WO2014154117A1

WO2014154117A1 - Isothiocyanate, preparation method therefor, and use against cancer

Info

Publication number: WO2014154117A1
Application number: PCT/CN2014/073923
Authority: WO
Inventors: 黄蓬; 文石军; 罗冰玲
Original assignee: Huang Peng; Wen Shijun; Luo Bingling
Priority date: 2013-03-25
Filing date: 2014-03-24
Publication date: 2014-10-02
Also published as: CN104072395A; CN104072395B

Abstract

An isothiocyanate having the following structural formula (I). The isothiocyanate synthesis processing technique is as follows: obtained by using conventional organic bases, carbon disulfide, and acetyl chloride to react in a low temperature or room temperature environment. The isothiocyanate has substantial damaging effect on tumor cells.

Description

Isothiocyanate, preparation method thereof and anticancer application

This invention relates to the field of chemical derivatives and, more particularly, to an isothiocyanate and a process for the preparation thereof. Background technique

Isothiocyanates are an important class of natural products found in cruciferous vegetables such as watercress and broccoli. These isothiocyanate-containing vegetables generally have a certain tumor prevention effect. A number of other groups and our team have done a lot of work on the anti-tumor effects of isothiocyanates, especially phenethyl isothiocyanate (PEITC). A growing body of research has shown that PEITC and its analogues can kill cancer cells efficiently, which has attracted the attention of scientists. The synthesis of novel isothiocyanates with higher antitumor activity has attracted our research interest.

At present, the synthesis method of isothiocyanate mostly uses highly toxic phosgene or expensive reagents, which limits the mass production of these compounds. It has recently been reported that isothiocyanate has been successfully prepared using p-tolyxyl chloride (TsCl) or dimethyldichlorosilane (J. Org. Chem. 2007, 3969-3971; CN1880302A). We used the two reagents in the initial study to prepare partial isothiocyanates, but they were also found to have some shortcomings during the experiment. In the preparation of less polar isothiocyanates, excessive p-tolyxyl chloride is too stable and has a low polarity, which makes it difficult to separate and purify less polar products, which tends to contaminate the product. Another reagent, dimethyldichlorosilane, is highly volatile and strongly stimulates the eyes and respiratory tract. Based on the potential antitumor activity of isothiocyanates and the limitations of current synthetic methods, it is important to find new and simple preparation methods for isothiocyanates.

Summary of the invention

The present invention is further illustrated by the combination of the reaction schemes, and the listed reaction schemes are only theoretically derived, which are not intended to limit the scope of the present invention.

The present invention provides a novel isothiocyanate having a structural formula of the formula (I):

(I)

At least one of R ¹ and R ² is hydrogen but not hydrogen at the same time.

The n is 1~3, The R ¹ and R ² are -H,

The R ³ is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentylhexyl, phenyl, 2-pyridyl, benzyl or phenethyl ,

The R4 is an oxygen atom, a carbon atom, a nitrogen methyl group, a nitrogen ethyl group or a nitrogen propyl group.

The R ₂ is preferably

or

The R is particularly selected from methyl, pentyl, ethyl, propyl, butyl or decyl _t

The R ⁴ is preferably a carbon atom, an oxygen atom or a nitrogen methyl group.

The following are the preferred 16 compounds:

Further, a method for preparing an isocyanate is provided, which comprises the following steps,

51. Dissolve the corresponding aromatic group-containing amine in an organic solvent, add a base, and then drip into an organic solvent in which carbon disulfide is dissolved.

52. After the temperature is 0 ° C, react for 5~15 minutes, stir at room temperature for 15~30 minutes.

53. At a temperature of 0 ° C, add Y, and then react at room temperature for 15 to 30 minutes.

54. Quenching reaction, purification, that is.

The base is trimethylamine, triethylamine or potassium carbonate; the solvent is tetrahydrofuran or dichloromethane; the oxime is acetyl chloride, acetic anhydride, benzoyl chloride or sulfonyl chloride, preferably acetyl chloride.

The reaction formula is derived as shown in the following reaction formula 1:

8) 032, « _s solvent

Ar.^ iNCS

, m

Sa

Y; AcC AcgO, BC , SO. _g C1⁄2

The aromatic group-containing amine is an amine corresponding to the above isothiocyanate, and has the general formula (11):

At least one of R ¹ and R ² is hydrogen but not hydrogen at the same time.

The n is 1~3,

The R ¹ and R ² are -H,

The R ³ is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, phenyl, 2-pyridyl, benzyl or benzene Ethyl,

The R ₄ is an oxygen atom, a nitrogen methyl group, a nitrogen ethyl group, or a nitrogen propyl group.

Further provided is the use of isocyanates in the preparation of anticancer drugs.

The use of the compound 1 for the preparation of a medicament for human pancreatic cancer, gastric cancer, intestinal cancer, nasopharyngeal carcinoma, liver cancer, glioma or leukemia.

The use of the compound 2 in the preparation of a medicament for human pancreatic cancer, gastric cancer, intestinal cancer, nasopharyngeal carcinoma, liver cancer, glioma, myeloma, lung cancer or leukemia.

The beneficial effects provided by the present invention are as follows:

1. Invented a series of new isothiocyanate compounds

2. Invented a new method for the synthesis of isothiocyanates with broad, rapid and high efficiency characteristics.

3. The invented compounds were found to have significant killing effects in a variety of tumor cells and have great potential for antitumor drug applications.

4. Compound 2 has the best overall inhibitory effect on human pancreatic cancer, gastric cancer, intestinal cancer, nasopharyngeal carcinoma, liver cancer, glioma, myeloma, lung cancer or leukemia, and has a good prospect in the application of antitumor drugs.

DRAWINGS

Figure 1: Drugs 1 μmol per liter of Compound 1, 2 and PEITC, respectively, after 72 hours of dosing A control map of DLD1 aggregation in human intestinal cancer cells.

Figure 2: A comparison of the drug 1, 2 and PEITC, respectively, causing apoptosis of human intestinal cancer cell DLD1 after 72 hours of dosing.

Specific actual IS^

The invention will be further described in detail below with reference to the drawings and specific embodiments. Unless otherwise stated, the reagents, devices, and methods employed in the present invention are conventionally commercially available reagents, equipment, and methods of routine use.

Example 1 Preparation of acetyl chloride-mediated phenylethyl isothiocyanate (PEITC) ^^,Α, CSs,* THF' -' 3⁄4γ- -: N:CS

| I , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , ,

, a) AcQ THF,

Triethylamine (0.71 mL, 5.45 mmol) and carbon disulfide (0.11 mL, 1.82 mmol) tetrahydrofuran (1.5) were added dropwise to a solution of phenethylamine (200 mg, 1.65 mmol) in tetrahydrofuran (2.5 mL). mL) solution. After reacting for 15 minutes in an ice water bath, the reaction solution was allowed to warm to room temperature and stirring was continued for 1 hour. Then, acetyl chloride (0.18 mL, 2.47 mmol) was added dropwise under ice water. After reacting at room temperature for 1 hour, the reaction was quenched by adding 1 M hydrochloric acid (5 mL) to a reaction mixture. The aqueous phase was extracted with EtOAc (15 mL×3), and the combined organic phase was washed with a saturated sodium chloride solution (5 mL×l) and then dried over anhydrous sodium sulfate. The dried organic phase was filtered, and the solvent was evaporated,jjjjjjjjjjjjjjjjj .

When acetic anhydride, benzoyl chloride and sulfonyl chloride were used as electrophiles, the reaction under the same conditions gave phenylethyl isothiocyanate yields of 96%, 42% and 80%, respectively (Table 1). From these data, it can be seen that, in addition to benzoyl chloride, acetyl chloride, acetic anhydride, and sulfonyl chloride give higher or comparable yields than the p-toluenesulfonyl chloride and dimethyldichlorosilane described in the literature or the patent. At the same time, we also used potassium carbonate as a base instead of triethylamine, and acetyl chloride as an electrophile, which also gave a reaction yield of 85%. This new invention provides an efficient and simple method of preparing isothiocyanates.

Table 1: Relative to reported p-toluenesulfonyl chloride and dimethyldichlorosilane, serial number 1 2 3 4 5 6

Reagent acetyl chloroacetic acid benzoyl chlorosulfonyl chloride p-toluenesulfonyl chloride dimethyl dichloride Yield 94% 96% 42% 80% 70% 79% Considering that acetyl chloride is easy to handle, we subsequently used acetyl chloride to prepare other isothiocyanates under the same conditions and achieved good results. (Table II). These data indicate that the electrophiles, especially acetyl chloride, in this patent can efficiently and simply prepare isothiocyanates, avoiding the problem of separation of low polarity products using p-toluenesulfonyl chloride and the use of dimethyl dichloride. Silicon is very exciting.

Table 2 Q: Preparation and yield of isothiocyanate Q ) cCU THF

; ^ 3 3 ^ 4 yield (%)

1 '- 93

2 95

3 into JW 96

Ϊ

4 f , 91

6 86

7 97

8, 91

9 87

i

, Ί V, ^:

10 80

Example 2, Synthesis of Compound 1

Compound 1 was prepared by the same conditions as in the synthesis of PEITC by using compound la, using acetyl chloride. The structure of Compound 1 is characterized by: 3⁄4 MR (400 Hz, CDCl3): δ 8.01 (d, 2H, J = 8.2 Hz,), 7.29 (d 2H, J = 8.2 Hz), 3.91 (s, 3H), 3.75 (t , 2H, J = 6.8 Hz), 3.04 (t, 2H, J = 6.8 Hz) ppm _; ¹³ C MR (100 Hz, CDC1 ₃ ): δ 166.9, 142.4, 130.2, 129.3, 129.0, 52.2, 46.0, 36.5 ppm. MS (ESI, m/z) 221.0 (M+H ⁺ ).

Preparation of compound la: 0.5 ml of acetyl chloride was added to 5 ml of methanol, and stirred at room temperature for 0.5 h, then 4-(2-aminoethyl)benzoate hydrochloride SI (500 mg, 2.48 mmol) was added. After refluxing at 65 ° C for 2 h, EtOAc (EtOAc m.)

Example 3, Synthesis of Compound 2

Oh,

Compound 2 was prepared by the same conditions as in the synthesis of PEITC using Compound 2a, using acetyl chloride. The structure of compound 2 is characterized by: ¹ H MR (400 Hz, CDCl ₃ ): δ 7.67 (d, 2H, J = 8.0 Hz), 7.2 l (d, 2H, J = 8.0 Hz), 6.04 (s, 1H), 3.68 (t, 2H, J=6.4Hz), 3.43(t, 2H, J=6.4Hz), 2.96(t, 2H, J=6.4Hz); 1.20(t, 3H, J=7.2Hz) ppm _; ¹³ C MR (100 Hz, CDCh): δ 167.1, 140.6, 134.0, 129.1, 127.5, 46.2, 36.4, 35.1, 15.0 ppm. MS (ESI, m/z) 235.1 (M+H+), 263.1 (M+H ₄ ⁺ ), 469.2 (2M+H+).

Preparation of compound 2a: 4-(2-Aminoethyl)benzoate hydrochloride SI (200 mg „ 0.99 mmol) and Boc ₂ OC 260 mg, 1.19 mmol) dissolved in 3 ml of tetrahydrofuran, 1 M added dropwise in an ice bath 2 ml of sodium hydroxide solution, reacted for 2 hours at room temperature, neutralized with 1 M hydrochloric acid to acidity, and extracted with ethyl acetate three times. The organic phase was collected and dried over anhydrous sodium sulfate, filtered and concentrated to give white solid 2c (240 mg, yield 91%).

2c was dissolved in 15 ml of dichloromethane, and HOBt (183 mg, 1.35 mmol) and triethylamine (0.25 ml, 1.98 mmol) were quickly added to the solution, and EDCI (259 mg, 1.35 mmol) was quickly added in an ice bath, and stirred at room temperature for 0.5 hour. Add 70% ethylamine solution (0.15ml, 1.81mmol) to the ice bath, stir at room temperature for 1 hour, then add saturated ammonium chloride solution, extract 3 times with dichloromethane, and wash once with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate (MgSO4). 2b was dissolved in 3 ml of dichloromethane, 0.3 ml of trifluoroacetic acid was added, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, The hydrazine was extracted 4 times (15 ml*4), EtOAc (EtOAc m.

Example 4, Synthesis of Compound 3 NsHCC3⁄4.

3 Compound 3 was prepared by the same conditions as in the synthesis of PEITC using Compound 3a using acetyl chloride. The structure of compound 3 is characterized by: 3⁄4 MR (400 Hz, CDCh): δ 7.68 (d, 2Η, J=8.0 Hz), 7.20 (d, 2H, J=8.0 Hz), 6.23(s, 1H), 3.68(t , 2H, J=6.8Hz), 3.34(dd, 2H, J=6.8, 13.6Hz): 2.95(t, 2H, J=6.8Hz), 1.57(dd, 2H, J=7.2, 14.4Hz), 0.9 l(t, 3H, J=7.2Hz) ppm; ¹³ C

MR (100 Hz, CDCh): δ 167.2, 140.6, 134.0, 129.1, 127.5, 46.2, 419, 36.4, 23.0, 11.5 ppm.

Preparation of compound 3a: 2c (200 mg, 0.75 mmol) was dissolved in 10 ml of dichloromethane, and HOBt (112 mg, 0.83 mmol) and triethylamine (0.22 ml, 1.66 mmol) were quickly added to the solution, and EDCI was quickly added to the ice bath. (217 mg, 1.13 mmol), stirred at room temperature for 0.5 hour; isopropylamine solution (0.14 ml, 1.66 mmol) was added dropwise in an ice bath, stirred at room temperature for 1 hour, then saturated ammonium chloride solution was added, and extracted with dichloromethane for 3 times, saturated The sodium chloride was washed once, and the organic layer was dried (jjjjjjjd 3b was dissolved in 3 ml of dichloromethane, added with 0.3 ml of trifluoroacetic acid, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, dichloromethane. The hydrazine was extracted 4 times (15 ml*4), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give white solid 3a (135 mg, yield 99%). Example 5, Synthesis of Compound 4

, HO3⁄4 3⁄4 . Off TFA.DO

. E L P

o

Ri)C% ₍ . THF

Compound 4 was prepared by the same conditions as in the synthesis of PEITC using Compound 4a using acetyl chloride. The structure of compound 4 is characterized by: iH MR OOHz DCh): δ 7.78 (d, 2H, J = 7.6 Hz), 7.57 (d 2H, J = 8.0 Hz), 7.32-7.07 (m, 5H), 3.70 (t, 2H, J=6.8Hz), 2.98(t, 2H, J=6.8Hz) ppm; ¹³ C MR (100Hz, CDC1 ₃ ): δ 165.5, 141.3, 138.0, 134.1, 129.4, 127.7, 124.8, 120.4, 46丄36.4 ppm.

Preparation of compound 4a: 2c (200 mg, 0.75 mmol) was dissolved in 10 ml of dichloromethane, and HOBt (112 mg, 0.83 mmol) and triethylamine (0.22 ml, 1.66 mmol) were quickly added to the solution, and EDCI was quickly added to the ice bath. (217mg, 1.13mmol), stirring at room temperature for 0.5 hours; adding aniline solution to the ice bath, stirring at room temperature for 1 hour, adding saturated ammonium chloride solution, extracting with dichloromethane for 3 times, washing with saturated sodium chloride, organic The mixture was dried over anhydrous sodium sulfate (MgSO4). 4b was dissolved in 4 ml of dichloromethane, added with 0.4 ml of trifluoroacetic acid, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, dichloromethane. The hydrazine was extracted 4 times (15 ml*4), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give white solid 4a (l lOmg, yield 60%).

Example 6 Synthesis of Compound 5

Compound 5 was prepared by the same conditions as in the synthesis of PEITC using Compound 5a using acetyl chloride. The structure of compound 5 is characterized by: ¹ H MR (400 Hz, CDCh): δ 8.68 (s, 1H), 8.39 (d, 1H, J = 8.4 Hz), 8.31 (d, 1H, J = 4.0 Hz), 7.93 ( d, 2H, J = 8.0Hz), 7.78 (dt, 1H, J = 1.6, 8.4Hz), 7.37 (d, 2H, J = 8.0Hz), 7.09 (m, 1H), 3.79 (t, 2H, J = 7.2 Hz), 3.08 (t, 2H, J = 7.2 Hz) ppm. MS (ESI, m/z) 284.1 (M+H+).

Preparation of compound 5a: 2c (100 mg, 0.38 mmol) and 2-aminopyridine (39 mg, 0.41 mmol) were dissolved in 4 ml of dichloromethane, and triethylamine (0.08 ml, 0.57 mmoi; ^P EDCI) was quickly added to the solution. 88mg _: After stirring at room temperature overnight, a saturated ammonium chloride solution was added, and the mixture was extracted three times with dichloromethane, washed once with saturated sodium chloride and dried over anhydrous sodium sulfate. Purification (ethyl acetate / petroleum ether = 1 : 1) gave 5b. 5b was dissolved in 2 ml of dichloromethane, 0.2 ml of trifluoroacetic acid was added, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, dichloromethane. The hydrazine was extracted 4 times (15 ml * 4), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give 5a.

Example 7 Synthesis of Compound 6

Compound 6 was prepared by the same conditions as in the synthesis of PEITC using Compound 6a using acetyl chloride. The structure of compound 6 is characterized by: iH MR OOHz^DCb): δ 7.67 (d, 2H, J=8.0Hz), 7.25-7.16(m, 7H), 6.39(s, 1H), 4.54(d, 2H, J =5.6 Hz), 3.64 (t, 2H, J = 6.8 Hz), 2.92 (t, 2H, J = 6.8 Hz) ppm _; ¹³ C MR (100 Hz, CDCh): δ 166.9, 140.8, 138.1, 133.4, 129.0, 128.8, 127.9, 127.6, 127.5, 46.1, 44.2, 36.3.

Preparation of compound 6a: 2c (300 mg, 1.13 mmol) was dissolved in 10 ml of dichloromethane, and HOBt (168 mg, 1.24 mmol) and triethylamine (0.44 ml, 3.39 mmol) were quickly added to the solution, and EDCI was quickly added to the ice bath. (325 mg, 1.70 mmol), stirred at room temperature for 0.5 hour; phenethylamine (0.25 ml, 2.26 mmol) was added dropwise in an ice bath, and stirred at room temperature for 2 hours, then saturated ammonium chloride solution was added, and extracted with dichloromethane. The organic phase was washed with anhydrous sodium sulfate and filtered, and then evaporated. . 6b was dissolved in 4 ml of dichloromethane, added with 0.4 ml of trifluoroacetic acid, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, The hydrazine/methanol (1/10) was extracted 4 times (15 ml*4). Example 8 Synthesis of Compound 7

Compound 7 was prepared by the same conditions as in the synthesis of PEITC using Compound 7a using acetyl chloride. The structure of compound 7 is characterized by: 3⁄4 MR (400 Hz, CDCb): δ 7.60 (d, 2H, J = 8.0 Hz), 7.27-7.16 (m, 7H), 6.13 (s, 1H), 3.68-3.62 (m, 4H), 2.94(t, 2H, J=6.8Hz), 2.86(t, 2H, J=6.8Hz) ppm _; ¹³ C MR (100Hz, CDCh): δ 167.2, 140.8, 139.0, 133.8, 129.1, 128.9 _: 128.8, 127.5, 126.7, 46.1, 41.3, 36.4, 35.8 ppm.

Preparation of compound 7a: 2c (300 mg, 1.13 mmol) was dissolved in 10 ml of dichloromethane, and HOBt (168 mg, 1.24 mmol) and triethylamine (0.44 ml, 3.39 mmol) were quickly added to the solution, and EDCI was quickly added to the ice bath. (325 mg, 1.70 mmol), stirred at room temperature for 0.5 hour; phenethylamine (0.29 ml, 2.26 mmol) was added dropwise in an ice bath, and stirred at room temperature for 2 hours, then a saturated ammonium chloride solution was added and extracted with dichloromethane. The organic phase was washed with EtOAc (EtOAc) (EtOAc m. . 7b was dissolved in 4 ml of dichloromethane, 0.4 ml of trifluoroacetic acid was added, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, The hydrazine/methanol (1/10) was extracted 4 times (15 ml*4).

Example 9 Synthesis of Compound 8:

Compound 8 was prepared by the same conditions as in the synthesis of PEITC using Compound 8a using acetyl chloride. The structure of compound 8 is characterized by:

δ 7.22-7.34(m, 4H), 3.72(t, 2H J=6.8Hz), 3.52(brs, 2H), 3.24(brs, 2H), 2.99(t, 2H, J=6.8Hz), 1.10-1.24 (m, 6H) ppm; ¹³ C MR (100Hz, CDC1 ₃ ): δ 171.1, 138.2, 136.3, 129.0, 126.9, 46.2, 43.4, 39.4, 36.3, 14.3, 13.0ppm.

Preparation of compound 8a: 2c (200 mg, 0.75 mmol) was dissolved in 10 ml of dichloromethane, and HOBt (112 mg, 0.83 mmol) and triethylamine (0.22 ml, 1.66 mmol) were quickly added to the solution, and EDCI was quickly added to the ice bath. (217 mg, 1.13 mmol), stirring at room temperature for 0.5 hour; dropwise addition of diethylamine solution (0.18 ml, 1.66 mmol) under ice-cooling, stirring at room temperature for 1 hour, adding saturated ammonium chloride solution and extracting 3 times with dichloromethane The organic phase was washed with anhydrous sodium sulfate and filtered, and then evaporated. ). 8b was dissolved in 3 ml of dichloromethane, added with 0.3 ml of trifluoroacetic acid, stirred at room temperature for 1 hour, then spun dry, added with 15 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, The hydrazine was extracted 4 times (15 ml*4), EtOAc (EtOAc m.

Example 10 Synthesis of Compound 9 The structure of Compound 9 was obtained by the same conditions as in the synthesis of PEITC using Compound 9a: iH MR OOHz DCls): δ 7.39 (d, 2H, J = 8.0 Hz) ), 7.26(d, 2H, J=8.0 Hz), 3.44-3.76(m, 10H), 3.02(t, 3H, J=6.4 Hz) ppm; ¹³ C MR (100Hz, CDC): 5170.1, 139.0, 134.3 , 129.0, 127.7, 66.9, 46.1, 36.2 ppm. MS (ESI, m/z) 277.2 (M+H ⁺ ).

Preparation of compound 9a: 2c (150 mg, 0.57 mmol) was dissolved in 10 ml of dichloromethane, and DMPA (10 mg) and triethylamine (0.25 ml, 1.87 mmol) were quickly added to the solution, and EDCI (163 mg, 0.85mmol), stirring at room temperature for 0.5 hours; adding morpholine (0.11ml, 1.24mmol) dropwise under ice bath, stirring at room temperature for 45min, adding saturated ammonium chloride solution, extracting with dichloromethane for 3 times, washing with saturated sodium chloride The organic phase was dried over anhydrous sodium sulfate, filtered, and then evaporated. m.jjjjjjjjjjjjjjjjjjj Add 0.3 ml of trifluoroacetic acid, stir at room temperature for 1 hour, spin dry, add 15 ml of dichloromethane and 1 ml of water, neutralize with alkaline sodium bicarbonate to alkaline, and extract 4 times with dichloromethane (15 ml*) 4), the organic phase is dried over anhydrous sodium sulfate, filtered and concentrated to give a white solid 9a (90 mg, yield 76.

.MHBoe /—,.

HO, ■ — W" 3⁄4H

',," EDCi.. DCM:

Compound 10 was prepared by the same conditions as in the synthesis of PEITC using Compound 10a using acetyl chloride. The structure of compound 10 is characterized by: 3⁄4 NMR (400 Hz, CDCl3): δ 7.16-7.30 (m, 4H), 3.38-3.72 (m, 4H), 3.65 (t, 2H, J = 6.8 Hz), 2.92 (t, 2H, J=6.8Hz), 2.32-2.41(m, 4H), 2.24(s, 3H) ppm _; ¹³ C MR (100Hz, CDC1 ₃ ): δ 170.2, 139.0, 134.8, 129.1, 127.8, 55.2, 46.2, 46.0, 36.4, 21.2 ppm.

Preparation of compound 10a: 2c (350mg, 1.32mmol) was dissolved in 10ml of dichloromethane, and HOBt (197mg, 1.45mmol) and triethylamine (0.85ml, 6.60mmol) were quickly added to the solution, and EDCI was quickly added to the ice bath. (379mg, 1.98mmol), stirring at room temperature for 0.5 hours; adding 1-methylpiperazine 219mg, 2.19mmol) aqueous solution under ice bath, stirring at room temperature for 2h, adding saturated ammonium chloride solution, extracting 3 times with dichloromethane The organic phase was washed with anhydrous sodium sulfate and filtered. 10b was dissolved in 8 ml of dichloromethane, added with 0.8 ml of trifluoroacetic acid, stirred at room temperature for 1 hour, then spun dry, added with 30 ml of dichloromethane and 1 ml of water, neutralized with sodium bicarbonate solid to alkaline, dichloromethane. The hydrazine was extracted 6 times (15 ml * 6), and the organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to give a white solid 10a (141 mg, yield 45%).

Example 12, Synthesis of Compound 11:

Compound 11 was prepared by the same conditions as in the synthesis of PEITC using compound la, using acetyl chloride. The structure of compound 11 is characterized by: _3⁄4 MR (400 Hz, CDCl ₃ ): δ 9.90 (s, 1H), 7.52 (d, 2H, J = 8.2 Hz), 7.19 (d, 2H, J = 8.2 Hz), 3.86 ( t, 2H, J=6.4 Hz), 2.89 (t, 2H, J=6.4 Hz), 2.03 ( s, 3H) ppm; ¹³ C MR (100 Hz, DMSO): 5167.9, 137.8, 131.8, 128.9, 118.8, 45.8 , 34.6, 23.7 ppm. MS (ESI, m/z) 2221. (M+H+), 441.1 (2M+H ⁺ ).

Preparation of compound 11a: 4-aminophenylacetonitrile (300 mg, 2.27 mmol), DMAP (30 mg), Et ₃ N (0.6 ml, 4.54 mmol) dissolved in 6 ml of dichloromethane. 0.33 ml, 3.40 mmol), after stirring for 5 min, it was warmed to room temperature, and was reacted for 3 h; 3 ml of water and 10 ml of dichloromethane were added, and the mixture was extracted three times. The organic phase was dried over anhydrous sodium sulfate. Ethyl acetate: petroleum ether = 1 : 1 ) 11c and Boc ₂ 0 were dissolved in 10 ml of ethyl acetate, palladium on carbon was added, and stirred under a hydrogen atmosphere at room temperature overnight to obtain l ib; l ib was added with 0.5 ml of TFA and 4 ml of DCM, stirred at room temperature for 2 h, spun dry, and added with 20 ml of dichloro Hyperthyroidism and 2ml water, fixed with sodium bicarbonate The mixture was neutralized to basic, and the dichloromethane was extracted 4 times (15 ml*4). The organic phase was dried over anhydrous sodium sulfate, filtered and

Example 13 Synthesis of Compound 12:

Compound 12 was prepared by the same conditions as in the synthesis of PEITC using Compound 12a, using acetyl chloride. The structure of compound 12 is characterized by: iH MR OOHz DCh): δ 7.42-7.96 (m, 4H), 3.92 (s, 3H), 3.76 (t, 2H, J = 6.8 Hz), 3.04 (t, 2H, J = 6.8 Hz) ppm; ¹³ C MR (100 Hz, CDC1 ₃ ): δ 166.8, 137.4, 133.4, 130.7, 129.8, 128.9, 128.5, 52.2, 46.1, 36.2 ppm. Preparation of compound 12a: 0.5 ml of acetyl chloride added to 5 ml of methanol After stirring at room temperature for 0.5 h, 3-carboxybenzaldehyde S3 was added. The mixture was refluxed at 65 ° C for 2 h, EtOAc (EtOAc)EtOAc. 12d, Mixing nitroformamidine, ethanol and water, adding sodium hydroxide (0.2 eq _U iv) solution to the ice bath, reacting for 2h, adding 1M hydrochloric acid solution to neutral, extracting 3 times with dichloromethane, organic phase Drying over anhydrous sodium sulfate and concentration by filtration afforded 12c. 12c (316mg, 1.40mmol) was dissolved in 4ml of acetic acid, added triethylamine (0.4ml, 2.81mmol), stirred at room temperature for 1h, extracted with ethyl acetate three times, dried over anhydrous sodium sulfate Purification by chromatography (ethyl acetate / petroleum ether = 1 :3) 12b, silica gel powder, isopropanol and dichloromethane were mixed, sodium borohydride was added under stirring, stirred for 30 min, 1 M HCl was added under ice bath, stirred for 20 min, filtered, and the filtrate was collected, and the filtrate was washed once with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate (MgSO4).

Example 14 Synthesis of Compound 13:

Compound 13 was prepared by the same conditions as in the synthesis of PEITC using Compound 13a, using acetyl chloride. The structure of compound 13 is characterized by: ¹ H MR (400 Hz, CDCl ₃ ): δ 7.69-7.33 (m, 4H), 6.57 (s, 1H), 3.71 (dd, 2H, J = 6.8, 11.2 Hz), 3.47 ( m, 2H), 2.99 (dd, 2H, J = 6.4, 11.2 Hz), 1.23 (t, 3H, J = 8.0 Hz) ppm _; ¹³ C MR (100 Hz, CDCh): δ 167.4, 137.5, 135.6, 131.8, 129.1, 127.6, 126.0, 46.3, 36.4, 35.1, 14.9 ppm.

Preparation of compound 13a: 3-carboxybenzaldehyde S3 was dissolved in 10 ml of dichloromethane, and HOBt and triethylamine were quickly added to the solution, and EDCI was quickly added in an ice bath, stirred at room temperature for 0.5 hour; and 70% was added dropwise in an ice bath. The amine solution was stirred at room temperature for 1 hour, and then a saturated ammonium chloride solution was added thereto, and the mixture was extracted three times with dichloromethane, washed once with saturated sodium chloride, and then dried over anhydrous sodium sulfate. (Ethyl acetate: petroleum ether = 1 : 1) gave a white solid 13d (see 2b). 13d, Mixing nitroformamidine (24 equiv), ethanol and water, adding sodium hydroxide (0.2 equiv) solution to the ice bath, reacting for 2 hours, adding 1M hydrochloric acid solution to neutral, and extracting 3 times with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered and evaporated. 13c (413 mg, 1.73 mmol) was dissolved in EtOAc (3 mL, EtOAc. Purification by silica gel column chromatography (EtOAc /EtOAcEtOAcEtOAc 13b, silica, isopropanol and dichloromethane were mixed, sodium borohydride was added under stirring, stirred for 30 min, 1 M HCl was added under ice bath, stirred for 20 min, filtered, and the filtrate was collected. The filtrate was washed once with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate (MgSO4)

Example 15 Synthesis of Compound 14 C:S

Compound 14 was prepared by the same conditions as in the synthesis of PEITC using compound 14a using acetyl chloride. The structure of compound 14 is characterized by: _3⁄4 MR (400 Hz, CDCl ₃ ): δ 7.71 (d, 2H, J = 8.0 Hz), 7.25 (d, 2H, J = 8.0 Hz), 6.10 (s, 1H), 3.53- 3.46 (m, 4H), 2.80 (t, 2H, J=6.8Hz), 2.03 (dd, 2H, J=6.8, 14.4Hz) 1.25 (t, 3H, J=7.2Hz) ppm; ¹³ C MR (100Hz , CDC1 ₃ ): 5167.4, 143.7, 133.0, 128.7, 127.4, 44.2, 35.0, 32.5, 31.2, 15.0 ppm. MS (ESI, m/z) 249.1 (M+H+), 497.2 (2M+H ⁺ ).

Preparation of compound 14a: N-ethyl-4-aldehydebenzamide (180 mg, 1.02 mmol) and triphenylphosphinoacetonitrile (367 mg, 1.22 mmol) were dissolved in 10 ml of dichloromethane and heated at 40 ° C. After refluxing for 3 h, the solvent was evaporated to dryness eluting with silica gel column (ethyl acetate: petroleum ether = 1:5 - 1:1) to give N-ethyl-4-(2-cyanovinyl)benzamide ( 160 mg, yield 79%). 10 ml of a solution of Boc2O (470 mg, 1.2 mmol) in tetrahydrofuran and 50 mg of Raney-Ni were added, and the mixture was stirred overnight under a hydrogen atmosphere at room temperature. Filtration, silica gel column chromatography (ethyl acetate: petroleum ether = 1:1) to give a white solid (124mg); dissolved in 3 ml of chloroform, trifluoroacetic acid 0.3 ml, stirred at room temperature for 1 hour, then Dry, add 15 ml of dichloromethane and 1 ml of water, neutralize to basic with sodium bicarbonate solid, extract 3 times with methylene chloride (15 ml * 3), dry the organic phase with anhydrous sodium sulfate, filter and concentrate to give 14a (71 mg, yield 100%).

Example 16: Synthesis of Compound 15

IS Compound 15 was prepared by the same conditions as in the synthesis of PEITC using Compound 15a, using acetyl chloride. The structure of compound 15 is characterized by: iH MR OOHz^DCb): 57.14-7.32 (m, 9H), 4.59 (s, 2H) ppm _; ¹³ C MR (100 Hz, CDC): 5136.0, 133.7, 131.8, 130.8, 129.7, 128.6, 126.6, 126.6, 47.3 ppm.

Preparation of compound 15a: According to the literature Buchwald, SL et al. Org. lett. 2002, 35. Synthesis of 15c using benzenethiol and p-iodobenzonitrile. 16c (394mg, 1.86mmol), Boc ₂ 0 (488mg, 2.24mmol) was dissolved in 8ml of THF, added 50mg Raney-Ni, under hydrogen atmosphere, stirred at room temperature overnight, filtered, spin-dried, purified by silica gel column (acetic acid B Ester: petroleum ether = 1 : 10), 15b; Add 0.3ml TFA and 3ml dichloromethane, stir at room temperature for 0.5h, spin dry, add 10ml dichloromethane and 1ml water, neutralize sodium bicarbonate to alkali The chloroform was extracted 3-4 times. The organic phase was dried over anhydrous sodium sulfate.

Example 17 Synthesis of Compound 16

Compound 16 was prepared by the same conditions as in the synthesis of PEITC using Compound 16a using acetyl chloride. The structure of compound 16 is characterized by: iH MR OOHz^DCb): 57.41-7.17 (m, 9H), 4.66 (s, 2H) ppm _; ¹³ C MR (100 Hz, CDCh): 5137.7, 135.3, 134.4, 132.0, 130.0, 129.6, 129.4, 128.9, 128.3, 128.2, 127.7, 126.8, 125.1 ppm.

Preparation of compound 16a: According to the literature Buchwald, SL et al. Org. lett. 2002, 35. Synthesis of 16c using benzenethiol and m-iodobenzonitrile. 16c (394mg, 1.86mmol), Boc ₂ 0 (488mg, 2.24mmol) was dissolved in 8ml of THF, added 50mg Raney-Ni, under hydrogen atmosphere, stirred at room temperature overnight, filtered, spin-dried, purified by silica gel column (acetic acid B Ester: petroleum ether = 1: 10), 16b; Add 0.3ml TFA and 3ml dichloromethane, stir at room temperature for 0.5h, spin dry, add 10ml dichloromethane and 1ml water, neutralize sodium bicarbonate to alkali The chloroform is extracted 3-4 times, the organic phase is dried over anhydrous sodium sulfate, filtered and evaporated

Example 18

Antitumor effects of compounds 1-16 obtained in Examples 2 to 17 respectively: Determination by in vitro MTT method Drug molecules on tumor cell killing effect: logarithmic growth of human tumor cells, was digested by centrifugation and counted in a cell density of about 3xl0 ⁴ th, cells were seeded in 96 well plates, i.e. seeded 3000 cells per well, to be After the cells are attached, add a culture medium containing different concentrations of DPI analogs (three replicate wells per drug concentration, another blank well for zero adjustment, and the concentration of DMSO in the DPI analog is controlled within 0.1%). After incubation for 72 hours in a carbon dioxide incubator, add 5 mg/ml of MTT. After 4 hours, add 200 ul of DMSO to dissolve purple formazan (the number of viable cells is proportional to the production of formazan), shake at room temperature for 10 minutes on a shaker. Thereafter, the absorbance of the 96-well plate at a wavelength of 570 nm was measured with a microplate reader, and the experiment was repeated three times. The cell survival rate was calculated as: the average absorbance value of the drug-treated group/the average absorbance value of the control group x100%, and the IC ₅ Q of 1-16 resulted in the drug concentration at the time of 50% tumor cell death, and IC _{5 was} calculated using Prism software. Value and plot the cell survival curve.

Compound 1-16 significantly killed human intestinal cancer DLD1, gastric cancer HGC27 and pancreatic cancer Panel in vitro MTT assay. In addition, Compound 1-2 also significantly kills human liver cancer in vitro MTT assay.

(SK-Hepl, Huh-7, HepG2), leukemia (ML-1, HL-60, Molml3), gastric cancer 7901 and glioma U87; Compound 2 also significantly killed human pancreatic cancer Apscl in in vitro MTT assay, Myeloma 8226, anti-cisplatin lung cancer A549/DDP and lung cancer NCI-H460.

Compared with the widely studied phenethyl isothiocyanate, most of the compounds of the present invention exhibit higher activity, in particular, compound 2 has the best antitumor activity, human intestinal cancer DLD1, esophageal cancer HGC27, and pancreatic cancer. The half-inhibitory concentration of HGC27, IC ₅ Q, can reach 0.63, 0.46, and 2.04 micromoles per liter, respectively. Table 3: Concentrations of compound 1-16 causing human intestinal cancer DLD 1, gastric cancer HGC27, and pancreatic cancer Panel 50% death (IC ₅ Q, unit μΜ), Note: PEITC was used as a control. Compound\Cell Cancer DLD1 HGC27 Panel

PEITC 3.55±0.55 2.95±0.41 10.80±0.25

1 1.23±0.04 1.09±0.07 4.55±0.32

2 0.63±0.09 0.46±0.02 2.04±0.21

3 0.47±0.04 0.15±0.01 2.43±0.38

4 0.86±0.27 0.34±0.00 2.78±0.72

5 1.50±0.18 0.77±0.18 4.70±0.27 6 0.91±0.16 0.72±0.08 2.14±0.79

7 1.45±0.13 0.82±0.06 4.06±0.57

8 0.81±0.04 0.62±0.04 2.25±0.21

9 2.77±0.03 1.68±0.00 3.76±0.05

10 3.92 ± 0.10 2.37 ± 0.43 6.24 ± 0.13

11 1.30±0.12 0.93±0.41 2.68±0.21

12 1.31±0.35 0.74±0.01 5.78±0.56

13 1.34±0.04 0.84±0.06 3.52±0.11

14 1.66±0.06 1.47±0.10 5.08±0.02

15 5.23±0.17 2.73±0.29 14.24±0.50

16 1.25±0.06 3.44±0.23 6.12±0.13 Table 4: Compound 1-2 causes human liver cancer (SK-Hepl, Huh-7, HepG2), leukemia (ML-1, HL-60, Molml3), gastric cancer 7901, nasopharyngeal Concentration of cancer CNE-2 and glioma U87 50% death (IC ₅₀ , unit μΜ), compound\cancer cell SK-Hepl Huh-7 HepG2 ML-1 HL-60 7901 Molml3 CNE-2 U87

1 2.55 10 14.99 3.69 3.02 1.38 10.7±2.69 8.19 7.71

2 0.62 2.89 3.48 1.57 1.15 5.68 4.44 ± 1.27 3.62 3.03 Table 5: Concentration of 50% death of human pancreatic cancer Apscl, myeloma 8226, anti-cisplatin lung cancer A549/DDP and lung cancer NCI-H460 (IC ₅ Q) , the unit is μΜ) Compound\Cell cancer Aspcl 8226 A549/DDP NCI-H460

2 2.48 5.18±1.15 5.33±0.00 4.12±0.59 Compounds 1 and 2 were also able to significantly affect the growth and aggregation of human intestinal cancer cell DLD1 and cause apoptosis of cancer cells.

Claims

An isothiocyanate characterized by a structural formula of the formula (I):

(I)

At least one of R1 and R2 is hydrogen but not hydrogen at the same time.

The n is 1~3,

The R ¹ and R ² are -H,

The R ⁴ is an oxygen atom, a carbon atom, a nitrogen methyl group, a nitrogen ethyl group or a nitrogen propyl group.

2. The isothiocyanate according to claim 1, wherein the R ² is

The isothiocyanate according to claim 1, wherein the R ³ is a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group or a hexyl group.

The isothiocyanate according to claim 1, wherein the R ⁴ is a carbon atom, an oxygen atom or a nitrogen methyl group.

5. The isothiocyanate according to claim 1, wherein

A method of producing an isocyanate according to any one of claims 1 to 5, which comprises the steps of

S1. Dissolving the corresponding aromatic group-containing amine in an organic solvent, adding a base, and then dropping the organic solvent in which the carbon disulfide is dissolved.

54. Quenching reaction, purification, that is.

The method for preparing an isocyanate according to claim 6, wherein the base is trimethylamine, triethylamine or potassium carbonate; the solvent is tetrahydrofuran or dichloromethane; the hydrazine is acetyl chloride, B Anhydride, benzoyl chloride or sulfonyl chloride.

8. Use of an isocyanate according to any one of claims 1 to 5 for antitumor.

The use of the isocyanate according to claim 5, wherein the compound 1 is used in human pancreatic adenocarcinoma, gastric cancer, intestinal cancer, nasopharyngeal carcinoma, liver cancer, glioma or leukemia.

The use of the isocyanate according to claim 5, wherein the compound 2 is in human pancreatic cancer, gastric cancer, intestinal cancer, nasopharyngeal carcinoma, liver cancer, glioma, myeloma, lung cancer or leukemia. application.

The use of the isocyanate according to claim 5, wherein the tumor is lymphoma, ovarian cancer, myeloma, human pancreatic cancer, gastric cancer, intestinal cancer, nasopharyngeal carcinoma, liver cancer, glioma , lung cancer or leukemia.