JPS63170324A - Vascular permeability accelerator - Google Patents

Abstract

PURPOSE:To obtain the titled pharmaceutical containing a tumor necrosis factor (TNF). CONSTITUTION:A tumor vascular permeability accelerator obtained by containing a tumor necrosis factor (TNF), e.g. a recombinant type human tumor necrosis factor, produced in Escherichia coli by genetic engineering and having 155 amino acid residues, etc.; cellulicidal effect providing 50% survival rate after incubating 10,000 L-M cells derived from mice at 37 deg.C for 48hr is taken as 1 unit (U) for activity units of the TNF, as an active ingredient. The content of the TNF is 10,000-10,000,000U/day administered once or in divided portions. Distribution of a carcinostatic agent into cancerous tissues can be enhanced by administering this accelerator about 1hr before administering the carcinostatic agent.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a vascular permeability enhancer. For more details,
The present invention utilizes tumor necrosis factor (Tumor Necrosis factor).
The present invention relates to a tumor vascular permeability enhancer characterized by containing Factor (hereinafter referred to as TNF).

In the present invention, TNF is defined as "a method in which one or more substances having a reticuloendothelial system activating effect is administered to a mammal, and then an endotoxin derived from Durham-negative bacteria is injected, or an endotoxin derived from a mammal-derived It is defined as a physiologically active substance that is induced by adding endotoxin derived from Durum-negative bacteria to a tissue culture system containing macrophages, and is a factor that causes necrosis of certain cancers when administered to tumor-bearing animals. (Unexamined Japanese Patent Publication No. 59-3
9829), and it is known that TNF not only causes necrosis in some types of cancer, but also that its action is not species-specific. For example, TNF obtained from rabbits can kill cancers in mice. Furthermore, T
It is known that NF has almost no harmful effect on normal cells in vitro and has the ability to kill certain cancer cells (for example, LM cells derived from mouse cancer). In this way, TNF has an anticancer effect,
It is expected to be an anticancer drug because it is nonspecific to species and does not act on normal cells.

[Problem that the invention seeks to solve]

Research and development of anticancer drugs has been actively conducted for a long time, and various anticancer drugs are used clinically to treat cancer. Although the results are improving year by year, the results are not necessarily satisfactory.

In addition, many attempts have been made to enhance the anticancer effect and reduce side effects by using combinations of various anticancer drugs, but none of these are definitive as chemotherapeutic agents for cancer. do not have.

While the development of newly regulated cancer drugs is awaited, it is an important issue to try to enhance the effectiveness of existing anticancer drugs.

Anticancer drugs exert their anticancer effect when they reach the cancer tissue, so inheriting the distribution of anticancer drugs into the cancer tissue is one of the best ways to reduce the side effects of anticancer drugs and enhance their anticancer effects. It is believed that there is. Based on this idea,
Research is underway into missile therapy, which uses anticancer drugs chemically bonded to antibodies that recognize cancer antigens, intratumoral administration, and intraarterial administration, but the results are still insufficient.

[Means to solve problems]

As the present inventors continued their research on the anticancer effect of TNF, they unexpectedly found that tumor necrosis effect was not observed before and after the administration of TNF, before and after the onset of tumor necrosis due to TNF. Even with low TNF doses,
The present inventors have discovered that TNF has the effect of enhancing vascular permeability at tumor sites, and have completed the present invention.

That is, the present invention provides a tumor vascular permeability enhancer characterized by containing TNF.

TNF used in the examples of the present invention is a recombinant human tumor necrosis factor produced in Escherichia coli by genetic engineering, and has 155 amino acid residues (Japanese Patent Application Laid-open No. 60-260
522, Method for Stabilizing Physiologically Active Substances Produced by Genetically Recombinant Plants), but the TNF referred to in the present invention is not limited thereto.

As the TNF activity unit used in the examples of the present invention, 10,000 mouse-derived LM cells were incubated at 37°C.
The cell-killing effect that leads to a survival rate of 50% after 48 hours of incubation is defined as 1 unit (U).
O-260522).

The content of TNF in the vascular permeability enhancer in the present invention is appropriately selected depending on the administration method, the symptoms to be treated, the age, etc., but is usually 10,000 to 0,000 per day.
o, ooo u, administered at once or in divided doses.

(Margins below) [Effects of the present invention] By administering the vascular permeability enhancer of the present invention approximately 1 hour before anticancer administration, the distribution of the anticancer agent to cancer tissues can be enhanced, and as a result, the anticancer agent For example, by intravenously injecting a low dose of TNF, which does not show an anticancer effect alone, into cancer-bearing mice, and then administering an anticancer drug such as cisplatin one hour later,
It can significantly enhance the effects of anticancer drugs.

(The following is a blank space) Hereinafter, embodiments of the present invention will be described in detail with reference to examples, but it goes without saying that the present invention is not limited to these examples.

Example 1 Eight-week-old BAL B/C female mice were infected with Colon-26 adenocarcin strain (Colon-26adenocarcin).
oma) A syngeneic tumor was transplanted subcutaneously (0.1 ml/3% cell suspension), and 3.0 OOU of TNF was intravenously injected on the 7th day. Thereafter, 1 ml of 1% Evans blue/physiological saline was intravenously injected at 0 to 4 hours. After 30 minutes, the tumor was observed from the outside, the carotid artery was immediately cut, and after exsanguination, the tumor was excised and fixed in formalin. Next, separate the epidermis and peritoneum, and proceed to II! The epidermis of the Ti tumor was observed from the back side. As shown in Table 1, Evans blue infiltration was observed in the tumor area 30 minutes to 1 hour after TNF administration, and it was observed that vascular permeability in the gyri was increased. Furthermore, no infiltration of Evans blue was observed in areas other than the tumor area.

(See margins below) Table 1 Example 2 In the same manner as in Example 1, the dose of TNF was varied from 300 to 30,0000. Note that Evans blue was administered for a period of time after the TNF administration. As shown in Table 2, an improvement in vascular permeability in the tumor area was observed with administration of TNF as low as 300 U. In this case as well, no infiltration of Evans blue was observed in areas other than the tumor area.

Table 2 Example 3 Syngeneic tumors of Colon-26 adenocarcinoma were implanted (0.1 ml/3% cell suspension) into 6-week-old BALB/C female mice, and 10 days after injection of 3.0 OOU. TNF was administered intravenously. One hour later, 2.5 μ/kg of cisplatin was administered intraperitoneally.

As a control group, a group to which TNF and cisplatin were not administered was also provided. Table 3 shows tumor weight (■) on day 17 after tumor transfer Vi.
[Y4 long axis (long axis) of tumor x (short axis (long axis) of tumor) 2] is shown. Significant inhibition of II tumor growth was observed only in the group receiving both TNF and cisplatin.

Table 3 *Average values of 5 animals per group Example 4 In the same manner as in Example 3, 8 days after tumor implantation, TNF was administered.
was administered, and one hour later, 40 mg/kg of cyclophosphamide was administered intraperitoneally. Cyclophosphamide was administered for 12 days, 16 days, and 40 days after tumor transplantation.
/kg was administered intraperitoneally. Table 4 shows the tumor weights 20 days after tumor implantation.

Significant tumor growth inhibition was observed only in the group receiving both TNF and cyclophosphamide.

Table 4 Patent distributor Hayashi Kiteitomerine Kawamasa (1 (spontaneous) August 28, 1985 Director General of the Patent Office Kunio Ogawa 1, Indication of the case 1988 Patent Application No. 748 2, Name of the invention: Vascular permeability advanced agent 3, Relationship to the person making the amendment Patent applicant address: 1-2-6 Eijimahama, Kita-ku, Osaka-shi, Osaka Name (
003) Asahi Kasei Kogyo Co., Ltd. 4, Agent Address: 3F Tamayagi Building, 3-1 Rokubancho, Chiyoda-ku, Tokyo Telephone: (261) 96366 Subject of amendment: ``Specification'' 7. Contents of amendment

Claims (1)

[Claims] A tumor vascular permeability enhancer characterized by containing TNF.