JP2007537246A - Cancer preventive and therapeutic agent containing nicotinic acid derivative as active ingredient - Google Patents

Abstract

  The present invention relates to nicotinic acid or a derivative thereof, and more particularly, a cancer preventive and therapeutic agent containing nicotinic acid or a derivative thereof is expressed by RUNX3 gene, which is a tumor suppressor gene inactivated by methylation To promote. The cancer preventive and therapeutic agent of the present invention is associated with inactivation of RUNX3 such as gastric cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder cancer, prostate cancer, pancreatic cancer, uterine cancer, esophageal cancer or breast cancer. Can be used for cancer.

Description

  The present invention relates to a cancer preventive and therapeutic agent comprising a nicotinic acid derivative as an active ingredient, and more specifically, promotes the expression of a RUNX3 gene, which is a cancer suppressor gene inactivated by methylation. The present invention relates to a cancer preventive and therapeutic agent comprising, as an active ingredient, a nicotinic acid derivative that suppresses protein deacetylation and increases the activity by inhibiting the degradation of the RUNX3 protein.

  Currently, there are three main types of treatment for cancer: surgical treatment, pharmacotherapy, and radiotherapy, but in reality, multidisciplinary therapy combining the above methods with laser therapy is widely performed. ing. However, considering the pain associated with the treatment and the possibility of metastasis, there is great expectation for drug therapy among the above methods. Therefore, many anticancer drugs that can respond to this have been developed and used, but most anticancer drugs have been developed based on the principle of obtaining an anticancer effect by selectively killing actively dividing cells. It is. Such anti-cancer drugs will kill normal cells such as hair follicle cells and root cells that actively divide normally in the body, so they have serious side effects and cannot be used for a long time. Problems have been pointed out. Therefore, development of a new anticancer agent based on elucidation of the cause of cancer onset is required.

  Oncogene activation, which induces cancer, induces abnormal cell growth, whereas tumor suppressor genes, on the other hand, suppress abnormal cell growth and sometimes cell death programs (cells). By activating a death program, the generation of cancer cells is blocked by killing specific cells. Even if a cell having an abnormally activated oncogene once the tumor suppressor gene shows normal activity, the cell cannot die and cannot die. Therefore, in order to become a cancer cell, not only the activation of the oncogene but also the inactivation of the tumor suppressor gene must appear simultaneously in one cell.

  Recently, it has been clearly clarified that the undifferentiated proliferation of cells characteristically appearing in cancer cells is due to activation of oncogenes and inactivation of tumor suppressor genes. Specifically, TGF-β-dependent apoptosis in cells plays a very important role in protecting normal cells from becoming cancerous cells, where tumor suppressor genes are inactivated. This increases cell division and suppresses TGF-β-dependent self-death, resulting in cancerous cells.

  One of the ways in which tumor suppressor gene inactivation occurs is abnormal DNA methylation to CpG islands (Jones and Laird, Nature genet. 1999, Vol. 21, pp. 163-167). The methylation of CpG islands is performed by an enzyme called DNA methylase, and when methylation is accumulated, proteins such as MeCP2 bind to the methylated DNA, thereby causing histone deacetylase ( hidtone deacethylase) will come together. Histone deacetylase again removes the acetyl group attached to the histone, resulting in a denser DNA structure in the neighborhood and suppression of gene expression. When gene expression is suppressed by DNA methylation, expression can be induced by a DNA methylase inhibitor or a histone deacetylase inhibitor.

  On the other hand, acetylation / deacetylation occurs not only with histones but also with several transcription factors such as p53 and RUNX3, but for example, acetylation of RUNX3 increases. For example, protein degradation due to ubiquitination is suppressed, and the activity of RUNX3 increases (Jin et al., J. Biol. Chem. 2004, 279, 29409-17). Thus, since increased RUNX3 activity can be induced by suppressing deacetylation, acetylation / deacetylation can act as an important initiation mechanism in protein function regulation. it can.

  In fact, RUNX3, which was clarified as a tumor suppressor gene by the present inventors, was reported to be suppressed by DNA methylation in various cancers of the human body. % (Li QL et al., Cell. 2002, 109 (1), 113-24; Oshimo Y. et al., Pathobiology. 2004, 71 (3), 137-43), 25- 40% (LI QL et al., BBRC. 2004, 314 (1), 223-8; Kim TY et al., Lab Invest. 2004, 84 (4), 479-84), in laryngeal cancer 62%, breast cancer 25% (Kim TY et al., Lab Invest. 2004, 84 (4), 479-84), pancreatic cancer 70% (Li J et al., J Clin Pathol. 2004, 57th) (3), 294-9; Wada M et al., Oncogene. 2004, 23 (13), 2401-7), 73% in hepatocellular carcinoma (Xiao WH and Liu WW, World J Gastroenterol. 2004) 10 (3), 376-80; Kim TY et al., Lab Invest. 2004, 84 (4), 479-84), 75% of cholangiocarcinoma (Wada M et al., Oncogene. 2004) 23 (13), 2401-7), 23 for prostate cancer % (Kang GH et al., J. Pathol. 2004, 202 (2), 233-40; Kim TY et al., Lab Invest. 2004, 84 (4), 479-84) in colon cancer 45% (Ku JL et al., Oncogene, 2004, Vol. 23, 6736-6742) are reported to be suppressed by methylation.

  In addition, activation of the RUNX3 gene has been proven to remarkably suppress the growth of cancer cells (Li et al., Cell, 2002, 109 (1), 113-24; Balmain, Nature 2002, 417 (6886), 235-7). Therefore, based on the above research results, it has become possible to expect cancer development and development of therapeutic agents by activation of the RUNX3 gene.

  In view of the above, as a result of efforts to search for a substance capable of activating the inactivated RUNX3 tumor suppressor gene in cancer cells in vivo, the nicotinic acid derivative was methylated. When the inactivated RUNX3 gene is activated to cause a quantitative increase in intracellular RUNX3 protein, the nicotinic acid derivative is effective in preventing and treating cancer when actually applied to an animal model. As a result, the present invention was completed.

  An object of the present invention is to activate a nicotinic acid derivative that activates the RUNX3 gene, which is a tumor suppressor gene inactivated by methylation in cancer cells, and increases the activity of the RUNX3 protein that is the product of this gene. It is providing the cancer preventive and therapeutic agent containing as.

  In order to achieve the above object, the present invention provides a cancer preventive and therapeutic agent comprising a nicotinic acid derivative as an active ingredient.

  The present invention will be described in detail below.

  The present invention provides a cancer preventive and therapeutic agent comprising a nicotinic acid derivative as an active ingredient.

  The present inventors expect that an active speaker capable of activating an inactivated RUNX3 tumor suppressor gene in vivo exhibits a anticancer effect, and therefore, a substance having an effect of activating the RUNX3 gene As a result of the search, it was confirmed that niacinamide can activate RUNX3.

  First, we confirmed whether RUNX3, a tumor suppressor gene, was methylated in the tissues of cancer patients, and investigated the relationship between methylation and cancer progression in cancer patients. As a result, the RUNX3 gene was methylated very frequently in cancer patient tissues. Moreover, it was shown that the patient group in which RUNX3 is methylated has a very high recurrence rate and progression rate of cancer (see Tables 1 to 3).

  The above results indicate that activating RUNX3 that has been inactivated by methylation is a method that can prevent cancer progression. Therefore, the present inventors tried to search for a substance capable of preventing the development and progression of cancer by searching for a substance having an activation effect on the RUNX3 gene, and as a result, niacinamide was inactivated by methylation. It was confirmed that the activated RUNX3 can be activated (see FIG. 2).

In experiments where we observed whether niacinamide induces a quantitative increase in RUNX3 protein, as RUNX3 is deacetylated by histone deacetylase, degradation is induced and Although the amount decreased, it was confirmed that administration of niacinamide increased acetylation of RUNX3 protein and increased the amount of protein (see FIGS. 3 and 4).
Subsequently, in order to confirm the anticancer effect of niacinamide in vivo, the cancer formation inhibitory effect was analyzed using a mouse in which cancer was induced by a carcinogen. As a carcinogen, N-butyl-N-4-hydroxybutylnitrosamine (BBN) was used. A cancer formation inhibitory effect analysis was performed by comparing the appearance of cancer in mice groups provided with BBN aqueous solution as drinking water for 20 weeks and mice groups provided with BBN and niacinamide aqueous solution as drinking water for 20 weeks (Figs. 5 and 5). (See Figure 6). As a result, in the BBN treatment group, RUNX3 activity was suppressed by inducing methylation of CpG island in the RUNX3 exon 1 site, and 100% bladder cancer occurred. Among them, it was confirmed that invasive cancer was 80% and superficial cancer was 20%. In addition, the group of mice treated with niacinamide with BBN not only significantly reduced the methylation of CpG islands in RUNX3 exon 1 site and reduced bladder cancer formation to 58%, but also the malignancy of the cancer. Decreased to 23% for invasive cancer and 77% for superficial cancer. The above results mean that niacinamide suppressed the inactivation of RUNX3 by BBN, and thereby the formation of cancer by BBN was also suppressed by niacinamide.

  From the above results, it can be confirmed that RUNX3 inactivated with niacinamide of the present invention can be reactivated and cancer growth can be remarkably suppressed.

  In addition to the above results, the tumor suppressor gene RUNX3 is actually inactivated by methylation in various human cancers such as gastric cancer, lung cancer, laryngeal cancer, breast cancer, pancreatic cancer, hepatocellular carcinoma, bile duct cancer, prostate cancer, colon cancer, etc. (Li QL et al., Cell. 2002, 109 (1), 113-24; Oshimo Y. et al., Pathobiology. 2004, 71 (3), 137-43); LI QL BBRC. 2004, 314 (1), 223-8; Kim TY et al., Lab Invest. 2004, 84 (4), 479-84; Li J et al., J Clin Pathol. 2004 57 (3), 294-9; Wada M et al., Oncogene. 2004, 23 (13), 2401-7; Xiao WH and Liu WW, World J Gastroenterol. 2004, 10 (3), 376-80; Kang GH et al., J. Pathol. 2004, 202 (2), 233-40; Ku JL et al., Oncogene, 2004, 23, 6736-6742 ) Activation of the RUNX3 gene has been shown to be able to significantly suppress cancer cell growth (Kim et al., Lab Invest. 2004, 84 (4), 479-84; Wada Et al., Oncogen, 2004 23 (13), 2401-7; Li et al., Biochem Biophys Res Commun, 2004, 314 (1), 223-8; Li et al., Cell, 2002, 109 (1) 113-24; Li et al., Cell, 2002, 109 (1), 113-24; Balmain, Nature, 2002, 417 (6886), 235-7). Therefore, the pharmaceutical composition comprising the nicotinic acid derivative of the present invention as an active ingredient reactivates the RUNX3 gene and is a cancer caused by inactivation of the RUNX3 gene, such as stomach cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder It can be used as a preventive and therapeutic agent for cancer, prostate cancer, pancreatic cancer, uterine cancer or breast cancer.

  Moreover, a nicotinic acid derivative can be used for the active ingredient of the cancer preventive and therapeutic agent of the present invention. The nicotinic acid derivatives are niacinamide, nicotinic acid, 6-aminonicotinamide, isonicotinamide, isonicotinohydrazide, N-methylnicotinamide. (N-Methylnicotinamide), Nicorandil, Pyridine-2-carboxamide, thionicotinamide or 6-Chloronicotinamide.

  The pharmaceutical composition containing the nicotinic acid derivative of the present invention as an active ingredient can be administered orally or parenterally at the time of clinical administration, and can be used in the form of a general pharmaceutical preparation.

  That is, the pharmaceutical composition of the present invention can be administered in various oral and parenteral dosage forms at the time of actual clinical administration, and in the case of formulating, a filler, a bulking agent, a binder, a moistening agent commonly used. It is prepared using a diluent or excipient such as an agent, a disintegrant, and a surfactant. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., such solid preparations include at least one excipient in one or more compounds of formula 1, For example, starch, calcium carbonate, sucrose or lactose, gelatin and the like are mixed to prepare. In addition to simple excipients, lubricants such as magnesium talc are also used. Liquid preparations for oral use include suspensions, liquids for internal use, emulsions, syrups, etc., and various excipients such as water and liquid paraffin, which are commonly used simple diluents, such as wetting agents, Sweeteners, fragrances, preservatives and the like can be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories. Nonaqueous solvents and suspending agents that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleic acid. As a base material for suppositories, hard fat (witepsol), macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

  The effective dose of the nicotinic acid derivative of the present invention is 1 to 50 mg / kg, preferably 10 to 30 mg / kg, and can be administered 1 to 3 times a day.

  As a result of an acute toxicity test on mice, the nicotinic acid derivative showed no toxicity change up to 2 g / kg as a result of an acute toxicity test on mice. It was confirmed that no side effects were observed.

  In addition, the present invention provides a method for preventing and / or treating cancer including a step of administering a nicotinic acid derivative to a subject, and provides a use for using the nicotinic acid derivative in the manufacture of a medicament for preventing and / or treating cancer. .

  Hereinafter, the present invention will be described in detail according to examples.

  However, the following examples merely illustrate the present invention, and the content of the present invention is not limited to the following examples.

Example 1: Correlation analysis between methylation of RUNX3 gene and carcinogenesis First, in order to confirm whether or not the RUNX3 gene is methylated in the tissue of cancer patients, the tissue of bladder cancer patients was extracted and the Tissue methylation was confirmed. Specifically, genomic DNA was isolated and purified from normal bladder tissue and bladder cancer tissue and treated with 3M NaSO ₂ (sodium bisulfite) at 50 ° C. for 16 hours. In the process, DNA that is not methylated is converted from cytosine to uracil.

  After the treatment, the DNA was purified again, and the RUNX3 genomic DNA CpG island primer sequence SEQ ID NO: 1 and SEQ ID NO: 2 were prepared based on the sequence of SEQ ID NO: 3 and SEQ ID NO: 3 Polymerase chain reaction (PCR) was performed using two kinds of primers of the DNA methylation primer set of SEQ ID NO: 4. The reaction conditions were 94 ° C for 15 seconds, 60 ° C for 50 seconds, 72 ° C for 58 seconds, and 40 times in total.

  When PCR is performed using the primers, the presence or absence of inactivation due to methylation of the RUNX3 gene can be confirmed. When DNA is amplified by the primer set of SEQ ID NO: 1 and SEQ ID NO: 2, RUNX3 is methylated It means that the DNA is amplified by the primer set of SEQ ID NO: 3 and SEQ ID NO: 4, meaning that RUNX3 is not methylated.

  As a result, as shown in FIG. 1, 20 normal bladder tissues had no methylation in the promoter region of the RUNX3 gene, and in 119 bladder cancer patients tissues, 88 individuals were methylated, 74 % Methylation degree. In addition, in the cancer recurrence rate of patients with bladder cancer, the cancer cell RUNX3 gene was methylated compared with 61.11% of the cancer cell RUNX3 gene methylated. Showed a very high recurrence rate in patients with a cancer cell RUNX3 gene methylation rate of 88% (Table 1).

  p = 0.0036, OR = 4.6667, 95% CI = 1.5774-13.8064

  As for the progression rate of cancer, as shown in Table 2 below, the progression rate was very high in cancer tissues in which RUNX3 was methylated. In other words, 94.74% of patients with advanced cancer showed that the RUNX3 gene in cancer cells was methylated, and methylation of the RUNX3 gene plays a very important role in the progression of cancer. I understand.

  p = 0.0155, OR = 9.00, 95% CI = 1.1236-72.0598

  In conclusion, intracellular RUNX3 gene methylation increased the incidence of cancer by a factor of 109, and even in cancer recurrence and progression, RUNX3 gene methylation was 5 times and It was confirmed that the increase was 9 times.

Example 2: Activation analysis of RUNX3 by niacinamide Based on the results confirmed in Example 1 above, the present inventors searched for a substance capable of suppressing methylation of the tumor suppressor gene RUNX3, and niacin It was confirmed that amide and its derivatives can suppress methylation of RUNX3 gene.

  That is, a culture solution of a cancer cell line in which the RUNX3 gene was inactivated by DNA methylation was treated with niacinamide for 4 days so that the final concentration was 1 mg / ml. Detection was performed by the reverse transcriptase polymerase chain reaction (RT-PCT) method using Ps-CA and the Ps-CB primer of SEQ ID NO: 6. As a result, as shown in FIG. 2, niacinamide is inactive in gastric cancer cell lines SUN16 and MKN74, lung cancer cell lines NCI-H226, NCI-H460 and Sq-1, and bladder cancer cell line T24. It was confirmed that the activated RUNX gene could be activated.

Example 3: Confirmation of RUNX3 protein degradation by deacetylation
In order to investigate the relationship between RUNX3 protein degradation and deacetylation, the cell lines were transduced with Sirt2 (type III Histone deacetylase) and RUNX3 expressing genes separately or together to immunize the effect. Analysis by immunoprecipitation and Western blotting (Janeway et al., Immunobiology. 5th ed. Garland Publishing, 2001).

  As a result, Sirt2 protein was detected by immunoprecipitation against RUNX3 (IP: RUNX3, IB: Sirt2), and RUNX3 protein was detected by immunoprecipitation against Sirt2 (IP: Sirt2, IB: RUNX3) (FIG. 3). ) We confirmed that RUNX3 and Sirt2 were physically combined. It was also confirmed that the amount of RUNX3 protein was decreased by Sirt2 (FIG. 3, IB: RUNX3).

Example 4: Confirmation of quantitative increase of RUNX3 protein by niacinamide In order to observe the quantitative change of RUNX3 protein by niacinamide, RUNX3, acetyltransferase (acetyltransferase, p300), diacetylase (deacetylase, Sirt2) Niacinamide was added to the cell medium after expression. As a result of analysis by immunoprecipitation and Western blotting, it was observed that the amount of RUNX3 protein decreased by the expression of Sirt2 was restored by administration of niacinamide (Fig. 4, lanes 1 and 2). (Fig. 4, lanes 3, 4, and 5).

Example 5: In vivo anticancer effect analysis of niacinamide In order to confirm the in vivo anticancer effect of niacinamide, a cancer formation inhibitory effect experiment was conducted using mice in which cancer was induced by a carcinogen. . Carcinogenesis inhibitory effect analysis was carried out using a group of mice provided with 0.05% (W / W) N-butyl-N-4-hydroxybutylnitrosamine (BBN) aqueous solution, which is a carcinogen, as drinking water for 20 weeks, 0.05% BBN and 1% This was done by comparing groups of mice that were provided with (W / W) aqueous niacinamide (10 mg / ml) as drinking water for 20 weeks. The mice used above are 6 week old C3H / HE mice. For the control group, water or a 1% (W / W) aqueous solution of niacinamide was administered for 20 weeks. On the final day of the experiment, mice were sacrificed to remove the bladder, and the methylation pattern of the BUNX3 gene was compared with the presence or absence of cancer and the size of the cancer.

  The appearance of bladder cancer in the experimental group could be confirmed with the naked eye and a microscope as shown in FIG. 5 and FIG. 6A. Specifically, BBN induces methylation of CpG island at the RUNX3 exon 1 site and suppresses RUNX3 activity, resulting in 100% bladder cancer in the group of mice treated with it. Cancers caused by BBN showed 80% invasive cancer and 20% superficial cancer (Table 3).

  In the group of mice treated with niacinamide together with BBN, methylation of CpG islands in RUNX3 exon 1 site was significantly reduced, not only reducing the incidence of bladder cancer to 58% (Figures 5 and 6). B) The malignancy of the cancer also decreased, with 23% invasive cancer and 77% superficial cancer (Table 4). The above results mean that niacinamide suppressed the inactivation of RUNX3 by BBN, and thereby the formation of cancer by BBN was also suppressed by niacinamide.

Example 6 In Vivo Methylation Inhibition Analysis of Nicotinic Acid Derivatives The present inventors analyzed the degree of in vivo methylation inhibition of nicotinic acid derivatives and showed the results as follows.

As a result of the experiment, 100% RUNX3 gene methylation occurred in the group of mice treated with BBN. In the group of mice administered with BBN and nicotinic acid derivatives together, methylation was suppressed as shown in Table 4. Therefore, the nicotinic acid derivative is considered to exhibit anticancer activity.
Example 7: Acute toxicity experiment for oral administration to mice A mouse (C3H / HE 6 mouse) was used to conduct an acute toxicity experiment of a pharmaceutical composition containing a nicotinic acid derivative of the present invention. Two mice per group were orally administered with the above pharmaceutical composition at a dose of 2 g / kg as drinking water every day for 5 months. Hematological and blood biochemical tests were performed by observing the presence or absence of moribundity, clinical symptoms, and changes in body weight of animals after administration of the test substance. The lesions were examined visually for abnormalities in the abdominal and thoracic organs.

  As a result of the test, there were no clinical symptoms or moribund animals that deserve special mention in all animals that received the test substance. No changes in toxicity were observed in body weight changes, blood tests, blood biochemical tests, autopsy findings, etc. As a result of the above, the tested pharmaceutical composition did not show toxic changes up to 2 g / kg in mice, and the minimum lethal dose (LD50) for oral administration was determined to be a safe substance of 5 g / kg or more. .

  The present invention relates to a cancer preventive and therapeutic agent comprising a nicotinic acid derivative as an active ingredient, and suppresses the methylation of the RUNX3 gene that is inactivated by methylation in cancer cells, thereby Provided is a cancer preventive and therapeutic agent comprising a nicotinic acid derivative to be activated as an active ingredient. The cancer preventive and therapeutic agent of the present invention is a solid cancer associated with inactivation of RUNX3 gene such as stomach cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder cancer, prostate cancer, pancreatic cancer, uterine cancer or breast cancer. Unlike other cancer drugs, which can be usefully used as a prophylactic and therapeutic agent for cancer, the anti-tumor gene of cancer cells is activated, unlike most anticancer drugs, by selectively killing actively dividing cells. Therefore, it can be used as a new anticancer agent with improved side effects of existing anticancer agents.

A is an electrophoretogram confirming the presence or absence of methylation of the RUNX3 gene in normal cells through PCR. M: Methylated DNA amplification U: Unmethylated DNA amplification B is RUNX3 gene methylation in bladder cancer cells Electrophoresis photograph confirming the presence or absence of methylation by PCR, M: Methylated DNA amplification U: Unmethylated DNA amplification It is the photograph which showed activation of the RUNX3 gene methylated by the niacinamide process. RUNX3 is a photograph showing that the amount of RUNX3 protein decreases as a result of binding of RUNX3 to histone deacetylase (Sirt2). A photograph showing that the decrease in RUNX3 protein level due to histone deacetylase (Sirt2) correlates with the decrease in RUNX3 protein acetylation, and that niacinamide administration increases intracellular RUNX3 protein acetylation and RUNX3 protein level. is there. Carcinogenicity of a group of mice that provided N-butyl-N-4-hydroxybutylnitrosamine (BBN) aqueous solution, a carcinogen, as drinking water for 20 weeks, and a group of mice that provided BBN and niacinamide aqueous solution as drinking water for 20 weeks It is the macroscopic observation photograph of the bladder cancer tissue which compared the aspect. A is a micrograph of the bladder tissue of a group of mice provided with BBN aqueous solution, which is a carcinogen, as drinking water for 20 weeks.

  B is a micrograph of the bladder tissue of a group of mice provided with BBN and an aqueous niacinamide solution as drinking water for 20 weeks.

SEQ ID NO: 1 and SEQ ID NO: 2 are methylated primer sets for DNA amplification,
SEQ ID NO: 3 and SEQ ID NO: 4 are unmethylated primer sets for DNA amplification,
SEQ ID NO: 5 is a Ps-CA primer,
SEQ ID NO: 6 is a Ps-CB primer.

Claims (16)

A cancer preventive and / or therapeutic agent comprising a nicotinic acid derivative having the following general formula or a pharmaceutically acceptable salt thereof as an active ingredient.

Where R ₁ is oxygen or sulfur and R ₂ is H, OH, C ₁ -C ₁₀ alkyl, NH ₂ , NH-NH ₂ , NH-C ₂ H ₄ -O-NO ₂ -or OR ₃ (R ₃ is C ₁ -C ₅ alkyl), X is H, NH ₂ or a halogen group atom, and Y is H or CH ₃ . The cancer preventive and / or therapeutic agent according to claim ₁ , wherein R ₁ is C ₁ -C ₅ alkyl.   The cancer preventive and / or therapeutic agent according to claim 1, wherein the halogen group atom is fluorine or chlorine. The cancer preventive and / or therapeutic agent according to claim 1, wherein R ₂ is CH ₃ or C ₂ H ₅ .   Nicotinic acid derivatives include niacinamide, nicotinic acid, 6-aminonicotinamide, isonicotinamide, isonicotinohydrazide, N-methylnicotinamide (N-Methylnicotinamide), Nicorandil, Pyridine-2-carboxamide, thionicotinamide or 6-Chloronicotinamide Item 2. The cancer preventive and / or therapeutic agent according to Item 1.   The cancer prevention and / or cancer according to claim 1, wherein the cancer is gastric cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder cancer, prostate cancer, pancreatic cancer, uterine cancer, esophageal cancer or breast cancer. Therapeutic agent.   A method for preventing and / or treating cancer comprising the step of administering the nicotinic acid derivative of claim 1 or a pharmaceutically acceptable salt thereof to an individual.   Nicotinic acid derivatives include niacinamide, nicotinic acid, 6-aminonicotinamide, isonicotinamide, isonicotinohydrazide, N-methylnicotinamide (N-Methylnicotinamide), Nicorandil, Pyridine-2-carboxamide, thionicotinamide or 6-Chloronicotinamide Item 8. The method for preventing and / or treating cancer according to Item 7.   The cancer prevention and / or cancer according to claim 7, wherein the cancer is stomach cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder cancer, prostate cancer, pancreatic cancer, uterine cancer, esophageal cancer or breast cancer. Method of treatment.   The use which uses the nicotinic acid derivative of Claim 1 or its pharmaceutically acceptable salt for manufacture of the pharmaceutical for cancer prevention and / or treatment.   Nicotinic acid derivatives include niacinamide, nicotinic acid, 6-aminonicotinamide, isonicotinamide, isonicotinohydrazide, N-methylnicotinamide (N-Methylnicotinamide), Nicorandil, Pyridine-2-carboxamide, thionicotinamide or 6-Chloronicotinamide Item 10. Use according to Item 10.   The use according to claim 10, wherein the cancer is stomach cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder cancer, prostate cancer, pancreatic cancer, uterine cancer, esophageal cancer and breast cancer.   A method of activating RUNX3 or a gene encoding RUNX3, comprising a step of administering the nicotinic acid derivative of claim 1 to an individual suffering from cancer.   14. The method according to claim 13, wherein RUNX3 is deacetylated in an individual who has cancer.   14. The method according to claim 13, wherein the individual having cancer is a gene in which a gene encoding RUNX3 is methylated.   14. The method according to claim 13, wherein the cancer is stomach cancer, lung cancer, liver cancer, laryngeal cancer, colon cancer, bladder cancer, prostate cancer, pancreatic cancer, uterine cancer, esophageal cancer or breast cancer.

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