JPH10324633A - Blood coagulation accelerant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a blood coagulation accelerant that contains sulfated lipid, as an active ingredient, promotes blood coagulation at the traumatic or subcutaneous bleeding sites or at an arbitrary site desirably to be clotted. SOLUTION: A sulfated lipid (preferably sulfated glycolipid because of its strength of blood coagulation and low antigenicity, for example, sulfated sphingoglycolipid as sulfated galactosylceramide monosulfate, sulfated glyceroglycolipid as galactosylalkylacylgrlycerol monosulfate) is used as an active ingredient to prepare a topical blood coagulation accelerant for topical application. When it is used as a hemostatic agent, it is given in the form of powder or liquid and directly sprinkled to the bleeding-halting site. In the case that it is used as a vascular occlusion, it is administered to the site to be clotted by using a vascular injector in an amount of 5-20 ml for 10-120 minutes. When it is used in the form of a liquid, the sulfated lipid or its salt is dissolved or dispersed in a solvent as physiological salt solution or glycerol in an amount of 0.01-50 mg/ml.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a local blood coagulation promoter which contains a sulfated lipid as an active ingredient and is intended to act locally. More specifically, by administering the sulfated lipid to the site of traumatic or subcutaneous bleeding, or any part of the blood vessel where occlusion is desired, it promotes blood coagulation at the site to inhibit bleeding or occlude the blood vessel. And a pharmaceutical composition for:

[0002]

2. Description of the Related Art Conventionally, by promoting blood coagulation,
There have been hemostatic agents that suppress the outflow of blood from blood vessels and vaso-occlusive agents that stop blood flow in blood vessels by occluding the blood vessels.

[0003] When a wound is caused in a living body, a naturally-occurring hemostasis mechanism, that is, various plasma proteins called platelets and blood coagulation factors are activated, and blood coagulates.
The function of blocking the damaged blood vessel wall suppresses the outflow of blood from the blood vessel, but if the wound is large, it causes serious problems such as shock due to blood loss and sometimes death, and thus bleeding control to prevent it Agents have been used. As these bleeding suppressants, those that strengthen the blood vessel wall, those that promote coagulation (hemostatic agents), and those that suppress the dissolution of the formed thrombus (such as antiplasmin agents) have been mainly used. Specifically, examples of the hemostatic agent include thrombin, collagen and sodium alginate.
It is also known that hemostasis is performed by applying chitosan to a trauma site (Japanese Patent Publication No. 5-4369).

[0004] In a condition such as cancer or esophageal varices,
Vascular occlusive agents are used to kill nutrient cancer cells and prevent varicose veins by selectively stopping blood flow in the site of cancer and in blood vessels that have varicose veins. Was. For example, a drug that causes blood coagulation by locally damaging vascular endothelial cells, or a type of sponge or gel near the site where blood vessel obstruction is to occur, and these substances are released there. By doing so, methods and devices for blocking blood vessels and blocking blood flow have been developed. For example, such as gelatin pledgets, coiled wires, and plastic materials. In addition, a vaso-occlusive agent that locally administers a liquid drug such as chitosan to a target site in a blood vessel and forms a clot in a blood vessel downstream from the administration site to stop blood flow is also known (Japanese Patent Publication No. 5-4370). ).

[0005]

However, in the case of the conventional bleeding suppressants, there is a problem that the effect is uncertain in many cases. In the case of the hemostatic agent, coagulation by a blood coagulation factor is the main function. For this reason, for example, thrombin extracted and purified from bovine blood, which is one of the factors, is used, but because it is a heterologous protein, it causes serious side effects such as anaphylactic shock,
The serious problem of biohazard due to prions remains. Extracted from human blood to overcome these problems,
Attempts have been made to use purified thrombin and to use recombinant thrombin. However, in the former, HI
There is a problem of biohazard due to V or HBV. The latter has a cost problem. Or, it is difficult to determine an appropriate dose to exert an effect, and sometimes, conversely, there is a problem of causing unnecessary thrombus formation (Today's therapeutic agent 96, PP.398-403 Nankodo). Among the substances conventionally used as a bleeding inhibitor, sodium alginate, which is considered to have the lowest antigenicity, has a low bleeding inhibitory activity, so a large amount of administration must be performed to obtain the desired effect, There was a problem that the burden on the patient was large.

Further, in the case of a conventional vaso-occlusive agent, it is impossible to cause blood coagulation only at an arbitrary place in a blood vessel or to stop thrombus dissolution to stop blood flow. There is a problem of serious side effects such as bleeding due to damage to the blood vessel wall itself (Today's Therapeutic Drug 96, PP.398-403 Nanko-do). Downstream it often causes problems such as malnutrition and necrosis. In addition, when a foreign body is administered, serious side effects are often caused by anaphylactic shock due to an antigen-antibody reaction, similarly to a bleeding inhibitor. For example, it is difficult to specifically occlude the target site of the conventional pledgets used for this purpose, so a large amount of gelatin pledget is injected into the blood vessel to occlude the target site. There is a need to.
In addition, chitosan used as a liquid substance, while flowing downstream from the point of injection into the bloodstream, fiberized chitosan is entangled with blood cell components to form a clot,
Blood vessels could not be accurately occluded at the administration site, and the formed clots were foreign substances that were not normally found in the living body, and had problems such as being antigenic and difficult to metabolize. .

[0007] Therefore, the antigenicity is low, the safety is high,
There has been a demand for a novel blood coagulation accelerator which has few side effects, has excellent blood coagulation activity, and can accurately coagulate blood at a target site.

[0008]

Means for Solving the Problems In order to solve the above problems,
As a result of repeated studies, the present inventors have found that in vivo, sulfated lipids activate coagulation factors present in blood and have a blood coagulation promoting effect, and by using this as a blood coagulation promoting agent, The present invention has been completed. That is,
The present invention has been completed by applying the action of rapidly coagulating blood when a sulfated lipid comes into contact with a foreign substance surface and blood to a hemostatic agent and a vaso-occlusive agent.

[0009] Sulfated lipids are widely found in mammals including humans, and have high commonality among animals. Therefore, allergic reactions due to heterologous proteins as observed in protein preparations are not observed. Although substances derived from cattle and sheep are at risk of prion contamination, such substances can be easily extracted from organs such as pigs in which prion problems have not occurred. Meanwhile,
The structure of many of these substances has already been determined, and it is also possible to artificially synthesize the same substance by chemical means. The sulfated lipid used in the present invention is considered to promote thrombus formation by activating blood coagulation factor XII.
Factor XII is one of endogenous blood coagulation factors, and is activated at a site called the contact phase such as skin or plastic surface (the latest system of internal medicine, 21, platelets, abnormal coagulation / fibrinolysis, p.157 (1992) Nakayama Shoten), unlike extrinsic blood coagulation factors, which are usually important for spontaneous hemostasis, it can be expected to form clots more efficiently on the contact phase. That is, since blood coagulation occurs due to binding to the contact phase, unlike conventional hemostatic agents that made thrombus anywhere, it is easy to specify the location and the generated thrombus is retained at a specific site Therefore, blood vessel occlusion by hemostatic thrombus or thrombus formation can be accurately performed where necessary.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION In the present specification, "acting locally" means that when a pharmaceutical composition is administered to a specific site where a pharmaceutical activity is desired to be obtained, the agent exerts an action only at the site. I do. The local blood coagulation promoter of the present invention is clearly distinguished from a drug that is effective when the administered drug moves by the blood flow in the body, such as the above-mentioned vaso-occlusive agent using chitosan.

[0011] In the present specification, "blood coagulation" refers to a complex of a blood cell component and a fiber produced from fibrin contained as a component of blood by a blood coagulation pathway originally provided by blood. And hardens, and is clearly distinguished from the case where a fibrous substance not derived from blood and a blood cell component form a complex to form an insoluble matter.

The sulfated lipid which is an active ingredient of the blood coagulation accelerator of the present invention is not particularly limited as long as it has a sulfate group, and examples thereof include sulfated glycolipid and cholesterol sulfate. Sulfated glycolipids are preferred because of their strength and low antigenicity. As sulfated glycolipids,
Sulfated glycosphingolipids and sulfated glyceroglycolipids, or derivatives thereof, are preferred.

As sulfated glycosphingolipids, for example, galactosylceramide monosulfate (eg, Gal (3SO ₄ ) β1-
1Cer), glucosylceramide monosulfate (eg, Glc (3S
O ₄ ) β1-1Cer), lactosylceramide monosulfate (for example,
Gal (3SO ₄ ) β1-4Glcβ1-1Cer), gangliotriaosylceramide monosulfate (eg, GalNAcβ1-4Gal (3SO ₄ ) β1-
4Glcβ1-1Cer), gangliotriaosylceramide-bisulfate (eg, GalNAc (3SO ₄ ) β1-4Gal (3SO ₄ ) β1-4Glcβ
1-1Cer), lactotriosylceramide monosulfate (eg, GlcNAc (6SO ₄ ) β1-3Galβ1-4Glcβ1-1Cer), trihexosylceramide monosulfate (eg, Gal (3SO ₄ ) 1-4Gal1-4Gl
c β1-1Cer), gangliotetraosylceramide-bisulfate (eg, Gal (3SO ₄ ) β1-3GalNAc β1-4Gal (SO ₄ ) β
1-4Glcβ1-1Cer), lactone neotetraosylceramide monosulfate (eg, Gal β1-4GlcNAc (6SO ₄ ) β1-3Galβ1-4G
lcβ1-1Cer), gangliotetraosylceramide monosulfate (eg, Gal (3SO ₄ ) β1-3GalNAc β1-4Galβ1-4Glcβ
1-1Cer), glucuronyl lactone neotetraosylceramide monosulfate (eg, GlcA (3SO ₄ ) β1-3Galβ1-4GlcNAcβ1
-3Galβ1-4Glcβ1-1Cer), glucuronyl lactone neohexaosylceramide monosulfate (eg, GlcA (3SO ₄ ) β1-3G
alβ1-4GlcNAc β1-3Galβ1-4GlcNAc β1-3Galβ1-4Glc
β1-1Cer), Sial (8SO ₄ ) α2-6Glcβ1-1Cer and Sial (8SO4)
₄ ) α2-6Glc1-8Sial α2-6Glcβ1-1Cer and the like (the symbols are as follows; Glc: glucose, Gal: galactose, GlcA: glucuronic acid, GlcNAc: N-acetylglucosamine, GalNAc: N-acetylgalactosamine) ,
Sial: sialic acid, Cer: ceramide). Among them, galactosylceramide monosulfate present in the human body,
Lactosylceramide monosulfate, glucuronyl lactoneotetraosylceramide monosulfate or glucuronyl lactoneohexosylceramide monosulfate is preferred.

Examples of the sulfated glyceroglycolipid include galactosylalkylacylglycerol monosulfate (eg, Gal (3SO ₄ ) β1-3alkylacylglycerol) and triglucosylalkylacylglycerol monosulfate (eg, Glc (6SO ₄ ) α1- 6Glcα1-6Glcα1-3alkylacylgl
ycerol) and the like are preferable.

Examples of the above-mentioned sulfated glycolipid derivatives include compounds in which the hydroxyl group of the monosaccharide constituting the sulfated glycolipid is sulfated, and compounds in which the glycerol or ceramide constituting the sulfated glycolipid is sulfated. However, compounds in which the constituent monosaccharides are sulfated are preferred.

The sulfated lipid, which is an active ingredient of the blood coagulation promoter of the present invention, can be used in the form of a pharmacologically acceptable salt or in a free form that does not form a salt. Specifically, the above-mentioned salt is extracted, purified, or supplied as a product in the form of a solution in the form of sulfated lipid (free form).
Is mixed with an alkali metal or alkaline earth metal hydroxide solution and, if desired, freeze-dried to obtain a powdery salt. Examples of the alkali metal and alkaline earth metal hydroxide used in the treatment include aqueous solutions of sodium hydroxide, magnesium hydroxide, potassium hydroxide, calcium hydroxide, and the like, and an aqueous sodium hydroxide solution is preferable. The resulting sulfated lipid salt becomes a sodium salt.

As a method of using the blood coagulation promoter of the present invention, a method of administering the blood coagulation promoter to a site where the action of the blood coagulation promoter is desired to be used is preferable. For example, the outflow of blood from a blood vessel due to trauma or the like is suppressed. As the hemostatic agent, the blood coagulation-promoting agent of the present invention may be made into a powdery or liquid form and sprayed directly onto the bleeding site to administer the bleeding site to the bleeding site. In addition, for example, when the blood coagulation promoter of the present invention is used as a vaso-occlusive agent that blocks blood vessels and stops blood flow, the blood coagulation promoter of the present invention is used as a solution, and the solution is used as, for example, a cannula, catheter, or syringe. A vascular infusion device such as the above is inserted into a desired site in a blood vessel to be occluded and administered. The blood coagulation accelerator solution of the present invention 5 to 20 from the tip of the device at the site
By administering 0 ml over 10 to 120 minutes, it becomes possible to form a thrombus accurately at a desired site in the blood vessel at the distal end of the device where the blood vessel occlusion is desired, thereby causing the blood vessel occlusion. The amount and duration of administration of the vaso-occlusive agent into the blood vessel are appropriately selected depending on the thickness and site of the blood vessel for which vascular occlusion is desired.

The dosage form of the blood coagulation promoter of the present invention includes, for example, a powdered solid preparation or a liquid preparation, and in addition to the sulfated lipid as an active ingredient, is pharmacologically acceptable according to the administration method and administration form. It may be a pharmaceutical composition to which a carrier is added. When using the blood coagulation promoter of the present invention as a liquid preparation, a sulfated lipid or a pharmacologically acceptable salt thereof, a pharmacologically acceptable solvent such as physiological saline,
0.04 in a suitable solvent such as glycerol or phosphate buffer
It is preferable to use it by dissolving or suspending it at a concentration of 1 to 50 mg / ml. The solid preparation such as a powder may be a solid consisting of only the above-mentioned pharmacologically acceptable salt, or may be a liquid obtained by drying the above liquid preparation.

[0019]

【Example】

1. Hemostatic activity against venous bleeding of the blood coagulation promoter of the present invention Among substances having no antigenicity, sodium alginate (Alto (registered trademark): Kaigen) which has been conventionally used as a hemostatic agent and the hemostatic agent (galactosylceramide) of the present invention (Monosulfuric acid). The rats were laparotomized under Nembutal anesthesia (25 mg / 0.5 ml / kg) to expose the abdominal vena cava. A 26-gauge needle was pierced into the vena cava in the manner of blood collection to cause bleeding, and the outflowing blood was absorbed by Kimwipe.
Galactosylceramide monosulfate was extracted and purified from porcine spinal cord (Hara and Radin, NS. Anal. Biochem, 100, 364-370).
(1979)). Galactosylceramide monosulfate was sprayed on the bleeding site immediately after bleeding, in the amount of 35 to 40 mg. Alto similarly applied 600-700 mg to the bleeding site. The effect of hemostasis was determined by measuring the amount of hemoglobin in the blood absorbed by Kimwipe using Hemoglobin Test B-Wako (Wako Pure Chemical Industries, Ltd.) and determining the amount of bleeding.

[0020]

[Table 1]

The hemorrhage when the positive control alto and galactosylceramide monosulfate were used as hemostatic agents was almost the same and was smaller than that in the non-treated group, but the amount of galactosylceramide monosulfate used was one of that of alto. / 10 or less.

2. Vascular occlusion experiment utilizing galactosylceramide monosulfate's thrombus-forming ability Rats were anesthetized with intraperitoneal injection of Nembutal, the left thigh was incised, and a polyethylene cannula (PE-10, Imamura) was placed 4 cm from the abdominal vena cava. Was inserted retrograde from the same site and fixed. After awakening of the rat, galactosylceramide monosulfate dissolved in phosphate buffered saline solution (PBS) was passed at a rate of 1.25 mg / kg / h through a syringe attached to an infusion pump (Model-22M, HARVARD). The administration was continued for 4 hours. Four hours later, the rat was opened, and the area around the polyethylene cannula was observed. As a control, a glycolipid having a negative charge like galactosylceramide monosulfate,
Ganglioside G _M1 (Sial α2-8Sial α2-3Galβ1-4Glc
β1-1Cer) and G _D3 (Galβ1-3GalNAc β1-4Gal (3-2αSia
l) β1-4Glcβ1-1Cer) (The symbols are as defined above.)
A similar experiment was carried out using No., but no thrombus was formed in the control group. When galactosylceramide monosulfate was used, a thrombus was formed only around the polyethylene cannula and occluded the blood vessel.

[0023]

Industrial Applicability According to the present invention, a blood coagulation promoting agent which is capable of accurately performing hemostasis or vascular occlusion at a desired site and which is safer is provided. In other words, as a hemostatic agent having no antigenicity and exhibiting an excellent effect, it can be applied to trauma, etc., and can be administered into a blood vessel using an administration device to form a thrombus accurately at a desired site. Thus, it became possible to occlude blood vessels and cut off blood flow.

Claims (7)

[Claims] 1. A local blood coagulation promoter comprising a sulfated lipid as an active ingredient. 2. The local blood coagulation promoter according to claim 1, wherein the sulfated lipid is sulfated glycolipid or cholesterol sulfate. 3. The local blood coagulation promoter according to claim 2, wherein the sulfated glycolipid is a sulfated glycosphingolipid, a sulfated glyceroglycolipid, or a derivative thereof. 4. The sulfated glycosphingolipid is galactosylceramide monosulfate, glucosylceramide monosulfate, lactosylceramide monosulfate, gangliotriaosylceramide monosulfate, gangliotriaosylceramide-bisulfate, lactotriosylceramide. Monosulfuric acid, trihexosylceramide monosulfuric acid, gangliotetraosylceramide-bisulfuric acid, lactone neotetraosylceramide monosulfuric acid, gangliotetraosylceramide monosulfuric acid, glucuronyl lactone neotetraosylceramide monosulfuric acid and The topical blood coagulation promoter according to claim 3, which is at least one selected from the group consisting of glucuronyl lactoneohexosylceramide monosulfate. 5. The topical blood coagulation promoter according to claim 3, wherein the sulfated glyceroglycolipid is galactosylalkylacylglycerol monosulfate or triglycosylalkylacylglycerol monosulfate. 6. The local blood coagulation promoter according to claim 1, which is a hemostatic agent. 7. The local blood coagulation promoter according to claim 1, which is a vaso-occlusive agent.