JP5237884B2 - Anti-cancer agent - Google Patents

Description

  The present invention relates to an anticancer agent, and more particularly to an anticancer agent containing a novel palmitic acid amide as an active ingredient.

  Cancer is the disease with the highest mortality rate today, and there are three major treatments: chemotherapy, radiation therapy, and surgery. Chemotherapy includes antitumor antibiotics such as mitomycin C and actinomycin D, antimetabolites such as 5-fluorouracil and methotrexate, alkylating agents such as cyclophosphamide and nimustine, hormones such as anastrozole and ethinyl estradiol Drugs, platinum preparations such as cisplatin and oxaliplatin, and anticancer agents such as plant alkaloids such as irinotecan and etoposide are used. However, anti-cancer drugs do not act selectively on cancer cells, but also on normal cells, so patients who receive anti-cancer drugs are vomiting, loss of appetite, nausea, hair loss, There are major problems such as bone marrow suppression, liver dysfunction, renal dysfunction, and cardiac dysfunction that can significantly reduce the quality of life (QOL), and it has excellent anticancer activity and toxicity to normal cells. Therefore, anticancer agents with low safety and high safety are always desired.

  Conventionally, there are various anticancer agents as described above, but there are proposals of antitumor agents containing fatty acids as active ingredients as highly safe anticancer agents (see Patent Document 1 and Patent Document 2). However, there has been no proposal for an anticancer agent containing as an active ingredient palmitic acid amide, which is a low molecular weight compound derived from in vivo lipids.

JP 2003-171272 A JP 2005-247754 A

  This invention makes it a subject to provide the anticancer agent which is excellent in anticancer activity and has high safety | security which uses a palmitic acid amide body as an active ingredient.

（式中、Ｒは3,4-ジヒドロイソキノリン-2(1H)-イル（3,4-dihydroisoquinolin-2(1H)-yl）又は3,4-ジヒドロキノリン-1(2H)-イル（3,4-dihydroquinolin-1(2H)-yl）を表す）で表されることを特徴とするパルミチン酸アミド体または薬理的に許容されるその塩を要旨とする。 The present invention that solves the above-described problems has the general formula (1).

Wherein R is 3,4-dihydroisoquinolin-2 (1H) -yl or 3,4-dihydroisoquinolin-2 (1H) -yl (3, 4-dihydroquinolin-1 (2H) -yl)), or a pharmacologically acceptable salt thereof.

（式中、Ｒはピペリジン-1-イル（piperidin-1-yl）又は3,4-ジヒドロイソキノリン-2(1H)-イル（3,4-dihydroisoquinolin-2(1H)-yl）又は3,4-ジヒドロキノリン-1(2H)-イル（3,4-dihydroquinolin-1(2H)-yl）を表す）で表されるパルミチン酸アミド体または薬理的に許容されるその塩を有効成分として含有することを特徴とする抗がん剤を要旨とする。
また、該抗がん剤の抗がん活性は大腸がんに対するものである。 Further, the present invention provides a compound represented by the general formula (2)

Wherein R is piperidin-1-yl or 3,4-dihydroisoquinolin-2 (1H) -yl or 3,4-dihydroisoquinolin-2 (1H) -yl Contains a palmitic acid amide compound represented by -dihydroquinolin-1 (2H) -yl (representing 3,4-dihydroquinolin-1 (2H) -yl) or a pharmaceutically acceptable salt thereof as an active ingredient The gist is an anticancer agent characterized by the above.
Moreover, the anticancer activity of the anticancer agent is for colorectal cancer.

The palmitic acid amide compound represented by the general formula ( 2 ) of the present invention or a pharmacologically acceptable salt thereof is excellent in anticancer activity and high in safety, so that it lowers the QOL of patients who have been administered. It is useful as a medicine that can prevent and effectively treat cancer, and a medicine that can prevent cancer and suppress recurrence.

It is explanatory drawing which shows the method to which the palmitic acid amide-form 1, 2, 3 (drug | medicine) is exposed to the human colon cancer cell line HT29 plated and cultured by 6-well plate. After exposing human colon cancer cell line HT29 seeded in 6-well plate in Fig. 1 to palmitic acid amide-1, 2, 3 (drug), the number of colonies of stained cancer cells It is explanatory drawing which shows the method to measure visually. FIG. 3 is a growth curve (dose response curve) of palmitic acid amides 1, 2, and 3 prepared based on the results measured in FIG. 2. Growth curve (dose response curve) prepared based on the results of measuring the survival rate of human colon normal cell line FHC, human colon cancer cell line HT29 and human colon cancer cell line HCT116 exposed to palmitic acid amide-3 It is.

The palmitic acid amide represented by the general formula ( 2 ) is a 1- (piperidin-1-yl) hexadecane represented by the following formula ( 3 ) when R is piperidin-1-yl. 1-one (1- (piperidin-1-yl) hexadecan-1-one) (hereinafter sometimes referred to as “palmitic acid amide-1”). When R is 3,4-dihydroisoquinolin-2 (1H) -yl (3,4-dihydroisoquinolin-2 (1H) -yl), the palmitic acid amide is represented by the following formula ( 4 ): 1- (3 , 4-Dihydroisoquinolin-2 (1H) -yl) hexadecan-1-one (hereinafter referred to as “palmitic acid amide”) Sometimes referred to as “body-2”.) R is 3,4-dihydro-quinoline -1 (2H) - yl case of (3,4-dihydroquinolin-1 (2H ) -yl), palmitic acid amide is 1- (3 shown by the following formula (5) , 4-Dihydroquinolin-1 (2H) -yl) hexadecan-1-one (1- (3,4-dihydroquinolin-1 (2H) -yl) hexadecan-1-one) (hereinafter referred to as “palmitic acid amide”) It may be called "body-3".)

The palmitic acid amides 1, 2, and 3 of the present invention can be produced by a known amide synthesis method.
For example, it can be produced by reacting palmitic acid with the amine component piperazine, 1,2,3,4-tetrahydroisoquinoline or 1,2,3,4-tetrahydroquinoline in the presence of a dehydration condensing agent such as carbodiimide. Alternatively, it can be produced by reacting palmitic acid halide, palmitic anhydride or palmitic acid ester with the amine component piperazine, 1,2,3,4-tetrahydroisoquinoline or 1,2,3,4-tetrahydroquinoline. After completion of the reaction, it can be isolated using known separation and purification means such as extraction, distribution, column chromatography and the like.

  The palmitic acid amides 1, 2, and 3 of the present invention can be free or pharmacologically acceptable salts or prodrugs. The salt may be an acid addition salt or a base addition salt. Acids that form acid addition salts are phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid and other inorganic acids, methanesulfonic acid, citric acid, fumaric acid, succinic acid, tartaric acid, oxalic acid, maleic acid, acetic acid, malic acid, lactic acid, Examples thereof include organic acids such as ascorbic acid. Bases that form base addition salts are inorganic bases such as ammonium hydroxide, alkali metal hydroxides, alkaline earth metal hydroxides, carbonates, bicarbonates, ethanolamine, triethylamine, tris (hydroxymethyl) aminomethane An organic base such as The palmitic acid amides 1, 2, and 3 of the present invention can be made into solvates. Examples of such solvates include hydrates and alcohol solvates.

  Palmitic acid amides 1, 2, and 3 of the present invention are colorectal cancer, stomach cancer, esophageal cancer, colon cancer, liver cancer, pancreatic cancer, breast cancer, lung cancer, gallbladder cancer, bile duct cancer, biliary tract Cancer, rectal cancer, ovarian cancer, uterine cancer, kidney cancer, bladder cancer, prostate cancer, osteosarcoma, brain tumor, leukemia, muscle tumor, skin cancer, malignant melanoma, malignant lymphoma, tongue It can be used for the treatment of cancer, myeloma, thyroid cancer, skin metastasis cancer, cutaneous melanoma, etc., but is not limited thereto, and can also be used for the treatment of precancerous lesions.

  The dosage form of the palmitic acid amides 1, 2, and 3 of the present invention as an anticancer agent is not particularly limited, and may be any oral or parenteral dosage form. Moreover, it can be made into a suitable dosage form according to the administration form, for example, oral preparations such as injections, capsules, tablets, granules, powders, pills, fine granules, rectal administration agents, and oily suppositories. It can be prepared into various preparations such as aqueous suppositories.

  Various preparations can be prepared by appropriately adding pharmacologically acceptable and commonly used additives such as excipients, binders, lubricants, disintegrants, surfactants, fluidity promoters, and the like. As excipients, for example, lactose, fructose, glucose, corn starch, sorbit, crystalline cellulose and the like as binders, for example, methyl cellulose, ethyl cellulose, gum arabic, gelatin, hydroxypropyl cellulose, polyvinyl pyrrolidone and the like as lubricants For example, talc, magnesium stearate, polyethylene glycol, hydrogenated vegetable oil, etc., as disintegrants, for example, starch, sodium alginate, gelatin, calcium carbonate, calcium citrate, dextrin, magnesium carbonate, synthetic magnesium silicate, etc. Examples of the activator include sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80, and the like. Examples of the fluidity promoter include light anhydrous silicic acid, dry aluminum hydroxide. Gel, synthetic aluminum silicate, etc. magnesium silicate, as the other additives, syrup, petrolatum, glycerin, ethanol, propylene glycol, citric acid, sodium chloride, sodium nitrite, and sodium phosphate.

  The dosage of palmitic acid amides 1, 2, and 3 of the present invention can be appropriately increased or decreased in consideration of usage, patient age, sex, symptom level, etc., but usually 200 to 400 mg per day for an adult, preferably 100 to 200 mg, which can be administered once or divided into several times a day.

  EXAMPLES Next, although an Example is given and this invention is demonstrated, this invention is not limited to a following example.

[Example 1] Synthesis of palmitic acid amide-1 Palmitic acid (250 mg, 1 mmol, Wako Pure Chemical Industries) and piperazine (100 mg, 1 mmol, Wako Pure Chemical Industries) were dissolved in anhydrous dichloromethane and stirred in an ice bath. N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (211mg, 1.1mmol, Wako Pure Chemical Industries, Ltd.) Was added. After stirring in an ice bath for 1 hour and further at room temperature for 8 hours, the reaction solution was poured into dilute hydrochloric acid solution and extracted with chloroform. The chloroform layer was concentrated to give 1- (piperidin-1-yl) hexadecan-1-one (1- (piperidin-1-yl) hexadecan-1-one).
C ₂₁ H ₄₁ NO: Molecular weight: 323, colorless oil, EIMS m / z (rel. Int.): 323 (M ⁺ , 8), 140 (21), 127 (100), 112 (7), ¹ H -NMR (400 MHz, CDCl ₃ ): δ 0.88 (3H, t, J = 6.8 Hz), 1.25 (24H, m), 1.52-1.64 (8H, m), 2.34 (2H, t, J = 8.0 Hz) , 3.92 (2H, t, J = 5.4 Hz), 3.53 (2H, t, J = 5.6 Hz).

[Example 2] Synthesis of palmitic acid amide-2 Anhydrous palmitic acid (250 mg, 1 mmol, Wako Pure Chemical Industries) and 1,2,3,4-tetrahydroisoquinoline (371 mg, 1 mmol, Wako Pure Chemical Industries) N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (211 mg, N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide) dissolved in dichloromethane and stirred in an ice bath 1.1 mmol, Wako Pure Chemical Industries, Ltd.) was added. After stirring in an ice bath for 1 hour and further at room temperature for 8 hours, the reaction solution was poured into dilute hydrochloric acid solution and extracted with chloroform. The chloroform layer was concentrated to 1- (3,4-dihydroisoquinolin-2 (1H) -yl) hexadecan-1-one (1- (3,4-dihydroisoquinolin-2 (1H) -yl) hexadecan-1- one).
C ₂₅ H ₄₁ NO: Molecular weight: 371, colorless oil, EIMS m / z (rel. Int.): 371 (M ⁺ , 35), 140 (21), 175 (19), 133 (100), ¹ H -NMR (400 MHz, CDCl ₃ ): δ 0.88 (3H, t, J = 7.2 Hz), 1.25 (26H, m), 1.69 (2H, m), 3.05 (2H, t, J = 7.8 Hz), 3.83 (2H, t, J = 6.0 Hz), 4.73 (2H, s), 7.18-7.26 (4H, m).

Example 3 Synthesis of Palmitic Acid Amide Form-3 Palmitic acid (250 mg, 1 mmol, Wako Pure Chemical Industries, Ltd.) and 1,2,3,4-tetrahydroquinoline (371 mg, 1 mmol, Wako Pure Chemical Industries, Ltd.) were anhydrous. N- (3-dimethylaminopropyl) -N′-ethylcarbodiimide hydrochloride (211 mg, 1.1 mmol, Wako Pure Chemical Industries, Ltd.), a dehydrating condensing agent, was added with stirring in an ice bath and stirring in an ice bath. After stirring in an ice bath for 1 hour and further at room temperature for 8 hours, the reaction solution was poured into dilute hydrochloric acid solution and extracted with chloroform. The chloroform layer was concentrated to give 1- (3,4-dihydroquinolin-1 (2H) -yl) hexadecan-1-one (1- (3,4-dihydroquinolin-1 (2H) -yl) hexadecan-1- one).
C ₂₅ H ₄₁ NO: Molecular weight: 371, colorless oil, EIMS m / z (rel. Int.): 371 (M ⁺ , 100), 188 (15), 175 (43), 160 (7), 132 ( 24), 117 (9), 104 (8), ¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.88 (3H, t, J = 7.2 Hz), 1.25 (24H, m), 1.62 (4H, m ), 2.49 (2H, t, J = 7.6 Hz), 2.70 (2H, t, J = 7.0 Hz), 3.78 (2H, t, J = 6.6 Hz), 7.15 (4H, m).

[Example 4] (Evaluation of anticancer activity of palmitic acid amide)
1. Preparation of Reagent Palmitic acid amides 1, 2, and 3 were prepared by dissolving the ones produced in Examples 1 to 3 in acetone and adjusting the concentration. The Giemsa staining solution was used by diluting 25 mL of “Sigma Cat. No. S128-4L” with 475 mL of distilled water.
2. Cell line culture Human colon normal cell line FHC, human colorectal cancer cell line HT29 and human colorectal cancer cell line HCT116 were each 10% fetal bovine serum (FBS) (BioWest: Cat.No. S1500) supplemented DMEM medium (Wako Pure Chemical Industries, Ltd .: Cat. No. 041-29775) was cultured under conditions of 37 ° C. and 5% CO ₂ . As a culture apparatus, a Napco 5400 CO ₂ incubator (Napco) was used.

3. (Evaluation of anticancer activity of palmitic acid amides 1, 2, and 3 against human colon cancer cell line HT29)
Human colon cancer cell line HT29 was seeded in a 6-well plate (Nippon Becton Dickinson: Cat. No. 353046) at a concentration of 500 cells / well and cultured overnight, and then palmitic acid amide-1 was added to 0, 5 , 10, 20, 40, 80 μM, palmitic acid amides-2, 3 were exposed to cancer cells at concentrations of 0, 6, 13, 25, 15, 100 μM, respectively, as shown in FIG. The cells were cultured for 5 days (acetone was added to 0.1% in 0 μM medium). After culturing, each well was washed twice with phosphate buffered saline (hereinafter abbreviated as “PBS”), and cancer cells were fixed with 2 mL of 100% methanol for 10 minutes per well. Air-dried, stained with 2 mL of Giemsa staining solution for each well for 30 minutes, and then washed with tap water. After air drying, the number of colonies of stained cancer cells was visually measured as shown in FIG.

Based on the results of the above-described cell proliferation assay, a growth curve (dose response curve) shown in FIG. 3 was prepared, and IC ₅₀ values of 50% growth inhibition rates of palmitic acid amides 1, 2, and 3 were measured. As a result, the IC ₅₀ values of the palmitic acid amide compound against the colon cancer cell line HT29 were 34 μM for palmitic acid amide-1, 35 μM for palmitic acid amide-2, and 6 μM for palmitic acid amide-3. Of the palmitic acid amide had anticancer activity. In particular, palmitic acid amide-3 showed a marked anticancer activity at a low concentration. The literature value of IC ₅₀ for 5-fluorouracil (5FU) used as first-line chemotherapy for colorectal cancer is 13 μM (Wiebke EA et al. J Surg Res 111: 63-69, 2003), palmitic It was suggested that acid amide-3 may have stronger anticancer activity than 5FU.

4). (Comparison of toxicity of palmitic acid amide-3 to human colon normal cell line and human colon cancer cell line)
3. above. Was examined for toxicity to human colon normal cell line FHC, human colon cancer cell line HT29 and human colon cancer cell line HCT116. Each cell line was seeded in a 6 cm dish (Nalge Nunc International: Cat. No. 150288) at a concentration of 2 × 10 ⁴ cells / dish and cultured overnight, and then palmitic acid amide-3 was added to each cell line from 0 to 0. The cells were exposed to cancer cells at a concentration of 25 μM and cultured for 3 days (acetone was added to 0.1% to 0 μM medium). After culturing, the medium was removed, 1 mL of a 0.25% trypsin-EDTA solution (Sigma: Cat. No. T4049) was added and incubated at 37 ° C. for 10 minutes, and then the cells were detached from the dish. 3 mL of 1 × PBS was added to the detached cells to loosen the cells, and the number of cells was measured with Coulter Counter Z1 (Becton Dickinson).

  Based on the results of the above cell proliferation assay, the proliferation curve shown in FIG. 4 was prepared. Measure the viability of human colorectal cancer cell line HT29 and human colorectal cancer cell line HCT116 under the exposure of palmitic acid amide-3, and simultaneously measure the cell viability of human colon normal cell line FHC, palmitic acid The cell viability when the concentration of amide 3 was 0 μM was defined as 100%, and the cell viability at each concentration of palmitic acid amide 3 was converted. As a result, when human colon cancer cell line HT29 and human colon cancer cell line HCT116 were killed by 85 to 90%, the concentration of palmitic acid amide-3 was 20 μM, and human colon normal cell line FHC at this concentration Cell viability was 100%. When human colon cancer cell line HT29 and human colon cancer cell line HCT116 were 100% killed, the concentration of palmitic acid amide-3 was 25 μM, and the cell viability of human colon normal cell line FHC at this concentration was 80 μm. %Met. Palmitic acid amide-3 was not toxic to human colon normal cell line FHC at anticancer activity concentrations up to 20 μM. Therefore, palmitic acid amide-3 does not exhibit toxicity against normal cells at an effective concentration that exhibits anticancer activity, exhibits potent anticancer activity, and is a novel non-toxic anticancer agent. Be expected.

  INDUSTRIAL APPLICABILITY The present invention is expected to be useful in the pharmaceutical industry as a novel anticancer agent that exhibits strong anticancer activity and has no toxicity.

Claims (3)

General formula (1)

Wherein R is 3,4-dihydroisoquinolin-2 (1H) -yl or 3,4-dihydroisoquinolin-2 (1H) -yl (3, 4-dihydroquinolin-1 (2H) -yl)), or a pharmacologically acceptable salt thereof. General formula (2)

Wherein R is piperidin-1-yl or 3,4-dihydroisoquinolin-2 (1H) -yl or 3,4-dihydroisoquinolin-2 (1H) -yl Contains a palmitic acid amide compound represented by -dihydroquinolin-1 (2H) -yl (representing 3,4-dihydroquinolin-1 (2H) -yl) or a pharmaceutically acceptable salt thereof as an active ingredient An anticancer agent characterized by this.   The anticancer agent according to claim 2, wherein the anticancer activity is against colorectal cancer.

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