JPS61171429A - Promoter for biological activity of normal tissue - Google Patents

Abstract

PURPOSE:The titled promoter obtained by blending at least fibrinogen with thrombin and calcium ion. CONSTITUTION:At least fibrinogen is blended with thrombin and calcium ion in a ratio of preferably 70-110mg; 4-1,000 IU; 10-80 millimol, respectively, and, if necessary, fibronectin, the factor XIII, plasminogen, aprotinin, albumin, anti-plasmin globulin, saccharide, organic acid, etc., to give a promoter for biological activity of normal tissue. The promoter can be further blended with an antitumor agent, antibiotic, antimicrobial agent, biological preparation, insecticide, anti-inflammatory drug, etc., or a substance obtained by making a focus part of tumor or tuberculous caseous focus innoxious or into a dead cell. This drug promotes biological activities such as phagocytotic action by macrophage, etc., detoxicating activity, immunological activity, divided germ formation activity, etc. Consequently, regeneration of tissue and function recovery are extremely improved.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] This invention relates to a drug used to advance the biological activity of normal tissues. The biological activity mentioned here includes, for example, phagocytosis activity, detoxification activity, immune activity B, and mitotic bud production activity. Such biological activity advances, for example, promote healing or regeneration of injured or injured organs or tissues of the body.

[Conventional technology]

Conventionally, as a drug applied to tissues, for example, α-cyanoacrylate monomer is commercially available.

This is a material (ie, a tissue adhesive) intended for adhering cut tissue (eg, bone).

However, since α-cyanoacrylate monomer is a synthetic substance, it has the disadvantage of poor affinity for tissues (and necrosis).Also, it is known that blood coagulation substances are used to stop bleeding.However, For example, drugs aimed at regenerating lost tissue have not yet been put into practical use.

[Description of the invention]

The inventor has surprisingly found that a drug comprising at least fibrinogen, thrombin, and calcium ions, and optionally a biological trace component, promotes the regeneration of tissue lost due to disease or resection, He completed this invention.

That is, this invention provides at least fibrinogen,
The present invention provides an advanced normal tissue biologically active agent characterized by containing thrombin and calcium ions. These are fibrinogen 7O-110q! Thrombin 4-1000IU: Calcium ion No-80
It is desirable that the molecules exist in millimolar combinations.

In addition to the above-mentioned fibrinogen, thoron, and calcium ions, the normal tissue biologically active advanced agents of this invention include, for example, fibronectin, factor x, plasminogen, aprotinin, albumin, antiplasmin globulin, sugar, Organic acids (e.g. It is possible to incorporate detoxified or dead cells of lesions such as malignant tumors (such as malignant tumors) or tuberculous caseous foci.

The fibrinogen, thrombin, and other biological substances are most preferably derived from humans, but from an industrial standpoint, those derived from mammals other than humans (eg, cows) are often used. As the calcium ion, calcium chloride is most preferable c, and the acid is pi-1
Used for mlI&.

Of the components listed above, at least fibrinogen 8 and thrombin are contained in separate containers or separate storage sections in one container (hereinafter collectively referred to as storage chambers), and mixed at the time of renewal. . Preferably, four storage chambers are used, of which the fibrinogen is contained in the y41 chamber, the fibrinogen solvent is contained in the second chamber, the thrombin is contained in the third chamber, and the thrombin is contained in the fourth chamber. It is appropriate to contain a solvent and a calcium ion, respectively. An example of a preferred ingredient formulation 2 aI range is as follows:
In the first chamber, fibrinogen 70-1101q, plasma fibronectin 2-9 cape, factor X 10-50U,
and 2°-80 μg of plasminogen, and put bovine aprotinin solution (1-4000 KIU/J) into the second chamber.
) or 1 yzl of distilled water for injection, put lyophilized bovine thrombin 4-100010 in the third chamber, and add calcium chloride solution (10-80 mmol/liter) Ig/ in the fourth chamber. put in. As the storage chamber, an independent ampoule or vial or a multi-chamber container is used. When using, the ingredients placed in the first chamber are transferred to the second chamber.
Dissolve the ingredients separately in the liquid in the third chamber, dissolve them in the liquid in the fourth chamber, and mix them.
Inject or apply it to the application site (affected area) and apply it closely.

〔effect〕

The normal tissue bioactivity-advanced agent of the present invention improves, for example, phagocytic activity, detoxification activity, immune activation, and mesobud production activity by macrophages and the like. That is, promotion of healing and regeneration of damaged or injured organs and tissues in the body of mammals (including humans), localization and occlusion of diseased areas such as various infections and cancer, fibrosis of affected areas, and blood vessels. It is highly effective in promoting endothelial formation, fibroblast formation, the birth and assembly of white blood cells, etc., as well as activating phagocytosis and promoting detoxification, as well as restoring injured organ wealth or tissue to its original form. has.

91E, it may be necessary to remove part or most of the liver lobe due to various liver diseases such as cirrhosis, chronic hepatitis, and liver cancer. It was found that when the drug was applied, the regeneration and functional recovery of the liver tissue in the above-mentioned areas (8) were dramatically improved compared to the conventional method in which restoration was impossible (see Test Example 1 below). .

〔Example〕

EXAMPLES Hereinafter, this invention will be explained in more detail by Examples, and the effects of this invention will be clarified by Test Examples.

Example 1 (formulation example) (A, l fibrinogen 100#
Plasma fibrocutin 71sy No. X1
ll factor 40U plasminogen 50μg (B) 1 box of bovine aprotinin (30U OV-IU/j?l) 1
vtl (shi) bovine thrombinsi dry quadrant 500I
UCD) Calcium chloride aqueous solution (40 mmol/liter) l ml
In use, (A) is dissolved in (B), CC) is separately dissolved in (1)), and both melting irons are mixed before use to prepare the drug of the invention.

6A, - Example 1 Rapid formation of regenerated liver and return to normal liver function (Method) Using m-sexual Wistar rats (overdosage 5), first, barbital drug @i (Somnoventil) 0.04 rx was administered.
l/body massage 10 (l was injected from the tail vein, anesthetized &!jJ1
Turn over and slightly pull out the largest lateral left liver lobe.
1, and an arbitrary section corresponding to 3/4 to 1/10 of the liver lobe was excised with an inverted V-shaped cut. During this time, the base of the liver lobe was compressed to stop the bleeding.

Immediately after excision, the preparation (mixture) of Example 1 was applied to the cut surface. Hemostasis of the cut surface was completed after about 3 minutes. The abdomen was sutured, and after awakening, the patient was given normal food.

As a control, known suture-based hemostasis methods, S and α
- Application of cyanoacrylate monomer (trade name Aron Alpha) was carried out separately.

(Results) After the above surgery, the animals were sacrificed in layers and the liver regeneration status was observed.As a result, when the preparation of Example 1 was used, liver lobe regeneration started 24 hours after the surgery; 168 hours (7
It was later allowed to fully restore its original condition.

The state of the liver (photographs with a magnification of 2.5 times) 6 hours, 72 hours, and 168 hours after surgery are shown in Figures 1, 2, and 3, respectively. 2. In Figure 3, the 7-shaped line indicates the position of the cut surface, and the inside of the 7 is the regenerated part.

These photographs show that when the formulation of Example 1 is used, an excellent regeneration promoting effect can be obtained.

On the other hand, when sutures were used, liver regeneration was observed, but the regenerated area was lumpy and not normal. Furthermore, when α-cyanoacrylate was used, no regeneration was observed, and the cut surface adhered to the ground tissue, resulting in necrosis.

In addition, when the specimen including the area near the cut surface was examined using a pathological microscope, it was found that when the preparation of Example 1 was used, new blood vessels and hepatic ducts were generated at the cut surface 72 hours after surgery, and the tip of the regenerated liver It was observed that hepatocytes were undergoing extremely vigorous regeneration in the area, and only traces of the cut surface remained after 168 hours. Figure 4 is a micrograph of a specimen stained with hematoxylin and niobium type 2 72 hours after surgery.The slightly #B diagonal line of So Nakao is the cut surface, the left side is the liver, and the right side is the regenerated liver. Show part. This photo shows that the regenerated liver is of the same quality as the original liver and has fused together as one.

On the other hand, when suture was used, hepatocyte formation was observed, but when α-cyanoacrylate monomer was used, no hepatocyte formation was observed, and both were lump-like and biliary duct formation. Regeneration of blood vessels and blood vessels is also extremely poor, and recovery of liver function is at 16.
Even after 2 weeks, which was well over 8 hours, it was not appropriate.

Test Example 2 Stopping proliferation of cancerous cells in cancerous lesions (method and results)
Approximately 1 x l O'6 melanoma skin cells were transplanted subcutaneously. After 10 days, the cancer mass had grown to a radius of approximately 7n. Mice were treated with a barbital anesthetic (somnopentyl)
The patient was anesthetized and the tumor mass was removed. Before removing the cancer mass, H-
Thymidine 5 μCi 10. The cells were labeled by tail vein injection of 1 ml of the solution. After removal, it was immediately returned to its original location and sutured. Two days later, a 14 cm thymidine 5 μCi/0.1 ml solution was injected from the tail vein, the cancer mass was removed again, and an autoradiograph was created. In this case, the distribution of H within the cancer lesion indicates cancer cell proliferation before surgery, and the distribution of C indicates proliferation after surgery. Under the above conditions, 3H114C also showed active proliferation of cancer cells.

If I forgot to cover it with the formulation of Example 1 during the first extraction, no 14C autoradiograph was obtained. Proliferation was observed on the surface, but when the preparation of Example 1 was mixed with melanized dead pharyngeal cells, uptake of 14C tidine, which indicates cancer cell proliferation, was observed even after several months. From these results, it was found that the growth of cancer cells was stopped by the formulation of Example 1, and that the combination of Formulation No. 1 and dead cancer cells effectively stopped the growth.

[Brief explanation of drawings]

Figures ml, 2g and 3 are photographs showing the state of a partially excised liver lobe of a rat 6.72 and 168 hours after application of the drug of this invention, respectively. FIG. 4 shows the head of the hematoxylin-niosif type 2 stained specimen after 72 hours. Patent originator Bioscience Research Co., Ltd. Agent Pu11 Riju Yamabo et al.18 Figure 1 Figure 2 Figure 3 Procedure amendment written form) May 20, 1985

Claims (9)

[Claims] (1) A normal tissue bioactivity enhancer, characterized in that it contains a mixture of at least fibrinogen, thrombin and calcium ions. (2) The agent according to claim 1, wherein the biological activity is phagocytosis activity, detoxification activity, immunoactivity, or mesobud production activity. (3) The agent according to claim 1, wherein the biological activity is aimed at promoting healing or regeneration of injured or injured organs or tissues in the body of mammals (including humans). (4) The agent according to claim 1, which is intended to promote healing of malignant tumors, benign tumors, or ulcers. (5) The agent according to claim 1, which contains an antitumor agent, an antibiotic, an antibacterial agent, a biological preparation, an anthelmintic agent, or an anti-inflammatory agent. (6) The agent according to claim 1, which is intended to prevent contact between the affected area and normal tissue. (7) A compound containing a detoxified or dead tumor or tuberculous caseous lesion, which is intended to regenerate normal tissue by bringing the lesion into close contact after at least a portion of the lesion has been excised. The agent according to claim 1, which is (8) The agent according to any one of claims 1 to 7, wherein the agent is contained in at least two storage chambers, and fibrinogen and thrombin are contained in separate chambers. (9) Contained in four storage chambers, the first chamber contains fibrinogen, the second chamber contains fibrinogen solvent, the third chamber contains thrombin, and the fourth chamber contains thrombin solvent. Claim 8 containing calcium ions
Agents listed in section.