JP2000247904A - Cerebral blood vessel twitching treatment agent and hematoma remover - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a treatment preparation that can inhibit cerebral blood vessel twitching after cerebral hemorrhage, particularly subarachnoid hemorrhage and can promote the removal of hematoma by using activated protein C (abbreviated to APC hereinafter) as an active ingredient. SOLUTION: This therapeutic agent includes APC as an active ingredient. In a preferred embodiment, the APC is particularly human blood-originating APC or human APC given by the gene technology. This blood originating APC is obtained by, for example, purifying protein C in affinity chromatography using anti-protein C-antibody, preferably the monoclonal antibody from human plasma and activating the purified protein C with human thrombin and purifying the activated protein C with the cationic ion-exchange chromatography. This preparation is preferably used as an injection preparation. The daily dose of this preparation is 0.5-10 mg/adult in the case of administration into the cistern.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an agent for treating cerebral vasospasm and an agent for removing hematoma.

[0002]

2. Description of the Related Art Subarachnoid hemorrhage is a disease with a poor prognosis in which the bleeding spreads to the subarachnoid space due to the rupture of a cerebral aneurysm or cerebral arteriovenous malformation, resulting in neurological deficits such as impaired consciousness and paralysis. About one week after the onset of the disease, cerebral vasospasm, which is a persistent and severe stenosis of the main artery, is said to be involved in the exacerbation of the disease state. That is, cerebral vasospasm reduces cerebral blood flow in the main artery and changes the properties of blood vessels and blood, resulting in circulatory disorders, which further exacerbates cerebral ischemia and causes severe neurological deficits. That is.

The details of the mechanism of the onset of cerebral vasospasm are unknown, but experimentally, blood is injected into the subarachnoid space of an animal to induce cerebral vasospasm, It has been suggested that there is a correlation between the amount of hematoma and the frequency of occurrence of cerebral vasospasm, and it is considered that the main cause is the presence of hematoma remaining in the subarachnoid space. So far, various physiologically active substances such as lipid peroxide, oxyhemoglobin, thromboxane A2, endothelin, and thrombin have been mentioned as direct causative substances of cerebral vasospasm, but they have not been specified.

For cerebral vasospasm after subarachnoid hemorrhage and cerebral ischemic symptoms associated therewith, treatment with systemic administration of a lipid peroxidation inhibitor, a thromboxane A2 synthase inhibitor, and a cerebral vasodilator is required. However, these treatments have not been satisfactory because they are symptomatic treatments in the presence of hematoma, which is the main cause of vasospasm.

[0005] In recent years, a therapeutic method by cisternal perfusion using a thrombolytic agent such as urokinase or tissue plasminogen activator has been attempted for the purpose of actively dissolving and removing hematomas due to the limited effectiveness of the above symptomatic treatment. ing. However, the optimal dose and administration method of these drugs are not clear, and bleeding complications such as intracerebral hemorrhage, subarachnoid hemorrhage, and epidural hemorrhage have been reported. It has been demanded.

[0006] In addition, since intracerebroventricular perfusion used after subarachnoid hemorrhage can be applied to the removal of hematoma at the time of cerebral hemorrhage, a safe and effective drug with less side effects of hemorrhage is also useful for the removal of hematoma after cerebral hemorrhage. It is extremely useful, and the development of such a drug is considered to be an important issue in this disease field.

On the other hand, activated protein C (hereinafter referred to as “AP
C ") is a double-stranded serine protease having an anticoagulant effect with a molecular weight of 61,000. This APC is usually present in the blood as its precursor, protein C (hereinafter abbreviated as “PC”), but once subjected to the coagulation system, it undergoes limited degradation by thrombin to exhibit enzyme activity.

APC suppresses thrombin production by inactivating activated factor VIII (hereinafter abbreviated as “FVIIIa”) and factor V (hereinafter abbreviated as “FVa”) in the cascade of coagulation factors. Show anticoagulant effect (Bioche
mistry, 16, 5824-5831, 1977, Biochem. Biophys. Act
a., 571, 333-342, 1979, Blood, 59, 1067-1072, 198
2, J. Biol. Chem., 258, 1914-1920, 1982, Biochemis
try, 19, 401-409, 1980). APC is in vitro
Inhibit plasminogen activator inhibitors, it is thought that they may indirectly add plasminogen activator to exhibit fibrinolytic activity (Proc. Natl. Acad. Sci. , 82, 1121-1125, 1985).
Generalized intravascular coagulation syndrome (hereinafter abbreviated as "DIC")
A strong anticoagulant effect is required in the state of hypercoagulability such as FVIIIa, but FVIIIa and APC that inactivate FVa suppress the production of thrombin, so that FVII
It blocks the production of Ia and FVa, ie, the positive feedback of the coagulation system, and is expected to have a strong anticoagulant effect. On the other hand, in a thrombus formation site such as DIC, the lack of thrombomodulin (hereinafter abbreviated as “TM”) due to damage to vascular endothelial cells is considered. In particular, when it is caused by endotoxin, TM activity is considered to decrease, and it is considered that conversion from PC to APC is not sufficient. Therefore, high-purity APC is expected as an effective therapeutic agent that directly suppresses the hypercoagulable state such as DIC.

[0009] However, it has not been known that APC suppresses cerebral vasospasm and is effective for removing intracranial hematoma.

[0010]

SUMMARY OF THE INVENTION The present inventors have conducted intensive studies to develop a therapeutic agent for cerebral vasospasm after subarachnoid hemorrhage and the like, and an agent for removing intracranial hematoma. The present inventors have found that the present invention can be applied clinically as a therapeutic agent and completed the present invention.

[0011]

That is, the present invention provides the following:
A therapeutic agent for cerebral vasospasm characterized by containing PC as an active ingredient or a hematoma removing agent characterized by containing APC as an active ingredient.

[0012]

DETAILED DESCRIPTION OF THE INVENTION APC used in the present invention is APC derived from blood obtained from human or other mammalian blood.
It includes PCs and APCs derived from mammals such as humans, which are produced by genetic recombination technology. As the APC used in the present invention, among these APCs, APC derived from human blood and human APC produced by a genetic recombination technique are preferable. In addition, as long as the effects of the object of the present invention as a therapeutic agent for cerebral vasospasm and a hematoma removing agent can be obtained, blood-derived APCs or APCs having physiologically equivalent activities to APCs produced by a genetic recombination technique can be used. Derivatives in which part of the entire amino acid sequence has been deleted, substituted, inserted, or added are also included in the APC of the present invention.

The APC of the present invention can be manufactured by a conventionally known method. For example, a method of activating PC isolated from human blood, a method of separating APC from human blood, a method of producing by genetic recombination technology, and the like can be mentioned.

Among them, the method of activating PC to APC is not particularly limited, and examples thereof include a method of activating with thrombin isolated from human or bovine blood or a method of activating with a synthetic peptide. Can be

The method for producing APC derived from blood includes the following method. For example, a method in which PC purified from human plasma by affinity chromatography using an anti-protein C antibody, preferably a monoclonal antibody, is activated with human thrombin, and then purified by cation exchange chromatography (Blood, 63, 115- 121,
1984) or PC obtained by Kisiel purification from human plasma by the steps of adsorption and elution of Ba citrate, ammonium sulfate fractionation, DEAE-Sephadex column chromatography, dextran sulfate agarose chromatography and polyacrylamide gel electrophoresis. To activate APC to APC (J. Clin. Invest. 64, 761-7
69, 1979) or a commercially available blood coagulation product containing PC was prepared by the method of Taylor et al. (J. Clin. Invest. 79, 918-925, 19).
87).

As a method for preparing APC using the gene recombination technique, for example, the methods described in JP-A-61-205487, JP-A-1-2338 or JP-A-1-85084, etc. is there.

In order to maintain the activity of the APC prepared by the above-described method to a maximum, the APC used in the present invention is fresh, or when stored at 4 ° C., used within 5 days after storage. Preferably, it is The APC used in the present invention may be a freeze-dried and stored APC together with a suitable stabilizer, or may be a frozen and stored APC solution.

The preparation of the present invention may contain APC as an active ingredient, and can be produced by any known pharmaceutical production method. The APC of the present invention may be any suitable carrier or medium generally used as a drug, such as sterile water or physiological saline, vegetable oil, mineral oil, higher alcohols, higher fatty acids, harmless organic solvents, etc. Forming agents, coloring agents, emulsifiers, suspending agents, surfactants, solubilizers, anti-adsorption agents, stabilizers, preservatives, humectants, antioxidants, buffers, tonicity agents, soothing agents, etc. Injectables suitable for effective administration to living organisms, nasal absorbents, or pharmaceutical preparations such as oral preparations, preferably injectables, can be prepared as appropriate. It is known. Injectable preparations can be provided, for example, in the form of a lyophilized product, a solution for injection, or a form enclosed in an osmotic pump.

The preparation of the present invention can be administered by arterial injection, intravenous injection or intracisternal injection. In addition, it is preferable to administer into the cisternal by puncture in the cistern because a more direct effect can be expected. In particular, a method of perfusing the cerebral cistern after insertion of drainage during craniotomy is optimal. For example, in the case of intracerebroventricular administration, an appropriate dose is 0.5 to 10 mg / person per day for general adults.

[0020]

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to Examples, but these Examples do not limit the scope of the present invention.

[Example 1] Rabbit subarachnoid hemorrhage (SAH)
According to the model, treatment of cerebral vasospasm was attempted by administering APC into the cistern after subarachnoid hemorrhage.

Using a rabbit, 3 ml of autologous blood was injected into the large tank and S
AH, APC 0.1mg, 0.5mg or solvent 0.5m after 15 minutes
The mixture was injected once into the large tank as l. The effect of the test drug was determined in DAY2 on the residual amount of hematoma in the subarachnoid space, basilar artery (hereinafter referred to as “B
A) and evaluated by the relaxation response (Emax) of BA to acetylcholine.

[0023] Residual hematoma in the subarachnoid space was observed mainly in the ventral part of the brainstem in the solvent group. Hematoma decreased in the APC 0.1 mg group and slightly remained in the ventral part of the brainstem, but hardly observed in the APC 0.5 mg group. BA blood vessel diameter was 77.5 ± 3.1% (n
= 7), compared with 97.2 ± 3.8% (n = 5) in the APC 0.5 mg group. The Emax of the relaxation response of BA to acetylcholine was 66.9 ± 6.3% in the solvent group and 106.9 ± 2.9% in the 0.5 mg group of APC, indicating an improvement in Emax in the APC administration group.

[0024]

The therapeutic agent containing the APC of the present invention as an active ingredient has the effect of suppressing cerebral vasospasm and promoting the removal of hematoma. Therefore, by using APC as an active ingredient, it can be used as a therapeutic agent for cerebral vasospasm or an agent for removing hematoma after cerebral hemorrhage, especially after subarachnoid hemorrhage. In addition, the APC protein of the present invention is a safe therapeutic agent for cerebral vasospasm and a hematoma removing agent because it has less side effects such as bleeding than existing drugs such as antithrombotic agents.

Continuation of front page (72) Inventor Kikuo Ohno 1-5-45 Yushima, Bunkyo-ku, Tokyo Tokyo Medical and Dental University Neurosurgery (72) Inventor Yusuke Mizuno 4-59-16 Chuo, Nakano-ku, Tokyo So Nakano In-hospital Neurosurgery (72) Inventor Tsunefumi Kobayashi 4-3-1 Asahigaoka, Hino-shi, Tokyo Teijin Inside Tokyo Research Center Co., Ltd. (72) Inventor Toyo 2-3-1-10 Kanda Surugadai, Chiyoda-ku, Tokyo Tokyo Medical and Dental University Biomaterials Engineering Laboratory F-term (reference) 4C084 AA02 BA44 DC03 MA56 MA66 NA06 ZA362 ZA542 ZC192

Claims (2)

[Claims] 1. A therapeutic agent for cerebral vasospasm, comprising activated protein C as an active ingredient. 2. A hematoma-removing agent comprising activated protein C as an active ingredient.