration of a trypsin inhibitor, aprotinin, to rats. It is shown by the animal experiment used. Also, O´Hagan et a
l. (Pharm. Res. 7,772 (1990)), Hussain
MA et al. (Pharm. Res. 6,186 (1989))
Is the absorption rate of peptides such as insulin, growth hormone, enkephalin, and protein pharmaceuticals in nasal fluid,
It has been shown by animal experiments using rats and sheep that the improvement by administration with aminopeptidase inhibitors such as amastatin and bestatin is improved.

However, these nasal solutions containing a protease inhibitor in combination have a problem that the effect of promoting the absorption of the protease inhibitor is low and, therefore, the absorption rate cannot be sufficiently improved. It was

[0006]

Means for Solving the Problems The present inventors have studied a nasal preparation of a peptide or protein drug, and as a result of studying a dosage form that significantly improves the absorption rate of the peptide or protein drug from the nasal mucosa, the results are surprising. In particular, they have found that the absorption promoting effect when gabexate mesylate is used together as a powder is significantly larger than the absorption promoting effect when it is used together as an aqueous solution, and the present invention has been accomplished.

That is, the present invention provides a peptide with improved absorbability, which comprises as powder one or more enzyme inhibitors selected from the group consisting of gabexate mesylate, nafamostat mesylate, and camostat mesylate. It is a protein drug nasal preparation.

The nasal preparation of the present invention contains one or more enzyme inhibitors selected from the group consisting of gabexate mesylate, nafamostat mesilate, and camostat mesylate as a powder.

[0009] Gabexate mesylate and nafamostat mesilate have serine protease inhibitory activity and are substances used for acute pancreatitis and general blood coagulation, and their safety to the living body has already been confirmed. However, it is chemically unstable and easily hydrolyzed, so it is used as an injectable preparation for business use. In addition, Takeyama et al. Of Kagawa Medical University
It has been shown that by applying an ointment containing these substances to the injection site during subcutaneous injection of insulin, the absorption rate of insulin is increased (Pharm. Res. 8 (1), 19).
91). On the other hand, camostat mesylate is a substance used as an oral preparation, although it has the same pharmacological effect as gabexate mesylate or nafamostat mesylate.
Morimoto et al. Of Osaka Pharmaceutical University have shown that iontophoretic transdermal absorption of salmon calcitonin is improved by the combined use of camostat mesylate and aprotinin (J.Phar
m, Pharmacol. 44 (3), 1992). However, in any case, a method for improving absorption in a nasal preparation is not known.

In the present invention, examples of the peptide or protein drug include a peptide or protein drug containing arginine or lysine in its amino acid sequence.
Calcitonins, somatostatin (GIF) and its derivatives, adrenocorticotropic hormone (ACTH) and its derivatives, parathyroid hormone (PTH), glucagon, calcitonin gene-related peptide (CGRP), endothelin, endorphin, galanin, gastrin releasing peptide ( GRP), gastrin release inhibitory peptide (GI
P), neurokinins, interleukins, insulins, insulin-like growth factor (IGF), neurotensin, platelet-derived growth factor (PDGF) s, substance P, growth hormone releasing hormone (GHRH) and its derivatives, luteinization Hormone stimulating hormone (LHRH)
And its derivatives, enkephalin and its derivatives, secretin, bradykinin and its derivatives, arginine vasopressin (AVP), atrial natriuretic peptide (ANP), interferons, erythropoietin,
The one or more peptides and proteinaceous drugs selected from the group consisting of granulocyte colony-stimulating factor (G-CSF), adrenocorticotropic hormone-releasing factor (CRF), VIPs, and angiotensins. Examples include peptides and protein drugs.

Among these, the peptides and protein pharmaceuticals of the present invention include calcitonin, somatostatin (GIF) and its derivatives, adrenocorticotropic hormone (A
CTH) and its derivatives, parathyroid hormone (PTH)
, Glucagon, calcitonin gene-related peptide (C
GRP), platelet-derived growth factor (PDGF) s, substance P, growth hormone releasing hormone (GHRH) and its derivatives, and luteinizing hormone stimulating hormone (LH).
RH) and derivatives thereof can be mentioned as preferred ones, and more preferred are calcitonins,
Mention may be made of growth hormone releasing hormone (GHRH) and its derivatives.

Among these drugs, calcitonins are peptide hormones that regulate calcium metabolism in the body, have a function of inhibiting bone resorption of calcium and reabsorbing calcium excreted by the kidney. However, salmon calcitonin, eel calcitonin, human calcitonin, pig calcitonin, chicken calcitonin, bovine calcitonin, sheep calcitonin, rat calcitonin, etc. are known, and in the present invention, any of these calcitonin and derivatives thereof are used. be able to.

Growth hormone releasing hormone (GHRH)
Is a growth hormone (G
H) is a peptide hormone, and human, porcine, rat, bovine, and ovine are known so far. In the present invention, any of these growth hormone-releasing hormones and their derivatives can be used. it can. For example, somatrelin acetate, which is an acetate of human growth hormone-releasing hormone, or a human growth hormone-releasing hormone derivative consisting of 29 amino acid residues on the N-terminal side of human growth hormone-releasing hormone can be used.

As the combination of the enzyme inhibitor of the present invention and the peptide or proteinaceous drug, gabexate mesylate, calcitonin such as salmon calcitonin as exemplified above, and human growth hormone releasing hormone are preferred. Growth hormone-releasing hormones such as and the like, and derivatives thereof are preferable, and above all, gabexate mesylate and peptides, calcitonin such as salmon calcitonin as a protein drug, growth hormone-promoting hormones such as human growth hormone-releasing hormone, and the like. And the like are preferable.

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, the amount of the enzyme inhibitor of the present invention is about 0.1 to 10 relative to the therapeutically effective amount of the peptide or proteinaceous drug.
It is preferably about 1 part by weight, particularly preferably about 1-5.
Parts by weight.

The nasal preparation of the peptide or protein drug of the present invention is a powder, and the powder is, for example, a finely powdered peptide obtained by freeze-drying, if necessary.
It is produced by mixing one or more species with the enzyme inhibitor of the present invention and, for example, 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 poorly water-soluble base is on the human nasal mucosa, or in an environment close to this,
That is, it means that it has a property of being absorbable and hardly soluble in water having a pH of about 7.4 and a temperature of about 36 to about 37 ° C.

Preferred specific examples include water-absorbent 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; gum arabic, tragacanth gum,
Water-absorbing and sparingly water-soluble gums such as glucomannan and derivatives thereof; cross-linked vinyl polymers such as polyvinyl polypyrrolidone, cross-linked polyacrylic acid and its salts, cross-linked polyvinyl alcohol, polyhydroxyethyl methacrylate and their derivatives Can be mentioned. Of 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 kinds of peptides and proteinaceous drugs, the absorption enhancer of the present invention and the above-mentioned bases, known lubricants, preservatives and preservatives as required. ,
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; preservatives such as stearic acid , Ascorbic acid stearate, etc .; as preservatives, for example, quaternary ammonium compounds such as benzalkonium chloride, paraoxybenzoic acid esters, phenol, chlorobutanol, etc .; as flavoring agents, for example, menthol, citrus flavors And so on.

The peptide or protein drug nasal preparation of the present invention is usually administered intranasally by the following administration methods. That is, for example, set a capsule filled with powder in a dedicated spray device equipped with a needle, penetrate the needle, thereby making minute holes at the top and bottom of the capsule, then send air with a rubber ball etc. There is a method to make it gush out.

Thus, the present invention provides a nasal preparation of a peptide or protein drug with improved absorbability, which is of great medical value.

[0023]

EXAMPLES The present invention will be described in detail below with reference to Examples, Comparative Examples, Control Examples, and Reference Experimental Examples, but these are provided for explaining the present invention and limit the present invention. Not a thing.

Reference Experimental Example 1 (1) A homogenate solution was prepared from a human nasal mucosa so that the amount of protein was 1 mg / ml, and 0.5 ml of this homogenate solution was used to prepare an aqueous solution of various peptides shown in Table 1 ( 0.5 mg (0.1 mg / ml) was added and incubated at 37 ° C., and then the peptide degradation product in the reaction solution was added to HP.
LC (L-6200 series (manufactured by Hitachi Ltd.), 0-5
Min: 100% H ₂ O, 5 to 55 min: 100% → 50% H
₂ O, 0-50% acetonitrile). The amino acid sequencer (Ap
The amino acid sequence was examined by plied Biosystems) to determine the decomposition site. The P ₁ site of degradation is shown in Table 1.

[Reference Experimental Example 2] A homogenate solution was prepared from human intranasal mucosa so that the protein amount was 1 mg / ml. To 0.5 ml of this homogenate solution, 250 μl of an aqueous solution of various peptides (72 mM) shown in Table 1 and an aqueous solution of gabexate mesylate (MG) of the present invention (72 M).
Mix 10 μl and incubate at 37 ° C. After a certain period of time, measure the amount of undegraded peptides and protein pharmaceuticals on HP.
The degradation inhibition rate (%) was calculated by LC. The results are also shown in Table 1.

[0026]

[Table 1]

In Table 1, the inhibition rate is (AB) / A × 100.
Where A is the ratio of the amount of degradation of various peptides after a certain period of time (one hour) in the absence of an inhibitor (in the presence of an inhibitor) to the initial amount of peptide. Represent SCT is salmon calcitonin, GIF is somatostatin, ACTH is adrenocorticotropic hormone, PTH is parathyroid hormone, and CGRP.
Is a calcitonin gene-related peptide, PDGF is a platelet-derived growth factor, S. P is substance P, GHR
H represents growth hormone releasing hormone, LHRH represents response-forming hormone stimulating hormone, and EDP represents endorphin. All of these peptides were manufactured by Bachem, and gabexate mesylate was manufactured by Wako Pure Chemical Industries.

As shown in Table 1, in the human nasal mucosa homogenate, most of the peptides were Cs of Lys or Arg.
It is easily decomposed on the end side, and such Lys or A
It was revealed that gabexate mesylate suppresses the degradation of the peptide that is degraded at the C-terminal side of rg.

[Example 1] 40 μg of salmon calcitonin,
Gabexate mesylate 100 μg, microcrystalline cellulose 3
0 mg and 15 μg of magnesium stearate were mixed in a mortar to prepare a calcitonin transnasal powder containing gabexate mesylate. Calcitonin amount 100 IU (20μ
g) and after being filled in No. 2 capsule, it was administered into the nasal cavity of Beagle dogs (male, 9.4 to 12.6 kg, 4 heads) with a pubizer (registered trademark, Teijin Ltd.). . After administration, 5 ml blood was collected from the leg vein at regular intervals,
The salmon calcitonin concentration in plasma was measured by the RIA method. The results are shown in Table 2 as bioavailability (absorption rate (%)).
Shown in.

Comparative Example 1 Gavexate 100 Mesylate
A calcitonin nasal dust formulation without gabexate mesylate was prepared in the same manner as in Example 1 except that the amount of calcitonin was 100 IU (20 μg). Similarly, after administration to beagle dogs (male, 9.4 to 12.6 kg, 4 dogs), salmon calcitonin concentration in plasma was measured after a certain period of time. The results are also shown in Table 2.

[Control Example 1] 40 μg of salmon calcitonin,
100 μg of gabexate mesylate was dissolved in 0.2 ml of distilled water to prepare a nasal calcitonin mesylate-added calcitonin preparation. Calcitonin amount 100 IU (20 μg)
Beagle dog (male, 9.4-12.
After administration to 6 kg, 4 heads), the salmon calcitonin concentration in plasma was measured after a certain period of time. The results are also shown in Table 2.

Control Example 2 A calcitonin nasal solution without gabexate mesylate was prepared in the same manner as in Control Example 1 except that gabexate mesylate was not added. Male, 9.4-12.
(6 kg, 4 animals), the salmon calcitonin concentration in plasma was measured after a certain period of time. The results are also shown in Table 2 as well.

[0033]

[Table 2]

From Table 2, when a mixture of gabexate mesylate and calcitonin was administered as a powder (Example 1)
The effect of gabexate mesylate as an absorption enhancer was remarkably improved. It was about 4.6 times that of the non-added powder (Comparative Example 1) and about 12 times that of the additive liquid agent (Control Example 1). It can be seen that the absorption rate is twice as excellent.

Example 2 Somatrain acetate (GHR)
H) 2.0 mg, gabexate mesylate 2.0 mg, microcrystalline cellulose 30 mg, magnesium stearate 1
5 μg was mixed in a mortar to prepare a GHRH transnasal powder containing gabexate mesylate. Weighed so that the amount of GHRH would be 1.0 mg, and filled in No. 2 capsule, and then beagle dog (male, male,
It was intranasally administered to 9.4 to 12.6 kg (4 animals). After administration, 5 ml of blood was collected from the forearm vein at regular intervals, and the GHRH concentration in plasma was measured by the RIA method. Table 3 shows the physiological utilization rate (absorption rate (%)).

Comparative Example 2 Gavexate Mesylate 2.0
A GHRH nasal powder without gabexate mesylate was prepared in the same manner as in Example 2 except that mg was not added.
After weighing so that the amount would be 1.0 mg and filling the No. 2 capsule, a beagle dog (male, 9.4 to 12.
(6 kg, 4 heads), GHR in plasma after a certain period of time
The H concentration was measured. The physiological utilization rate is shown in Table 3.

[Comparative Example 3] Somatrain acetate (GHR
H) 2.0 mg and gabexate mesylate 2.0 mg were dissolved in 0.2 ml of distilled water to prepare a calcitonin nasal solution containing gabexate mesylate. GHRH amount 1.0 mg
Beagle dog (male, 9.4-12.
(6 kg, 4 heads), GHR in plasma after a certain period of time
The H concentration was measured. The physiological utilization rate is also shown in Table 3.

[Comparative Example 4] Gabexate mesylate 2.0
A GHRH nasal powder without gabexate mesylate was prepared in the same manner as in Example 3 except that mg was not added.
Beagle dog (male, 9.4 to 12.6 kg, 4
After administration, the GHRH concentration in plasma was measured after a fixed time. The physiological utilization rate is also shown in Table 3.

[0039]

[Table 3]

From Table 3, gabexate mesylate and GHR
When the mixture of H was administered as a powder (Example 2), the action of gabexate mesylate as an absorption enhancer was remarkably improved, and it was about 6 times that of the non-added powder (Comparative Example 2). 1 compared to the case of the additive liquid (control example 3)
It can be seen that the absorption rate is twice as excellent.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Office reference number FI technical display location A61K 38/22 38/28 38/26 38/23 38/11 38/27 38/21 // ( A61K 38/00 31: 215) A61K 9/14 U 37/24 37/26 37/28 37/30 37/32 37/34 37/36 37/66 (A61K 37/02 31: 215) (72) Invention Kazuki Suzuki 4-3-2 Asahigaoka, Hino-shi, Tokyo Inside Teijin Limited Tokyo Research Center

Claims (7)

[Claims] 1. A peptide with improved absorbability, which comprises at least one enzyme inhibitor selected from the group consisting of gabexate mesylate, nafamostat mesilate, and camostat mesylate as a powder, and a proteinaceous drug agent. Nasal formulation. 2. The peptide or protein drug nasal preparation according to claim 1, wherein the enzyme inhibitor is gabexate mesylate. 3. Peptides and protein drugs are calcitonin, somatostatin (GIF) and its derivatives, adrenocorticotropic hormone (ACTH) and its derivatives, parathyroid hormone (PTH), glucagon, calcitonin gene-related peptide (CGRP). ), Endothelin, endorphin, galanin, gastrin-releasing peptide (GR
P), gastrin release inhibitory peptide (GIP), neurokinins, interleukins, insulins, insulin-like growth factor (IGF) s, neurotensin, platelet-derived growth factor (PDGF) s, substance P, growth hormone releasing hormone (GHRH) and its derivatives,
Luteinizing hormone stimulating hormone (LHRH) and its derivatives, enkephalin and its derivatives, secretin, bradykinin and its derivatives, arginine vasopressin (AVP), atrial natriuretic peptide (AN).
P), one or more peptides selected from the group consisting of interferons, erythropoietin, granulocyte colony-stimulating factor (G-CSF), adrenocorticotropic hormone releasing factor (CRF), VIPs, and angiotensins.
The peptide or nasal proteinaceous drug preparation according to claim 1, which is a proteinic drug. 4. Peptides and protein pharmaceuticals are calcitonin, somatostatin (GIF) and its derivatives, adrenocorticotropic hormone (ACTH) and its derivatives, parathyroid hormone (PTH), glucagon, calcitonin gene-related peptide (CGRP). ), Platelet-derived growth factor (PDGH), substance P, growth hormone releasing hormone (GHRH) and its derivatives, and luteinizing hormone stimulating hormone (LHRH) and its derivatives, The nasal preparation according to claim 1, which is a pharmaceutical product. 5. The nasal preparation according to claim 1, wherein the peptide or proteinaceous drug is calcitonin, growth hormone releasing hormone (GHRH), or a derivative thereof. 6. The method according to claim 1, wherein the peptide or proteinaceous drug is one or more calcitonins selected from salmon, eel, human, pig, chicken, bovine, sheep, rat calcitonin and derivatives thereof. Nasal preparations of the described peptides and proteinaceous drugs. 7. A peptide, proteinaceous drug is used as a growth hormone-releasing hormone (GH) of human, pig, rat and sheep.
RH) and derivatives thereof, the nasal peptide or protein drug preparation according to claim 1 or 2.