WO2011147254A1 - Phenylbutyryl curcumin derivatives and uses for preparing anti-tumor drugs thereof - Google Patents

Phenylbutyryl curcumin derivatives and uses for preparing anti-tumor drugs thereof Download PDF

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WO2011147254A1
WO2011147254A1 PCT/CN2011/073844 CN2011073844W WO2011147254A1 WO 2011147254 A1 WO2011147254 A1 WO 2011147254A1 CN 2011073844 W CN2011073844 W CN 2011073844W WO 2011147254 A1 WO2011147254 A1 WO 2011147254A1
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compound
curcumin
group
pharmaceutically acceptable
cancer
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PCT/CN2011/073844
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French (fr)
Chinese (zh)
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许建华
刘洋
吴丽贤
林燕芳
吴枝娟
郭晓丹
吴敏
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福建医科大学
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Priority to GB1223251.8A priority Critical patent/GB2494595B/en
Publication of WO2011147254A1 publication Critical patent/WO2011147254A1/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/21Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/216Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/40Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino groups bound to carbon atoms of at least one six-membered aromatic ring and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/42Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino groups bound to carbon atoms of at least one six-membered aromatic ring and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton with carboxyl groups linked to the six-membered aromatic ring, or to the condensed ring system containing that ring, by saturated carbon chains
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • A61P35/02Antineoplastic agents specific for leukemia
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/14Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof
    • C07C227/18Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters

Definitions

  • the invention belongs to the field of pharmacy, and in particular relates to a phenylbutyryl curcumin derivative and a preparation method thereof, and the use thereof in preparing an antitumor drug.
  • Curcumin (Cur) is an active ingredient extracted from the rhizome of the turmeric, turmeric, turmeric, and turmeric of the genus Zingiberaceae. It has anti-tumor, anti-inflammatory, anti-human immunodeficiency virus, anti-cholesterol, anti-oxidation and so on. A pharmacological effect has a good clinical application potential.
  • Cur is unstable, rapid metabolism in the body, low bioavailability, and it is difficult to achieve an effective concentration in the body, which seriously restricts the development of curcumin into an effective anticancer drug.
  • Curcumin derivatives synthesized through structural transformation, improve bioavailability, prolonged biological t 1/2 elimination half-life is important.
  • One of the objects of the present invention is to provide 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin and 4,4'-[bis(2-chloroethyl:)amino]bisphenylbutanoyl turmeric And its pharmaceutically acceptable salt.
  • a second object of the present invention is to provide 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin and 4,4'-[bis(2-chloroethyl:)amino]bisphenylbutanoyl turmeric And a method for preparing the pharmaceutically acceptable salt thereof.
  • the pharmaceutically acceptable salts include alkali metal salts, alkaline earth metal salts, organic base-containing salts, and organic base-containing salts.
  • the acceptable salts include calcium salts, magnesium salts, ammonium salts, triethylamine salts, and ethanolamine salts.
  • the preparation method of the curcumin derivative and the salt of the present invention is specifically as follows: The curcumin is dissolved in the dried dichloromethane, and the catalytic amount of DMAP ( ⁇ , ⁇ -4-dimethylaminopyridine) is added. Then, 4-[bis(2-chloroethyl:)amino] phenylbutyric acid which has been dissolved in methylene chloride is added for esterification, and then the product is separated and purified by column chromatography.
  • a dehydrating agent DCC (dicyclohexylcarbodiimide) or EDCI (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) may be added first.
  • DCC diclohexylcarbodiimide
  • EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
  • the curcumin derivatives of the compound 1 and the compound 2 synthesized by the present invention are all obtained by nuclear magnetic resonance spectroscopy (NMR), infrared spectroscopy (IR), ultraviolet spectroscopy (UV), and liquid chromatography-mass spectrometry (HPLC-Mass). After the identification of Compound 1 and Compound 2, a suitable pharmaceutically acceptable dosage form is prepared as needed.
  • the present invention is a pharmaceutical composition or preparation comprising a medicament for protecting Compound 1 or Compound 2 or a pharmaceutically acceptable salt thereof, or a medicament containing Compound 1 or Compound 2 or a pharmaceutically acceptable salt thereof.
  • the carrier is a conventional pharmaceutical carrier in the pharmaceutical field, such as a diluent, an excipient, a filler, a binder, a disintegrant, a surfactant, a lubricant, etc.;
  • Various dosage forms for oral administration and other modes of administration such as oral liquids, suspensions, capsules, tablets, pills, granules, powder injections, etc.; prepared by conventional production methods in the pharmaceutical field, which are selected to contain weight percentages It is 0.1% to 99.5% active ingredient of Compound 1 or Compound 2.
  • Pharmaceutical compositions which may also contain from 0.1% to 99.5% of Compound 1 or Compound 2; are those which can be practiced by one of ordinary skill in the art.
  • the dosage of the present invention may be changed according to the route of administration, the age and weight of the patient, the type and severity of the disease to be treated, and the dose may be 0.001 - 10 g / k g body weight, which may be given one or more times. medicine.
  • the third object of the present invention is to 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin and 4,4'-[bis(2-chloroethyl)amino]bisphenylbutanoyl curcumin, For the preparation of anti-tumor drugs.
  • Curcumin derivatives can be used, but are not limited to, to prepare drugs for the treatment of leukemia, skin cancer, gastric cancer, colon cancer, liver cancer, breast cancer or prostate cancer.
  • Leukemia is preferably human chronic myeloid leukemia.
  • the compound 1 curcumin 4- [bis(2-chloroethyl)amino] phenylbutyrate of the present invention is significantly inhibited in various animal tumor cell transplantation models in vivo, especially
  • the inhibitory effect on human chronic myeloid leukemia K562 cells in nude mice can be as high as 60%, and the weight of nude mice in the drug-administered group did not decrease significantly, and no animal died.
  • FIG. 1 is a graph showing the inhibitory effect of intravenous administration of Compound 1 on mouse liver cancer H22 xenografts.
  • Figure 2 is a graph showing the inhibitory effect of oral administration of compound 1 on liver cancer H22 xenografts in mice.
  • Figure 3 is a graph showing the growth curve of compound 1 in human chronic myeloid leukemia K562 cells transplanted in nude mice.
  • Figure 4 is a diagram showing the effect of Compound 1 on human chronic myeloid leukemia K562 cells transplanted in nude mice.
  • Figure 5 and Figure 6 are the gross anatomy of mouse 1 in mouse model of human chronic myeloid leukemia and the mouse bone marrow bcr-abl gene. Expression map
  • Figure 7 and Figure 8 are the peripheral blood images of NS group and compound 1 administered to human chronic myeloid leukemia model mice.
  • Figure 9 is a graph showing the inhibitory effect of Compound 2 on mouse liver cancer H22 xenografts
  • H22 mouse xenografts were established, and the physiological saline (control group) of the mice was intravenously administered, Compound 1 : 50, 70 mg/kg (administration group).
  • the tumor inhibition rate is as high as 60%.
  • H22 mouse xenografts were established and administered orally to mice with physiological saline (control group).
  • Compound 1 50, 75, 100 mg/kg (administration group), curcumin 50 mg/kg (curcumin) Control group).
  • the tumor inhibition rates of the drug-administered group were 41.21%, 52.93%, and 75.06%, respectively.
  • the anti-tumor rate of the curcumin control group (equal to the compound lOOmg/kg) was only 16.58%, and the anti-tumor effect of the compound was significantly stronger than that of curcumin.
  • a nude mouse xenograft model of human chronic myeloid leukemia K562 cells was established and administered orally to nude mice (control group), compound 1: 40, 60 mg/kg (administration group).
  • control group nude mice
  • compound 1 40, 60 mg/kg
  • the length, width and height of the mouse tumor were measured with vernier calipers.
  • Calculated tumor volume length> ⁇ width X height xl/2
  • percentage of growth (%) (average tumor volume of the experimental group)
  • the mean tumor volume of the control group was ⁇ 100%.
  • the growth of tumors in the drug-administered group was significantly inhibited.
  • a nude mouse xenograft model of ⁇ 562 cells was established and orally administered to nude mice (control group), compound 1: 40, 60 mg/kg (administration group).
  • the tumor inhibition rates were 66.6% and 56.7%, respectively.
  • a human chronic myeloid leukemia model was constructed using NOD-SCID mice.
  • the control group was grossly dissected, and a large amount of bloody ascites was seen in the abdominal cavity, and solid tumors appeared.
  • the characteristic gene bcr-abl of human chronic myeloid leukemia was detected by RT-PCR, indicating that human chronic myeloid leukemia cell K562 homing to the bone marrow of NOD-SCID mice.
  • NOD-SCID mouse human chronic myeloid leukemia model was orally administered to normal saline (control group), compound 1: 60 mg/kg (administered group), peripheral blood group of the drug-administered group.
  • the naive cells were also significantly less than the control group.
  • the physiological saline (control group) of the mice was orally administered, Compound 2: 35, 50, 75, 115 mg/kg (administration group), curcumin 50 mg/kg (curcumin control group).
  • the tumor inhibition rates of the drug-administered group were 27.43%, 32.25%, 62.60%, and 58.39%, respectively.
  • Curcumin group (compound 100m g / kg equimolar) inhibition rate was 13.42%, visible inhibitory effect of Compound 2 significantly stronger than curcumin.
  • EDCI 1 92g (10mmol) was dissolved in anhydrous dichloromethane 200ml, stirred in an ice bath and added with curcumin 7.36g (20mmol), DMAP 0. 244g (2mmol), then slowly added to dissolve 4- [Bis(2-chloroethyl)amino] phenylbutyric acid 3. 04 g (10 mmol) of dichloromethane (200 ml), and the reaction was further stirred at room temperature for 6 hours. After washing with distilled water, the organic phase was dried over anhydrous magnesium sulfate, and then filtered and evaporated to give a crude product.
  • EDCI 1 92g (10mmol) was dissolved in 100ml of anhydrous dichloromethane, stirred in an ice bath and added with curcumin 1.84g (5mmol), DMAP 0. 122g (lmmol), then slowly added to dissolve 4- [Bis(2-chloroethyl)amino] phenylbutyric acid 3. 04 g (10 mmol) of dichloromethane (200 ml), and the reaction was further stirred at room temperature for 6 hours. After washing with distilled water, the organic phase was dried over anhydrous magnesium sulfate, and then filtered and evaporated to give a crude product.
  • mice 8 ⁇ 12 weeks old Kunming healthy mice, female, weighing (20 ⁇ 2) g; mice were provided by Experimental Animal Center of Fujian Medical University (certificate No. SCXK ( ⁇ :) 200420002). Mouse liver cancer cell line H22.
  • H22 tumor cells were passed for more than two generations. The number of cells was adjusted to 10 7 /ml, 0.2 mL/only inoculated into the right forelimb of mice, and 2 inoculated. After about two weeks, the tumor was stripped, homogenized, and inoculated again.
  • a total of 3 groups The mice after 24 h inoculation of tumors were randomly divided into 3 groups: Group I was a saline control group, and Compound 1 was prepared by the method of the above examples. The same group II was compound 1 low. The dose group (Compound 1 50 mg/kg), Group III was Compound 1 high dose group (70 mg/kg), and the administration method was tail vein injection, 0.1 ml/10 g.
  • mice A total of 5 groups: 24 hours after inoculation of tumors, the mice were randomly divided into 5 groups: group I was saline group (ie, tumor control group), group II was compound 1 low dose group (dose was 50 mg/kg) , III group compound 1 dose (a dose of 75mg / kg), IV group compound high dose groups (dose 100m g / k g), V group Cur group (at a dose of 50mg / kg, with a high dose Group IV equimolar), the method of administration is gavage,
  • the tumor inhibition rate (%) the average tumor quality of the control group - the average tumor mass of the experimental group / the average tumor mass of the control group ⁇ %.
  • Compound 1 has a significant inhibitory effect on the transplant rate of mouse liver cancer H22 (see Table 1, Figure 1 of the accompanying drawings).
  • Table 1 Compound 1 intravenous administration to mouse liver cancer H22 xenograft inhibition agent number of rats tumor weight (g) tumor inhibition rate group other mg/kg > ⁇ day before / after (%)
  • the antitumor effect of oral administration for 6 days was observed. It can be seen that the tumor inhibition rate increases with the increase of dose, showing a good dose-effect relationship.
  • the anti-tumor rate of the drug-administered group (100m g /k g ) was significantly higher than that of the equimolar control drug curcumin (50 mg/kg), and no mice died in the large dose. Compound 1 was safer. (See Table 2 and Figure 2 of the drawing) Instruction manual
  • K562 cells were passed for more than two generations, and the number of cells was adjusted to 5 ⁇ 10 7 /ml, 0.2 mL/only inoculated into the right forelimb of nude mice, and 30 cells were inoculated.
  • mice After 6 days of inoculation of tumors, the mice were randomly divided into 3 groups according to the tumor size of the mice: Group I was the saline control group, and Group II was the compound 1 low dose group. 40 mg/kg), group III was compound 1 high dose group (60 mg/kg), and the administration method was intragastric administration, 0.1 ml/10 g.
  • the tumor inhibition rate (%) (the average tumor mass of the control group - the average tumor mass of the experimental group) / the average tumor mass of the control group X 1 oo %.
  • Fig. 3 of the accompanying drawings The dynamic changes of tumor growth in each group of tumor-bearing mice are shown in Fig. 3 of the accompanying drawings, and it can be seen that the tumor growth rate of the drug-administered group is significantly lower than that of the NS control group.
  • mice Both female and male, 4 to 6 weeks old, weighing 18 to 22 g, provided by the Laboratory Animal Research Institute of the Chinese Academy of Medical Sciences, license number: SCXK (Beijing) 2005-0013.
  • SCXK Beijing 2005-0013.
  • NOD-SCID mice were housed in a SPF laboratory laminar box with a cover mouse (in accordance with SPF standards:). Standard pellet feed, drinking water, litter and all contact with the mouse are sterilized.
  • NOD-SCID mice received whole body irradiation with 2.0 Gy X-rays, and K562 cells in logarithmic growth phase were used for transplantation the next day, and l xlO 7 cell/mouse was injected once in the tail vein.
  • mice After transplanting K562 cells into NOD-SCID mice, they were randomly divided into normal saline control group and drug-treated group, with four groups in each group.
  • the blank control group was given 0.2 ml/supply of normal saline, and the treatment group was administered with compound 1 60 mg/kg, and the administration was started two weeks after the transplantation, and the administration was continued for 5 days, and the treatment group was stopped for 5 days. Continuous medication for 8 days.
  • the general condition, body weight and food intake of the mice were closely observed during the administration, and the toxicity of the drug and the tolerance of the animal to the drug were evaluated to adjust the administration schedule.
  • the survival time of the mice in each experimental group was recorded for three months. The survival of K562 cells after transplantation for more than two months was long-term survival.
  • mice in each experimental group were counted before the inoculation and 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks after the inoculation, and the total number of white blood cells was counted in the tail vein blood, and blood smears were prepared. , stained by Wright-Gimsa, sorted and counted under oil mirror.
  • mice in the saline control group were 32.5 ⁇ 9.0 days after transplantation of human chronic myeloid leukemia cell K562.
  • Peripheral blood leukocytes increased significantly in the three weeks after transplantation, which was 2-10 times before transplantation, and a large number of immature cells were seen in peripheral blood smears.
  • the dead mice were grossly dissected, and a large amount of bloody ascites was seen in the abdominal cavity, and solid tumors appeared, and the splenomegaly was not obvious (see Figure 5 of the specification).
  • the saline-controlled group of NOD-SCID mouse bone marrow cells detected the human bcr-abl gene by RT-PCR (see Figure 6 of the specification), indicating that human chronic myeloid leukemia cell K562 homing to the bone marrow of NOD-SCID mice.
  • the human chronic granulocyte white blood model was successfully constructed.
  • Compound 2 was prepared by the method of the above examples, and the test method of this example was the same as Compound 1.
  • Compound 2 was formulated to the desired concentration prior to use.
  • the mice with good tumor growth were sacrificed by cervical dislocation, the tumor pieces were removed under aseptic conditions, and the tumor tissues with good growth state were taken, and the single cell suspension was prepared by homogenization filtration, and the number of cells was adjusted to 1.
  • OX IOVml each inoculation 0. 2ml was placed in the right axilla of the mouse to establish a H22 mouse xenograft model.
  • mice were randomized by body weight, 9 mice per group, and the pre-dose weight was recorded.
  • Tumor inhibition rate (%) [analytical tumor mass of the control group - mean tumor mass of the experimental group] / average tumor mass of the control group X 100%.
  • the present invention can also be verified in the same manner as the above examples, including Compound 1 or Compound 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing Compound 1 or Compound 2 or a pharmaceutically acceptable salt form thereof having similar properties.
  • the pharmaceutically acceptable salt include an alkali metal salt such as a sodium salt or a potassium salt, an alkaline earth metal salt such as a calcium salt or a magnesium salt, a salt containing an organic base such as an ammonium salt, or a salt of an organic base such as triethylamine.

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Abstract

Provided are phenylbutyryl curcumin derivatives, their preparation methods, pharmaceutical compositions containing said derivatives and uses thereof for preparing anti-tumor drugs. Said curcumin derivatives are specifically 4-[bis(2-chloroethyl)amino]phenylbutyryl curcumin and 4,4'-bis[bis(2-chloroethyl)amino]phenylbutyryl curcumin, and their pharmaceutically acceptable salts. The curcumin derivatives are effective on inhibiting various animal tumor cell transplantation models in vivo, and do not have serious toxicity in mouse.

Description

说 明 书  Description
苯丁酰基姜黄素衍生物及其在制备抗肿瘤药物中的应用 Phenylbutyryl curcumin derivative and its application in preparing antitumor drugs
技术领域 Technical field
本发明属制药领域, 尤其涉及苯丁酰基姜黄素衍生物及其制备方法, 以及 其在制备抗肿瘤药物中的应用。  The invention belongs to the field of pharmacy, and in particular relates to a phenylbutyryl curcumin derivative and a preparation method thereof, and the use thereof in preparing an antitumor drug.
背景技术 Background technique
姜黄素(Curcumin, 简称 Cur)是从姜科姜黄属植物姜黄、 莪术、 郁金等的 根茎中提取的有效成分, 具有抗肿瘤、 抗炎、 抗人类免疫缺陷病毒、 抗胆固醇、 抗氧化等多种药理作用,具有良好的临床应用潜力。 但 Cur不稳定, 在体内代谢 迅速, 生物利用度低, 体内难以达到有效浓度, 严重制约了姜黄素开发成有效 的抗癌药。 通过结构改造合成姜黄素衍生物, 提高生物利用度、 延长生物消除 半衰期 t1/2有重要意义。 Curcumin (Cur) is an active ingredient extracted from the rhizome of the turmeric, turmeric, turmeric, and turmeric of the genus Zingiberaceae. It has anti-tumor, anti-inflammatory, anti-human immunodeficiency virus, anti-cholesterol, anti-oxidation and so on. A pharmacological effect has a good clinical application potential. However, Cur is unstable, rapid metabolism in the body, low bioavailability, and it is difficult to achieve an effective concentration in the body, which seriously restricts the development of curcumin into an effective anticancer drug. Curcumin derivatives synthesized through structural transformation, improve bioavailability, prolonged biological t 1/2 elimination half-life is important.
发明内容 Summary of the invention
本发明的目的之一在于提供 4- [双 (2-氯乙基:)氨基]苯丁酰基姜黄素和 4,4'- [双 (2-氯乙基:)氨基]双苯丁酰基姜黄素及其药学上可接受的盐。 本发明的目的之二在于提供 4- [双 (2-氯乙基:)氨基]苯丁酰基姜黄素和 4,4'- [双 (2-氯乙基:)氨基]双苯丁酰基姜黄素及其药学上可接受的盐的制备方法。 为实现本发明的目的采用的技术方案如下: 本发明所述的 4- [双 (2-氯乙基:) 氨基]苯丁酰基姜黄素其结构式如下列化合物 1 所示; 4,4'- [双 (2-氯乙基)氨基] 双苯丁酰基姜黄素其结构式如下列化合物 2所示, 以及本发明所述的姜黄素衍 生物的盐类是下式结构的化合物 1和化合物 2在药学上可接受的盐类:  One of the objects of the present invention is to provide 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin and 4,4'-[bis(2-chloroethyl:)amino]bisphenylbutanoyl turmeric And its pharmaceutically acceptable salt. A second object of the present invention is to provide 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin and 4,4'-[bis(2-chloroethyl:)amino]bisphenylbutanoyl turmeric And a method for preparing the pharmaceutically acceptable salt thereof. The technical scheme adopted for the purpose of the present invention is as follows: 4-[bis(2-chloroethyl:)amino]phenylbutyryl curcumin as described in the present invention has the structural formula shown as the following compound 1; 4, 4'- [Bis(2-chloroethyl)amino] bis-butyroyl curcumin having the structural formula shown in the following compound 2, and the salt of the curcumin derivative of the present invention is a compound 1 and a compound 2 of the following formula Pharmaceutically acceptable salts:
Figure imgf000003_0001
说 明 书
Figure imgf000003_0001
Instruction manual
Figure imgf000004_0001
Figure imgf000004_0001
在药学上可接受盐包括碱金属盐、 碱土金属盐、 含有机碱的盐、 含有机碱 的盐。 该可接受盐包括钙盐、 镁盐、 铵盐、 三乙基胺盐、 乙醇胺盐。 The pharmaceutically acceptable salts include alkali metal salts, alkaline earth metal salts, organic base-containing salts, and organic base-containing salts. The acceptable salts include calcium salts, magnesium salts, ammonium salts, triethylamine salts, and ethanolamine salts.
本发明所述的姜黄素衍生物和盐类的制备方法, 具体歩骤如下: 将姜黄素 溶于干燥过的二氯甲垸, 加入催化量 DMAP (Ν,Ν-4-二甲基氨基吡啶), 而后加 入已溶解于二氯甲垸的 4- [双 (2-氯乙基:)氨基]苯丁酸进行酯化反应, 而后将产物 经过柱层析分离纯化。在上述本发明的方法中可以先加脱水剂 DCC (二环己基 碳二亚胺) 或 EDCI ( 1 -乙基 -3-(3-二甲氨基丙基)碳二亚胺盐酸盐), 再加化学 领域的通用催化量的 DMAP。  The preparation method of the curcumin derivative and the salt of the present invention is specifically as follows: The curcumin is dissolved in the dried dichloromethane, and the catalytic amount of DMAP (Ν, Ν-4-dimethylaminopyridine) is added. Then, 4-[bis(2-chloroethyl:)amino] phenylbutyric acid which has been dissolved in methylene chloride is added for esterification, and then the product is separated and purified by column chromatography. In the above method of the present invention, a dehydrating agent DCC (dicyclohexylcarbodiimide) or EDCI (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) may be added first. A general catalytic amount of DMAP in the chemical field is added.
本发明所合成的化合物 1和化合物 2结构的姜黄素衍生物, 均获得核磁共 振光谱 (NMR)、 红外线光谱 (IR)、 紫外线光谱 (UV)、 液相色谱-质谱法 (HPLC-Mass) 鉴定, 在鉴定后的化合物 1和化合物 2, 依据需要而制成合适 的药物学上的剂型。  The curcumin derivatives of the compound 1 and the compound 2 synthesized by the present invention are all obtained by nuclear magnetic resonance spectroscopy (NMR), infrared spectroscopy (IR), ultraviolet spectroscopy (UV), and liquid chromatography-mass spectrometry (HPLC-Mass). After the identification of Compound 1 and Compound 2, a suitable pharmaceutically acceptable dosage form is prepared as needed.
本发明是保护化合物 1或化合物 2的药物或其药学上可接受的盐, 或含有 化合物 1或化合物 2的药物或其药学上可接受的盐组成的药物组合物或制剂。  The present invention is a pharmaceutical composition or preparation comprising a medicament for protecting Compound 1 or Compound 2 or a pharmaceutically acceptable salt thereof, or a medicament containing Compound 1 or Compound 2 or a pharmaceutically acceptable salt thereof.
在制备药物学上的剂型中, 其载体为药学领域常规的药物载体, 例如稀释 剂、 赋形剂、 填充剂、 粘合剂、 崩解剂、 表面活性剂、 润滑剂等; 其可以制成 口服和其他给药方式的各种剂型, 如口服液、 混悬液、 胶囊、 片剂、 丸剂、 颗 粒剂、 及粉针注射液等; 按药学领域的常规生产方法制备, 其选用含有重量百 分比为 0.1 %— 99.5 %活性成分的化合物 1或化合物 2。 也可含有 0.1 %— 99.5 % 的化合物 1或化合物 2的药物组合物; 为同领域一般技术人员能实现的技术。  In the preparation of a pharmaceutically acceptable dosage form, the carrier is a conventional pharmaceutical carrier in the pharmaceutical field, such as a diluent, an excipient, a filler, a binder, a disintegrant, a surfactant, a lubricant, etc.; Various dosage forms for oral administration and other modes of administration, such as oral liquids, suspensions, capsules, tablets, pills, granules, powder injections, etc.; prepared by conventional production methods in the pharmaceutical field, which are selected to contain weight percentages It is 0.1% to 99.5% active ingredient of Compound 1 or Compound 2. Pharmaceutical compositions which may also contain from 0.1% to 99.5% of Compound 1 or Compound 2; are those which can be practiced by one of ordinary skill in the art.
本发明的使用量可根据用药途径, 患者的年龄、 体重、 所治疗的疾病的类 型和严重程度等变化, 其服用量可以是 0. 001— 10g / kg体重, 可以一次或多 次给药。 本发明目的之三在于 4- [双 (2-氯乙基:)氨基]苯丁酰基姜黄素和 4,4'- [双 (2-氯 乙基)氨基]双苯丁酰基姜黄素, 用于制备抗肿瘤药物的应用。 姜黄素衍生物, 可用于但不局限于制备治疗白血病、 皮肤癌、 胃癌、 结肠 癌、 肝癌、 乳腺癌或前列腺癌药物。 白血病优选人慢性粒细胞白血病。 说 明 书 本发明的有益效果: 本发明所述的化合物 1姜黄素 4- [双 (2-氯乙基)氨基]苯 丁酸酯在体内对多种动物肿瘤细胞移植模型均有明显抑制, 尤其是对人慢性粒 细胞白血病 K562细胞裸鼠移植瘤的抑制作用可高达 60%, 而且给药组裸鼠体 重无明显下降, 无动物发生死亡。 化合物 1对人慢性粒细胞白血病 K562细胞 株接种 NOD-SCID小鼠构建人慢性粒细胞白血病模型, 生命延长率也提高了, 且未见有对小鼠的严重毒性。对小鼠肝癌 H22在体抑瘤率为 40— 75 %, 同样化 合物 2由于结构相似, 实验证明也有同样的效果。 因此, 该类化合物具有比姜 黄素更强的抑瘤作用。 附图说明 图 1 是化合物 1静脉给药对小鼠肝癌 H22移植瘤的抑制作用图 The dosage of the present invention may be changed according to the route of administration, the age and weight of the patient, the type and severity of the disease to be treated, and the dose may be 0.001 - 10 g / k g body weight, which may be given one or more times. medicine. The third object of the present invention is to 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin and 4,4'-[bis(2-chloroethyl)amino]bisphenylbutanoyl curcumin, For the preparation of anti-tumor drugs. Curcumin derivatives can be used, but are not limited to, to prepare drugs for the treatment of leukemia, skin cancer, gastric cancer, colon cancer, liver cancer, breast cancer or prostate cancer. Leukemia is preferably human chronic myeloid leukemia. DESCRIPTION OF THE INVENTION Advantageous Effects of the Invention: The compound 1 curcumin 4- [bis(2-chloroethyl)amino] phenylbutyrate of the present invention is significantly inhibited in various animal tumor cell transplantation models in vivo, especially The inhibitory effect on human chronic myeloid leukemia K562 cells in nude mice can be as high as 60%, and the weight of nude mice in the drug-administered group did not decrease significantly, and no animal died. Compound 1 inoculated NOD-SCID mice to human chronic myeloid leukemia K562 cell line to construct a human chronic myeloid leukemia model, and the life extension rate was also improved, and no serious toxicity to mice was observed. The tumor inhibition rate of mouse liver cancer H22 was 40-75%, and the same compound 2 proved to have the same effect due to similar structure. Therefore, such compounds have a stronger antitumor effect than curcumin. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the inhibitory effect of intravenous administration of Compound 1 on mouse liver cancer H22 xenografts.
图 2 是化合物 1口服给药对小鼠肝癌 H22移植瘤的抑制作用图  Figure 2 is a graph showing the inhibitory effect of oral administration of compound 1 on liver cancer H22 xenografts in mice.
图 3 是化合物 1对人慢性粒细胞白血病 K562细胞裸鼠移植瘤生长曲线 图  Figure 3 is a graph showing the growth curve of compound 1 in human chronic myeloid leukemia K562 cells transplanted in nude mice.
图 4 是化合物 1对人慢性粒细胞白血病 K562细胞裸鼠移植瘤的作用图 图 5和图 6 是化合物 1对人慢性粒细胞白血病小鼠模型的小鼠大体解剖 和小鼠骨髓 bcr-abl基因表达图  Figure 4 is a diagram showing the effect of Compound 1 on human chronic myeloid leukemia K562 cells transplanted in nude mice. Figure 5 and Figure 6 are the gross anatomy of mouse 1 in mouse model of human chronic myeloid leukemia and the mouse bone marrow bcr-abl gene. Expression map
图 7和图 8 是 NS组和化合物 1给药对人慢性粒细胞白血病模型小鼠外 周血象图  Figure 7 and Figure 8 are the peripheral blood images of NS group and compound 1 administered to human chronic myeloid leukemia model mice.
图 9是化合物 2对小鼠肝癌 H22移植瘤的抑制作用图;  Figure 9 is a graph showing the inhibitory effect of Compound 2 on mouse liver cancer H22 xenografts;
如图 1所示, 建立 H22小鼠移植瘤, 分别静脉给予小鼠生理盐 (对照组), 化合物 1 : 50、 70mg/kg (给药组)。 抑瘤率高达 60%。  As shown in Fig. 1, H22 mouse xenografts were established, and the physiological saline (control group) of the mice was intravenously administered, Compound 1 : 50, 70 mg/kg (administration group). The tumor inhibition rate is as high as 60%.
如图 2所示, 建立 H22小鼠移植瘤, 分别口服给予小鼠生理盐 (对照组), 化合物 1 : 50、 75、 100mg/kg (给药组), 姜黄素 50 mg/kg (姜黄素对照组)。 给药组抑瘤率分别为 41.21%、 52.93%、 75.06%。 姜黄素对照组 (与化合物 lOOmg/kg等摩尔)抑瘤率仅为 16.58%,可见化合物抑瘤作用明显比姜黄素要强。  As shown in Figure 2, H22 mouse xenografts were established and administered orally to mice with physiological saline (control group). Compound 1: 50, 75, 100 mg/kg (administration group), curcumin 50 mg/kg (curcumin) Control group). The tumor inhibition rates of the drug-administered group were 41.21%, 52.93%, and 75.06%, respectively. The anti-tumor rate of the curcumin control group (equal to the compound lOOmg/kg) was only 16.58%, and the anti-tumor effect of the compound was significantly stronger than that of curcumin.
如图 3所示, 建立人慢性粒细胞白血病 K562细胞裸鼠移植瘤模型, 分别 口服给予裸鼠生理盐 (对照组), 化合物 1 : 40、 60mg/kg (给药组) 。 于第 0、 4、 8、 11天用游标卡尺测小鼠瘤的长、 宽、 高, 计算瘤体积 =长><宽 X高 x l/2, 增长百分率 (%) = (实验组平均瘤体积一对照组平均瘤体积) I对照组平均瘤 体积 χ 100%。 从图 3可见, 给药组肿瘤的生长受到明显抑制。  As shown in Fig. 3, a nude mouse xenograft model of human chronic myeloid leukemia K562 cells was established and administered orally to nude mice (control group), compound 1: 40, 60 mg/kg (administration group). On the 0th, 4th, 8th and 11th day, the length, width and height of the mouse tumor were measured with vernier calipers. Calculated tumor volume = length><width X height xl/2, percentage of growth (%) = (average tumor volume of the experimental group) The mean tumor volume of the control group) The average tumor volume of the control group was χ100%. As can be seen from Fig. 3, the growth of tumors in the drug-administered group was significantly inhibited.
如图 4所示, 建立 Κ562细胞裸鼠移植瘤模型, 分别口服给予裸鼠生理盐 (对照组), 化合物 1 : 40、 60mg/kg (给药组)。 抑瘤率分别为 66.6%、 56.7%。  As shown in Fig. 4, a nude mouse xenograft model of Κ562 cells was established and orally administered to nude mice (control group), compound 1: 40, 60 mg/kg (administration group). The tumor inhibition rates were 66.6% and 56.7%, respectively.
如图 5、 图 6所示, 用 NOD-SCID小鼠构建人慢性粒细胞白血病模型, 对 照组小鼠大体解剖, 腹腔内见大量血性腹水, 并出现实体瘤。 小鼠骨髓细胞 说 明 书 As shown in Fig. 5 and Fig. 6, a human chronic myeloid leukemia model was constructed using NOD-SCID mice. The control group was grossly dissected, and a large amount of bloody ascites was seen in the abdominal cavity, and solid tumors appeared. Mouse bone marrow cells Description
RT-PCR均检到人人慢性粒细胞白血病的特征基因 bcr-abl, 说明人慢性粒细胞 白血病细胞 K562归巢至 NOD-SCID小鼠骨髓。 The characteristic gene bcr-abl of human chronic myeloid leukemia was detected by RT-PCR, indicating that human chronic myeloid leukemia cell K562 homing to the bone marrow of NOD-SCID mice.
如图 7、 图 8所示, NOD-SCID小鼠人慢性粒细胞白血病模型, 分别口服 给予小鼠生理盐水 (对照组), 化合物 1 : 60mg/kg (给药组) , 给药组外周血 象幼稚细胞亦明显少于对照组。  As shown in Fig. 7 and Fig. 8, NOD-SCID mouse human chronic myeloid leukemia model was orally administered to normal saline (control group), compound 1: 60 mg/kg (administered group), peripheral blood group of the drug-administered group. The naive cells were also significantly less than the control group.
如图 9所示, 分别口服给予小鼠生理盐(对照组), 化合物 2: 35、 50、 75、 115mg/kg (给药组), 姜黄素 50 mg/kg (姜黄素对照组)。 给药组抑瘤率分别为 27.43%、 32.25%、 62.60%、 58.39%。 姜黄素对照组 (与化合物 100mg/kg等摩 尔) 抑瘤率仅为 13.42%, 可见化合物 2的抑瘤作用明显比姜黄素要强。 具体实施方式 As shown in Fig. 9, the physiological saline (control group) of the mice was orally administered, Compound 2: 35, 50, 75, 115 mg/kg (administration group), curcumin 50 mg/kg (curcumin control group). The tumor inhibition rates of the drug-administered group were 27.43%, 32.25%, 62.60%, and 58.39%, respectively. Curcumin group (compound 100m g / kg equimolar) inhibition rate was 13.42%, visible inhibitory effect of Compound 2 significantly stronger than curcumin. detailed description
下列结合附图和实例对本发明进行详细说明 The present invention will be described in detail below with reference to the accompanying drawings and examples.
实施例 1 4- [双 (2-氯乙基:)氨基]苯丁酰基姜黄素 (化合物 1 ) 的合成 Example 1 Synthesis of 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin (Compound 1)
EDCI 1. 92g (10mmol)溶于无水二氯甲垸 200ml中, 在冰浴下搅拌并加入姜 黄素 7. 36g (20mmol), DMAP 0. 244g ( 2mmol ) , 再缓慢滴加溶有 4- [双(2-氯乙 基)氨基]苯丁酸 3. 04g (10mmol) 的二氯甲垸 200ml, 室温下继续搅拌反应 6小 时。 用蒸馏水洗涤, 有机相用无水硫酸镁干燥后, 过滤浓缩得粗品, 粗品用中 压制备色谱分离,反向 C18柱, 流动相甲醇: 水 50%→100% (体积百分比) 梯度 洗脱, 得到 4- [双 (2-氯乙基)氨基]苯丁酰基姜黄素纯品 1. 30g,产率 20%, 化学 结构式如下: EDCI 1. 92g (10mmol) was dissolved in anhydrous dichloromethane 200ml, stirred in an ice bath and added with curcumin 7.36g (20mmol), DMAP 0. 244g (2mmol), then slowly added to dissolve 4- [Bis(2-chloroethyl)amino] phenylbutyric acid 3. 04 g (10 mmol) of dichloromethane (200 ml), and the reaction was further stirred at room temperature for 6 hours. After washing with distilled water, the organic phase was dried over anhydrous magnesium sulfate, and then filtered and evaporated to give a crude product. The crude product was purified by medium pressure preparative chromatography, reversed C18 column, mobile phase methanol: water 50%→100% (volume percent) gradient elution, Obtaining 4-[bis(2-chloroethyl)amino]phenylbutanoyl curcumin pure product 1. 30 g, yield 20%, the chemical structural formula is as follows:
Figure imgf000006_0001
分子式 C35H37C12N07, mpl00-102°C
Figure imgf000006_0001
Molecular formula C 35 H 37 C1 2 N0 7 , mpl00-102°C
1H MR (D6-DMSO): 51.88(t,2H, -COCH2CH2CH2Ph), 2.51(t,2H, -COCH2CH2CH2Ph), 2.58(m,2H,-COCH2CH2CH2Ph), 3 62(t,4H,-NCH2CH2Cl),3.71 (t,4H,-NCH2CH2Cl),3.85 1H MR (D6-DMSO): 51.88 (t, 2H, -COCH 2 CH 2 CH 2 Ph), 2.51 (t, 2H, -COCH 2 CH 2 CH 2 Ph), 2.58 (m, 2H, -COCH 2 CH 2 CH 2 Ph), 3 62(t,4H,-NCH 2 CH 2 Cl), 3.71 (t,4H,-NCH 2 CH 2 Cl), 3.85
(s,6H,2Ar-OCH3), 6.14 (s, 1H, -COCH=C(OH)-), 6.70 (d, 2H, 2-CH=CH-Ar), 6.78-6.99(m,2H): 7.13-7.19(m,2H) , 7.33(m,2H), 7.51 (d,2H),7.58 (d,2H),7.62 (d,2H ,2Ar-CH=),9.72 (s,lH , Ar-OH);HPLC-MS m/z: 654.2(M+1) 说 明 书 实施例 2 4,4 ' - [双 (2-氯乙基:)氨基]双苯丁酰基姜黄素 (化合物 2) 的合 成 (s,6H,2Ar-OCH3), 6.14 (s, 1H, -COCH=C(OH)-), 6.70 (d, 2H, 2-CH=CH-Ar), 6.78-6.99 (m, 2H) : 7.13-7.19(m,2H), 7.33(m,2H), 7.51 (d,2H), 7.58 (d,2H), 7.62 (d,2H,2Ar-CH=), 9.72 (s,lH, Ar- OH);HPLC-MS m/z: 654.2 (M+1) Description of the Invention Example 2 Synthesis of 4,4 ' - [bis(2-chloroethyl:)amino] bis-butyroyl curcumin (Compound 2)
EDCI 1. 92g (10mmol)溶于无水二氯甲垸 100ml中, 在冰浴下搅拌并加入姜 黄素 1. 84g (5mmol), DMAP 0. 122g ( lmmol ) , 再缓慢滴加溶有 4- [双(2-氯乙 基)氨基]苯丁酸 3. 04g (10mmol) 的二氯甲垸 200ml, 室温下继续搅拌反应 6小 时。 用蒸馏水洗涤, 有机相用无水硫酸镁干燥后, 过滤浓缩得粗品, 粗品用中 压制备色谱分离,反向 C18柱, 流动相甲醇: 水 50%→100% (体积百分比) 梯度 洗脱,得到 4, 4 ' - [双 (2-氯乙基)氨基]双苯丁酰基姜黄素纯品(化合物 2 ) 1. 88g, 产率 40%, 化学结构式如下: EDCI 1. 92g (10mmol) was dissolved in 100ml of anhydrous dichloromethane, stirred in an ice bath and added with curcumin 1.84g (5mmol), DMAP 0. 122g (lmmol), then slowly added to dissolve 4- [Bis(2-chloroethyl)amino] phenylbutyric acid 3. 04 g (10 mmol) of dichloromethane (200 ml), and the reaction was further stirred at room temperature for 6 hours. After washing with distilled water, the organic phase was dried over anhydrous magnesium sulfate, and then filtered and evaporated to give a crude product. The crude product was purified by medium pressure preparative chromatography, reversed C18 column, mobile phase methanol: water 50%→100% (volume percent) gradient elution, Obtained 4, 4 ' - [bis(2-chloroethyl)amino] bis-butyroyl curcumin pure product (Compound 2) 1. 88 g, yield 40%, the chemical structural formula is as follows:
Figure imgf000007_0001
Figure imgf000007_0001
C49H54C14N2O8, mp58-60°C,1H MR (CDC13): 52.04(m,4H, -COCH2CH2CH2Ph), 2.29(t,4H, -COCH2CH2CH2Ph),2.60(m,4H,-COCH2CH2CH2Ph),3.63(t,4H,-NCH2CH2Cl),3.70(t,4H,-NCH2C H2Cl),3.87(s,6H,2Ar-OCH3),6.58(d,2H,J=16Hz,2-CH=CH-Ar),6.63(d,4H,J=8Hz),6.66(s, 1H,-C0 CH=C(OH)-),7.03(s,2H),7.05(m,2H),7.08(m,2H),7.12(d,4H,J=8Hz),7.17(m,2H),7.51(d,2H),7.53 (d,2H),7.62 (d,2H ,J=16Hz,2Ar-CH=); HPLC-MS m/z: 941.3(M+1) 实施例 3 化合物 1对小鼠肝癌 H22移植瘤模型的作用 3.1 材料 C49H54C14N2O8, mp58-60°C, 1H MR (CDC1 3 ): 52.04 (m, 4H, -COCH 2 CH 2 CH 2 Ph), 2.29 (t, 4H, -COCH 2 CH 2 CH 2 Ph), 2.60 (m) , 4H, -COCH 2 CH 2 CH 2 Ph), 3.63 (t, 4H, -NCH 2 CH 2 Cl), 3.70 (t, 4H, -NCH 2 CH 2 Cl), 3.87 (s, 6H, 2Ar-OCH3 ), 6.58 (d, 2H, J = 16 Hz, 2-CH=CH-Ar), 6.63 (d, 4H, J = 8 Hz), 6.66 (s, 1H, -C0 CH=C(OH)-), 7.03 (s, 2H), 7.05 (m, 2H), 7.08 (m, 2H), 7.12 (d, 4H, J = 8 Hz), 7.17 (m, 2H), 7.51 (d, 2H), 7.53 (d, 2H) ), 7.62 (d, 2H, J = 16 Hz, 2Ar-CH=); HPLC-MS m/z: 941.3 (M+1) Example 3 Effect of Compound 1 on Mouse Hepatoma H22 Transplanted Tumor Model 3.1 Materials
8〜12周龄昆明种健康小鼠, 雌性, 体重为 (20±2) g;小鼠由福建医科大学实 验动物中心提供 (合格证号 SCXK(闽:) 200420002)。 小鼠肝癌细胞株 H22。 化合物8~12 weeks old Kunming healthy mice, female, weighing (20±2) g; mice were provided by Experimental Animal Center of Fujian Medical University (certificate No. SCXK (闽:) 200420002). Mouse liver cancer cell line H22. Compound
1使用前用配成所需浓度。 1 Use the required concentration before use.
3.2 方法 3.2 Methods
3.2.1荷瘤小鼠模型的建立  3.2.1 Establishment of tumor-bearing mouse model
H22肿瘤细胞传两代以上,调整细胞数至 107/ml, 0.2mL/只接种于小鼠右前 肢皮下,接种 2只, 待约两周, 剥瘤, 匀浆, 再接种 80只。 说 明 书 H22 tumor cells were passed for more than two generations. The number of cells was adjusted to 10 7 /ml, 0.2 mL/only inoculated into the right forelimb of mice, and 2 inoculated. After about two weeks, the tumor was stripped, homogenized, and inoculated again. Instruction manual
3.2.2 分组 3.2.2 Grouping
A: 共分为 3组: 接种瘤株 24 h后的小鼠随机分成 3组: I组为生理盐水对照组, 化合物 1采用上述实施例方法制得的, 以下同, II组为化合物 1低剂量组 (化 合物 1 50mg/kg), III组为化合物 1高剂量组 (70mg/kg), 给药方法为尾静脉 注射, 0.1ml/10g。 A: A total of 3 groups: The mice after 24 h inoculation of tumors were randomly divided into 3 groups: Group I was a saline control group, and Compound 1 was prepared by the method of the above examples. The same group II was compound 1 low. The dose group (Compound 1 50 mg/kg), Group III was Compound 1 high dose group (70 mg/kg), and the administration method was tail vein injection, 0.1 ml/10 g.
B: 共分为 5组: 接种瘤株 24 h后的小鼠随机分成 5组: I组为生理盐水组 (即 肿瘤对照组), II组为化合物 1低剂量组(剂量为 50mg/kg), III组为化合物 1中 剂量组 (剂量为 75mg/kg), IV组为化合物 1高剂量组 (剂量为 100mg/kg), V 组为 Cur 组 (剂量为 50mg/kg, 与高剂量 IV组等摩尔), 给药方法为灌胃,
Figure imgf000008_0001
B: A total of 5 groups: 24 hours after inoculation of tumors, the mice were randomly divided into 5 groups: group I was saline group (ie, tumor control group), group II was compound 1 low dose group (dose was 50 mg/kg) , III group compound 1 dose (a dose of 75mg / kg), IV group compound high dose groups (dose 100m g / k g), V group Cur group (at a dose of 50mg / kg, with a high dose Group IV equimolar), the method of administration is gavage,
Figure imgf000008_0001
3.2.3抑瘤率 处死小鼠后剥瘤称瘤质量, 计算抑瘤率. 肿瘤抑制率 (%) =对照组平均 瘤质量 -实验组平均瘤质量 /对照组平均瘤质量 χΐοο%。  3.2.3 tumor inhibition rate After the mice were sacrificed, the tumor mass was measured and the tumor inhibition rate was calculated. The tumor inhibition rate (%) = the average tumor quality of the control group - the average tumor mass of the experimental group / the average tumor mass of the control group χΐοο%.
3.3 结果  3.3 Results
3.3.1 化合物 1静脉给药的抑瘤作用  3.3.1 Compound 1 antitumor effect of intravenous administration
观察不同给药剂量的抑瘤作用, 可见化合物 1对小鼠肝癌 H22移植瘤率具有 明显的抑制作用 (见表 1, 说明书附图的图 1 )。  Observing the antitumor effect of different doses, it can be seen that Compound 1 has a significant inhibitory effect on the transplant rate of mouse liver cancer H22 (see Table 1, Figure 1 of the accompanying drawings).
表 1 化合物 1静脉给药对小鼠肝癌 H22移植瘤抑制作用 剂 里 鼠 数 瘤 重(g ) 抑 瘤 率 组 另 mg/kg > <day 前 /后 (%)  Table 1 Compound 1 intravenous administration to mouse liver cancer H22 xenograft inhibition agent number of rats tumor weight (g) tumor inhibition rate group other mg/kg > <day before / after (%)
NS 0χ 10 10/10 1.01±0.47 0  NS 0χ 10 10/10 1.01±0.47 0
低剂量组 50 χ7 10/9 0.40士 0.15 60.84** 高剂量组 70 χ5 10/6 0.335士 0.16 66.83** 注: NS (生理盐水对照组), 与 NS组相比, * P<0.05;** P<0.01.  Low dose group 50 χ7 10/9 0.40 ± 0.15 60.84** High dose group 70 χ 5 10/6 0.335 ± 0.16 66.83 ** Note: NS (normal saline control group), compared with NS group, * P < 0.05; * P<0.01.
3.3.2 化合物 1口服给药的抑瘤作用 3.3.2 Compound 1 anti-tumor effect of oral administration
观察口服给药 6天的抑瘤作用。 可见随剂量的增加抑瘤率逐渐提高, 呈现 良好的量效关系。 给药组 (100mg/kg) 比等摩尔的对照药姜黄素 (50 mg/kg) 抑瘤率有显著提高, 而且大剂量未见有小鼠死亡, 可见化合物 1较安全。 (见表 2 及说明书附图的图 2) 说 明 书 The antitumor effect of oral administration for 6 days was observed. It can be seen that the tumor inhibition rate increases with the increase of dose, showing a good dose-effect relationship. The anti-tumor rate of the drug-administered group (100m g /k g ) was significantly higher than that of the equimolar control drug curcumin (50 mg/kg), and no mice died in the large dose. Compound 1 was safer. (See Table 2 and Figure 2 of the drawing) Instruction manual
表 2口服给药对小鼠肝癌 H22移植瘤的抑制作用 Table 2 Inhibition of oral administration of mouse liver cancer H22 xenografts
鼠 数 体 重 ( ± s) 瘤 重 (g ) 抑瘤率 组 另 J  Rat number body weight (± s) tumor weight (g) tumor inhibition rate group
前 /后 前 1 后 (%) Before/after 1 before (%)
NS 10/10 20.26±1.68/ 26.43±4.00 1.60±0.24 0 化合物 1 ( 50mg/kg ) 10/10 20.79±1.63/ 22.12±2.48 0.94±0.33 NS 10/10 20.26±1.68/ 26.43±4.00 1.60±0.24 0 Compound 1 ( 50mg/kg ) 10/10 20.79±1.63/ 22.12±2.48 0.94±0.33
化合物 1 ( 75mg/kg ) 10/10 21.02±1.54/ 22.00±3.98 0.76±0.26 52.93** 化合物 1 ( 100mg/kg ) 10/10 21.10±1.57/ 18.64±1.91 0.40±0.15 75.06** 姜黄素 50mg/kg 10/10 21.17±1.62/ 26.61±3.67 1.34±0.25 16.58 注: 与 NS组相比, * P<0.05;** P<0.01. Cur为姜黄素  Compound 1 (75 mg/kg) 10/10 21.02±1.54/ 22.00±3.98 0.76±0.26 52.93** Compound 1 (100mg/kg) 10/10 21.10±1.57/ 18.64±1.91 0.40±0.15 75.06** Curcumin 50mg/ Kg 10/10 21.17±1.62/ 26.61±3.67 1.34±0.25 16.58 Note: Compared with NS group, * P<0.05; ** P<0.01. Cur is curcumin
实施例 4 化合物 1对人慢性粒细胞白血病 K562细胞裸鼠移植瘤模型的作用 4.1 材料 Example 4 Effect of Compound 1 on human chronic myeloid leukemia K562 cell xenograft model in nude mice 4.1 Materials
4-6周龄 SPF级裸鼠,雄性,购自由上海斯莱克实验动物有限责任公司提供(许 可证号: SCXX (沪) 2007-0005 ),在 SPF实验室饲养。人类慢性粒细胞白血病 K562 细胞购自中国科学院上海细胞生物学研究所细胞库。化合物 1使用前配成所需浓 度。  4-6 weeks old SPF nude mice, male, purchased from Shanghai Slack Laboratory Animals Co., Ltd. (license number: SCXX (Shanghai) 2007-0005), raised in SPF laboratory. Human chronic myeloid leukemia K562 cells were purchased from the Cell Bank of the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. Compound 1 was formulated to the desired concentration prior to use.
4.2 方法 i)s  4.2 Methods i)s
* * * *
4.2.1荷瘤小鼠模型的建立 4.2.1 Establishment of tumor-bearing mouse model
K562细胞传两代以上,调整细胞数至 5 X 107/ml, 0.2mL/只接种于裸鼠右前 肢皮下, 接种 30只。 K562 cells were passed for more than two generations, and the number of cells was adjusted to 5×10 7 /ml, 0.2 mL/only inoculated into the right forelimb of nude mice, and 30 cells were inoculated.
4.2.2 分组 共分为 3组: 接种瘤株 6天后, 根据小鼠肿瘤大小, 将小鼠分层随机随机 分成 3组: I组为生理盐水对照组, II组为化合物 1低剂量组 (40mg/kg), III组 为化合物 1高剂量组 (60mg/kg), 给药方法为灌胃给药, 0.1ml/10g。 4.2.2 Grouping into 3 groups: After 6 days of inoculation of tumors, the mice were randomly divided into 3 groups according to the tumor size of the mice: Group I was the saline control group, and Group II was the compound 1 low dose group. 40 mg/kg), group III was compound 1 high dose group (60 mg/kg), and the administration method was intragastric administration, 0.1 ml/10 g.
4.2.3瘤增长百分率 分别于第 0、 4、 8、 11天用游标卡尺测小鼠瘤的长、 宽、 高, 计算瘤体积 = 长 X宽 X高 χ 1/2, 增长百分率 (%) = (实验组平均瘤体积一对照组平均瘤体积) I对照组平均瘤体积 χ 100%。 说 明 书 4.2.3 Percentage of tumor growth The length, width and height of the mouse tumor were measured with vernier calipers on days 0, 4, 8, and 11, respectively. Calculated tumor volume = length X width X height χ 1/2, percentage of growth (%) = (Average tumor volume of the experimental group - mean tumor volume of the control group) I The average tumor volume of the control group was χ 100%. Instruction manual
4.2.4抑瘤率 处死小鼠后剥瘤称瘤质量, 计算抑瘤率. 肿瘤抑制率 (%) = (对照组平均 瘤质量一实验组平均瘤质量) /对照组平均瘤质量 X 1 oo%。 4.2.4 tumor inhibition rate after the mice were sacrificed, the tumor mass was measured, and the tumor inhibition rate was calculated. The tumor inhibition rate (%) = (the average tumor mass of the control group - the average tumor mass of the experimental group) / the average tumor mass of the control group X 1 oo %.
4.3 结果  4.3 Results
4.3.1瘤增长百分率  4.3.1 Percentage of tumor growth
各组荷瘤小鼠肿瘤生长的动态变化见说明书附图的图 3, 可见给药组的肿瘤 增长速率明显比 NS对照组要低。  The dynamic changes of tumor growth in each group of tumor-bearing mice are shown in Fig. 3 of the accompanying drawings, and it can be seen that the tumor growth rate of the drug-administered group is significantly lower than that of the NS control group.
4.3.2 口服给药的抑瘤作用 观察口服给药 11天的抑瘤作用。 结果显示化合物 1能明显抑制 K562细胞裸 鼠移植瘤的生长,其抑瘤率分别为 66.6%、 56.7%, 经统计学处理 (t检验) 具有 显著性差异, 具有一定的量效关系 (见表 3, 说明书附图的图 4)。 与生理盐水组 比较,给药组裸鼠体重无明显下降, 无动物发生死亡, 提示化合物在提高药物抗 癌作用的同时并没有增加药物的毒性。试验表明化合物 1对人慢性粒细胞白血病 K562细胞裸鼠移植瘤模型具有显著的抑瘤作用。 表 3 口服给药对人慢性粒细胞白血病 K562细胞裸鼠移植瘤的抑制作用 鼠 数 体 重( ± s) 瘤 重 (g ) 抑瘤率 组 另 J  4.3.2 Antitumor effect of oral administration The antitumor effect of oral administration for 11 days was observed. The results showed that Compound 1 could significantly inhibit the growth of K562 cells transplanted in nude mice, and the tumor inhibition rate was 66.6% and 56.7%, respectively. There was significant difference in statistical analysis (t test), which had a certain dose-effect relationship (see Table). 3, Figure 4) of the drawing of the specification. Compared with the saline group, the weight of the nude mice in the drug-administered group did not decrease significantly, and no animal died. This suggests that the compound does not increase the toxicity of the drug while improving the anticancer effect of the drug. The test showed that Compound 1 has a significant anti-tumor effect on human chronic myeloid leukemia K562 cell xenograft model. Table 3 Inhibition of human chronic myeloid leukemia K562 cells transplanted into nude mice by oral administration. Rat body weight (± s) tumor weight (g) tumor inhibition rate group
前 /后 前 1 后 (%) Before/after 1 before (%)
NS 6/6 19.9±1.8 1 21.1±1.0 1.36±1.19 0 化合物 1 ( 40mg/kg ) 6/6 19.6±1.9 1 20.6±1.6 0.46±0.17 65.90** 化合物 1 ( 60mg/kg ) 6/6 20.7±1.6 1 21.5±1.2 0.50±0.40 57.70** 注: 与 NS组相比, * P<0.05;** P<0.01. 实施例 5 化合物 1对人慢性粒细胞白血病小鼠模型的作用 5.1 材料 NS 6/6 19.9±1.8 1 21.1±1.0 1.36±1.19 0 Compound 1 (40mg/kg) 6/6 19.6±1.9 1 20.6±1.6 0.46±0.17 65.90** Compound 1 (60mg/kg) 6/6 20.7± 1.6 1 21.5 ± 1.2 0.50 ± 0.40 57.70** Note: * P < 0.05; ** P < 0.01 compared with the NS group. Example 5 Effect of Compound 1 on a mouse model of human chronic myeloid leukemia 5.1 Materials
5.1.1 NOD-SCID小鼠  5.1.1 NOD-SCID mice
雌性和雄性兼用, 4 〜6周龄, 体重 18~22g, 中国医学科学院实验动物研究 所提供, 许可证编号: SCXK (京) 2005-0013。 NOD-SCID小鼠在 SPF实验室层 流架内带盖鼠盒中饲养 (符合 SPF标准:) 。标准颗粒饲料,饮水,垫料及一切与鼠接 触物品均经灭菌处理。  Both female and male, 4 to 6 weeks old, weighing 18 to 22 g, provided by the Laboratory Animal Research Institute of the Chinese Academy of Medical Sciences, license number: SCXK (Beijing) 2005-0013. NOD-SCID mice were housed in a SPF laboratory laminar box with a cover mouse (in accordance with SPF standards:). Standard pellet feed, drinking water, litter and all contact with the mouse are sterilized.
5.1.2 人白血病细胞株(K562) 说 明 书 购自中国科学院上海细胞生物学研究所细胞库, 本室常规传代培养, 采用 对数生长期的细胞做动物体内移植。 5.1.2 Human leukemia cell line (K562) The instructions were purchased from the Cell Bank of the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. The cells were routinely subcultured, and the cells in the logarithmic growth phase were used for animal transplantation.
5.1.3 化合物 1使用前配成所需浓度。 5.2 方法  5.1.3 Compound 1 is formulated to the desired concentration before use. 5.2 Method
5.2.1小鼠慢性粒细胞白血病模型的建立 5.2.1 Establishment of a mouse model of chronic myeloid leukemia
NOD-SCID小鼠接受 2.0 Gy X射线全身照射, 次日取对数生长期 K562细 胞用于移植, 尾静脉一次性注射 l xlO7 cell/ 鼠。 NOD-SCID mice received whole body irradiation with 2.0 Gy X-rays, and K562 cells in logarithmic growth phase were used for transplantation the next day, and l xlO 7 cell/mouse was injected once in the tail vein.
5.2.2化合物 1对 NOD-SCID小鼠慢性粒细胞白血病模型作用 5.2.2 Compound 1 Effect on NOD-SCID Mouse Chronic Myeloid Leukemia Model
NOD-SCID小鼠移植 K562细胞后, 随机分组, 分设生理盐水对照组及给 药治疗组, 每组四只。 空白对照组给予生理盐水 0.2ml/只灌胃, 给药治疗组给 予化合物 1 60mg/kg灌胃, 于移植后两周开始给药, 连续给药 5天, 给药治疗 组停药 5天后再连续用药 8天。给药期间密切观察小鼠一般情况、体重及食量, 评估药物毒性及动物对药物的耐受程度以调整给药方案。 记录各实验组小鼠生 存时间, 观察时间为三个月, K562细胞移植后存活两个月以上者为长期生存。 After transplanting K562 cells into NOD-SCID mice, they were randomly divided into normal saline control group and drug-treated group, with four groups in each group. The blank control group was given 0.2 ml/supply of normal saline, and the treatment group was administered with compound 1 60 mg/kg, and the administration was started two weeks after the transplantation, and the administration was continued for 5 days, and the treatment group was stopped for 5 days. Continuous medication for 8 days. The general condition, body weight and food intake of the mice were closely observed during the administration, and the toxicity of the drug and the tolerance of the animal to the drug were evaluated to adjust the administration schedule. The survival time of the mice in each experimental group was recorded for three months. The survival of K562 cells after transplantation for more than two months was long-term survival.
5.2.3外周血涂片及白细胞计数 各实验组小鼠分别于接种前和接种后 1周、 2周、 3周、 4周、 5周, 取尾 静脉血计数白细胞总数, 并制备血涂片, 经瑞氏-吉姆萨染色, 油镜下分类、 计 数。 5.2.3 Peripheral blood smear and white blood cell count The mice in each experimental group were counted before the inoculation and 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks after the inoculation, and the total number of white blood cells was counted in the tail vein blood, and blood smears were prepared. , stained by Wright-Gimsa, sorted and counted under oil mirror.
5.3结果 5.3 Results
5.3.1 NOD-SCID小鼠人慢性粒细胞白血病模型构建 5.3.1 Construction of a chronic myeloid leukemia model in NOD-SCID mice
生理盐水对照组小鼠移植人慢性粒细胞白血病细胞 K562后, 自然存活时间 为 32.5±9.0天。 动物皮毛起皱、 萎糜少动、 歩态不稳,体重逐渐减轻。 移植后三 周外周血白细胞明显升高, 可达移植前 2-10倍, 外周血涂片见大量幼稚细胞。 死亡小鼠大体解剖, 腹腔内见大量血性腹水, 并出现实体瘤, 脾大不明显 (见 说明书附图 5 )。 生理盐水对照组 NOD-SCID小鼠骨髓细胞 RT-PCR均检到人 bcr-abl基因 (见说明书附图 6), 说明人慢性粒细胞白血病细胞 K562归巢至 NOD-SCID小鼠骨髓。 证明人慢性粒细胞白血模型构建成功。  The physiological survival time of the mice in the saline control group was 32.5±9.0 days after transplantation of human chronic myeloid leukemia cell K562. Animal fur wrinkles, wilting and less movement, unstable state, and gradually reduced weight. Peripheral blood leukocytes increased significantly in the three weeks after transplantation, which was 2-10 times before transplantation, and a large number of immature cells were seen in peripheral blood smears. The dead mice were grossly dissected, and a large amount of bloody ascites was seen in the abdominal cavity, and solid tumors appeared, and the splenomegaly was not obvious (see Figure 5 of the specification). The saline-controlled group of NOD-SCID mouse bone marrow cells detected the human bcr-abl gene by RT-PCR (see Figure 6 of the specification), indicating that human chronic myeloid leukemia cell K562 homing to the bone marrow of NOD-SCID mice. The human chronic granulocyte white blood model was successfully constructed.
5.3.2 口服给药对小鼠慢性粒细胞白血病作用  5.3.2 Effect of oral administration on chronic myeloid leukemia in mice
给药治疗组截至结束为止 4只小鼠中有 2只尚存活, 生存时间已达 85天, 实 现长期生存, 其余小鼠生存时间较生理盐水对照组明显延长, 外周血白细胞数 明显低于生理盐水对照组, 外周血象幼稚细胞亦明显少于对照组。 (见表 4、 5, 及说明书附图 7、 8), 试验表明实施例 1所合成的化合物 1对小鼠慢性粒细胞白血 说 明 书 病有作用。 In the drug-treated group, 2 of the 4 mice survived, and the survival time has reached 85 days, achieving long-term survival. The survival time of other mice was significantly longer than that of the saline control group, and the number of peripheral white blood cells was significantly lower than that of the physiological group. In the saline control group, the peripheral blood like immature cells were also significantly less than the control group. (See Tables 4 and 5, and Figures 7 and 8 of the specification). The experiment showed that Compound 1 synthesized in Example 1 was used to treat chronic granulocyte white blood in mice. Explain that book disease has a role.
表 4 口服给药对慢性粒细胞白血病小鼠生存时间影响 剂 量 给药前后体重 长期生存 生存时间 (d) 生存时间 组 别 前 1后 延长率  Table 4 Effect of oral administration on survival time of mice with chronic myeloid leukemia. Dosage Before and after administration, body weight, long-term survival, survival time (d) survival time, group, before and after, elongation rate
(mg/kg) ( ± S) ( ±5) (%) (mg/kg) ( ± S) ( ±5) (%)
NS 0 26.3±2.5/ 28 0/4 32.5±9.0 0 化合物 1 60 25.5±2.1/ 23.3±0.6 2/4 68.5±24.4 110.77* NS 0 26.3±2.5/ 28 0/4 32.5±9.0 0 Compound 1 60 25.5±2.1/ 23.3±0.6 2/4 68.5±24.4 110.77*
*: P<0.05, VS NS组。 *: P < 0.05, VS NS group.
表 5 各实验组小鼠外周血白细胞数 (X 107L) 组 别 K562细胞接种后时间 (周) Table 5 Peripheral blood leukocyte counts of each experimental group (X 107L) Group K562 cells after inoculation time (weeks)
0 1 2 3 4 5 0 1 2 3 4 5
NS 4.02±1.02 3.31±1.34 4.68±1.52 14.03±5.52 8.91±2.66 死亡 化合物 1 3.92±0.34 2.53±0.70 2.52±1.27 7.86±2.52 7.35±3.11 6.47±3.86 NS 4.02±1.02 3.31±1.34 4.68±1.52 14.03±5.52 8.91±2.66 Death Compound 1 3.92±0.34 2.53±0.70 2.52±1.27 7.86±2.52 7.35±3.11 6.47±3.86
实施例 6 化合物 2对小鼠肝癌 H22移植肿瘤的抑制作用 Example 6 Inhibition of Compound 2 on Mouse Hepatoma H22 Transplanted Tumor
化合物 2为上述实施例方法制得的, 本实例的测试方法同化合物 1。 将化 合物 2使用前配成所需浓度。 取肿瘤生长良好的种鼠脱颈处死, 无菌条件下剥 取瘤块, 取生长状态良好的肿瘤组织, 匀浆过滤制成单细胞悬液, 调整细胞数 为 1. O X IOVml , 每只接种 0. 2ml于小鼠右腋下, 建立 H22小鼠移植瘤模型。接 种次日, 将小鼠按体重随机分组, 每组 9 只小鼠, 并且记录给药前体重。 按 0. 2ml/10g体重给药, 每天一次, 共 8次口服给药。 于末次给药后 24h脱颈处 死, 剥瘤拍照并称重(见表 6和说明书附图 9), 计算肿瘤平均重量及肿瘤抑制 率。肿瘤抑制率(%) = [对照组平均瘤质量一实验组平均瘤质量] /对照组平均瘤 质量 X 100 %。 说 明 书 Compound 2 was prepared by the method of the above examples, and the test method of this example was the same as Compound 1. Compound 2 was formulated to the desired concentration prior to use. The mice with good tumor growth were sacrificed by cervical dislocation, the tumor pieces were removed under aseptic conditions, and the tumor tissues with good growth state were taken, and the single cell suspension was prepared by homogenization filtration, and the number of cells was adjusted to 1. OX IOVml , each inoculation 0. 2ml was placed in the right axilla of the mouse to establish a H22 mouse xenograft model. On the next day of inoculation, mice were randomized by body weight, 9 mice per group, and the pre-dose weight was recorded. It was administered at a dose of 0.2 ml/10 g body weight once a day for 8 times orally. The neck was sacrificed 24 hours after the last administration, and the tumor was taken and weighed (see Table 6 and Figure 9 of the specification) to calculate the average tumor weight and tumor inhibition rate. Tumor inhibition rate (%) = [analytical tumor mass of the control group - mean tumor mass of the experimental group] / average tumor mass of the control group X 100%. Instruction manual
 Group
表 6 化合物 2口服给对小鼠肝癌 H22移植瘤的抑制作用 Table 6 Compound 2 Orally administered to mice liver cancer H22 xenografts
体: ( s) 瘤重 (g)
Figure imgf000013_0001
Body: ( s) tumor weight (g)
Figure imgf000013_0001
¾7后 (x±s)  After 3⁄47 (x±s)
NS 9/9 20.08士 1.03/28.9士 3.15 1.24士 0.64  NS 9/9 20.08士 1.03/28.9士 3.15 1.24士 0.64
19.83±0.96/30.08士 3.77 1.08士 0.33 13.42 19.83±0.96/30.08士 3.77 1.08士 0.33 13.42
50mg/kgX8  50mg/kgX8
阳性对照  Positive control
9/9 19.96士 1.19/21.56士 3.28 0.90士 0.20 27.25 20mg/kgX8  9/9 19.96 士 1.19/21.56士 3.28 0.90士 0.20 27.25 20mg/kgX8
化合物 2  Compound 2
20.52±1.23/26.23±3.41 0.90±0.36 27.43 35mg/kgX8  20.52±1.23/26.23±3.41 0.90±0.36 27.43 35mg/kgX8
化合物 2  Compound 2
20.38士 1.24/23.74士 1.85 0.84士 0.42 32.25 50mg/kgX8  20.38士 1.24/23.74士 1.85 0.84士 0.42 32.25 50mg/kgX8
化合物 2  Compound 2
20.56士 1.82/21.08士 1.01 0.46士 0.26 62.60** 75mg/kgX8  20.56士 1.82/21.08士 1.01 0.46士 0.26 62.60** 75mg/kgX8
化合物 2  Compound 2
9/9 20.14±1.52/18.69士2.55 0.52士 0.16 58.39** 115mg/kgX8  9/9 20.14±1.52/18.69士2.55 0.52士 0.16 58.39** 115mg/kgX8
注: 与 NS组相比, 叩 < 0.05; **P< 0.01 。 本发明以上述实施例同样的方式也可验证包括化合物 1或化合物 2或其在 药学上可接受盐以及含有化合物 1或化合物 2或其在药学上可接受盐形式的药 物组合物具有类似的特性;药学上可接受盐的例子包括碱金属盐如钠盐或钾盐, 碱土金属盐如钙盐或镁盐, 含有机碱的盐如铵盐, 或舍有机碱的盐如三乙基胺 Note: Compared with the NS group, 叩 < 0.05; **P < 0.01. The present invention can also be verified in the same manner as the above examples, including Compound 1 or Compound 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing Compound 1 or Compound 2 or a pharmaceutically acceptable salt form thereof having similar properties. Examples of the pharmaceutically acceptable salt include an alkali metal salt such as a sodium salt or a potassium salt, an alkaline earth metal salt such as a calcium salt or a magnesium salt, a salt containing an organic base such as an ammonium salt, or a salt of an organic base such as triethylamine.

Claims

权 利 要 求 书  Claims
Figure imgf000014_0001
Figure imgf000014_0001
2、 权利要求 1所述的化合物 1或化合物 2的制备方法, 其特征在于: 将姜黄素 溶于干燥过的二氯甲垸,加入催化量 DMAP,而后加入已溶解于二氯甲垸的 4- [双 (2-氯乙基)氨基]苯丁酸进行酯化反应获得化合物 1或化合物 2产物,而后将化合 物 1或化合物 2产物经过柱层析分离纯化得到纯化的化合物 1或化合物 2产物。 The method for producing the compound 1 or the compound 2 according to claim 1, wherein: the curcumin is dissolved in the dried methylene chloride, the catalytic amount of DMAP is added, and then the dissolved tetrachloromethane is added. - [Bis(2-chloroethyl)amino] phenylbutyric acid is esterified to obtain the product of compound 1 or compound 2, and then the product of compound 1 or compound 2 is purified by column chromatography to obtain purified compound 1 or compound 2 .
3、 根据权利要求 2的化合物 1或化合物 2的制备方法, 其特征在于所述的二氯 甲垸溶解脱水剂 DCC或 EDCI后再溶解姜黄素, 而后加入催化量的 DMAP。 A process for the preparation of a compound 1 or a compound 2 according to claim 2, characterized in that the methylene chloride dissolves the dehydrating agent DCC or EDCI and then dissolves curcumin, and then a catalytic amount of DMAP is added.
4、 权利要求 1所述的化合物 1或化合物 2的姜黄素衍生物及其药学上可接受的 盐在制备抗肿瘤药物中的应用。 The use of the curcumin derivative of the compound 1 or the compound 2 according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of an antitumor drug.
5、 含有权利要求 1所述的化合物 1或化合物 2姜黄素衍生物或其药学上可接受 的盐组合成的药学上可接受的药物组合物或制剂在制备抗肿瘤药物的应用。 A use of a pharmaceutically acceptable pharmaceutical composition or preparation comprising the compound 1 or the compound 2 curcumin derivative of claim 1 or a pharmaceutically acceptable salt thereof for the preparation of an antitumor drug.
6、 根据权利要求 4或 5所述的应用, 其特征在于: 所述的抗肿瘤药物用于制备 治疗白血病、 淋巴瘤、 皮肤癌、 胃癌、 结肠癌、 肝癌、 乳腺癌、 前列腺癌或其 权 利 要 求 书 6. The use according to claim 4 or 5, wherein: the antitumor drug is used for the preparation of leukemia, lymphoma, skin cancer, gastric cancer, colon cancer, liver cancer, breast cancer, prostate cancer or Claim
他恶性肿瘤的药物。 His drugs for malignant tumors.
7、 一种含有权利要求 1所述的药物或其药学上可接受的盐, 或含有权利要求 1 所述的药物或其药学上可接受的盐组成的药物组合物或制剂。  A pharmaceutical composition or preparation comprising the medicament according to claim 1 or a pharmaceutically acceptable salt thereof, or the medicament according to claim 1 or a pharmaceutically acceptable salt thereof.
8、权利要求 1所述的 4- [双 (2-氯乙基:)氨基]苯丁酰基姜黄素的单酯或双酯或其药 学上可接受的盐或含有 4- [双 (2-氯乙基:)氨基]苯丁酰基姜黄素的单酯或双酯及其 药学上可接受的盐组合成的药物组合物或制剂在制备抗肿瘤药物中的应用。  A monoester or diester of 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin according to claim 1 or a pharmaceutically acceptable salt thereof or 4- [double (2-) Use of a pharmaceutical composition or formulation of a combination of a monoester or a diester of chloroethyl:)amino] phenylbutyryl curcumin and a pharmaceutically acceptable salt thereof for the preparation of an antitumor drug.
9、 根据权利要求 4或 5或 8所述的应用, 其特征在于: 所述的肿瘤为白血病、 皮肤癌、 胃癌、 结肠癌、 肝癌、 乳腺癌或前列腺癌药物。  9. Use according to claim 4 or 5 or 8, characterized in that the tumor is a leukemia, skin cancer, gastric cancer, colon cancer, liver cancer, breast cancer or prostate cancer drug.
10、 根据权利要求 9所述的的应用, 其特征在于: 白血病为人慢性粒细胞白血  10. The use according to claim 9, wherein: leukemia is human chronic granulocyte white blood
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