JPH0160007B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an anti-cancer substance sustained-release embolic agent, and more specifically, it can be suitably used in cancer or tumor treatment and diagnosis methods, such as vasoocclusive therapy or puncture therapy. The present invention relates to an anti-cancer substance sustained-release embolic agent.

In recent years, vascular occlusion therapy has been increasingly recognized as an effective means for treating liver cancer, breast cancer, and the like. Vascular occlusion therapy involves reaching the feeding artery of the cancer or tumor tissue with the tip of the vascular catheter, and injecting embolic material from the other end to occlude the feeding artery and stop blood flow. The purpose of this treatment is to cut off nutritional supply to the body and cause necrosis of these tissues. It has been reported that the wound protection material on which blood coagulation factors and thrombin are immobilized, which was previously proposed by the applicant (Japanese Patent Application Laid-Open No. 135214/1983), can be effectively used as an embolic material used in vascular occlusion therapy. However, this therapy has the disadvantage that it cannot be used concurrently with chemotherapy, which is an effective means of treating cancer or tumors. In other words, after vascular occlusion therapy, even if an anticancer substance is administered orally or by injection, the feeding artery is occluded, so it can no longer reach the cancer or tumor tissue, and the administration is meaningless. Become. Therefore, when administering vascular occlusion therapy, a two-step method is used: either abandoning chemotherapy or injecting an anticancer substance through a vascular catheter before occlusion. However, the amount of anticancer substances injected using this method is not sufficient, and because it is usually injected in the form of a solution and the feeding artery has not yet been occluded, the anticancer substance is only available for a very short period of time. However, it is difficult to expect an effect because it will spread to areas other than the target cancer or tumor tissue.
As described above, the problem with vasoocclusive therapy, that is, the inability to concurrently use chemotherapy, remains unresolved.

On the other hand, along with vascular occlusion therapy, puncture is a therapy that is expected to be a new cancer treatment method. Puncture originally refers to inserting a hollow needle into the body to suck out internal fluid, but the puncture method referred to here particularly refers to methods for examining and treating cancer or tumor tissue.

The testing method generally refers to what is called a puncture biopsy method, which is a method of collecting tissue to be tested from a living body and testing it by scraping tissue that is thought to be cancer or tumor with the tip of a needle. The tissue attached to the needle tip is taken out of the body and examined. It has been pointed out that a problem with this needle biopsy method is that there is a risk that cancer or tumor tissue attached to the needle tip may be scattered onto other normal tissue when the needle is removed. No countermeasures have been taken. It has been pointed out that another problem is that bleeding tends to occur and it is often difficult to stop the bleeding. In other words, unlike vascular occlusion therapy, which involves inserting a catheter through a blood vessel, the puncture method involves inserting a needle directly from the body surface to reach the target tissue, which results in bleeding from the area injured by the needle. . If the injured area is normal, even if bleeding occurs, it will stop spontaneously, the injured blood vessel will be repaired in a relatively short period of time, and there will be no adverse effects after the puncture procedure. However, when puncturing cancer or tumor tissue, typically
Not only the cancer or tumor tissue itself, but also nearby tissues often have insufficient hemostatic ability and ability to repair damaged blood vessels, resulting in increased bleeding tendency, difficulty in hemostasis, and the possibility of large amounts of bleeding. In such cases, the patient may have no choice but to abandon the puncture procedure. In particular, puncture of the viscera is currently not performed very often due to bleeding. Therefore, when performing puncture, hemostasis is a major problem, and it is often necessary to remove the needle while stopping the bleeding. The method of hemostasis in this case is usually to remove the needle while gradually injecting an aqueous solution of thrombin, but thrombin itself has insufficient hemostasis ability, and thrombin is in liquid form. Hemostasis has not been achieved satisfactorily because the bleeding tends to flow from the area requiring repair to other areas. On the other hand, the treatment method refers to a treatment method in which an anticancer substance is directly injected onto a target cancer or tumor tissue through a puncture needle. Currently, when injecting drugs through a puncture needle, the puncture needle used is usually 19G or more.
Because it is extremely thin, about 23G, it is only used to inject solutions such as antibiotics. However, when an anticancer substance is injected in the form of a solution, sufficient effects cannot be expected because it immediately flows out to areas other than the target tissue and the significance of direct local administration is lost. For this reason, it is possible to spray on the target tissue and near the puncture needle path through a puncture needle with a diameter that is normally used.
Moreover, there is a long-awaited demand for a preparation that can adhere to the sprayed site and remain there for a long time to release anticancer substances in a sustained manner.

In view of the above-mentioned current situation, the present inventors have proposed that when used in vascular occlusion therapy, the present inventors can rapidly occlude blood vessels.
It can be performed reliably and has the ability to effectively cause anti-cancer substances to act on cancer or tumor tissue for a long time, and when used for puncture, it quickly and reliably prevents bleeding during the procedure. It can suppress and repair damaged blood vessels at an early stage, and is anti-cancer not only for the cancer or tumor tissue itself but also for cancer or tumor tissue that is likely to be dispersed near the puncture needle route during the biopsy procedure. As a result of intensive research with the aim of developing a drug that has the ability to effectively cause cancer-causing substances to act for a long period of time, surprisingly, a structure immobilized with an anti-cancer substance and a blood coagulant was found to be effective against blood vessels. The fact that it has excellent occlusion ability, hemostasis ability, and damaged blood vessel repair ability.Furthermore, when it is injected from outside the body and reaches the body, it quickly attaches to the area it reaches, that is, cancer or tumor tissue, or their nearby areas, and stays there. The present invention was achieved based on the discovery that anti-cancer substances are not washed away by body fluids and body fluids, and that anti-cancer substances are sustainedly released over a long period of time.

That is, the present invention is a structure in which an anticancer substance and a blood coagulant are immobilized on a structure made of a bioabsorbable substance and having a shape such as a fiber aggregate, sponge, powder, monofilament, film, or microcapsule. It is an anti-cancer substance sustained release embolic agent.

In the present invention, an embolic agent refers to a preparation that is injected into a blood vessel to stop blood flow, and a preparation that fills a damaged area of a blood vessel to stop bleeding and repair the damaged blood vessel. Since it is necessary to quickly pass through the puncture needle, it is preferable that the material can be finely suspended in a medium such as water.

In the present invention, the materials constituting the structure are often injected into the body and cannot be recovered outside the body. In addition, foreign substances that remain in the body have a negative effect on the cure of cancer or tumors, so they must be bioabsorbable, and in the present invention, for example, gelatin, chitin, collagen, polyglycolic acid, -Lactic acid copolymer, polyglutamic acid, amylose, etc., among which gelatin, amylose, and chitin are preferably used.

The anticancer substance as used in the present invention refers to a substance generally called an anticancer agent, an anticancer agent, or an antitumor agent, and a substance generally called an immunostimulant or an immunostimulant, and the former substance includes: For example, nitrogen mustard, nitromine, chlorambucil, cyclophosphamide, melphalan, uracil mustard, mannomustine, dopan, etc.
BCNU, triethylenemelamine, thio-TEPA,
Aza-TEPA, Trenimon, Isoprocyone,
Alkylating agents such as busulfan, dimethylmyleran, piposulfan, ethoglucide, epoxypropidine, epoxypiperazine, hexamethylmelamine, dibromomannitol, pipobroman, aminopritene, methotrexate, guanine, 8-azaganine, 6-mercaptopurine, azathioprine, uracil , 5-fluorouracil, cytarabine, azaserine, antimetabolites such as diazomycin, actinomycin D, cyclomycin, mitomycin C, daunomycin,
Antibiotics such as pleomycin, chromomycin, cardinophyllin, synthetic agents such as 5-HP, IQ-1, plant ingredients such as thioteba, cyclophosphamide, doxorubicin, daunorubicin, neocarcinostane, Hg-hematoporphyrin, Co―
Examples include protoporphyrin, stilbestrol, hydroxyurea, procarbazine, methylglyoxal-bis-guanylhydrazone, L-asparaginase, and these may be used alone or in combination of two or more. A common method is to select and use a combination of one alkylating agent, one antimetabolite, and one antidepressant; a combination of endoxan, 5-fluorouracil, mitomycin, or pleomycin is common. are particularly common.
Examples of the latter substances include chimic hormone and its related substances, BCG, cell wall skeleton and its methanol-insoluble fraction, bacteria and bacterial components such as Corynebacterium parvum and OK-432, pisibanil, reentinan, SPG, mannan,
Polysaccharides such as levan and glucan, muramyldipept and its derivatives, levamisole, bestatin,
Vaccines such as isoprinosine, NPT15392, azimecline, transfer factor, lymphokine, immunoRNA, interferon and its inducer, Maruyama vaccine, etc. may be used alone, or two or more of these may be used. It is generally used in combination with the aforementioned anticancer agents or substances called anticancer agents or antitumor agents.

The blood coagulant used in the present invention includes, for example, blood coagulation factor,
Examples include factor factor, factor 1, factor 1, factor 1, factor 1, factor 1, factor XI, factor XII, factor 1, prekallikrene, polymeric kininogen, thrombin, and the like. These can be used alone or in combination of two or more. In the present invention, blood coagulation factor (hereinafter abbreviated as F) and thrombin are particularly preferably used. F is called a fibrin stabilizing factor, and is a factor that promotes the production of stabilized fibrin due to isopeptide bonds between fibrin molecules. F is isolated from the blood or placenta of humans, cows, etc., but when applied to humans, it is preferable to use human-derived F. Thrombin is a proteolytic enzyme that can convert fibrinogen to fibrin. Thrombin is isolated from blood of humans, cows, pigs, etc., but when applied to humans, it is preferable to use human thrombin.

The anticancer substance and blood coagulant used in the present invention are:
binding or adsorption to the structure;
Alternatively, it can be immobilized by encapsulating it. The attachment of anti-cancer substances and blood clotting agents to structures is described, for example, in O. Zaborsky, “Immobilized
Conventionally known covalent bonding methods and ionic bonding methods such as those described in "Enzymes" CRC Press. 1973 can be used, and for adsorption,
Similarly, a physical adsorption method or an encapsulation method can be adopted, and for encapsulation, a method of microencapsulating using a known microencapsulation method using the material constituting the structure as an outer wall can be adopted. can.

When producing the embolic agent of the present invention, an anticancer substance and a blood coagulant can be bound to the structure, for example, in the following manner. In other words, when an anticancer substance or a blood coagulant has a functional group capable of forming a covalent bond or an ionic bond, such as an amino group or a carboxyl group, a solution containing these functional groups can form a covalent bond or an ionic bond. By treating a structure having a functional group capable of ionically bonding, it is possible to perform immobilization through desired bonding. In addition, in this case, if the structure has no or only a few functional groups that can covalently or ionically bond with the functional groups of the anticancer substance or blood coagulant, those functional groups are added to the structure in advance. After being introduced by chemical reaction, anti-cancer substances and blood clotting agents can be attached to the structure. If the anticancer substance or blood coagulant does not have a functional group, it is possible to use it after introducing a functional group through a chemical reaction as in the case described above, but in many cases this is not possible. This method is preferably not employed, since the drug properties of the anticancer substance or blood coagulant will be damaged due to the chemical reaction. In the case of covalent bonding, it is preferable to use a dehydration condensation agent such as dicyclohexylcarbodiimide, 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide-meth-p-toluenesulfonate.

Furthermore, when producing the embolic agent of the present invention, an anticancer substance and a blood coagulant can be adsorbed onto a structure by a physical adsorption method, an entrapment method, or the like as follows. That is, by dissolving or suspending an anticancer substance and a blood coagulant in a solvent that can wet the structure, and treating the structure with this solution, the anticancer substance and blood coagulant can be physically adsorbed. can. The entrapment method is a method in which anticancer substances and blood coagulants are encapsulated in a fine gel lattice so that they cannot be released. This adsorption method and entrapment method are effective for any combination of anticancer substances, blood coagulants, and structures, and are preferred in the present invention because they are simple and rarely impair the properties as a drug.

In order to produce the embolic agent of the present invention, in addition to the method of binding or adsorbing an anticancer substance and a blood coagulant to a structure as described above, first, the anticancer substance and the blood coagulant are added to the material itself before being processed into a structure. It can also be manufactured by combining or adsorbing the anti-cancer substance and the blood coagulant, and then processing the material to which the anti-cancer substance and the blood coagulant are combined or adsorbed into a structure. For example, the embolic agent of the present invention can be produced by obtaining a structure using a polymeric substance in which an anticancer substance is bound or adsorbed in advance.

When immobilizing an anticancer substance and a blood coagulant by any of the above methods, the anticancer substance and the blood coagulant may be immobilized at the same time, or the anticancer substance may be immobilized first. After that, the blood coagulant may be subsequently immobilized, or vice versa.

When producing the embolic agent of the present invention, in addition to anticancer substances and blood coagulants, protease inhibitors such as antiplasmin, albumin, and α ₂ -macroglobulin, and blood proteins such as ceruloplasmin, haptoglobin, and cold insoluble globulin are used. Proteins, fibronectin, antibiotics, etc. can be immobilized on the structure. Antiplasmin is an inhibitor of plasmin, which is a fibrinolytic enzyme, and therefore exerts its effect by inhibiting plasmin. In the present invention, as antiplasmin, for example, ε-aminocaproic acid, tranexamic acid, etc. are preferably used.

The embolic agent of the present invention is preferably used in the treatment of cancer or tumors, and is particularly preferably used in vascular occlusion methods, puncture methods, and the like.

The embolic agent of the present invention can be applied to vascular occlusion therapy by, for example, passing the tip of a vascular catheter through a blood vessel to the feeding artery to the target cancer or tumor tissue, and suspending the embolic agent in physiological saline or the like from the other end. It is made by injecting. At this time, a portion of the embolic agent reaches the target tissue and its neighboring tissues, adheres to the target tissue, and remains there, and a portion remains inside the feeding blood vessel, quickly occluding the blood vessel. Also, occluded blood vessels do not recanalize. Furthermore, the embolic agent that remains on the target tissue, its neighboring tissue, and the vascular occlusion acts as a sustained release agent for anticancer substances, and the anticancer substance is sustainedly released from this agent and localized to the cancer or tumor tissue. It acts over a long period of time to accelerate and ensure necrosis.
This means that when the embolic agent of the present invention is used, it is possible to perform vascular occlusion therapy quickly and accurately, and at the same time, since this embolic agent acts as a sustained release agent for anticancer substances, it is possible to perform vascular occlusion therapy quickly and accurately. This means that chemotherapy, which could not be performed in parallel, can now be performed in parallel.

Application of the embolic agent of the present invention to the puncture method is, for example, after injecting a required amount of embolic agent suspended in physiological saline through a puncture needle that has reached the target cancer or tumor tissue, and then This is done by removing the needle while gradually injecting this suspension.
This allows the embolic agent to be dispersed onto the target tissue and the tissue near the path of the puncture needle and remains attached to the site. The embolic agent that remains on the target tissue immediately stops bleeding and repairs the vascular damage site, while also acting as a sustained release agent for anti-cancer substances, allowing them to be effectively released against cancer or tumor tissue for a long time. It will work. On the other hand, the embolic agent injected at the time of needle removal, dispersed and adhered to the tissue near the puncture needle path, immediately suppresses bleeding from the damaged site of the nearby blood vessel and repairs it.
Moreover, it acts effectively for a long period of time on cancer or tumor tissue dispersed near the needle route.

As described above, the embolic agent of the present invention is used in procedures such as vascular occlusion therapy and puncture, and has both excellent performance as an embolic agent and performance as an anticancer substance sustained release preparation.

Furthermore, the embolic agent of the present invention exhibits great effects even when used simply as an anticancer substance sustained release agent.
In other words, even if conventional sustained-release anti-cancer agents are administered locally by being sprayed onto cancerous tissue exposed through incisional surgery, the sustained-release agents may not be effective because of blood, body fluids, etc. After being sprayed, it immediately washes away from the local area, making it difficult to expect an effect, but if the embolic agent of the present invention is similarly administered, it will not stick to the local area immediately and will not wash away.

The present invention will be explained in more detail with reference to Examples below.

Example 1 200 mg of powdered Zelform (absorbable powdered gelatin, manufactured by Nippon Upjiyon) was dissolved in 4 ml of water with 1 bottle of fibrogamin aqueous solution [1 bottle of concentrated dry preparation of human F (manufactured by Hoechst). ] 4 ml, physiological saline solution of thrombin (1 bottle of concentrated dry human thrombin preparation (Midori Juji) dissolved in 5 ml of physiological saline.] 4 ml and mitomycin C aqueous solution [20 mg/4 ml] 4 ml mixed solution at room temperature. After soaking for 5 minutes, freeze-dry at -30℃ for 15 hours and
An immobilized powder Zelform on which thrombin and mitomycin C were immobilized was obtained.

On the other hand, make a loop using a 34 cm Siriyun medical tube with an inner diameter of 4 mm, and place it in a room at 2°C and add 2 ml of ACD preserved blood to 10 wt% aqueous solution of Cacl ₂ .
ml was added, and then 20 mg of the previously prepared immobilized powder Zelform was added, and the mixture was rotated at 16 revolutions/min on a rotary plate having an inclination of 23 degrees. One minute after the start of rotation, formation of a clot was observed in the blood. At this point, stop the rotation, and 1 hour after stopping, use a paper disk (manufactured by Toyo Seisakusho, paper disk for antibiotic testing, diameter 8 mm).
Thoroughly immerse or adhere to the blood or clot in the loop, use this as a sample, and use it as a test bacterium.
Using Bacillus Subtilis ATCC6633, the size of the inhibition circle produced by the cylindrical plate method was determined, and the blood concentration of mitomycin C at 1 hour was determined from the size of the inhibition circle, and was found to be 10 μg/mg. Similarly, 5th hour, 10th hour, 24th hour, 1.5th day,
The blood concentrations on the second and third days were 15 μg/mg, 21 μg/mg, 41 μg/mg, and 82 μg/mg, respectively.
mg, 112 μg/mg, and 198 μg/mg.

Comparative Example 1 The same rotating loop as in Example 1 was carried out using 20 mg of mitomycin-immobilized Zelform obtained in the same manner as in Example 1 except that F and thrombin were not used. A blood coagulation test and measurement of mitomycin concentration in the blood revealed that no clot was formed even after 3 hours had passed after the start of rotation, and the blood concentration of mitomycin C at this point was 600 μg/ml. This result shows that most mitomycin C
This indicates that the drug is released into the blood within a certain period of time, meaning that it does not have sustained release properties.

Example 2 200 mg of powdered Zelform (absorbable powdered gelatin, manufactured by Nippon Upjiyon) was dissolved in 8 ml of water with 1 bottle of fibrogamin aqueous solution [1 bottle of concentrated dry preparation of human F (manufactured by Hoechst). ]3 to 4 ml at room temperature
After soaking for 1 minute, freeze-drying was performed at -30°C for 15 hours. Next, this material was treated with 5-fluorouracil.
After immersing 25 mg in 4 ml of dimethylformamide at room temperature for 5 minutes, freeze-drying at -30°C for 15 hours to obtain an immobilized powder of F and 5-fluorouracil. I got it. When a blood coagulation test using a rotating loop was conducted using 20 mg of this material in the same manner as in Example 1, formation of a blood clot was observed 1.5 minutes after the start of rotation. At this point, the rotation is stopped, and 5 hours after stopping,
At 10th hour, 24th hour, 1.5th day, 2nd day, and 3rd day, sample 50 μg of blood or clot, freeze-dry each sample, and burn in an oxygen flask using a mixture of sodium hydroxide and water as an absorption liquid. The amount of fluorine contained in each sample is determined by the
-The weight of fluorouracil was measured and the blood concentration of 5-fluorouracil at each sampling was determined to be 120 μg/mg, 170 μg/mg, respectively.
380μg/mg, 780μg/mg, 1010μg/mg, 1620μ
g/mg.

Example 3 Immobilized powdered chitin on which F, thrombin, and mitomycin C were immobilized was obtained in the same manner as in Example 1, except that powdered chitin (manufactured by Kyowa Yushi Co., Ltd., molecular weight: 1 million) was used instead of powdered Zelform. When 20 mg of this product was used and blood coagulation tests and blood concentration measurements were performed in the same manner as in Example 1, a clot was formed one minute after the start of rotation, and the fluidity of the blood in the tube was lost. Also, the 1st hour, 5th hour, 10th hour,
The concentration of mitomycin C in the blood at 24 hours, 1.5 days, 2 days, and 30 days was 8μ, respectively.
g/ml, 14 μg/ml, 19 μg/ml, 38 μg/ml,
They were 78 μg/ml, 108 μg/ml, and 178 μg/ml.

Example 4 One sheet (2.5 cm x 5 cm x 0.5 cm) of Sponzel (absorbable sponge-like gelatin, manufactured by Yamanouchi Pharmaceutical) was mixed with a physiological saline solution of thrombin (1 bottle dissolved in 10 ml).
After immersing in a solution of 200 mg of bleomycin and 20 mg of 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide-meth-p-toluenesulfonate dissolved in 10 ml at room temperature for 5 minutes, -30
Freeze-drying was performed at ℃ for 15 hours to obtain immobilized sponzel on which thrombin and bleomycin were immobilized. Using 20 mg of this product, a blood coagulation test using a rotating loop and measurement of the blood concentration of pleomycin were performed in the same manner as in Example 1, and the formation of a clot was observed 1 minute after the start of rotation. After stopping the rotation, the blood concentration of bleomycin at 1 hour, 5 hours, 10 hours, 24 hours, 1.5 days, 2 days, and 3 days was 5 μg/mg, 7 μg/mg, respectively.
11μg/mg, 21μg/mg, 41μg/mg, 62μg/mg,
It was 123 μg/mg.

Claims (1)

[Claims] 1. An anticancer substance in which an anticancer substance and a blood coagulant are immobilized on a structure made of a bioabsorbable substance in the shape of a fiber aggregate, sponge, powder, monofilament, film, microcapsule, etc. Sustained-release embolic agents.