WO2009039773A1 - New medicine use of 1-substituted aryl -2(1h)-pyridone - Google Patents

New medicine use of 1-substituted aryl -2(1h)-pyridone Download PDF

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Publication number
WO2009039773A1
WO2009039773A1 PCT/CN2008/072406 CN2008072406W WO2009039773A1 WO 2009039773 A1 WO2009039773 A1 WO 2009039773A1 CN 2008072406 W CN2008072406 W CN 2008072406W WO 2009039773 A1 WO2009039773 A1 WO 2009039773A1
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Prior art keywords
pirfenidone
hours
flufenidone
proliferation
cancer cells
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PCT/CN2008/072406
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French (fr)
Chinese (zh)
Inventor
Lijian Tao
Gaoyun Hu
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Central South University
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Priority to CN2008800110076A priority Critical patent/CN101652138B/en
Publication of WO2009039773A1 publication Critical patent/WO2009039773A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4412—Non condensed pyridines; Hydrogenated derivatives thereof having oxo groups directly attached to the heterocyclic ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a novel pharmaceutical use of a 1-substituted aryl-2(1H)-pyridone compound, in particular a novel pharmaceutical use of a fluorosubstituted phenyl 2-1(1H)-pyridone.
  • Pyridone compounds are a large family with a wide range of physiological activities and different physiological activities due to the different groups substituted on the pyridine ring. Including insecticidal, anti-inflammatory, anti-fibrotic and the like.
  • the drug is an oral active small molecule drug approved by the US FDA.
  • the treatment of pulmonary, renal and hepatic fibrotic diseases has entered Phase II clinical trials.
  • the inventors of the present application have disclosed the compound 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone (Journal of Central South University (Medical Edition) (2004, 29(2)).
  • AKF-PD AKF-PD
  • PCT application WO2004/110245A2 discloses the use of pirfenidone for the treatment of cancer.
  • pirfenidone When pirfenidone is used in cancer treatment, it is administered in combination with a therapeutic amount of IP-10.
  • IP-10 When pirfenidone is used in cancer treatment, it is administered in combination with a therapeutic amount of IP-10.
  • the literature states that a therapeutic amount of pirfenidone in combination with a therapeutic amount of IP-10 can reduce tumors by 20%. Further, it may be administered in combination with IFN-a, an anti-proliferative agent, or as an adjuvant therapeutic agent.
  • IFN-a an anti-proliferative agent
  • the above prior art discloses a process for the preparation of the above compounds. Summary of the invention
  • the object of the present invention is to provide a new medical use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridone (abbreviated as flufenidone).
  • the technical solution provided by the present invention is: The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone for the preparation of antitumor drugs.
  • the tumor is lung cancer.
  • the tumor is breast cancer.
  • the tumor is colon cancer.
  • the tumor is gastric cancer.
  • the tumor is liver cancer.
  • said tumor is prostate cancer.
  • the present invention discloses that 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone has a stronger antitumor effect than the pirfenidone disclosed in the prior art.
  • Triflurane inhibits proliferation of lung cancer cells
  • Cell Human lung cancer cell (A549)
  • MTT kit is American R&D Company Second, the method:
  • a thiazole blue (MTT) method cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 ⁇ L per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 20, 44, 68, 92, and 116 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved. The OD value was measured by microplate reader. . Statistical methods, using one-way analysis of variance.
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.01).
  • 500 ⁇ ⁇ / ⁇ 1 of flufenidone and pirfenidone inhibited the proliferation of human lung cancer cells, but flufenidone 500 ⁇ ⁇ / ⁇ 1 was stronger than the same dose of pirfenidone.
  • 100 ⁇ ⁇ / ⁇ 1 of flufenidone inhibited the proliferation of human lung cancer cells, while the same dose of pirfenidone did not inhibit the proliferation of human lung cancer cells.
  • Triflurane inhibits proliferation of breast cancer cells
  • Cells Human breast cancer cells (MCF7 cells).
  • the cells were cultured in a DMEM medium containing 10% calf serum using a thiazole blue ( ⁇ ) method, and the cells were made into a lx10 5 /ml cell suspension, and 100 ⁇ L per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 20 and 44 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.05)
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.01).
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.001)
  • Triflurane inhibits proliferation of colon cancer cells ( ⁇ -29)
  • the cells were cultured in a DMEM medium containing 10% calf serum using a thiazole blue ( ⁇ ) method, and the cells were made into a lx10 5 /ml cell suspension, and 100 ⁇ L per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 24, 48, and 72 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance. Third, the experimental results: MTT results of flufenidone inhibiting proliferation of colon cancer cells
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.05)
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.01).
  • ⁇ ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.001)
  • Flufenidone 100ug/ml inhibited the proliferation of human colon cancer cells at 72 hours (p ⁇ 0.05 compared with the control group); pirfenidone lOOug/ml did not inhibit the proliferation of human colon cancer cells at 72 hours.
  • Flufenidone 200ug/ml inhibited the proliferation of human colon cancer cells at 48 and 72 hours (p ⁇ 0.01 compared with the control group; p ⁇ 0.001 at 72 hours); pirfenidone 200ug/ml dose at 48 The effect of inhibiting the proliferation of human colon cancer cells was inhibited by the hour, and the proliferation of human colon cancer cells was inhibited at 72 hours (p ⁇ 0.01 compared with the control group). 3.
  • Flufenidone 400ug/ml inhibited the proliferation of human colon cancer cells at 24, 48, 72 hours (p ⁇ 0.05 compared with the control group at 24 hours; p ⁇ 0.001 at 48 and 72 hours); pirfenidone 400ug/ Ml did not inhibit the proliferation of human colon cancer cells at 24 hours, inhibited the proliferation of human colon cancer cells at 48 and 72 hours (p ⁇ 0.001 compared with the control group); flufenidone 400 ug/ml dose group at 72 hours The inhibition of human colon cancer cells is greater than the equivalent dose of pirfenidone
  • Flufenidone 800ug/ml, 1600ug/ml and pirfenidone 800ug/ml, 1600ug/ml can inhibit the proliferation of human colon cancer cells at 24, 48, 72 hours (p ⁇ 0.001 compared with the control group) And with the increase of time, the inhibition rate increased; fenfluramine 800ug/ml, 1600ug/ml inhibited colon cancer cells at 24, 48, 72 hours than the same dose of pirfenidone (with the same dose) Pirfenidone was compared at 24 hours p ⁇ 0.05; 48, 72 hours p ⁇ 0.001). Fourth, the conclusion:
  • Triflurane inhibits proliferation of gastric cancer cells (BGC-803)
  • MTT kit is American R&D Company
  • a thiazole blue (MTT) method cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 ⁇ L per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 24, 48, and 72 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.05)
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.001)
  • 1, flufenidone and pirfenidone 1600ug / ml dose group can inhibit the proliferation of human gastric cancer cells (BGC-803) at 24, 48, 72 hours (compared with the control group ⁇ 0.001), and over time Prolonged, increased inhibition rate;
  • flufenidone and pirfenidone 800ug / ml dose group can inhibit the proliferation of human gastric cancer cells (BGC-803) at 72 hours (compared with the control group, flufenidone ⁇ 0.01, pirfenidone? ⁇ 0.001).
  • flufenidone and pirfenidone 400ug / ml dose group can inhibit the proliferation of human gastric cancer cells (BGC-803) at 72 hours (p ⁇ 0.05 compared with the control group). 4.
  • the flufenidone 1600ug/ml dose group inhibited colon cancer cells (BGC-803) at 72 hours more than the same dose of pirfenidone (p ⁇ 0.001).
  • Triflurane inhibits proliferation of gastric cancer cells (MGC-803)
  • MTT kit is American R&D Company
  • a thiazole blue (MTT) method cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 ⁇ L per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 24, 48, and 72 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
  • flufenidone and pirfenidone 400ug / ml dose group can inhibit the proliferation of human gastric cancer cells (MGC-803) at 72 hours (compared with the control group flufenidone p ⁇ 0.001, pirfenidone 2 ⁇ 0.05).
  • the flufenidone 800ug/ml dose group inhibited gastric cancer cells (MGC-803) at 72 hours more than the equivalent dose of pirfenidone (p ⁇ 0.05).
  • Triflurane inhibits proliferation of hepatoma cells (Bel-7402)
  • Cell Human liver cancer cell (Bel-7402)
  • MTT kit is American R&D Company
  • a thiazole blue (MTT) method cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 ⁇ L per well was seeded in a 96-well plate. After the cells were attached to the wall, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the culture containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was performed. Base, 5 replicate holes per concentration. After 24 and 48 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
  • ⁇ is statistically significant compared with the same dose group of pirfenidone ( ⁇ 0.05)
  • the flufenidone and pirfenidone 400ug/ml dose groups inhibited the proliferation of human hepatoma cells (Bel-7402) at 48 and 72 hours.
  • flufenidone and pirfenidone 800ug / ml, 1600ug / ml dose group can suppress people at 24, 48, 72 hours Proliferation of liver cancer cells (Bel-7402).
  • flufenidone 1600ug / ml dose group inhibited human hepatoma cells (Bel-7402) at 72 hours greater than the same dose of pirfenidone (p ⁇ 0.05).

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Abstract

The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridone in manufacture of a anti-tumor medicament is provided in the present invention, wherein the said tumor is lung cancer, breast cancer, colon cancer, stomach cancer, liver cancer and prostate cancer. According to the present invention, the effect of the said compound is better than that of the pirfenidone in the prior arts.

Description

说 明 书  Description
1-取代芳基- 2(1H)-吡啶酮化合物的新医药用途 技术领域  New medical use of 1-substituted aryl-2(1H)-pyridone compounds
本发明涉及 1-取代芳基- 2(1H)-吡啶酮化合物的新医药用途，具体地是氟取代苯基 _2( 1H) -吡啶酮的新医药用途。 背景技术  The present invention relates to a novel pharmaceutical use of a 1-substituted aryl-2(1H)-pyridone compound, in particular a novel pharmaceutical use of a fluorosubstituted phenyl 2-1(1H)-pyridone. Background technique
吡啶酮类化合物是一个庞大的家族， 具有广泛生理活性， 由于吡啶环上所取代的基团 不同而具有不同的生理活性。 包括具有杀虫， 抗炎， 抗纤维化等。  Pyridone compounds are a large family with a wide range of physiological activities and different physiological activities due to the different groups substituted on the pyridine ring. Including insecticidal, anti-inflammatory, anti-fibrotic and the like.
德国专利 DE 4343528 ， 欧洲专利 EP259048 、 EP367410 、 EP398499 , 欧洲专利 EP 216541 , 欧洲专利 EP488220, 美国专利 US3839346A, US4052509A, US4042699以及 USP3839346 ( 1974. 10. 1公布） 均公开了涉及上述吡啶酮类化合物的制备和用途。  The preparation of the above pyridone compounds is disclosed in German Patent No. DE 4, 343, 528, European Patent No. EP 2 590 486, EP 367 410, EP 398 499, European Patent No. EP 216 541, European Patent No. EP 228 420, European Patent No. EP 488,220, U.S. Patent No. 3,839, 346 A, U.S. Patent No. 4, 520, 091, U.S. And use.
美国专利 US 5,310,562在 1994年第一次公布了 1-苯基 -5-甲基 -2(1H)-吡啶酮，  U.S. Patent 5,310,562 first published 1-phenyl-5-methyl-2(1H)-pyridone in 1994.
[5-methyl- 1 -phenyl-2( 1 H)-pyridone] 又称吡非尼酮 （Pirfenidone, PFD)具有抗纤维化的生物 活性， EP0383591中描述了吡非尼酮在预防和治疗纤维化疾病方面有广泛的应用。该药物是 一种口服的活性小分子药物， 已被美国 FDA批准， 对肺、 肾和肝的纤维化疾病的治疗已进入 II期临床。 [5-methyl- 1 -phenyl-2( 1 H)-pyridone] Also known as pirfenidone (PFD) has anti-fibrotic biological activity, and EP0383591 describes pirfenidone in the prevention and treatment of fibrosis There are a wide range of applications for diseases. The drug is an oral active small molecule drug approved by the US FDA. The treatment of pulmonary, renal and hepatic fibrotic diseases has entered Phase II clinical trials.
申请号为 CN200510031445. 7和 CN02114190. 8中涉及了 1_芳基 _5_甲基 _2 (1H) -吡啶酮化 合物在制备抗纤维化药物中的用途。  Application Nos. CN200510031445. 7 and CN02114190. 8 relate to the use of a 1-aryl-5-methyl-2(1H)-pyridone compound for the preparation of an anti-fibrotic drug.
本申请发明人曾在 《中南大学学报》 （医学版） （2004， 29 ( 2 ) ) 上公开了化合物 1- ( 3-氟苯基） -5-甲基 -2 ( 1H) -吡啶酮 （AKF-PD) 对肾成纤维细胞的细胞实验， 显示其具 有抑制肾成纤维细胞生长的作用。  The inventors of the present application have disclosed the compound 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone (Journal of Central South University (Medical Edition) (2004, 29(2)). AKF-PD) Cellular experiments on renal fibroblasts have been shown to inhibit the growth of renal fibroblasts.
除此之外， PCT申请 W02004/110245A2公开了吡非尼酮 的在治疗癌症中的用途。 将吡 非尼酮用于癌症治疗时， 与治疗量的 IP-10联合用药。 该文献中称， 将治疗量的吡非尼酮与 治疗量的 IP-10联合用药可以减少肿瘤 20%。进一步还可以与 IFN-a， 抗增生剂联合用药， 还 可以是作为辅助治疗药。 上述现有技术中公开了上述化合物的制备方法。 发明内容 In addition, PCT application WO2004/110245A2 discloses the use of pirfenidone for the treatment of cancer. When pirfenidone is used in cancer treatment, it is administered in combination with a therapeutic amount of IP-10. The literature states that a therapeutic amount of pirfenidone in combination with a therapeutic amount of IP-10 can reduce tumors by 20%. Further, it may be administered in combination with IFN-a, an anti-proliferative agent, or as an adjuvant therapeutic agent. The above prior art discloses a process for the preparation of the above compounds. Summary of the invention
本发明的目的是提供 1- ( 3-氟苯基） -5-甲基 -2 ( 1H) -吡啶酮 （简称氟非尼酮） 的新 医药用途。  The object of the present invention is to provide a new medical use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridone (abbreviated as flufenidone).
本发明提供的技术方案是： 1- ( 3-氟苯基） -5-甲基 -2 ( 1H) -吡啶酮用于制备抗肿瘤 药物中的应用。  The technical solution provided by the present invention is: The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone for the preparation of antitumor drugs.
根据本发明的实施例， 优选所说的肿瘤是肺癌。  According to an embodiment of the invention, preferably the tumor is lung cancer.
根据本发明的实施例， 优选所说的肿瘤是乳腺癌。  According to an embodiment of the invention, it is preferred that the tumor is breast cancer.
根据本发明的实施例， 优选所说的肿瘤是结肠癌。  According to an embodiment of the invention, preferably the tumor is colon cancer.
根据本发明的实施例， 优选所说的肿瘤是胃癌。  According to an embodiment of the invention, it is preferred that the tumor is gastric cancer.
根据本发明的实施例， 优选所说的肿瘤是肝癌。  According to an embodiment of the invention, it is preferred that the tumor is liver cancer.
根据本发明的实施例， 优选所说的肿瘤是***癌。  According to an embodiment of the invention, preferably said tumor is prostate cancer.
本申请人通过研究发现， 1- ( 3-氟苯基） -5-甲基 -2 ( 1H) -吡啶酮较现有技术中公开 的吡非尼酮有更强的抗肿瘤作用。 通过对实验结果的统计学分析， 显示 1- ( 3-氟苯基） -5- 甲基 -2 ( 1H) -吡啶酮与吡非尼酮的抗肿瘤效果的差异具有显著性。  The Applicant has found through research that 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone has a stronger antitumor effect than the pirfenidone disclosed in the prior art. Statistical analysis of the experimental results showed that the difference in antitumor effect between 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridone and pirfenidone was significant.
本发明公开了 1- ( 3-氟苯基） -5-甲基 -2 ( 1H) -吡啶酮较现有技术中公开的吡非尼酮 有更强的抗肿瘤作用。 具体实施方式  The present invention discloses that 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone has a stronger antitumor effect than the pirfenidone disclosed in the prior art. detailed description
下面通过具体实施方案进一步说明本发明。 所述实施例仅用于说明或解释本发明的实 施方案， 而不能限制本发明的保护范围。 本发明实施例所涉及的细胞均可以从 ATCC公司商 业化产品中获得。  The invention is further illustrated by the following specific embodiments. The examples are only intended to illustrate or explain the embodiments of the invention, and are not intended to limit the scope of the invention. The cells involved in the examples of the present invention are all available from ATCC commercial products.
实施例 1 Example 1
氟非尼酮抑制肺癌细胞增殖的试验  Triflurane inhibits proliferation of lung cancer cells
一、 材料： First, the material:
1.细胞： 人肺癌细胞 (A549)  1. Cell: Human lung cancer cell (A549)
2.试剂： MTT试剂盒为美国 R&D公司 二、 方法： 2. Reagents: MTT kit is American R&D Company Second, the method:
用噻唑蓝 （MTT) 方法， 细胞用含 10%小牛血清的 DMEM培养基培养， 将细胞制成 lxl05/ml的细胞悬液， 每孔 lOOul接种于 96孔板中。 待细胞贴壁后换无血清 DMEM培养基， 24小时后弃去无血清培养基， 换含不同浓度氟非尼酮和吡非尼酮的含 10%小牛血清的培养 基， 每个浓度设 5个复孔。 分别于加药后 20、 44、 68、 92、 116小时后， 每孔加 MTTlOul, 4 小时后将 MTT吸出， 每孔加 MTT溶解液 100ul， 15min后待 MTT完全溶解， 酶标仪测 OD值。 统计学方法， 采用单因素方差分析。 Using a thiazole blue (MTT) method, cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 μL per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 20, 44, 68, 92, and 116 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved. The OD value was measured by microplate reader. . Statistical methods, using one-way analysis of variance.
三、 实验结果： Third, the experimental results:
氟非尼酮抑制肺癌细胞增殖的 MTT结果  MTT results of flufenidone inhibiting proliferation of lung cancer cells
Figure imgf000004_0001
Figure imgf000004_0001
* 表示与对照组比较有统计学意义 （p<0.05)  * indicates statistically significant comparison with the control group (p<0.05)
** 表示与对照组比较有统计学意义 （p<0.01 )  ** indicates statistical significance compared with the control group (p<0.01)
*** 表示与对照组比较有统计学意义 （p<0.001 )  *** indicates statistical significance compared with the control group (p<0.001)
Δ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.05) Δ indicates statistical significance compared with the same dose group of pirfenidone (ρ<0.05)
ΔΔ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.01 ) 在第 3、 4天 500μ§/ηι1的 氟非尼酮和吡非尼酮均能抑制人肺癌细胞的增殖， 但氟非尼酮 500μ§/ηι1的作用强于同等剂量的吡非尼酮。 在第 5天 100μ§/ηι1的 氟非尼酮能够抑制人肺癌 细胞的增殖， 而同等剂量的吡非尼酮不能抑制人肺癌细胞的增殖。 ΔΔ is statistically significant compared with the same dose group of pirfenidone (ρ<0.01). On day 3 and 4, 500 μ § / ηι1 of flufenidone and pirfenidone inhibited the proliferation of human lung cancer cells, but flufenidone 500 μ § / ηι1 was stronger than the same dose of pirfenidone. On day 5, 100 μ § / ηι1 of flufenidone inhibited the proliferation of human lung cancer cells, while the same dose of pirfenidone did not inhibit the proliferation of human lung cancer cells.
四、 结论： 氟非尼酮和吡非尼酮均能够抑制人肺癌细胞的增殖， 但氟非尼酮的作用强于吡 非尼酮。 实施例 2 4. Conclusions: Both fenfenidone and pirfenidone can inhibit the proliferation of human lung cancer cells, but flufenidone is more effective than pirfenidone. Example 2
氟非尼酮抑制乳腺癌细胞增殖的试验  Triflurane inhibits proliferation of breast cancer cells
一、 材料： First, the material:
1.细胞： 人乳腺癌细胞 （MCF7细胞）。  1. Cells: Human breast cancer cells (MCF7 cells).
2.试剂： ΜΤΤ试剂盒为美国 R&D公司  2. Reagents: ΜΤΤ kit is the US R&D company
二、 方法： Second, the method:
用噻唑蓝 （ΜΤΤ) 方法， 细胞用含 10%小牛血清的 DMEM培养基培养， 将细胞制成 lxl05/ml的细胞悬液， 每孔 lOOul接种于 96孔板中。 待细胞贴壁后换无血清 DMEM培养基， 24小时后弃去无血清培养基， 换含不同浓度氟非尼酮和吡非尼酮的含 10%小牛血清的培养 基， 每个浓度设 5个复孔。 分别于加药后 20、 44小时后， 每孔加 MTTlOul, 4小时后将 MTT 吸出， 每孔加 MTT溶解液 100ul， 15min后待 MTT完全溶解， 酶标仪测 OD值。 统计学方法， 采用单因素方差分析。 The cells were cultured in a DMEM medium containing 10% calf serum using a thiazole blue (ΜΤΤ) method, and the cells were made into a lx10 5 /ml cell suspension, and 100 μL per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 20 and 44 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
三、 实验结果： Third, the experimental results:
氟非尼酮抑制乳腺癌细胞增殖的 MTT结果  MTT results of flufenidone inhibiting proliferation of breast cancer cells
Figure imgf000005_0001
* 表示与对照组比较有统计学意义 （p<0.05 )
Figure imgf000005_0001
* indicates statistically significant compared with the control group (p<0.05)
** 表示与对照组比较有统计学意义 （p<0.01 )  ** indicates statistical significance compared with the control group (p<0.01)
*** 表示与对照组比较有统计学意义 （p<0.001 )  *** indicates statistical significance compared with the control group (p<0.001)
Δ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.05 ) Δ is statistically significant compared with the same dose group of pirfenidone (ρ<0.05)
ΔΔ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.01 ) ΔΔ is statistically significant compared with the same dose group of pirfenidone (ρ<0.01).
ΔΔΔ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.001 ) ΔΔΔ is statistically significant compared with the same dose group of pirfenidone (ρ<0.001)
在 24小时， 2500μ§/ηι1的 氟非尼酮能抑制人乳腺癌细胞的增殖， 在 48小时， 1000μ§/ηι1、 2500μ§/ηι1的 氟非尼酮和吡非尼酮均能抑制人乳腺癌细胞的增殖， 但氟非尼酮 2500μ§/ηι1 的作用强于同等剂量的吡非尼酮。 At 24 hours, 2500μ § /ηι1 of flufenidone inhibited the proliferation of human breast cancer cells. At 48 hours, 1000μ § /ηι1, 2500μ § /ηι1 of flufenidone and pirfenidone inhibited human mammary gland The proliferation of cancer cells, but the effect of flufenidone 2500μ § / ηι1 is stronger than the same dose of pirfenidone.
四、 结论： 氟非尼酮和吡非尼酮均能够抑制人乳腺癌细胞的增殖， 氟非尼酮的作用强于吡 非尼酮。 实施例 3 4. Conclusions: Both fenfenidone and pirfenidone can inhibit the proliferation of human breast cancer cells. The effect of flufenidone is stronger than that of pirfenidone. Example 3
氟非尼酮抑制结肠癌细胞 (ΗΤ-29)增殖的试验  Triflurane inhibits proliferation of colon cancer cells (ΗΤ-29)
一、 材料： First, the material:
1.细胞： 人结肠癌细胞 (ΗΤ-29)  1. Cell: Human colon cancer cells (ΗΤ-29)
2.试剂： ΜΤΤ试剂盒为美国 R&D公司  2. Reagents: ΜΤΤ kit is the US R&D company
二、 方法： Second, the method:
用噻唑蓝 （ΜΤΤ) 方法， 细胞用含 10%小牛血清的 DMEM培养基培养， 将细胞制成 lxl05/ml的细胞悬液， 每孔 lOOul接种于 96孔板中。 待细胞贴壁后换无血清 DMEM培养基， 24小时后弃去无血清培养基， 换含不同浓度氟非尼酮和吡非尼酮的含 10%小牛血清的培养 基， 每个浓度设 5个复孔。 分别于加药后 24、 48、 72小时后， 每孔加 MTTlOul, 4小时后 将 MTT吸出， 每孔加 MTT溶解液 100ul， 15min后待 MTT完全溶解， 酶标仪测 OD值。 统计 学方法， 采用单因素方差分析。 三、 实验结果： 氟非尼酮抑制结肠癌细胞增殖的 MTT结果 The cells were cultured in a DMEM medium containing 10% calf serum using a thiazole blue (ΜΤΤ) method, and the cells were made into a lx10 5 /ml cell suspension, and 100 μL per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 24, 48, and 72 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance. Third, the experimental results: MTT results of flufenidone inhibiting proliferation of colon cancer cells
Figure imgf000007_0001
Figure imgf000007_0001
* 表示与对照组比较有统计学意义 （ρ<0.05 )  * indicates statistically significant compared with the control group (ρ<0.05)
** 表示与对照组比较有统计学意义 （ρ<0.01 )  ** indicates statistical significance compared with the control group (ρ<0.01)
*** 表示与对照组比较有统计学意义 （ρ<0.001 )  *** indicates statistical significance compared with the control group (ρ<0.001)
Δ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.05 ) Δ is statistically significant compared with the same dose group of pirfenidone (ρ<0.05)
ΔΔ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.01 ) ΔΔ is statistically significant compared with the same dose group of pirfenidone (ρ<0.01).
ΔΔΔ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.001 ) Δ ΔΔ is statistically significant compared with the same dose group of pirfenidone (ρ<0.001)
1. 氟非尼酮 100ug/ml在 72小时可抑制人结肠癌细胞的增殖（与对照组比较 p<0.05 ); 吡非尼酮 lOOug/ml在 72小时无抑制人结肠癌细胞增殖的作用。  1. Flufenidone 100ug/ml inhibited the proliferation of human colon cancer cells at 72 hours (p<0.05 compared with the control group); pirfenidone lOOug/ml did not inhibit the proliferation of human colon cancer cells at 72 hours.
2. 氟非尼酮 200ug/ml在 48、 72小时可抑制人结肠癌细胞的增殖 （与对照组比较 48 小时 p<0.01 ; 72小时 p<0.001 ); 吡非尼酮 200ug/ml剂量在 48小时无抑制人结肠癌细胞增 殖的作用， 在 72小时可抑制人结肠癌细胞的增殖 （与对照组比较p<0.01 )。 3. 氟非尼酮 400ug/ml能在 24、 48、 72小时抑制人结肠癌细胞的增殖（与对照组比较 24小时 p<0.05; 48、 72小时 p<0.001 )； 吡非尼酮 400ug/ml在 24小时无抑制人结肠癌细 胞增殖的作用， 在 48、 72小时能抑制人结肠癌细胞的增殖 （与对照组比较p<0.001 )； 氟 非尼酮 400ug/ml 剂量组在 72 小时对人结肠癌细胞的抑制作用大于同等剂量的吡非尼酮2. Flufenidone 200ug/ml inhibited the proliferation of human colon cancer cells at 48 and 72 hours (p<0.01 compared with the control group; p<0.001 at 72 hours); pirfenidone 200ug/ml dose at 48 The effect of inhibiting the proliferation of human colon cancer cells was inhibited by the hour, and the proliferation of human colon cancer cells was inhibited at 72 hours (p<0.01 compared with the control group). 3. Flufenidone 400ug/ml inhibited the proliferation of human colon cancer cells at 24, 48, 72 hours (p<0.05 compared with the control group at 24 hours; p<0.001 at 48 and 72 hours); pirfenidone 400ug/ Ml did not inhibit the proliferation of human colon cancer cells at 24 hours, inhibited the proliferation of human colon cancer cells at 48 and 72 hours (p<0.001 compared with the control group); flufenidone 400 ug/ml dose group at 72 hours The inhibition of human colon cancer cells is greater than the equivalent dose of pirfenidone
(ρ<0·001 )。 (ρ<0·001).
4. 氟非尼酮 800ug/ml、 1600ug/ml和吡非尼酮 800ug/ml、 1600ug/ml均能在 24、 48、 72小时抑制人结肠癌细胞的增殖 （与对照组比较 p<0.001 )， 并且随着时间的延长， 抑制 率增加； 氟非尼酮 800ug/ml、 1600ug/ml在 24、 48、 72小时对结肠癌细胞的抑制作用均大 于同等剂量的吡非尼酮（与同等剂量的吡非尼酮比较 24小时 p<0.05; 48、72小时 p<0.001 )。 四、 结论：  4. Flufenidone 800ug/ml, 1600ug/ml and pirfenidone 800ug/ml, 1600ug/ml can inhibit the proliferation of human colon cancer cells at 24, 48, 72 hours (p<0.001 compared with the control group) And with the increase of time, the inhibition rate increased; fenfluramine 800ug/ml, 1600ug/ml inhibited colon cancer cells at 24, 48, 72 hours than the same dose of pirfenidone (with the same dose) Pirfenidone was compared at 24 hours p < 0.05; 48, 72 hours p < 0.001). Fourth, the conclusion:
氟非尼酮和吡非尼酮均能够抑制人结肠癌细胞的增殖， 氟非尼酮的抑制作用强于同等 剂量吡非尼酮。 实施例 4  Both fenfenidone and pirfenidone inhibited the proliferation of human colon cancer cells, and flufenidone was more potent than the same dose of pirfenidone. Example 4
氟非尼酮抑制胃癌细胞 (BGC-803)增殖的试验  Triflurane inhibits proliferation of gastric cancer cells (BGC-803)
一、 材料： First, the material:
1.细胞： 人胃癌细胞 (BGC-803)  1. Cell: Human gastric cancer cell (BGC-803)
2.试剂： MTT试剂盒为美国 R&D公司  2. Reagents: MTT kit is American R&D Company
二、 方法： Second, the method:
用噻唑蓝 （MTT) 方法， 细胞用含 10%小牛血清的 DMEM培养基培养， 将细胞制成 lxl05/ml的细胞悬液， 每孔 lOOul接种于 96孔板中。 待细胞贴壁后换无血清 DMEM培养基， 24小时后弃去无血清培养基， 换含不同浓度氟非尼酮和吡非尼酮的含 10%小牛血清的培养 基， 每个浓度设 5个复孔。 分别于加药后 24、 48、 72小时后， 每孔加 MTTlOul, 4小时后 将 MTT吸出， 每孔加 MTT溶解液 100ul， 15min后待 MTT完全溶解， 酶标仪测 OD值。 统计 学方法， 采用单因素方差分析。 Using a thiazole blue (MTT) method, cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 μL per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 24, 48, and 72 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
三、 实验结果： 氟非尼酮抑制胃癌细胞 (BGC-803)增殖的 MTT结果 Third, the experimental results: MTT results of flufenidone inhibiting proliferation of gastric cancer cells (BGC-803)
Figure imgf000009_0001
Figure imgf000009_0001
* 表示与对照组比较有统计学意义 （ρ<0.05 ) * indicates statistically significant compared with the control group (ρ<0.05)
** 表示与对照组比较有统计学意义 （ρ<0.01 )  ** indicates statistical significance compared with the control group (ρ<0.01)
*** 表示与对照组比较有统计学意义 （ρ<0.001 )  *** indicates statistical significance compared with the control group (ρ<0.001)
Δ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.05 ) Δ is statistically significant compared with the same dose group of pirfenidone (ρ<0.05)
ΔΔΔ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.001 ) ΔΔΔ is statistically significant compared with the same dose group of pirfenidone (ρ<0.001)
1、 氟非尼酮和吡非尼酮 1600ug/ml 剂量组均能在 24、 48、 72 小时抑制人胃癌细胞 (BGC-803)的增殖 （与对照组比较 <0.001 )， 并且随着时间的延长， 抑制率增加；  1, flufenidone and pirfenidone 1600ug / ml dose group can inhibit the proliferation of human gastric cancer cells (BGC-803) at 24, 48, 72 hours (compared with the control group <0.001), and over time Prolonged, increased inhibition rate;
2、 氟非尼酮和吡非尼酮 800ug/ml剂量组均能在 72小时抑制人胃癌细胞 (BGC-803)的 增殖 （与对照组比较， 氟非尼酮 <0.01， 吡非尼酮？<0.001 )。  2, flufenidone and pirfenidone 800ug / ml dose group can inhibit the proliferation of human gastric cancer cells (BGC-803) at 72 hours (compared with the control group, flufenidone <0.01, pirfenidone? <0.001).
3、 氟非尼酮和吡非尼酮 400ug/ml剂量组均能在 72小时抑制人胃癌细胞 (BGC-803)的 增殖 （与对照组比较 p<0.05)。 4、 氟非尼酮 1600ug/ml剂量组在 72小时对结肠癌细胞 (BGC-803)的抑制作用大于同等 剂量的吡非尼酮 （p<0.001 )。 3, flufenidone and pirfenidone 400ug / ml dose group can inhibit the proliferation of human gastric cancer cells (BGC-803) at 72 hours (p<0.05 compared with the control group). 4. The flufenidone 1600ug/ml dose group inhibited colon cancer cells (BGC-803) at 72 hours more than the same dose of pirfenidone (p<0.001).
四、 结论：  Fourth, the conclusion:
氟非尼酮和吡非尼酮均能够抑制人胃癌细胞 (BGC-803)的增殖； 氟非尼酮 1600ug/ml的 抑制作用强于同等剂量吡非尼酮。 实施例 5  Both fenfenidone and pirfenidone inhibited the proliferation of human gastric cancer cells (BGC-803); flufenidone 1600ug/ml was more potent than the same dose of pirfenidone. Example 5
氟非尼酮抑制胃癌细胞 (MGC-803)增殖的试验  Triflurane inhibits proliferation of gastric cancer cells (MGC-803)
一、 材料：  First, the material:
1.细胞： 人胃癌细胞 (MGC-803)  1. Cell: Human gastric cancer cell (MGC-803)
2.试剂： MTT试剂盒为美国 R&D公司  2. Reagents: MTT kit is American R&D Company
二、 方法：  Second, the method:
用噻唑蓝 （MTT) 方法， 细胞用含 10%小牛血清的 DMEM培养基培养， 将细胞制成 lxl05/ml的细胞悬液， 每孔 lOOul接种于 96孔板中。 待细胞贴壁后换无血清 DMEM培养基， 24小时后弃去无血清培养基， 换含不同浓度氟非尼酮和吡非尼酮的含 10%小牛血清的培养 基， 每个浓度设 5个复孔。 分别于加药后 24、 48、 72小时后， 每孔加 MTTlOul, 4小时后 将 MTT吸出， 每孔加 MTT溶解液 100ul， 15min后待 MTT完全溶解， 酶标仪测 OD值。 统计 学方法， 采用单因素方差分析。 Using a thiazole blue (MTT) method, cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 μL per well was seeded in a 96-well plate. After the cells were attached, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the medium containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was changed. 5 duplicate holes. After 24, 48, and 72 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
三、 实验结果：  Third, the experimental results:
氟非尼酮抑制胃癌细胞 (MGC-803)增殖的 MTT结果  MTT results of flufenidone inhibiting proliferation of gastric cancer cells (MGC-803)
Figure imgf000010_0001
吡非尼酮 100ug/ml
Figure imgf000010_0001
Pirfenidone 100ug/ml
0.245 ±0.0 0.456±0.013 0·559±0·021  0.245 ±0.0 0.456±0.013 0·559±0·021
吡非尼酮 200ug/ml Pirfenidone 200ug/ml
0.244±0.014 0.453 ±0.029 0.593 ±0.035  0.244±0.014 0.453 ±0.029 0.593 ±0.035
吡非尼酮 400ug/ml Pirfenidone 400ug/ml
0.235 ±0.020 0.447 ±0.020 0.493 ±0.028* 吡非尼酮 800ug/ml  0.235 ±0.020 0.447 ±0.020 0.493 ±0.028* pirfenidone 800ug/ml
0.184±0.015*** 0.353 ±0.045*** 0.444 ±0.029*** 0.184±0.015*** 0.353 ±0.045*** 0.444 ±0.029***
* 表示与对照组比较有统计学意义 （ρ<0.05) * indicates statistically significant compared with the control group (ρ<0.05)
** 表示与对照组比较有统计学意义 （ρ<0.01 ) ** indicates statistical significance compared with the control group (ρ<0.01)
*** 表示与对照组比较有统计学意义 （ρ<0.001 ) *** indicates statistical significance compared with the control group (ρ<0.001)
Δ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.05) Δ indicates statistical significance compared with the same dose group of pirfenidone (ρ<0.05)
1、 氟非尼酮和吡非尼酮 800ug/ml均能在 24、 48、 72小时抑制人胃癌细胞 (MGO803) 的增殖 （与对照组比较 <0.001 )。  1. Both florfenidone and pirfenidone 800ug/ml inhibited the proliferation of human gastric cancer cells (MGO803) at 24, 48, and 72 hours (<0.001 compared with the control group).
2、 氟非尼酮和吡非尼酮 400ug/ml剂量组均能在 72小时抑制人胃癌细胞 (MGC-803)的 增殖 （与对照组比较氟非尼酮 p<0.001， 吡非尼酮2<0.05)。  2, flufenidone and pirfenidone 400ug / ml dose group can inhibit the proliferation of human gastric cancer cells (MGC-803) at 72 hours (compared with the control group flufenidone p < 0.001, pirfenidone 2 <0.05).
3、 氟非尼酮 800ug/ml剂量组在 72小时对胃癌细胞 (MGC-803)的抑制作用大于同等剂 量的吡非尼酮 （p<0.05)。  3. The flufenidone 800ug/ml dose group inhibited gastric cancer cells (MGC-803) at 72 hours more than the equivalent dose of pirfenidone (p<0.05).
四、 结论： Fourth, the conclusion:
氟非尼酮和吡非尼酮均能够抑制人胃癌细胞 (MGC-803)的增殖， 氟非尼酮 800ug/ml 的 抑制作用强于同等剂量吡非尼酮。 实施例 6  Both fenfenidone and pirfenidone inhibited the proliferation of human gastric cancer cells (MGC-803), and the inhibitory effect of flufenidone 800ug/ml was stronger than that of the same dose of pirfenidone. Example 6
氟非尼酮抑制肝癌细胞（Bel-7402)增殖的试验  Triflurane inhibits proliferation of hepatoma cells (Bel-7402)
一、 材料： First, the material:
1.细胞： 人肝癌细胞 （Bel-7402)  1. Cell: Human liver cancer cell (Bel-7402)
2.试剂： MTT试剂盒为美国 R&D公司  2. Reagents: MTT kit is American R&D Company
二、 方法： Second, the method:
用噻唑蓝 （MTT) 方法， 细胞用含 10%小牛血清的 DMEM培养基培养， 将细胞制成 lxl05/ml的细胞悬液， 每孔 lOOul接种于 96孔板中。 待细胞贴壁后换无血清 DMEM培养基， 24小时后弃去无血清培养基， 换含不同浓度氟非尼酮和吡非尼酮的含 10%小牛血清的培养 基，每个浓度设 5个复孔。分别于加药后 24、 48小时后，每孔加 MTTlOul, 4小时后将 MTT 吸出， 每孔加 MTT溶解液 100ul， 15min后待 MTT完全溶解， 酶标仪测 OD值。 统计学方法， 采用单因素方差分析。 Using a thiazole blue (MTT) method, cells were cultured in DMEM medium containing 10% calf serum, and the cells were made into a lx10 5 /ml cell suspension, and 100 μL per well was seeded in a 96-well plate. After the cells were attached to the wall, the serum-free DMEM medium was changed. After 24 hours, the serum-free medium was discarded, and the culture containing 10% calf serum containing different concentrations of flufenidone and pirfenidone was performed. Base, 5 replicate holes per concentration. After 24 and 48 hours after dosing, MTTlOul was added to each well. After 4 hours, MTT was aspirated. MTT solution was added to each well for 100 ul. After 15 min, MTT was completely dissolved, and the OD value was measured by a microplate reader. Statistical methods, using one-way analysis of variance.
三、 实验结果：  Third, the experimental results:
氟非尼酮抑制肝癌细胞（Bel-7402)增殖的 MTT结果  MTT results of flufenidone inhibiting proliferation of hepatoma cells (Bel-7402)
Figure imgf000012_0001
Figure imgf000012_0001
* 表示与对照组比较有统计学意义 （ρ<0.05 ) * indicates statistically significant compared with the control group (ρ<0.05)
** 表示与对照组比较有统计学意义 （ρ<0.01 )  ** indicates statistical significance compared with the control group (ρ<0.01)
*** 表示与对照组比较有统计学意义 （ρ<0.001 )  *** indicates statistical significance compared with the control group (ρ<0.001)
Δ表示与相同剂量组吡非尼酮比较有统计学意义 （ρ<0.05 ) Δ is statistically significant compared with the same dose group of pirfenidone (ρ<0.05)
1、氟非尼酮和吡非尼酮 400ug/ml剂量组均能在 48、72小时抑制人肝癌细胞 (Bel-7402) 的增殖。  1. The flufenidone and pirfenidone 400ug/ml dose groups inhibited the proliferation of human hepatoma cells (Bel-7402) at 48 and 72 hours.
2、 氟非尼酮和吡非尼酮 800ug/ml 、 1600ug/ml剂量组均能在 24、 48、 72小时抑制人 肝癌细胞 （Bel-7402) 的增殖。 2, flufenidone and pirfenidone 800ug / ml, 1600ug / ml dose group can suppress people at 24, 48, 72 hours Proliferation of liver cancer cells (Bel-7402).
3、氟非尼酮 1600ug/ml剂量组在 72小时对人肝癌细胞（Bel-7402)的抑制作用大于同 等剂量的吡非尼酮 （p<0.05)。  3, flufenidone 1600ug / ml dose group inhibited human hepatoma cells (Bel-7402) at 72 hours greater than the same dose of pirfenidone (p <0.05).
四、 结论： 氟非尼酮和吡非尼酮均能够抑制人肝癌细胞的增殖， 氟非尼酮 1600ug/ml 的抑制作用强于同等剂量吡非尼酮。  IV. Conclusion: Both fenfenidone and pirfenidone can inhibit the proliferation of human hepatoma cells. The inhibitory effect of flufenidone 1600ug/ml is stronger than that of the same dose of pirfenidone.

Claims

权 利 要 求 书 Claim
1、 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤药物中的应用。1. The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone for the preparation of antitumor drugs.
2、 根据权利要求 1所述的 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤 药物中的应用， 优选所说的肿瘤是肺癌。 The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone according to claim 1 for the preparation of an antitumor drug, preferably said tumor is lung cancer.
3、 根据权利要求 1所述的 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤 药物中的应用， 优选所说的肿瘤是乳腺癌。  3. Use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone according to claim 1 for the preparation of an antitumor drug, preferably said tumor is breast cancer.
4、 根据权利要求 1所述的 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤 药物中的应用， 优选所说的肿瘤是结肠癌。  The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone according to claim 1 for the preparation of an antitumor drug, preferably said tumor is colon cancer.
5、 根据权利要求 1所述的 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤 药物中的应用， 优选所说的肿瘤是胃癌。  The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone according to claim 1 for the preparation of an antitumor drug, preferably wherein the tumor is gastric cancer.
6、 根据权利要求 1所述的 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤 药物中的应用， 优选所说的肿瘤是肝癌。  The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone according to claim 1 for the preparation of an antitumor drug, preferably wherein the tumor is liver cancer.
7、 根据权利要求 1所述的 1- (3-氟苯基） -5-甲基 -2 (1H) -吡啶酮用于制备抗肿瘤 药物中的应用， 优选所说的肿瘤是***癌。  7. The use of 1-(3-fluorophenyl)-5-methyl-2(1H)-pyridinone according to claim 1 for the preparation of an antitumor drug, preferably said tumor is prostate cancer.
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