JPH07267880A - Sustained release suppressive agent for cerebral vasospasm - Google Patents

Abstract

PURPOSE:To obtain a sustained release suppressive agent for cerebral vasospasm, containing a suppressive substance for the cerebral vasospasm on a sustained release carrier, usable by the intracephalic embedding, capable of manifesting effects on a morbid state in which therapeutic effects cannot be expected by an intravenous administration, a drainage methods, etc., and excellent in persistence of pharmacodynamic effects. CONSTITUTION:This sustained release suppressive agent for cerebral vasospasm usable by the intracephalic embedding is obtained by supporting (B) a suppressive agent for the cerebral vasospasm such as a calcitonin-related peptide or maxadilan on (A) a sustained release carrier containing a water-soluble polymer such as hydroxypropyl cellulose, fats and oils such as hardened oils or waxes and a fatty acid such as stearic acid. Furthermore, the suppressive agent is preferably a tablet type for arranging in the subarachnoid space and/or sulci on the cerebral surface.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sustained release cerebral vasospasm inhibitor comprising a sustained release carrier carrying a cerebral vasospasm inhibitor. Specifically, by implanting this preparation in the brain, a brain-embedded sustained-release cerebral blood vessel that can show effects on disease states for which therapeutic effects cannot be expected by intravenous administration, drainage method, etc. It relates to a spasm suppressant.

[0002]

2. Description of the Related Art Sustained-release preparations for subcutaneous administration, oral administration, and injection administration are well known when a drug is administered in vivo, but sustained-release preparations for implantation in the brain for cerebral vasospasm have been heretofore known. It was unknown.

As a conventional treatment method for cerebral vasospasm, a method of removing adhering blood cleanly at the time of craniotomy for subarachnoid hemorrhage, a tube such as drainage is attached at the time of this operation, and this tube is postoperatively used. There was a method of flushing the blood remaining in the brain by flushing physiological saline or the like, a method of administering a calcium antagonist through this tube, and a method of intravenously administering the calcium antagonist. However, none of them are sufficiently effective, and at present, no method for obtaining a reliable therapeutic effect has been developed.

[0004]

During administration of the drug, since the cerebrospinal fluid circulates in the special structure of the brain, that is, the subarachnoid space and the groove of the brain surface, the drug is dispersed in the brain. As shown in the property of being easy, the environment is significantly different from subcutaneous, intestinal, blood vessels, etc., so it is necessary to keep in mind that the drug is not rapidly dispersed throughout the brain. Further, cerebral vasospasm occurs on average 3 to 7 days after the occurrence of subarachnoid hemorrhage. Therefore, by maintaining the dosage form, it is necessary to reduce the waste of the drug, devise the formulation so that the drug acts on the local area for a long time and does not exert unnecessary effects on other parts. . Moreover, there is a demand for a formulation which has bioabsorbability so that it does not need to be removed after release, and does not cause changes such as exfoliation in cells in contact with tablets as much as possible.

[0005] In such a situation, although there are conventionally preparations that release the drug immediately after administration, preparations in which the release timing and the release period of the drug can be adjusted, for example, drug release after one day or more after administration Until now, there has been no preparation which has been started and has sustained drug release over a long period of 10 days or more.

[0006]

Therefore, as a result of intensive studies by the present inventors, they found that the above problems can be solved by supporting a cerebral vasospasm-suppressing substance on a sustained-release carrier, and completed the present invention. I have arrived.

That is, according to the present invention, 1) a sustained release cerebral vasospasm inhibitor for implanting in the brain, characterized in that 1) a sustained release carrier carrying a cerebral vasospasm inhibitor, 2 3. The sustained release cerebral vasospasm inhibitor according to 1, which is a calcitonin gene-related peptide, 3. The sustained release cerebral vasospasm inhibitor according to 1, wherein the cerebral vasospasm inhibitor is maxadilan, 4. The cerebral vasospasm inhibitor is GSIL. Modified sustained release cerebral vasospasm inhibitor according to 1, which is a modified maxadilan, 5 A tablet according to any one of 1-4, which is a tablet type for placement in the subarachnoid space and / or groove of the brain surface 6. Slow-release cerebral vasospasm inhibitor, 6. Slow-release cerebral vasospasm inhibitor according to any one of 1-5, which has a cerebral vasospasm inhibitor sustained-release ability in the cerebrospinal fluid for 2 to 14 days. Agent, 7 Holds the dosage form for 7 days or longer in cerebrospinal fluid To 1-6
7. The sustained-release cerebral vasospasm inhibitor according to any one of 8), 8 which disappears rapidly after the release of the cerebral vasospasm inhibitor 1-7
9. The sustained release cerebral vasospasm inhibitor according to any one of 9 to 9, 9 any one of 5 to 8 wherein the tablet has a thickness of 5 mm or less.
Item 10. Sustained release cerebral vasospasm inhibitor according to any one of the above items, 10) The sustained release cerebral vasospasm inhibitor according to any one of 5 to 9, wherein the swelling ratio of the tablet in the cerebrospinal fluid is 200% or less. 11. The release of the cerebral vasospasm inhibiting substance in the cerebrospinal fluid is 1/5 or less of the total content in the sustained-release preparation on the first day after administration, according to any one of 1-10. 12. Slow-release cerebral vasospasm inhibitor, 12. The sustained-release cerebral vasospasm according to any one of 1 to 11, wherein the sustained-release carrier is a carrier containing a water-soluble polymer, oil or wax, and fatty acid. Inhibitor, 13 Sustained-release cerebral vasospasm inhibitor comprising 13 saccharides, 14 Sustained release according to any one of 12 to 13 wherein the water-soluble polymer having a cellulose skeleton is hydroxypropyl cellulose Cerebral vasospasm inhibitor, 15 In any one of 12-14, wherein the fat and oil is hardened oil Placing a slow release Kano vasospasm inhibitors, slow release Kano vasospasm inhibitor according to any one of 12-14 16 fatty acid is stearic acid, 17 saccharide is lactose and / or glucose 1
The sustained release cerebral vasospasm inhibitor according to any one of 3-16, 18 The sustained release carrier according to any one of 1-11, wherein the sustained release carrier is a water-soluble polymer having a cellulose skeleton and a saccharide. 20. The sustained-release cerebral vasospasm inhibitor, 19. The sustained-release cerebral vasospasm inhibitor according to 18, wherein the water-soluble polymer having a cellulose skeleton is hydroxypropyl cellulose, 20. The saccharide is lactose and / or glucose.
21. The sustained release cerebral vasospasm inhibitor according to 8; 21. The sustained release cerebral vasospasm inhibitor according to any one of 1-11, wherein the sustained release carrier is a carrier containing crystalline cellulose and Eudragit. 22 Fatty acid 21. A sustained-release cerebral vasospasm inhibitor according to 21, wherein waxes and / or oils and fats are added, 23. A sustained-release cerebral vasospasm inhibitor according to 22, wherein the fats and oils are hardened oils, 24. 25. The sustained release cerebral vasospasm inhibitor according to 22. 25. The sustained release carrier according to any one of 1-11, wherein the sustained release carrier is a carrier containing a polyion complex of hyaluronic acid and a cationic polyacrylic acid derivative. A sustained release cerebral vasospasm inhibitor is provided.

The water-soluble polymer having a cellulose skeleton used in the present invention includes hydroxypropyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose and the like. Of these, hydroxypropyl cellulose is preferred. The amount thereof is not limited and is determined in consideration of the release start time and the release duration, but generally 10% by weight-90% by weight, preferably 40% by weight-based on the total weight of the preparation. 60% by weight
Is.

The fatty acids include stearic acid, lauric acid, myristic acid, isostearic acid, palmitic acid,
A saturated or unsaturated carboxylic acid having 12 to 22 carbon atoms such as behenic acid, linoleic acid, linolenic acid and oleic acid is used. Of these, stearic acid is preferred. The compounding amount is not limited, and is also determined in consideration of the release start time and the release duration time, and is generally 1% by weight to 30% by weight, preferably 1% by weight based on the total weight of the preparation.
It is 0% to 20% by weight.

As sugars, sucrose, lactose, glucose, fructose, maltose, dextrin, trehalose, pullulan and the like are used. Of these, lactose and glucose are preferable. The sustained release rate can be adjusted by the addition of sugars, and the release rate will be accelerated if a large amount is added. The content of the saccharide is not limited and is determined in consideration of the release start time and the release duration, and is generally 1% by weight-70% by weight, preferably 10% by weight-based on the total weight of the preparation.
It is 50% by weight.

The blending amount of the crystalline cellulose used in the present invention is not limited and is determined in consideration of the release initiation time and the release duration, but generally 10% by weight-90% based on the total weight of the preparation. % By weight, preferably 40% -80% by weight.

Eudragit used in the present invention is a kind of ester derivative of polyacrylic acid.
All Eudragits of this type can be used. By adding Eudragit, it is possible to prevent the excessive adsorption of the cerebral vasospasm inhibitor to the crystalline cellulose and to release it efficiently and slowly. The amount of Eudragit used is not limited as long as it is an amount for achieving the object of the present invention, but is generally 0.
01% by weight-10% by weight, preferably 0.1% by weight-5
% By weight.

As the oil and fat, hardened oil, cacao butter, beef tallow, lard and the like are used. Of these, hydrogenated oils are preferred.
And the blending amount thereof is not limited, and is also determined in consideration of the release start time and the release duration time, and is generally 1% by weight to 30% by weight, preferably 10% by weight, based on the total weight of the preparation. -25% by weight.

As waxes, beeswax, carnauba wax, white wax and the like are used. And the blending amount thereof is not limited, and is also determined in consideration of the release start time and the release duration time, and is generally 1% by weight to 30% by weight, preferably 10% by weight, based on the total weight of the preparation. -25% by weight
Is.

Hyaluronic acid is a kind of sugar acid, but it can be used as a natural product or a product obtained by genetic engineering.

Specific examples of the cationic polyacrylic acid derivative include Eudragit E and Eudragit RS manufactured by Reem, Germany, with Eudragit E being particularly preferred. In the present invention, these cationic polyacrylic acid derivatives may be used alone or in combination of two or more.

The polyion complex of hyaluronic acid and the cationic polyacrylic acid derivative used here is a novel compound and can be obtained by reacting hyaluronic acid and the cationic polyacrylic acid derivative under aqueous conditions.

When the cerebral vasospasm inhibiting substance and the polyion complex of hyaluronic acid and a cationic polyacrylic acid derivative are mixed, the preparation of the present invention can be obtained. However, each component is sufficiently ground and a powder having a small particle size is obtained. It is preferable to arrange them. Specifically, the particle size is 500 microns or less, preferably between 300 microns and 10 microns, and more preferably 200 microns or less. A usual crusher is used for crushing.

Other sustained-release bases include copolymers of lactic acid and glycolic acid (PLGA), collagen and / or gelatin, Eudragit (Eudragit E, L, S,
RS, etc.), Eudragit (preferably a mixture of Eudragit L and RS) and silicates (preferably magnesium aluminometasilicate, synthetic hydrotalcite, etc.)
Etc. can be used, and tablets can be formed in the same manner as other examples.

Examples of cerebral vasospasm inhibitors include calcitonin gene-related peptide, maxadilan, deferoxamine, methylprednisolone, nicorandil, nicarabene, magnesium sulfate, actinomycin D, 21-aminosteroid, isoproterenol, tPA, nimodipine, hydrocortisone, nicardipine. , Nifedipine, diltiazem, dilazep, teprotide, AA86
1, papaverine, OKY1581, Amyl nitrite, erythrityl tetranitrate (Er
ythrityl tetranitrate), isosorbide dinitrate, nitroglycerin, pentaerythritol tetronitrate (Pentaerythritol te)
tronitrate), VIP, vasopressin, bradkin,
PACAP, SOD, CATALASE, Bepridil, Nadololol, Felodipine, Isradipine, Verapamil, Atenolol,
Metoprolol, Propranolol (Pr
opranolol) or the like is used.

The blending amount varies depending on the drug used, but is generally 1 × 10 ⁻¹² wt% −5 based on the total weight of the preparation.
% By weight.

As Maxadilan in the above exemplified compounds, EA. Lerner et al Internation
Examples thereof include the vasodilator protein described in al Patent Publication No. WO 91/00293, its analogs, or active fragments. In the present invention, this is called natural type maxadilan.

As another maxadilan, a modified maxadilan can be mentioned. This modified type of maxadilan is a fruit fly (Lutzomyia longipalpi).
s) a protein and / or peptide containing the amino acid sequence of the active site of a protein having a vasodilatory action derived from the salivary gland, and is, for example, EA. Lerner et al.
J. Biol. Chem., 267: 1063 (1992). GIL modified maxadilan.

In addition, in the present invention, in addition to the GIL-modified maxadilan, a modified maxadilan having another amino acid sequence at the N-terminal of a natural-type maxadilan,
For example, M. U. of Tajima et al. S. P. Application No.0
Mention may be made of the vasodilatory proteins described in 8/102757, their analogues or active fragments. Among them, those having an amino acid sequence represented by the following structural formula are particularly preferable.

[0025]

[Chemical 1]

The modified maxadilan represented by the above structural formula is Gly-Ser-Ile-Leu (GS-I-
The peptide having the amino acid sequence of L) is fused to the N-terminus of natural maxadilan. Such a modified maxadilan is referred to as a GSIL modified maxadilan in the present invention. The amino acid sequence of the N-terminal of native type maxadilan is Cys-Asp-Ala-Thr (CDA-
T), the N-terminal amino acid sequence of GSIL-modified maxadilan is Gly-Ser-Ile-Leu-
Cys-Asp-Ala-Thr (G-S-I-L-C
-D-A-T).

It has been confirmed that such GSIL-modified maxadilan has higher biological activity than that of natural-type maxadilan, and the biological activity of the modified maxadilan in the skin erythema reaction is at least 10 times that of natural-type maxadilan. Has been done.

The modified maxadilan used in the present invention can be chemically synthesized, but can be obtained by applying a gene recombination technique. In the method, the desired modified maxadilan may be produced by direct gene recombination, but a modified maxadilan fusion in which a certain peptide at the N-terminal of the modified maxadilan, for example, a peptide having a thrombin cleavage site is fused The desired modified maxadilan can be obtained by obtaining a protein and then cleaving it with thrombin.

For formulation, known formulation techniques can be applied. Then, it may be formulated as it is, but conventionally known disintegration regulators, stabilizers, antioxidants, wetting agents, binders,
Add a lubricant, etc. and formulate into tablets, pills, capsules, etc.

The sustained release cerebral vasospasm inhibitor containing the cerebral vasospasm inhibitor thus obtained is not only an epoch-making drug which can be used by implanting it in the brain, but also for 7 days or more,
In most cases, the drug is sustained for more than 10 days while maintaining the dosage form, so the drug is distributed only in the administered part and does not disperse throughout the brain. It has the feature that it does not exert unnecessary action. Furthermore, without damaging the cells at the administration site,
It has an ideal property that the drug is stably released over a long period of 2 days to 2 weeks after administration, and thereafter the base is absorbed by the living body. When administered as a tablet, the subarachnoid space and / or
Alternatively, it is desirable to place it in a groove on the surface of the brain. Further, in order to be embedded in the brain without trouble, the thickness is preferably 5 mm or less, and if the expansion rate of the tablet is too large, it may cause damage such as cell detachment to surrounding cells and tissues after administration. Therefore, the expansion rate is preferably 200% or less.

Since this preparation does not sufficiently release the drug on the first day of administration, if it is necessary to release the drug during that period, it is advisable to use another formulation effective for a short period of time.

[0032]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto, and modifications and improvements made by those skilled in the art are also within the scope of the present invention. It must be understood that it is included.

[Example 1] Production of sustained-release preparation (I) Stearic acid (10 g) and hydrogenated oil (10 g) were mixed with 0.4% calcitonin gene-related peptide (CGRP).
2.0 g of solution (equal to 8 mg of CGRP) and lactose 2
0 g is added and mixed, to which 60 g hydroxypropyl cellulose is added. After thoroughly mixing this, flat tablets having a diameter of 13 mm were manufactured by a KBr tablet molding machine (150 kg, 1 minute).

[Example 2] Production of sustained-release preparation (II) 20 g of stearic acid and 20 g of hydrogenated oil were mixed, and 0.4% calcitonin gene-related peptide (CGRP) was added to the mixture.
2.5 g of the solution (equivalent to 10 mg of CGRP) and 20 g of lactose are added and mixed, and 40 g of hydroxypropyl cellulose is added thereto. After mixing this thoroughly, Correct 19K tablet molding machine (KIKUSUICLEAN PRES
It was pressure molded with S) (6 mmΦ × 2 mm).

[Example 3] Production of sustained-release preparation (II
I) 15 g of palmitic acid and 15 g of beeswax were mixed, and 0.4% calcitonin gene-related peptide (CGRP) was added to the mixture.
2.0 g of the solution (equal to 8 mg of CGRP) was added and mixed, to which 70 g of hydroxypropyl cellulose was added. After mixing this well, Correct 19
It was pressure molded with a K tablet molding machine (KIKUSUI CLEAN PRESS) (6 mmΦ x 2 mm).

[Example 4] Production of sustained-release preparation (IV) Stearic acid (10 g) and hydrogenated oil (10 g) were mixed, and 0.4% calcitonin gene-related peptide (CGRP) was added.
2.0 g of solution (equal to 8 mg of CGRP) and glucose 2
0 g is added and mixed, to which 60 g hydroxypropyl cellulose is added. After mixing well with this, Correct 19K tablet molding machine (KIKUSUI CLEAN PRESS)
(6 mmΦ x 2 mm).

Example 5 Drug Release Test (I) 5 ml of Hartmann's solution (Midori Cross) is aseptically placed in a 15 ml tube. Thereafter, the tablets obtained in Example 1 are similarly aseptically put and shaken at 37 ° C. and 120 rpm. The drug release amount was determined by sampling on days 1, 2, 3, 7, 11, and 14 and quantifying by high performance liquid chromatography. The result is shown in FIG.

The numbers in the figure indicate the disintegrated state of the tablets, and their meanings are as follows.

Collapsed state 1: No change.

Swelling of 2: 20% is seen.

3: There are small cracks.

4: There is a large crack.

5: Shattered.

Example 6 Drug Release Test (II) The formulation obtained in Example 2 was tested in the same manner, and the results are shown in FIG.
Shown in. The numbers in the figure have the same meanings as above.

Example 7 Drug Release Test (III) The formulation obtained in Example 4 was tested in the same manner, and the results are shown in FIG.
Shown in. The numbers in the figure have the same meanings as above.

The conditions of high performance liquid chromatography are as follows. Hereinafter, the same conditions are used unless otherwise specified.

Detector: UV 214 nm Flow rate: 1.5 ml / min Mobile phase: A: 0.1% TFA B: 0.085% TFA / acetonitrile At an initial concentration of 20% of B, linearly up to 60% in 20 minutes. Change the concentration.

Column: Shiseido CAPCELL PA
K SG300 (6 mm × 35 mm) Device: Shimadzu LC-6AD [Example 8] Production of sustained-release preparation (V) Stearic acid 10 g and hydrogenated oil 10 g were mixed, and 1.6% calcitonin gene-related peptide ( CGRP)
2.5 g of the solution (corresponding to 40 mg of CGRP) and 20 g of lactose were added and mixed, and 60 g of hydroxypropyl cellulose was added thereto. After mixing this thoroughly, Correct 19K tablet molding machine (KIKUSUICLEAN PRES
It was pressure molded with S) (6 mmΦ × 2 mm).

Example 9 Production of sustained-release preparation (VI) Stearic acid (10 g) and hydrogenated oil (10 g) were mixed and mixed with 1 g of the mixture.
2.5 g of 0% calcitonin gene-related peptide (CGRP) solution (equivalent to 250 mg of CGRP) and lactose 2
0 g is added and mixed, to which 60 g hydroxypropyl cellulose is added. After mixing this thoroughly, Correct 19K tablet molding machine (KIKUSUICLEAN PRESS)
(6 mmΦ x 2 mm).

Example 10 Drug Release Test (IV) Using the tablets obtained in Example 2, an in vivo test for the drug release effect of 3 rabbits was conducted by the method described below. The result is shown in FIG.

[Test method] <Method of embedding tablet in brain> A rabbit (2.5-3.0 kg) after pentobarbital anesthesia was fixed on the prone position,
The occipital periosteum (dura) was exposed by incision, the occipital bone was drilled, and the dura was exposed more widely. Thereafter, the dura and arachnoid membrane were incised by about 8 mm, and 2 out of 3 animals were embedded with the tablet of the sustained release preparation of Example 2 and 1 was embedded with a placebo tablet.
The dura, muscle and skin were sutured, and an appropriate amount of antibiotic was administered to the incision site.

Cerebrospinal fluid was collected daily from the rabbit by the following method, and CGRP in the cerebrospinal fluid was measured by the method described below.
The concentration (nM) was measured.

<Cerebrospinal Fluid Sampling Method> The above rabbits after pentobarbital anesthesia were fixed on the prone position, the occipital periosteum (dural membrane) was exposed by incision, and the occipital periosteum was incised, and the cerebrospinal fluid was collected therefrom.

<Method for measuring the concentration of physiologically active substance in cerebrospinal fluid> The concentration was measured by the following radioimmunoassay method.

The labeled compound [2- (
¹²⁵ I-iodohistidyl ¹⁰ ) CGRP] 4000 cpm,
Separately, synthetic CGRP (manufactured by Bachem) was used as a standard substance for 1, 2, 5, 10, 50, 100, 500, 10
100 μl of a standard solution of 00 fmol was prepared. In a tube containing 100 μl of test sample, standard solution and water, 100 μl of antibody (Amersham RPN1841 dissolved in 2 ml and further diluted to 12.5 ml)
And analysis buffer [50 mM sodium phosphate (pH
7.4), 0.3% bovine serum albumin, 10 mM EDT
A] Insert 600 μl, close the tube cover, and 4 ℃
It was left still for 5 days. Add dextran / activated carbon solution [50 mM sodium phosphate (pH 7.4), 0.2 to the tube.
5% gelatin, 10 mM EDTA] 250 μl was added,
Centrifugation was performed immediately at 2000 × g for 20 minutes. Measure both the precipitate and the supernatant with a γ-counter for 200 seconds,
The concentration of the physiologically active substance (CGRP) in cerebrospinal fluid was measured from the standard curve obtained from the standard substance solution.

Further, using the tablets obtained in Example 2, an in vivo study on the disintegration state of tablets by rabbits other than the above
I ran the test. The method of embedding the tablet in the brain was performed in the same manner as above, and the disintegrated state of the tablet was observed by craniotomy. The results are shown below. The following numerical values show the disintegration state of the tablet, and the meaning thereof is the same as above.

1st day 5th day 10th day after implantation Disintegration state 2 2 4 From these results, the sustained release cerebral vasospasm inhibitor of the present invention is
It can be seen that the dosage form is retained in vivo 10 days after implantation in the brain.

[Example 11] Vasodilation test (I) Using the tablets obtained in Examples 8 and 9 and tablets (placebo tablets) containing no calcitonin gene-related peptide (CGRP), which is a physiologically active peptide, were used. A vasodilation test was conducted by the method described below. The results are shown in Example 8.
FIG. 5 shows the tablets of No. 2 and FIG. 6 about the tablets of Example 9.

[Method for producing placebo tablet] Production of sustained-release preparation of Example 8 (V) and production of sustained-release preparation of Example 9 except that calcitonin gene-related peptide (CGRP) was replaced with hydroxypropylcellulose. A sustained-release preparation for placebo was produced in the same manner as in (VI).

[Vasodilation test] Rabbit weighs 2.5 to 3
8 kg for the tablets of Example 8, Example 9
With respect to the tablets of 7 above, an experiment was carried out by the following procedure using 7 animals.

(1) After a rabbit basilar artery radiograph, a known method [DG Vollmer et.al. Neurosurgery 28: 27-
32 (1991)], a subarachnoid hemorrhage model was prepared. (Day 0) (2) After 24 hours (Day 1), anesthesia was performed with pentobarbital, and the occipital region was incised along the life from the occipital bone to the first cervical vertebra.

(3) The muscles attached to the occipital bone, the first cervical vertebra and the occipital periosteum were carefully peeled off with a scalpel so as not to damage the vein.

(4) The exposed occipital bone was scraped from the lower portion by about 2 to 5 mm with a surgical drill.

(5) After that, the occipital periosteum is 8 to 1 with a scalpel.
An incision of about 0 mm was made.

(6) From this site, the tablets were placed under the arachnoid membrane with tweezers.

(7) After inserting the tablet, the occipital periosteum was sutured with silk thread.

(8) Along with the suture, the wound was further closed with Aron Alpha (registered trademark).

(9) Then, the muscle and the skin were sutured with a silk thread, and an appropriate amount of an antibiotic was administered.

(10) The blood vessel diameter was evaluated daily by angiography immediately after the administration to the fifth day (Day 6).

[Comparative Example 1] Drug release test (V) and vasodilation test (II) Calcitonin was compared with one rabbit as a comparative example by the cisternal puncture method according to the known method in (1) above. The gene-related peptide (CGRP) aqueous solution was administered, and the calcitonin gene-related peptide (CGRP) concentration (nM) in the cerebrospinal fluid was measured over time by the same method as in Example 10. The result is shown in FIG. 7.

Also, a rabbit model (two for CGRP aqueous solution, in which clear basilar artery spasm was clearly observed after subarachnoid hemorrhage by the cisternal puncture method according to the known method of (1) above, As a comparative example, an aqueous solution of calcitonin gene-related peptide (CGRP) and distilled water were administered to distilled water (4 animals), and the blood vessel diameter was evaluated in the same manner as in Example 11. The results are shown in Fig. 8.

Example 12 Production of sustained-release preparation (VI
I) Stearic acid (10 g) and hydrogenated oil (10 g) were mixed, and thereto was added 0.6% GSIL-modified maxadilan solution (2.5 g) (equivalent to GSIL-modified maxadilan (15 mg)) and lactose (20 g). 60 g of hydroxypropyl cellulose is added thereto. After mixing this well, Correct 19
Pressed with a K tablet molding machine (KIKUSUI CLEAN PRESS),
It was crushed and mixed, and pressure-molded again with the same molding machine (6 mmΦ
X 2 mm).

Example 13 Production of sustained-release preparation (VI
II) 10 g of stearic acid and 10 g of hydrogenated oil are mixed, and 2.5 g of a GSIL-modified maxadilan solution of 1.0% represented by the above structural formula (25 mg of GSIL-modified maxadilan and the like) and 20 g of lactose are added. 60 g of hydroxypropyl cellulose is added thereto. After mixing this well, Correct 19
Pressed with a K tablet molding machine (KIKUSUI CLEAN PRESS),
It was crushed and mixed, and pressure-molded again with the same molding machine (6 mmΦ
X 2 mm).

Example 14 Production of sustained-release preparation (I
X) Mix 10 g of stearic acid and 10 g of hardened oil, and add 1 to it.
2.5 g of GSIL modified maxadilan solution represented by the above structural formula of 0% (GSIL modified maxadilan 2
50 mg and the like) and 20 g of lactose are added and mixed,
To this is added 60 g hydroxypropyl cellulose.
After thoroughly mixing this, it was pressure-molded with a Correct 19K tablet molding machine (KIKUSUI CLEAN PRESS), crushed and mixed, and pressure-molded again with the same molding machine (6 mmΦ × 2
mm).

Example 15 Production of Sustained Release Formulation (X) 15 g of palmitic acid and 15 g of beeswax were mixed with 0.4 g of a GSIL-modified maxadilan solution of 0.4% represented by the above structural formula. (Equal to 10 mg of GSIL-modified maxadilan) is added and mixed, and 70 g of hydroxypropyl cellulose is added thereto. After mixing this well, Correct 19K tablet molding machine (KIKUSUI)
CLEAN PRESS) was used for pressure molding (6 mmΦ x 2 mm).

Example 16 Drug Release Test (VI) The preparation obtained in Example 12 was tested in the same manner as in the drug release test (I), and the results are shown in FIG. The numbers in the figure have the same meanings as above.

Example 17 Drug Release Test (VII) The preparation obtained in Example 13 was tested in the same manner as the drug release test (I), and the results are shown in FIG. The numbers in the figure have the same meanings as above.

Example 18 Drug Release Test (VII
I) The preparation obtained in Example 14 was tested in the same manner as in the drug release test (I), and the results are shown in FIG. The numbers in the figure have the same meanings as above.

Example 19 Vasodilation Test (III) Using the tablet obtained in Example 13 and the tablet (placebo tablet) containing no GSIL-modified maxadilan represented by the above structural formula, which is a physiologically active peptide, A vasodilation test was performed in the same manner as in the method described in Example 11. The result is shown in FIG. The placebo tablets were produced in the same manner as in Example 13 except that GSIL-modified maxadilan was replaced with hydroxypropyl cellulose.

Example 20 Production of sustained-release preparation (X
I) 0.4% calcitonin gene-related peptide (CGRP)
2.0 g of solution (equal to 8 mg of CGRP) and lactose 2
Mix 0 g well and add 80 g hydroxypropyl cellulose to it. After thoroughly mixing this, flat tablets having a diameter of 13 mm were produced with a KBr tablet molding machine (150 kg, 1 minute).

Example 21 Production of sustained-release preparation (XI
I) 0.4% calcitonin gene-related peptide (CGRP)
A solution (2.5 g) (equivalent to 10 mg of CGRP) and 20 g of glucose were mixed well, and 80 g of methyl cellulose was added thereto. After mixing this well, KBr tablet press (1
A flat plate tablet having a diameter of 13 mm was produced at 50 kg for 1 minute.

Example 22 Drug Release Test (IX) 5 ml of Hartmann's solution is aseptically placed in a 15 ml tube. Thereafter, the tablets obtained in Example 20 are similarly put aseptically and shaken at 37 ° C. and 120 rpm. 1, 2,
The drug release amount was determined by sampling on days 3, 7, 11, and 14 and quantifying by high performance liquid chromatography. The result is shown in FIG. The meaning of the numbers is the same as above.

Example 23 Production of sustained-release preparation (XI
II) 0.4% calcitonin gene-related peptide (CGRP)
Solution 2.5g (equivalent to 10mg CGRP) and 13g Eudragit were added and mixed, then crystalline cellulose 87
Add g. After thoroughly mixing these, flat tablets having a diameter of 13 mm were produced with a KBr tablet molding machine (150 kg, 1 minute).

Example 24 Production of sustained-release preparation (XI
V) 17 g of stearic acid and 17 g of hydrogenated oil (hydrogenated castor oil) are mixed, and 2.0 g of 0.4% calcitonin gene-related peptide (CGRP) solution (equivalent to 8 mg of CGRP)
And 0.3 g of Eudragit L30D-5.5 are added and mixed, then 66 g of crystalline cellulose is added. After mixing them well, KBr tablet molding machine (150 kg, 1 minute)
A flat-plate tablet having a diameter of 13 mm was produced by.

Example 25 Drug Release Test (X) 5 ml of Hartmann's solution is aseptically placed in a 15 ml tube. Thereafter, the tablets obtained in Example 23 are similarly put aseptically and shaken at 37 ° C. and 120 rpm. 1, 2, 4,
The drug release amount was determined by sampling on days 7, 11, and 14 and quantifying by high performance liquid chromatography. The result is shown in FIG. The meaning of the numbers is the same as above.

Example 26 Drug Release Test (XI) In the same manner as in Example 25, a release test of the preparation of Example 24 was carried out. The result is shown in FIG. The meaning of the numbers is the same as above.

Reference Example 1 Preparation of Polyion Complex of Hyaluronic Acid and Eudragit E 100 g of a 2% aqueous solution of hyaluronic acid and 100 g of a 2% aqueous solution of Eudragit E are reacted at room temperature for 2 hours while stirring. The reaction product is centrifuged at 3000 rpm for 10 minutes to recover the product, and the product is vacuum dried to obtain a polyion complex of hyaluronic acid and Eudragit E.

The obtained solid is pulverized and classified to obtain a powder having a particle size of 150 microns or less.

Example 27 Production of sustained-release preparation (X
V) 100 g of polyion complex of hyaluronic acid and Eudragit E obtained in Reference Example 1, 0.75 g of 0.4% calcitonin gene-related peptide (CGRP) solution (C
GRP3mg is mixed with the phase) and collect (Correct)
Pressed and molded with a 19K tablet molding machine (KIKUSUI CLEAN PRESS), crushed and mixed, and pressed again with the same molding machine (6m
mΦ × 2mm). Formulations obtained by performing two replicates (respectively,
HA-1 and HA-2 for CGRP in vitr
o The results of the release test are shown in Figure 16. [Example 28] Drug release test (XII) Using the tablets obtained in Example 2, an in vivo test relating to the drug release effect was performed by the method described below. In this drug release test, 30 rabbits (Normal, day
5 animals were used for 1, day 2, day 3, day 4, and day 5). The result is shown in FIG.

[Test Method] <Method of embedding tablet in brain> A rabbit (2.5-3.0 kg) after pentobarbital anesthesia was fixed on the prone position,
The occipital periosteum (dura) was exposed by incision, the occipital bone was drilled, and the dura was exposed more widely. Thereafter, the dura and arachnoid were incised by about 8 mm, and the tablets of the sustained release preparation of Example 2 were embedded in 25 rabbits, the dura, muscle and skin were sutured, and an appropriate amount of antibiotic was administered to the incised site. did. In addition, five rabbits without embedded tablets were prepared.

Using the rabbits in which the above-mentioned tablets were embedded, the following collection method was carried out.
Day (day2), day 3 (day3), day 4 (day4),
5 rabbits each on the 5th day
Cerebrospinal fluid (CSF) was collected from the animals, and the CGRP concentration (nM) in the cerebrospinal fluid (CSF) was measured by the following measuring method. Also, rabbits without embedded tablets (Normal)
Cerebrospinal fluid (CSF) was also collected from 5 animals, and the CGRP concentration was measured in the same manner.

<CGRP in rabbit cerebrospinal fluid (CSF)
Concentration measurement method> 1. Anesthesia was anesthetized with pentobarbital (50 mg / kg) on the day of cerebrospinal fluid (CSF) collection.

2. After anesthesia, the trachea was exposed on the back and the intubation tube was fixed in the trachea. 3. In that state, the incision was advanced toward the brain stem while paying attention to the blood vessels from under the jaw.

4. When the dura was reached, a careful incision was made to expose the arachnoid.

5. A 22G needle was inserted into the subarachnoid space to prevent blood from entering, and the cerebrospinal fluid (CS
F) was aspirated and collected.

6. Immediately after collection, it was frozen and stored at -20 ° C.

7. After the stored sample was dissolved, the CGRP concentration (nM) was measured in the same manner as in the radioimmunoassay described in Example 10 using a rabbit (Normal) without a tablet embedded therein as a control.

[Example 29] Test for disappearance effect of sustained-release preparation (tablet) (I) Using the tablet obtained in Example 2, a sustained-release preparation (tablet) in the brain was prepared by the following method. Disappearance effect in vi
I did a vo test. In this test, 18 rabbits (1st day, 2nd day, 3rd day, 4th day, 5th day,
Two mice each were used for the 10th day, the 1st month, the 3rd month, and the 6th month). The results are shown below. [Test method] <Method of implanting tablet into brain> A rabbit (2.5 to 3.0 kg) after pentobarbital anesthesia was fixed on the prone position,
The occipital periosteum (dura) was exposed by incision, the occipital bone was drilled, and the dura was exposed more widely. Thereafter, the dura and arachnoid were incised by about 8 mm, and the tablets of the sustained release preparation of Example 2 were embedded in 18 rabbits, the dura, muscle and skin were sutured, and an appropriate amount of antibiotic was administered to the incised site. did.

<Measurement Method and Evaluation Criteria> 1 day, 2nd day, 3rd day, 4th day, 5th day, 10th day, 1st month, 3rd month, 6th month after the above-mentioned rabbit was implanted with tablets. Two rabbits were exposed to the eyes at the position where the tablets were implanted by craniotomy and visually observed.
The evaluation criteria were as follows.

-: Almost disappeared.

+: About half of the tablets have disappeared.

++: Most of the tablets have disappeared.

+++: The tablet disappeared completely.

<Test results> ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ Evaluation ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ ━ ━ ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ 5th day + 10th day + 1st month ++ 3rd month +++ 6th month +++ ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ ━ None of the rabbits died during the period when the elimination effect test of this sustained-release preparation (tablet) was conducted.

[0105]

INDUSTRIAL APPLICABILITY The sustained release cerebral vasospasm inhibitor according to the present invention can be used by implanting it in the brain, and can not only stably release the medicinal component over a long period of time but also disperse it in the brain. Since it is not present, it is a drug that exerts its action efficiently.

[Brief description of drawings]

FIG. 1 is a graph showing the% CGRP detection over time of the preparation of Example 1.

FIG. 2 is a diagram showing% of CGRP detected over time in the preparation of Example 2.

FIG. 3 is a diagram showing% of CGRP detected over time in the preparation of Example 4.

FIG. 4 is a diagram showing nM of CGRP detected over time in the preparation of Example 2.

FIG. 5 is a view showing the time-dependent vasodilatory effect% of CGRP of the preparation of Example 8.

FIG. 6 is a view showing the time-dependent vasodilatory effect% of CGRP of the preparation of Example 9.

FIG. 7: Concentration n detected with time of the CGRP aqueous solution of Comparative Example 1
It is a figure which shows M.

FIG. 8 is a diagram showing the time-dependent vasodilatory effect% of the aqueous solution of CGRP of Comparative Example 1.

FIG. 9 shows the% detection of GSIL-modified maxadilan of the preparation of Example 12 over time.

FIG. 10 shows the% detection of GSIL-modified maxadilan of the preparation of Example 13 over time.

FIG. 11 shows the% detection of GSIL-modified maxadilan in the preparation of Example 14 over time.

FIG. 12 is a graph showing the time-dependent vasodilatory effect% of GSIL-modified maxadilan in the preparation of Example 13.

FIG. 13:% CGRP detection over time for the formulation of Example 20
FIG.

FIG. 14:% CGRP detection over time for the formulation of Example 23
FIG.

FIG. 15:% CGRP detection over time for the formulation of Example 24
FIG.

FIG. 16:% CGRP detection over time for the formulation of Example 27.
FIG.

FIG. 17 is a graph showing a time-dependent change in human α-CGRP concentration in CSF after the CGRP preparation in Example 28 was embedded in a large tank. The arrow indicates the time of implantation of the CGRP formulation. The average value shown as nmol / L in the graph is based on n = 5 at each measurement point. Vertical lines represent standard deviation. The statistically significant difference from the 0th day (day 0) is indicated by # (#: P <0.05;##: P <
0.01).

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI technical display location A61K 38/23 45/00 ABN

Claims (25)

[Claims] 1. A sustained-release cerebral vasospasm inhibitor for implantation in the brain, which comprises a sustained-release carrier carrying a cerebral vasospasm inhibitor. 2. The sustained-release cerebral vasospasm inhibitor according to claim 1, wherein the cerebral vasospasm inhibitor is a calcitonin gene-related peptide. 3. The sustained-release cerebral vasospasm inhibitor according to claim 1, wherein the cerebral vasospasm inhibitor is maxadilan. 4. The sustained-release cerebral vasospasm inhibitor according to claim 1 or 2, wherein the cerebral vasospasm inhibitor is GSIL-modified maxadilan. 5. The sustained-release cerebral vasospasm inhibitor according to any one of claims 1 to 4, which is a tablet type for placement in the subarachnoid space and / or a groove on the brain surface. 6. The sustained release cerebral vasospasm inhibitor according to any one of claims 1 to 5, which has a sustained release ability of the cerebral vasospasm inhibitor in cerebrospinal fluid for 2 to 14 days. 7. The sustained-release cerebral vasospasm inhibitor according to any one of claims 1 to 6, which has the ability to retain the dosage form in the cerebrospinal fluid for 7 days or longer. 8. The sustained release cerebral vasospasm inhibitor according to any one of claims 1 to 7, which disappears rapidly after the release of the cerebral vasospasm inhibitor. 9. The tablet having a thickness of 5 mm or less.
The sustained release cerebral vasospasm inhibitor according to any one of -8. 10. The expansion coefficient of the tablet in cerebrospinal fluid is 200.
% Or less, The sustained release cerebral vasospasm inhibitor according to any one of claims 5 to 9. 11. The release of the cerebral vasospasm inhibiting substance in cerebrospinal fluid is 1/5 or less of the total content in the sustained release preparation on the first day after administration. The sustained release cerebral vasospasm inhibitor according to item 1. 12. The sustained-release cerebral vasospasm inhibitor according to any one of claims 1 to 11, wherein the sustained-release carrier is a carrier containing a water-soluble polymer, an oil or a wax, and a fatty acid. 13. The sustained-release cerebral vasospasm inhibitor according to claim 12, which further comprises a saccharide. 14. The sustained release cerebral vasospasm inhibitor according to any one of claims 12 to 13, wherein the water-soluble polymer having a cellulose skeleton is hydroxypropyl cellulose. 15. The fat or oil is hydrogenated oil according to claim 12-14.
The sustained release cerebral vasospasm inhibitor according to any one of 1. 16. The fatty acid is stearic acid.
The sustained release cerebral vasospasm inhibitor according to any one of 2-14. 17. The sustained-release cerebral vasospasm inhibitor according to any one of claims 13 to 16, wherein the saccharide is lactose and / or glucose. 18. The sustained release cerebral vasospasm inhibitor according to any one of claims 1 to 11, wherein the sustained release carrier is a carrier containing a water-soluble polymer having a cellulose skeleton and a saccharide. 19. The sustained release cerebral vasospasm inhibitor according to claim 18, wherein the water-soluble polymer having a cellulose skeleton is hydroxypropyl cellulose. 20. The sustained release cerebral vasospasm inhibitor according to claim 18, wherein the saccharide is lactose and / or glucose. 21. The sustained release carrier is a carrier containing crystalline cellulose or Eudragit.
The sustained release cerebral vasospasm suppressant according to the item. 22. The sustained release cerebral vasospasm inhibitor according to claim 21, which further comprises fatty acids, waxes and / or oils and fats. 23. The sustained release cerebral vasospasm inhibitor according to claim 22, wherein the fat is a hardened oil. 24. The fatty acid is stearic acid.
The sustained release cerebral vasospasm inhibitor according to 2. 25. The sustained release cerebral vasospasm inhibitor according to any one of claims 1 to 11, wherein the sustained release carrier is a carrier containing a polyion complex of hyaluronic acid and a cationic polyacrylic acid derivative.