WO2011147254A1

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

Info

Publication number: WO2011147254A1
Application number: PCT/CN2011/073844
Authority: WO
Inventors: 许建华; 刘洋; 吴丽贤; 林燕芳; 吴枝娟; 郭晓丹; 吴敏
Original assignee: 福建医科大学
Priority date: 2010-05-26
Filing date: 2011-05-09
Publication date: 2011-12-01
Also published as: GB2494595B; CN101830819B; GB201223251D0; GB2494595A; CN101830819A

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

Phenylbutyryl curcumin derivative and its application in preparing antitumor drugs

Technical field

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.

Background technique

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.

Summary of the invention

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:

Instruction manual

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 (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.

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.

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.

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.

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;

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%.

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.

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.

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.

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

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.

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.

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

The present invention will be described in detail below with reference to the accompanying drawings and examples.

Example 1 Synthesis of 4-[bis(2-chloroethyl:)amino]phenylbutanoyl curcumin (Compound 1)

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:

Molecular formula C ₃₅ H ₃₇ C1 ₂ N0 ₇ , mpl00-102°C

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) Description of the Invention Example 2 Synthesis of 4,4 ' - [bis(2-chloroethyl:)amino] bis-butyroyl curcumin (Compound 2)

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:

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 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 Use the required concentration before use.

3.2 Methods

3.2.1 Establishment of tumor-bearing mouse model

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 Grouping

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: 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,

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 Results

3.3.1 Compound 1 antitumor effect of intravenous administration

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).

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

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 Compound 1 anti-tumor effect of oral administration

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

Table 2 Inhibition of oral administration of mouse liver cancer H22 xenografts

Rat number body weight (± s) tumor weight (g) tumor inhibition rate group

Before/after 1 before (%)

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

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

Example 4 Effect of Compound 1 on human chronic myeloid leukemia K562 cell xenograft model in nude mice 4.1 Materials

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 Methods i)s

* *

4.2.1 Establishment of tumor-bearing mouse model

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 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 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 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 Results

4.3.1 Percentage of tumor growth

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 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

Before/after 1 before (%)

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 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. 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 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 Compound 1 is formulated to the desired concentration before use. 5.2 Method

5.2.1 Establishment of a mouse model of chronic myeloid leukemia

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 Compound 1 Effect on NOD-SCID Mouse Chronic Myeloid Leukemia Model

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 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 Results

5.3.1 Construction of a chronic myeloid leukemia model in NOD-SCID mice

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 Effect of oral administration on chronic myeloid leukemia in mice

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.

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) (%)

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 group.

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

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

Example 6 Inhibition of Compound 2 on Mouse Hepatoma H22 Transplanted Tumor

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

Table 6 Compound 2 Orally administered to mice liver cancer H22 xenografts

Body: ( s) tumor weight (g)

After 3⁄47 (x±s)

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

50mg/kgX8

Positive control

9/9 19.96 士 1.19/21.56士 3.28 0.90士 0.20 27.25 20mg/kgX8

Compound 2

20.52±1.23/26.23±3.41 0.90±0.36 27.43 35mg/kgX8

Compound 2

20.38士 1.24/23.74士 1.85 0.84士 0.42 32.25 50mg/kgX8

Compound 2

20.56士 1.82/21.08士 1.01 0.46士 0.26 62.60** 75mg/kgX8

Compound 2

9/9 20.14±1.52/18.69士2.55 0.52士 0.16 58.39** 115mg/kgX8

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

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 .

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.

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.

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. 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

His drugs for malignant tumors.

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.

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. 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. The use according to claim 9, wherein: leukemia is human chronic granulocyte white blood