JP2010024170A - Local delivery-reinforcing agent for anti-malignant tumor agent to tumor tissue - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a local delivery-reinforcing agent for an anti-malignant tumor agent to a tumor tissue. <P>SOLUTION: Provided is the local delivery-reinforcing agent for reinforcing the delivery of the effect of the anti-malignant tumor agent to the tumor tissue, using a nitrogen monoxide (NO) donor such as nitroglycerol or isosorbide dinitrate as an active ingredient together with the anti-malignant tumor agent. In more detail, the nitrogen monoxide donor is especially coated and administered on a malignant tumor site as an external preparation such as an ointment. Thereby, the leakiness of an agent from the lumens of tumor blood vessels into the tumor tissue is enhanced to reinforce the local delivery of the agent such as the anti-malignant tumor agent, namely reinforcing the accumulation of the agent in the tumor tissues. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a local delivery enhancer for tumor tissue containing a nitric oxide (NO) donor as an active ingredient. More specifically, the present invention relates to an agent that enhances local delivery of an antineoplastic agent to tumor tissue by enhancing local delivery of the antineoplastic agent, that is, accumulation of the agent in tumor tissue.

  The inventors of the present invention have previously accumulated substances and oils containing high biocompatible polymer drugs, micellized preparations, and liposomal preparations in tumor tissues as compared with normal tissues, and have increased for a long time. It was discovered that the concentration was maintained (Enhanced Permeability and Retention effect, EPR effect) (Non-patent Document 1). That is, in tumor tissue, (1) increase in intravascular tumor density due to vigorous neovascularization, (2) increase in vascular permeability by various vasoactive factors, and (3) retrograde properties of macromolecular substances through tumor blood vessels. (4) Biocompatible high-molecular substances (such as the above-mentioned drugs) and oil-based ultrafine particles administered into the blood as a result of failure to recover macromolecules due to failure of lymphatic recovery Becomes highly accumulated in the tumor tissue. In order to utilize this principle, it is possible to greatly extend the blood half-life by polymerizing the drug, and as a result, the drug is selectively and locally accumulated in the tumor site very efficiently. On the other hand, accumulation in a normal normal tissue is extremely low. It has been clarified that this makes it possible to exert a stronger anti-malignant tumor action at the same time as significantly reducing side effects (Non-patent Document 2).

  On the other hand, nitric oxide (NO), one of the endogenous substances that control blood pressure, that is, endothelial-derived relaxing factor (EDRF), guanylate cyclase of vascular smooth muscle. It is known to relax blood vessels by activation (Non-patent Document 3).

Since the present inventors immediately react with NO and superoxide (O ₂ ⁻ ) and consume NO, the NO level in the blood is lowered, it becomes difficult to maintain normal blood pressure, and high blood pressure may be caused. It is clear.

Further, it has been found that NO synthesized by NO synthase (NOS) induces tumor blood vessel permeability (enhanced leakage) in solid tumors (Non-patent Document 4). Also, EPR effect is an inhibitor of ^NOS L-N ω - nitro -L- arginine methyl ester (L-NAME) and NO is the scavenger 2-phenyl-4,4,5,5-tetramethyl-imidazoline It has also been found that it is suppressed by administration of -1-oxyl 3-oxide (PTIO) (Non-patent Documents 5 and 6).

  Tumors, on the other hand, produce angiogenic factors such as vascular endothelial growth factor and fibroblast growth factor, and produce tumor blood vessels, while these factors (EDRF) increase NO production and increase vascular permeability, As a result, it has vigorous blood vessel formation ability to acquire nutrition, oxygen and the like through blood (Non-patent Document 7).

By proceeding with this concept, GTN, which is NO (and its donor), can be applied from the surface of the tumor by application to relax excess tumor blood vessels and accumulate many drugs locally in the tumor. Thought.
Matsumura, Y. and Maeda, H .: Cancer Res. 46: 6387-92, 1989 Maeda H. J Controlled Release, 19, 315-324, 1992 Moncada S et al. Pharm Rev, 43, 109-142, 1991 Doi K et al. Cancer 77, 1598-604, 1996, Br J Cancer, 80, 1945-54, 1999 Maeda H et al. Jpn J Cancer Res 85, 331334, 1994, Wu J et al. Cancer Res 58, 159-65, 1998 Iyer AK et al. Drug Discov Today, 11, 812-818, 2006 Folkman J, Nat Med 1: 27, 1995

  Malignant tumors are currently one of the most difficult diseases to treat and cure among human diseases, and various anti-neoplastic agents have been developed as drugs, but there is no definitive one. It is. In addition, side effects of antineoplastic agents are always a problem. Under such circumstances, enhancing the deliverability of a powerful anti-neoplastic agent to tumor tissue is not only one effective means in the treatment and treatment of malignant tumors, but also the amount of anti-neoplastic agent This reduces the side effects of antineoplastic agents. Accordingly, there is a need for an agent that enhances the delivery of excellent antineoplastic agents to tumor tissue.

  Against this background, when the present inventors applied a nitric oxide (NO) donor such as nitroglycerin together with an antineoplastic agent, surprisingly, the antineoplastic agent malignant tumor We have found that delivery to tissues is enhanced. More particularly, the present invention enhances local delivery of antineoplastic agents, ie, the accumulation of agents in tumor tissue, by applying a nitric oxide (NO) donor together with the antineoplastic agent. can do. Such an effect of enhancing delivery of a malignant tumor agent to a tumor site has been found by the present inventors for the first time.

  Accordingly, it is an object of the present invention to provide an agent for enhancing local delivery of an antineoplastic agent containing a nitric oxide (NO) donor as an active ingredient to a malignant tumor tissue.

  Hereinafter, the present invention will be described in more detail.

  In the present specification, “nitrogen monoxide (NO) donor (donor)” is a general term for drugs that can continuously release nitric oxide (NO) in a living body, of which nitro group is released, It also includes those that are reduced to NO by enzymes in the body. Typical examples include nitrate esters such as nitroglycerin (GTN), glyceryl mononitrate, glyceryl dinitrate, isosorbide mononitrate (ISMN), and isosorbide dinitrate (ISDN). Others include, for example, sodium nitroprusside, molsidomine and the like. In the present invention, the most preferable is a nitroglycerin preparation.

  In practicing the present invention, nitric oxide (NO) donors are usually applied in the form of external preparations such as ointments, gel ointments, creams, solutions, patches, suppositories, sprays, intranasal, etc. Any dosage form may be used as long as it is an external preparation applicable depending on the site of the tumor. A preferable application form includes applying to the affected area as an ointment or spraying.

  When applying as an ointment, the ointment is prepared by a conventional method. Ointments and external preparations for skin are semi-solid preparations, which are composed of an active ingredient and a base. The active ingredient is present in the base, and the active ingredient is dissolved in the base. Is distributed. The base includes a hydrophobic base that is an oleaginous base and a hydrophilic base such as an emulsion base, a water-soluble base, and a suspending base. Hydrophobic bases include petrolatum, paraffin, plastibase, silicone, vegetable oil, waxes, and emulsion bases include polyethylene glycol esters, lanolin, and hydrophilic petrolatum. , Such as tuna gogol, and the suspending bases include gels, jellies and gels based on cellulose.

  In the case of nitroglycerin, which is a representative example of the nitric oxide (NO) donor used in the present invention, an ointment based on petrolatum and lanolin can be easily obtained. That is, there is a nitroglycerin ointment that is a treatment / prevention agent for acute heart failure, angina pectoris, myocardial infarction, for example, Bathorator (Sanwa Chemical).

  Furthermore, as described above, the external preparation containing a nitric oxide (NO) donor used in the present invention can be used in a different dosage form depending on the site of the malignant tumor to be treated / treated. Sprays, patches, suppositories, liquids, and the like can also be used. For these, various commercially available patch agents (for example, nitroderm, myris, sydrene, batholator, Meditrans, minitro, etc.) can be used.

  There is no particular limitation as long as it is a malignant tumor to which an external preparation containing a nitric oxide (NO) donor used in the present invention can be applied, but for example, breast cancer, esophageal cancer, colon, and rectum are usually applied. Gastrointestinal cancer such as colorectal cancer including cancer, urinary cancer such as bladder cancer, bronchopulmonary cancer, respiratory tract cancer, genital cancer such as uterine cancer, vaginal cancer, skin cancer, Examples include superficial tumors such as cancer on the eyes, abdomen, and chest wall.

  Examples of breast cancer include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and breast lobular non-invasive carcinoma in situ.

  Examples of cancers of the respiratory tract include, but are not limited to, small cell lung cancer, non-small cell lung cancer, and bronchial adenoma and pleuropulmonary blastoma.

  Male reproductive organ tumors include, but are not limited to, prostate and testicular cancer. Tumors of the female reproductive tract include, but are not limited to, endometrium, cervix, ovary, vagina, and vulva cancer, and uterine sarcoma.

  Gastrointestinal tumors include, but are not limited to, anus, large intestine, colorectal, esophagus, gallbladder, stomach, pancreas, rectum, small intestine, and salivary gland cancer.

  Tumors of the urinary tract include, but are not limited to, bladder, penis, kidney, kidney disc, ureter, urethra, and human papillary kidney cancer.

  Eye cancers include, but are not limited to intraocular melanoma and retinoblastoma.

  Examples of liver cancer include hepatocellular carcinoma (hepatocellular carcinoma with or without a fiber lamina), bile duct cancer (intrahepatic cholangiocarcinoma), and hepatocellular carcinoma, bile duct Including, but not limited to, mixed types.

  Skin cancers include, but are not limited to squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancer.

  Head and neck cancers include, but are not limited to, pharynx, hypopharynx, nasopharynx, oropharyngeal cancer, lip and oral cancer, and squamous cells. Hematological cancers include, but are not limited to, various types of leukemia as well as lymphomas such as AIDS-related lymphoma, non-Hodgkin lymphoma, cutaneous T-cell lymphoma, Burkitt lymphoma, Hodgkin's disease, and central nervous system lymphoma .

  Although these diseases have been characterized in humans, there are similar etiologies in other mammals and can be treated by administering a local delivery enhancer of the antineoplastic agent of the present invention. Agents for local delivery of antineoplastic agents include not only human but also mammalian applications.

  When using the agent for local delivery of an anti-neoplastic agent of the present invention to a tissue of a malignant tumor, the anti-neoplastic agent for treating / treating the malignant tumor is not particularly limited from the mechanism of the present invention. It includes not only currently used antineoplastic agents but also drugs that are under development or that will be developed in the future. There may be one anti-neoplastic agent in carrying out the present invention, or two or more.

  The anti-neoplastic agent in carrying out the present invention may be any drug as described above, and is not particularly limited. Among drugs having an anti-malignant tumor action, a tumor of an anti-neoplastic agent is a polymer drug. It shows the effect of enhancing delivery to a site. For example, the effect is shown in the case of a drug such as PEG-ZnPP used in Experimental Example 5 below. PEG-ZnPP (polyethylene glycol-linked zinc protoporphine) is a drug that is expected to be effective in Gleevec-resistant chronic myeloid leukemia (CML), mast cell leukemia (MCL), breast cancer, colon cancer, etc. It is a drug that also has an action of enhancing the antineoplastic effect of an antineoplastic agent in combination with an antineoplastic agent. Furthermore, these high molecular weight drugs include PEGylated interferon, micellized preparation, and liposome preparation, but any other high molecular weight anticancer agent may be used.

  Examples of polymer drugs include, for example, NK012 (Nippon Kayaku), which is a PEG-poly (asp) block copolymer micelle containing the active part (SN38) of Smanx (registered trademark), doxorubicin (Doxol), and camtracin. Conjugated polyglutamic acid [Xyoptax, abbreviation PG-TXL CODE CT-2103] (Cell Therapeutic) and the like can be mentioned, but they are not limited, and are not limited to, but are being developed or are expected to be developed in the future. It may be a malignant tumor agent.

  Representative examples of currently used antineoplastic agents include alkylating agents such as ifosfamide, cyclophosphamide, dacarbazine, temozolomide, melphalan, busulfan, and nimustine, enocitabine, capecitabine, carmofur, Antimetabolites such as gemcitabine, cytarabine, tegafur uracil, tegafur, gimeracil, oteracil potassium, fluorouracil methotrexate, irinotecan, etoposide, sobuzoxan, docetaxel, nogitecan, paclitaxel, vinorelbine, vinsulfine, vinsulfine, vindecine D, aclarubicin, idarubicin, epirubicin, daunorubicin, doxorubicin, pirarubicin, bleomycin, pep Antineoplastic antibiotics such as mycin, mitomycin C, mitoxantrone, platinum preparations such as oxariblastin, carboplastin, cisplastin, nedaplastin, anastrozole, exemestane, itinylelastradiol, tamoxifen, letrozole, etc. Hormonal agents, interferon α, interferon β, interferon γ, interleukin 2, dry BCG, lentinan and other biological response modifiers, imatinib, gefitinib, temivarotene, trastuzumab, tretinoin, gemtuzumab ozogamicin, portezomib, Molecular targeting drugs such as rituximab, high molecular weight preparations such as Smanx, PEGylated interferon, liposome preparations such as doxil and NK012 (active part of camtracin (SN38)) Examples thereof include micelle preparations such as PEG-poly (asp) block copolymer micelles), and polymer anticancer agents such as OPAXIO (capritaxel-bound polyglutamic acid).

  These antineoplastic agents are appropriately selected and used depending on the type, site, severity, physical strength, age, etc. of the malignant tumor. For example, for lung cancer, cisplatin and carboplatin and other platinum drugs combined with other antineoplastic agents, for esophageal cancer, cisplatin and oluracil, etc., for gastric cancer, cisplatin, Irinotecan is a combination of fluorouracil, irinotecan, oxaplatin, and tegafur uracil for colorectal cancer, gemcitabine for pancreatic cancer, and combinations of cyclophosphamide, metrequidate, fluorouracil, doxorubicin, etc. for breast cancer. For cervical cancer, a combination based on cisplastin is used, and for endometrial cancer, a combination of cisplastin, doxorubicin, cyclophosphamide, and the like is used.

  Any of the above-mentioned antineoplastic agents shows an effect of enhancing the delivery of the antineoplastic agent to the tumor tissue. For example, in the case of an anti-neoplastic agent such as an anti-neoplastic antibiotic, a drug having a molecular weight of 848.33 It is.

  Aclarubicin binds to DNA of malignant tumor cells and is said to strongly inhibit nucleic acid synthesis, especially RNA synthesis, and is an anti-neoplastic agent useful for the treatment and treatment of breast cancer, stomach cancer, lung cancer, ovarian cancer, etc. is there.

The dosage of these antineoplastic agents takes into account the severity of the malignant tumor, the appearance of side effects, the patient's physical strength, age, etc., but the dosage per body surface area is usually determined. For example, in the case of CPT-P therapy, Irinotekarin 60 mg / ^{m 2} (body surface area) per day, has a cisplatin 60 mg / ^{m 2,} in the CAP therapy, cyclophosphamide 500 mg / ^{m 2,} doxorubicin It has a 30 to 50 mg / ^{m 2,} in the case of CP therapy, cyclophosphamide 800~900mg / ^{m 2,} and has a cisplatin 60~75mg / ^{m 2.}

There is no particular limitation on the dose of the “nitrogen monoxide (NO) donor” of the present invention. It may vary depending on the severity of the malignant tumor, the appearance of side effects, the patient's physical strength, age, and the like. When nitroglycerin is used, a commercially available nitroglycerin-containing ointment is applied directly to the site of malignant tumor development. In this case, for surface cancers such as breast cancer, skin cancer, colorectal cancer, uterine cancer, and bladder cancer, the purpose can be achieved by directly applying such ointment and cream. If esophageal cancer, colorectal cancer, etc. cannot be applied directly, such drugs can be administered directly to the affected area using an instrument such as an endoscope, or by spraying. When nitroglycerin is used in patients with possible hypotension, careful attention should be paid to its dosage. The ratio of use of the present invention with the anticancer agent used by the nitric oxide (NO) donor is not related and cannot be generally stated, but, for example, 0.01 to 1.0 mg / cm ² is applied per tumor surface. That is.

  Nitroglycerin-containing ointment is, for example, a batholator ointment, which is a yellowish white ointment containing lactose hydrate, hydrolanolin, white petrolatum as additives, and 20 mg of nitroglycerin effective per 1 g Contains as a component.

  When preparing such an ointment, it can be carried out by an ordinary method for producing an ointment. For example, an appropriate amount of 20 g of nitroglycerin, white petrolatum of pharmacopoeia and lanolin is added, and an ointment containing nitroglycerin as an active ingredient is prepared as 1000 g. can do.

  In addition, external preparations other than ointments, solutions, lotions, gels, sprays (sprays), patches, creams, and the like can be easily prepared by methods that are usually performed in this technical field.

  When the external preparation of the present invention is administered, it may be administered at the same time as various appropriate antineoplastic agents, or may be administered at a slightly different time interval before or after the administration of the anticancer agent. Good.

  Furthermore, the combination of the antineoplastic agent according to the present invention and a nitric oxide (NO) donor may be used as needed by applying an external preparation such as an ointment according to the present invention to an anti-neoplastic agent and a drug pack, kit, or patient pack. It may be provided in the form.

  The present invention uses various nitric oxide (NO) donors, such as nitroglycerin, together with an antineoplastic agent, whereby various anti-neoplastic agents, particularly micelles, liposomes, high-molecular-weight drug-binding, and protein-binding anticancers. It enhances the delivery of the agent to the tumor site, resulting in enhanced accumulation within the tumor tissue. These effects are extremely useful for the treatment and treatment of malignant tumors, and can reduce the dosage of various anti-neoplastic agents, and can significantly reduce the side effects peculiar to anti-neoplastic agents. Therefore, the value of the present invention is high.

  Furthermore, especially this invention has a higher effect by using together with the high molecular agent with an antineoplastic effect. It has been found that enhancing the effect of local delivery of antineoplastic agents increases the drug delivery of nitric oxide donors such as GTN to surface tumors and enhances the therapeutic effect of malignant tumors. high.

  In order to prove the above effect, the results of animal experiments conducted by the present inventors are shown below.

  Hereinafter, the present invention will be specifically described with reference to examples and experimental examples of the present invention. However, the present invention is not limited to these examples.

Example 1: GTN of Sarcoma 180 (Sarcoma 180) of coating group and non-applied group into tumors Evans Blue - the dorsal subcutaneous tumor tissue Accumulation difference 6 week-old male ddY mice albumin, 2 × 10 ⁶ cells Mouse sarcoma Sarcoma 180 (S-180) tumor cells were transplanted to produce solid tumors. About 2 weeks later, a mouse having a tumor diameter of 8 to 15 mm was selected, the tumor part was shaved, and 2% GTN ointment (Basolator Ointment ^{(registered trademark)} , Sanwa Chemical, Nagoya) was applied and administered as GTN. After applying to the tumor only with a curved plastic with a hole of the same diameter as the tumor diameter in an amount of 1.0 mg), 0.1 ml of Evans Blue (EB) physiological saline solution is immediately tailored to 10 mg / kg. It was injected intravenously. After administration of EB, 30 minutes, 1, 4, 6 and 24 hours later, the mouse was sacrificed under anesthesia, the tumor was excised, and its weight was measured, followed by extraction of EB. The amount of EB accumulated was quantified by absorbance at 620 nm after adding about 4 ml of formamide in a test tube and elution extraction at 60 ° C. for 48 hours. In addition, in order to examine the concentration correlation between the applied dose of GTN and the amount of tumor tissue accumulation, 2% GTN ointment was diluted with white petrolatum so that the applied dose of GTN was 0.1, 0.01 and 0.001 mg. A GTN ointment was made. 50 mg of these ointments were applied to only the tumor site as described above, then administered at 10 mg / kg EB, sacrificed 4 hours later, EB extracted, and compared. The effect of increasing vascular permeability was evaluated by comparing the amount of EB-albumin complex accumulated in the tumor tissue in the GTN application group and the non-application group. FIG. 1 shows the effect of increasing the accumulation of GTN in Evans Blue in S-180 tumor tissue. As shown in FIG. 1, in the GTN application group, the tumor tissue accumulation of the drug EB increased about 2.8 times (p <0.05) over 30 minutes to 6 hours as compared with the non-application group. Among them, it accumulated most in the tumor tissue 4 hours after EB administration. For this reason, in the future study, the amount of tissue accumulation in the tumor at 4 hours after administration of EB was compared.
Based on the above results, the concentration correlation between the dose of GTN applied and the amount of EB accumulated in the tumor tissue was examined at 4 hours after EB administration. FIG. 2 shows the GTN concentration-dependent effect of increasing tumor tissue accumulation. In FIG. 2, ^* indicates P <0.05, and ^** indicates P <0.01 (GTN application group with respect to the non-application group (control group)). As is clear from FIG. 2, EB tumor tissue accumulation increased depending on the concentration of GTN. Even when the applied dose of GTN was 1.0 μg, the tumor tissue accumulation was significantly increased by about 1.6 times (p <0.05).

Example 2: GTN of Meth-A coating group and non-applied group into tumors Evans Blue - the dorsal subcutaneous tumor tissue Accumulation difference 6-week-old female BALB / c mice albumin, 2 × 10 ⁶ cells Mouse fibrosarcoma Meth-A tumor cells were transplanted to produce solid tumors. About 10 days after transplantation, a mouse having a tumor diameter of 5 to 13 mm was selected, and 2% GTN ointment 50 mg (1.0 mg per topical GTN application amount) and white petrolatum prepared according to Example 1 were used. 50 mg of GTN ointment diluted with GTN ointment (0.001 mg per local area of GTN applied) was applied only to the tumor site according to Example 1, and then 0.1 ml of physiological saline solution so that EB would be 10 mg / kg. Were injected into the tail vein. After 4 hours, the mice were sacrificed and evaluated in the same manner as in Example 1. FIG. 3 shows the effect of increasing the accumulation of GTN in Evans Blue in Meth-A tumor tissue. ^{* Indicates} P <0.05, and ^** indicates P <0.01 (GTN application group relative to the non-application group (control group)). As is clear from FIG. 3, the GTN-applied dosage of 1.0 and 0.001 mg of the GTN ointment-applied group showed an increase of about 2.0 times the tumor tissue accumulation compared to the non-applied group (p. <0.05).

Example 3 Difference in Evans Blue-Albumin Accumulation in Tumor Tissue between GTN Applied to Colon 38 Tumor and Unapplied Group Approximately 6 mg of mouse colon cancer subcutaneously in the back of 6-week-old male C57 / BL6 mice Colon 38 (C38) tumor pieces were transplanted using a trocar needle to create a solid tumor. About 12 days after transplantation, mice having a tumor diameter of 6 to 15 mm were selected, GTN was applied according to Example 1, EB was administered from 10 mg / kg tail vein, and evaluation was carried out in Example 1. According to the same procedure. FIG. 4 shows the effect of increasing the accumulation of GTN in Evans Blue in Colon 38 tumor tissue. In FIG. 4, ^* indicates P <0.05, and ^** indicates P <0.01 (GTN application group with respect to the non-application group (control group)). As is clear from FIG. 4, the GTN-applied group, like S-180 and Meth-A, showed an increase in tumor tissue accumulation of about 1.6 times compared to the non-applied group (p <0. 05).

Example 4: Difference in accumulation in tumor tissue of polymer drug Evans blue-albumin between applied and non-applied groups in rat breast cancer caused by administration of GTN chemical carcinogen 7,12-dimethylbenzanthracene (DMBA) Seven-week-old female SD rats were gavaged with DMBA in corn oil at 10 mg / ml using a sonde, and solid tumors developed in about 15 weeks. Select a rat with a tumor diameter of 6 to 15 mm, and add GTN ointment diluted with 100 mg of 2% GTN ointment (2.0 mg per topical GTN application) and white petrolatum prepared according to Example 1. After 100 mg (0.2 and 0.02 mg per GTN applied dose per local area) was applied only to the tumor site according to Example 1, 0.5 ml of physiological saline solution was injected into the tail vein so that EB would be 10 mg / kg. Injected. Evaluation was performed according to Example 1. FIG. 5 shows the effect of increasing the accumulation of GTN Evans Blue in DMBA-induced rat breast cancer tumor tissue. ^{* Indicates} P <0.05 ^** indicates P <0.01 (GTN application group relative to the non-application group (control group)). As is apparent from FIG. 5, similar to transplanted tumors such as S-180, the GTN ointment dosage groups of 0.02, 0.2, and 2.0 mg were compared with the non-application group by about A 1.9, 2.5 and 2.4-fold increase in tumor tissue accumulation was observed (p <0.05).

Example 5: Combined effect of GTN and high-molecular-weight drug on S-180 tumor 2 × 10 ⁶ S-180 tumor cells were transplanted subcutaneously into the back of 6-week-old male ddY mice, and reared for 7 days. Mice in which a solid tumor of 5 to 7 mm was formed were selected and used for experiments. The high molecular drug used in the experiment was polyethylene glycol (PEG) -linked zinc-type protoporphyrin IX (PEG-ZnPP), apparently having a molecular weight of around 130,000, chronic myeloid leukemia (CML) cells resistant to Gleevec and obesity It has a strong cytostatic effect on cellular leukemia (MCL) (Sahoo SK et al. Bioconj Chem 13, 1031-1038, 2002, Fang J et al. Cancer Res 63, 3567-3574, 2003). As test groups, (a) control group (no treatment), (b) GTN control group (GTN application only), (c) 1 mg / kg PEG-ZnPP group (PEG-ZnPP administration), (d) 5 mg / kg PEG-ZnPP There were six groups (group) (PEG-ZnPP administration), (e) 1 mg / kg PEG-ZnPP combined with GTN, and (f) 5 mg / kg PEG-ZnPP combined with GTN. After applying 50 mg of 2% GTN ointment (1 mg per local concentration of GTN) only to the tumor site according to Example 1, PEG-ZnPP, which is a polymer drug, was administered daily from the tail vein for 3 days. The tumor size was measured using calipers, and the tumor size was measured by a conventional method. That is, the tumor volume (V) is V = L × W2 / 2, where long diameter (L) × width (W). As described above, as a control group, a treatment group by a non-treatment group and a non-GTN application group was provided. FIG. 6 shows the combined effect of GTN on the S-180 tumor with the high molecular weight drug PEG-ZnPP, and shows the tumor volume after tumor inoculation in the case of the 6 groups described above. In FIG. 6, x is (a) control group (no treatment), ○ is (b) GTN control group (only GTN application), ■ is (c) 1 mg / kg PEG-ZnPP group (PEG-ZnPP administration) , Δ (triangle filled in black) is (d) 5 mg / kg PEG-ZnPP group (PEG-ZnPP administration), □ is (e) 1 mg / kg PEG-ZnPP and GTN combined, Δ is (f) 5 mg / kg The kgPEG-ZnPP and GTN combination groups are shown. ↓ indicates PEG-ZnPP administration, and ▽ (black inverted triangle) indicates GTN application. Data were expressed as mean ± SE (error bar). ^{* Indicates} P <0.05, ^** indicates P <0.01 (PEG-ZnPP or PEG-ZnPP and GTN combined administration group with respect to the control group). From FIG. 6, it is clear that significant antitumor activity is exhibited by single administration of PEG-ZnPP, but still stronger antitumor activity is exhibited by administration in combination with GTN.

Example 6: Combined effect of GTN and aclarubicin hydrochloride on C38 tumor A 50 mg C38 tumor piece was transplanted subcutaneously into the back of a 6-week-old male C57BL / 6 mouse using a trocar needle, and after 7 days of breeding, 5-7 mm Mice with solid tumors were selected and used for experiments. As test groups, (a) control group (no treatment), (b) GTN control group (only GTN application), (c) 5 mg / kg aralubicin hydrochloride group (administration of aralubicin hydrochloride manufactured by Mercian Co., Ltd.) and (d) Four groups of 5 mg / kg aclarubicin hydrochloride and GTN combination group were provided. As described above, as a control, a treatment group with no treatment group and a non-GTN application group were provided. After applying 50 mg of 2% GTN ointment (1 mg per GTN applied dose locally) only to the tumor site, aclarubicin hydrochloride dissolved in physiological saline was administered from the tail vein for 3 consecutive days. The size of the tumor was measured using calipers, and the tumor size was measured by a conventional method. That is, the tumor volume (V) is V = L × W2 / 2, where long diameter (L) × width (W). FIG. 7 shows the combined effect of GTN on the C38 tumor with the low-molecular drug aclarubicin hydrochloride. In FIG. 7, x is (a) control group (no treatment), ○ is (b) GTN control group (GTN application only), ■ is 5 mg / kg aclarubicin hydrochloride treatment group, □ is (d) 5 mg The / kg combination treatment group of aclarubicin hydrochloride and GTN is shown. ↓ indicates administration of aclarubicin hydrochloride, and ▽ (black inverted triangle) indicates GTN application. Data were expressed as mean ± SE (error bar). ^** indicates P <0.01 (combined administration group of GTN and aclarubicin hydrochloride relative to the control group). From FIG. 7, it is clear that the combination treatment group of GTN and aclarubicin hydrochloride shows significant tumor growth suppression compared with the aclarubicin hydrochloride alone and non-treatment groups.

  In the above animal experiments, it has been proved that the combination with an antineoplastic agent, particularly with a high molecular weight drug, enhances delivery to a tumor site. Therefore, nitric oxide donors, especially GTN, can enhance drug delivery to superficial tumors and benefit treatment.

It is a figure which shows the accumulation increasing effect in the S-180 tumor tissue of Evans blue of GTN. It is a figure which shows the GTN density | concentration dependence tumor tissue accumulation increasing effect. It is a figure which shows the accumulation increasing effect in the Meth-A tumor tissue of Evans blue of GTN. It is a figure which shows the accumulation increasing effect in Colon 38 tumor tissue of Evans blue of GTN. It is a figure which shows the accumulation increasing effect in DMBA induction rat breast cancer tumor tissue of Evans blue of GTN. It is a figure which shows the combined use effect with respect to S-180 tumor with the high molecular agent PEG-ZnPP of GTN. It is a figure which shows the combined use effect with respect to C38 tumor with the low molecular weight drug aclarubicin hydrochloride of GTN.

Claims (15)

  An agent for enhancing local delivery of an antineoplastic agent containing a nitric oxide (NO) donor as an active ingredient to a tumor tissue.   The agent for enhancing local delivery of an antineoplastic agent according to claim 1 to the tumor tissue by enhancing the accumulation of the drug in the tumor tissue.   The tumor tissue of the antineoplastic agent according to claim 1 or 2, wherein the nitric oxide (NO) donor is selected from the group consisting of nitroglycerin, glyceryl mononitrate, glyceryl dinitrate, isosorbide mononitrate and isosorbide dinitrate. Topical delivery enhancer.   The enhancer for local delivery of an antineoplastic agent to tumor tissue according to claim 1 or 2, wherein the nitric oxide (NO) donor is nitroglycerin.   The agent for locally delivering an antineoplastic agent to tumor tissue according to claim 1 or 2, wherein the drug containing a nitric oxide (NO) donor as an active ingredient is an external preparation.   The topical delivery enhancer for an anti-neoplastic agent according to claim 5, wherein the external preparation is an ointment, gel ointment, cream, liquid, patch, suppository, spray, or intranasal external preparation.   The topical delivery enhancer for tumor tissue of an antineoplastic agent according to claim 5, wherein the external preparation is an ointment.   The agent for local delivery of an antineoplastic agent to a tumor tissue according to claim 5, wherein the external preparation is a spray.   The malignant tumor is a superficial tumor, The local delivery enhancer to the tumor tissue of the antineoplastic agent according to claim 1-9.   Superficial tumors include breast cancer, esophageal cancer, stomach cancer, colorectal cancer, urological cancer, bronchopulmonary cancer, respiratory tract cancer, genital cancer, skin cancer, head and neck cancer, eye cancer, and abdominal / chest wall. The agent for local delivery of an antineoplastic agent according to claim 9 to a tumor tissue, which is a superficial tumor selected from the group consisting of cancer.   The agent for enhancing local delivery of an antineoplastic agent to a tumor tissue according to claim 1, wherein the antineoplastic agent is aclarubicin.   The agent for locally delivering an antineoplastic agent to a tumor tissue according to claim 1, wherein the antineoplastic agent is a polymer drug.   The agent for local delivery of an antineoplastic agent to a tumor tissue according to claim 12, wherein the high molecular agent is PEG-ZnPP, PEGylated interferon, micellized preparation, liposome preparation or other high molecular weight antitumor agent.   A drug kit, drug pack, or patient pack comprising an anti-neoplastic agent and an agent for enhancing local delivery of the anti-neoplastic agent according to claim 1 to tumor tissue.   The drug kit, drug pack or patient pack according to claim 14, wherein the agent for locally delivering an antineoplastic agent to a tumor tissue is an ointment.

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