CN106928235A - The LSD1 of triazole containing pyrimido inhibitor, its preparation method and application - Google Patents

Abstract

The invention belongs to the field of medicinal chemistry, and discloses a class of pyrimidotriazole compounds, a preparation method thereof and the use of lysine-specific demethylase (hereinafter referred to as LSD1) as a target in the preparation of antitumor drugs Applications. The general formula of the compound of the present invention is shown in I. The LSD1 enzyme inhibitory activity experiment in vitro proves that this type of compound has obvious inhibitory and killing effects on a variety of tumor cells by inhibiting the activity of LSD1, and can be used as a lead compound for further development and applied to the preparation of anti-tumor drugs.

Description

Pyrimidotriazole-containing LSD1 inhibitor, its preparation method and application

technical field

The invention belongs to the field of medicinal chemistry, and specifically relates to pyrimidotriazole compounds, their preparation methods and their use in antitumor drugs targeting histone lysine specific demethylase (hereinafter referred to as LSD1) application.

Background technique

Tumor is a disease that seriously endangers human health and is difficult to overcome. There are many anti-tumor drugs that have been marketed, but there are still some problems in these drugs, such as high toxicity, weak targeting, and easy drug resistance. Therefore, the research and development of new anticancer drugs is particularly important.

Histone covalent modification is an important epigenetic mode, including histone acetylation, methylation, phosphorylation, and ubiquitination, among which acetylation and methylation are more frequently studied for the mechanism of histone modification. histone modification. Before 2004, histone methylation was considered to be irreversible, and lysine-specific demethylase 1 (LSD1) was the first to be discovered that can catalyze histone H3K4me1/2 and H3K9me1/ 2 Demethylation-specific demethylases, thereby regulating the transcription of downstream target genes.

The expression of LSD1 in various tumor cells is significantly higher than that in normal cells, such as neuroblastoma, eye cancer, prostate cancer, breast cancer, lung cancer, bladder cancer, etc. Experiments have shown that reducing the expression of LSD1 or reducing the activity of LSD1 at the cellular level through RNAi technology or small molecule inhibitors can inhibit cell proliferation and induce the expression of some cell differentiation-related genes; under the action of small molecule monoamine oxidase inhibitor PCPA, it can also Inhibits the growth of various tumor cells and solid tumors. Therefore, LSD1 inhibitors can not only be used as epigenetic research tools to elucidate biological functions, but also can be used as epigenetic drugs for the prevention and treatment of tumors. hotspot.

At the same time, pyrimidines and triazole heterocyclic compounds have a wide range of biological activities, such as antiviral, antibacterial, anti-inflammatory and anti-tumor. However, there are few reports on the combination of new compounds combining pyrimidine and triazole structures with the anti-tumor effect based on the LSD1 target, so such research is of great value.

Contents of the invention

In order to develop and utilize existing clinical drug resources, the purpose of the present invention is to provide a class of derivatives with pyrimidotriazole as the mother nucleus, thereby opening up a new approach for finding a new class of anti-tumor drugs based on the LSD1 target; Another object of the present invention is to provide its preparation method and its application in the preparation of antitumor drugs and LSD1 inhibitors.

In order to achieve the purpose of the present invention, the general structural formula of the pyrimidotriazole LSD1 inhibitor described in the present invention is as follows:

In general formula I, R ₂ is C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, Any one of them, wherein, R is halogen, H, -OH, -NO ² , -OCH ³ , C1-4 acyl, n=1~6; MsO- is methylsulfonic acid group, TsO- is toluenesulfonic acid base;

R ₁ is any one of the following groups: C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, hydrogen atom, wherein, X is nitrogen, oxygen or sulfur atom, R is halogen, H, -OH, -NO ₂ , -OCH ₃ , C1-4 acyl, n=1~6;

R ₃ is any one of the following groups:

Wherein, R is halogen, H, -OH, -NO ₂ , -OCH ₃ , CF ₃ , C1-4 alkyl or C1-4 acyl.

Preferred in general formula I:

R ₂ is C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, Any one of them; n=1～3;

R ₁ is any one of the following groups: C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, hydrogen atom, wherein, X is a sulfur atom, n=1~3; _R3 is any one of the following groups:

Wherein, R is C1-4 alkyl, chlorine, bromine, H.

More preferably one of the following compounds:

8:R ¹ =propyl-S-,

9:R ¹ =propyl-S-,

10:R ¹ =propyl-S-,

11:R ¹ =propyl-S-,

12:R ¹ =propyl-S-,

13:R ¹ =propyl-S-,

14:R ¹ =propyl-S-,

15:R ¹ =propyl-S-,

16:R ¹ =propyl-S-,

17:R ¹ =propyl-S-,

18:R ¹ =propyl-S-,

19:R ¹ =propyl-S-,

20:R ¹ =propyl-S-,

21:R ¹ =propyl-S-,

The preparation method of the pyrimidotriazole LSD1 inhibitor of the present invention is mainly obtained through the following steps:

1. The preparation method of general formula 2:

In a solvent, compound 1 and brominated hydrocarbon were stirred and reacted at room temperature under the action of an alkaline substance. After the reaction was completed, the compound 2 was obtained by filtering, washing and drying. In this reaction, the solvent used may be one or a mixture of two or more of acetone, methanol, ethanol, propanol, isopropanol, tetrahydrofuran, acetonitrile, water, DMF, methylene chloride, chloroform, and dioxane. The alkaline substance used can be one of triethylamine, diisopropylethylamine, pyridine, sodium hydroxide, potassium hydroxide. The brominated hydrocarbon corresponds to the R1 substituent in the general formula;

2. The preparation method of general formula 3:

In acetic acid, add nitrating reagent, then add series compound 2 in batches, stir for reaction, after the reaction, pour into water, suction filter, wash and dry to obtain series compound 3. In this reaction, the nitrating reagent is selected from fuming nitric acid and concentrated nitric acid. The preferred temperature is 15-60°C.

3. The preparation method of general formula 4:

In the solvent, add a chlorination reagent, then add a series of compound 3 in batches, add an organic base dropwise, reflux reaction, after the reaction, cool to room temperature, hydrolyze, extract with an organic solvent, wash with water, neutralize, and dry the organic phase to obtain a series of compounds 4 Crude product (the next step can be directly carried out without further optimization, and the pure product can be obtained by column chromatography). In this reaction, the chlorination reagent can be phosphorus oxychloride, phosphorus pentachloride, solvent can be toluene, dioxane, THF, ethyl acetate etc., and described organic base can be triethylamine, N, N-dimethylaniline, dimethylacetamide (DMA), N,N-diethylaniline or pyridine, etc. The preferred temperature is 40-120°C.

4. The preparation method of general formula 5:

Dissolve series compound 4 in a mixed solvent of ethanol and acetic acid, then add reduced iron powder in batches, heat up to reflux for reaction, filter with suction, spin dry the solvent, extract with organic solvent, wash with water, and dry the organic phase to obtain series compound 5 Crude and pure products can be obtained by column chromatography.

5. the preparation method of general formula 6:

The series compound 5 and the amine compound are dissolved in the solvent, then add an organic base, reflux reaction, after the reaction, evaporate the solvent, add ethyl acetate, wash with water, and dry the organic phase to obtain the crude product of the series compound 6 (without further purified and used directly in the next reaction). In this reaction, the solvent used in the reaction can be methanol, ethanol, isopropanol, DMF, dioxane, THF, acetonitrile, etc., and the organic base can be triethylamine, pyridine or diisopropylethylamine, etc. The preferred reaction temperature is 60-120°C. The amine compound corresponds to the substituent R in the general _formula ;

6. The preparation method of general formula 7:

The series compound 6 was dissolved in the mixed solution of acetic acid and water, and the aqueous solution of sodium nitrite was added dropwise under ice bath for reaction. After the reaction was completed, ethyl acetate and water were added, the layers were separated, washed with water, neutralized, and the organic phase was dried to obtain The crude product of series compound 7 was directly carried out to the next reaction without further purification.

7. The preparation method of compound 8～21 in the general formula:

Dissolve the series compound 7 and the mercapto aromatic ring compound in the solvent, add an acid-binding agent, react at room temperature or reflux, after the reaction, evaporate the solvent to dryness, dissolve in ethyl acetate, wash with water, dry, pass through the column, and the mobile phase is Different proportions of petroleum ether and ethyl acetate. In this reaction, the solvent is methanol, ethanol, isopropanol, THF, acetonitrile, DMF, dioxane, etc., and the acid-binding agent is triethylamine, pyridine, isopropylethylamine, etc. The preferred reaction temperature is 20-100°C.

The pyrimidotriazole derivatives described in the present invention have a good inhibitory effect on LSD1 through LSD1 enzyme activity experiments. Therefore, the pyrimidotriazepine derivatives provided by the present invention have opened up another effective way for the development of novel antitumor drugs, drug combinations and novel LSD1 inhibitor drugs. The synthetic design of these compounds is reasonable, the reaction conditions are mild, and the operation The method is simple, the reaction yield is high, and the total yield reaches more than 60%. If it is developed into a new drug, it will have a good market application prospect.

detailed description

In order to better illustrate the present invention, special examples are as follows:

Compound 8 of Example 1, R ₁ =propyl-S-, preparation of

(1) Preparation of Compound 2 (R ₁ =propyl-S-)

Barbituric acid (3g, 1eq) and triethylamine (2.9ml, 1eq) were added to 30ml of methanol, under reflux, bromopropane (1.8ml, 1eq) was slowly added dropwise, and reflux was continued for 1 hour after the addition , cooled, and suction filtered to obtain 3.7g of pink solid compound 2b with a yield of 97%.

(2) Preparation of Compound 3 (R ₁ =propyl-S-)

Under ice bath, carefully dissolve 3ml of fuming nitric acid in 6ml of acetic acid, then add 2.9g of compound 2b in batches, after the addition, continue to stir for 2 hours, then add the reaction solution to 18ml of ice water, filter with suction, After washing with water, a dark red powder compound 3b was obtained with a yield of 77.5%.

(3) Preparation of Compound 4 (R ₁ =propyl-S-)

Compound 3b (12.4g, 1eq) was dissolved in 50ml of phosphorus oxychloride, and DMA (12ml, 1.8eq) was slowly added dropwise, then heated to reflux, and reacted for 5 hours. Cool to room temperature, hydrolyze, then extract with EA, wash with water, and then wash with saturated sodium carbonate solution, and dry the organic phase to obtain 13 g of crude brown compound 4b with a yield of 90.2%.

(4) Preparation of Compound 5 (R ₁ =propyl-S-)

Dissolve compound 4b (0.5g, 1eq) in 4ml of methanol and 2ml of acetic acid, then add reduced iron powder (0.3g, 3eq) in batches, reflux for 2 hours, cool to room temperature, filter with suction, evaporate the filtrate to dryness, dissolve In ethyl acetate, washed with saturated sodium carbonate solution, washed with water, dried the organic phase, and evaporated to dryness to obtain 0.44 g of crude compound 5b, with a yield of 95%.

(5) Compound 6 (R ₁ =propyl-S-, ) preparation

Compound 5b (3.9g, 1eq), ethanolamine (1.0g, 1eq) and triethylamine (3ml, 1.3eq) were dissolved in ethanol, refluxed for 48 hours, evaporated to dryness, dissolved in ethyl acetate, and then washed with dilute hydrochloric acid Neutralized to neutral, washed with water three times, dried, and evaporated to dryness to obtain the crude compound 6b, which was directly carried out to the next step without further purification.

(6) Compound 7 (R ₁ =propyl-S-, ) preparation

Dissolve the crude product of compound 6 in the previous step in a mixture of acetic acid and water, add an aqueous solution of sodium nitrite (1.13g, 1eq) dropwise under an ice bath at 0°C, keep the temperature not exceeding 10°C, and then continue to stir for reaction 1 hour, the reaction solution was dissolved in ethyl acetate, washed three times with water, and then neutralized to neutral with saturated sodium bicarbonate solution, washed with water, dried organic phase, and evaporated to dryness to obtain the crude product of compound 7b, which was directly carried out without purification Next step.

(7) Preparation of Compound 8

The intermediate compound 7b (100mg, 1eq), mercaptobenzothiazole (61mg, 1eq) and triethylamine (37mg, 1eq) were dissolved in ethanol, heated to reflux for several hours, and the reaction was monitored by TLC (PE/EA). After the end, the solvent was evaporated to dryness, dissolved in ethyl acetate, washed with water, dried, and passed through the column to obtain 89 mg of white solid with a yield of 60%. ¹ HNMR (400MHz, DMSO-d ₆ , ppm): δ8.23-8.25 (d, J = 7.6Hz, 1H), 8.10-8.12 (d, J = 8.0Hz, 1H), 7.55-7.63 (m, 2H ),4.95-4.97(t,J=5.8Hz,1H),4.64-4.67(t,J=5.4Hz,2H),3.92-3.96(m,2H),2.95-2.99(t,J=7.0Hz, 2H), 1.48-1.57(m, 2H), 0.77-0.80(t, J=7.4Hz, 3H). ¹³ C NMR (100MHz, DMSO-d ₆ , ppm): δ169.45, 159.12, 155.22, 152.07, 149.55, 137.35, 131.15, 127.31, 126.72, 123.39, 122.66, 59.26, 50.35, 33.09, 22.33, 13.44...

Compound 9 of Example 2, R ₁ =propyl-S-, preparation of

The hydroxyethylamine protected by methanesulfonyl chloride was replaced by ethanolamine, and the same method as in Example 7 was used to prepare 9. ¹ HNMR (400MHz, CDCl ₃ , ppm): δ8.12-8.14 (d, J=8.0Hz, 1H), 7.97- 7.99(d, J=7.6Hz, 1H), 7.55-7.59(m, 1H), 7.49-7.53(m, 1H), 4.95-4.97(t, J=5.2Hz, 2H), 4.79-4.81(t, J＝5.2Hz, 2H), 3.00(s, 3H), 1.57-1.62(m, 2H), 0.85-0.89(t, J＝7.4Hz, 3H). ¹³ CNMR(100MHz, CDCl ₃ , ppm): δ171 .34, 160.36, 154.99, 152.17, 149.24, 137.33, 131.06, 126.62, 126.08, 123.47, 121.31, 65.21, 46.06, 37.95, 33.50, 22.17, 13.26. Yield 70%.

Example 3 Compound 10, R ₁ =propyl-S-, preparation of

Benzoyl-protected hydroxyethylamine was used instead of ethanolamine, and ^10.1 HNMR (400MHz, CDCl ₃ , ppm) was prepared by the same method as in Example 7: δ8.11-8.13 (d, J=8.0Hz, 1H), 7.96- 7.98(d,J=7.6Hz,1H),7.91-7.94(m,2H),7.54-7.58(m,2H),7.48-7.52(m,1H),7.40-7.44(m,2H),5.02- 5.05(t, J=5.2Hz, 2H), 4.83-4.85(t, J=5.2Hz, 2H), 2.87-2.90(t, J=7.2Hz, 2H), 1.50-1.59(m, 2H), 0.83 -0.86(t, J＝7.4Hz, 3H). ¹³ CNMR (100MHz, CDCl ₃ , ppm): δ171.04, 166.04, 160.02, 155.33, 152.08, 149.33, 137.23, 133.37, 131.01, 129.75, 129.20, 128.50, 125.99, 123.40, 121.26, 62.34, 46.12, 33.32, 22.12, 13.27... Yield 63%.

Example 4 Compound 11, R ₁ =propyl-S-, preparation of

Glycolamine was used instead of ethanolamine, and 11.1 HNMR (400MHz, _{CDCl 3} ^, ppm) was prepared by the same method as in Example 7: δ8.08-8.10 (d, J=8.0Hz, 1H), 7.94-7.96 (d ,J=8.0Hz,1H),7.52-7.56(m,1H),7.45-7.49(m,1H),4.78-4.81(t,J=5.2Hz,2H),4.03-4.06(t,J=5.2 Hz, 2H), 3.65(m, 2H), 3.58-3.60(t, J=4.2Hz, 2H), 2.91-2.95(t, J=7.4Hz, 2H), 1.52-1.58(m, 2H), 0.80 -0.84(t, J＝7.4Hz, 3H). ¹³ CNMR(100MHz, CDCl ₃ , ppm): δ170.83, 160.15, 155.16, 152.20, 149.21, 137.36, 131.09, 126.58, 126.04, 123.45, 121.28, 72.557, 68 61.61, 46.90, 33.47, 22.19, 13.24... Yield 65%.

Compound 12 of Example 5, R ₁ =propyl-S-, preparation of

Using p-toluenesulfonyl-protected ethylene glycol amine instead of ethanolamine, the same method as in Example 7 was used to prepare 12. ¹ HNMR (400MHz, CDCl ₃ , ppm): δ8.10-8.12 (d, J=8.0Hz, 1H) ,7.95-7.97(d,J=7.6Hz,1H),7.73-7.75(d,J=8.4Hz,2H),7.53-7.57(m,1H),7.46-7.50(m,1H),7.32-7.34 (d, J=8.4Hz, 2H), 4.70-4.73(t, J=5.6Hz, 2H), 4.05-4.08(t, J=4.6Hz, 2H), 3.99-4.02(t, J=5.6Hz, 2H), 3.64-3.66(t, J=4.6Hz, 2H), 2.96-3.00(t, J=7.2Hz, 2H), 2.44(s, 3H), 1.56-1.64(m, 2H), 1.24-1.28 (t, J=7.2Hz, 3H). ¹³ C NMR (100MHz, CDCl ₃ , ppm): δ170.72, 159.91, 155.39, 152.15, 149.21, 144.94, 137.30, 131.03, 129.88, 127.90, 126.54, 125.97, 121.42 , 68.88, 68.86, 68.55, 68.41, 46.49, 33.47, 22.21, 13.27... Yield 72%.

Example 6 Compound 13, R ₁ =propyl-S-, preparation of

Using furfurylamine instead of ethanolamine, the same method as in Example 7 was used to prepare ^13.1 HNMR (400MHz, CDCl ₃ , ppm): δ8.08-8.10 (d, J=8.0Hz, 1H), 7.94-7.96 (d, J =8.0Hz,1H),7.52-7.56(m,1H),7.46-7.49(m,1H),7.36-7.37(d,J=1.2Hz,1H),6.48-6.49(d,J=3.2Hz, 1H),6.35-6.36(m,1H),5.76(s,2H),3.00-3.03(t,J=7.4Hz,2H),1.61-1.66(m,2H),0.89-0.93(t,J= 7.4Hz,3H). ¹³ CNMR(100MHz,CDCl ₃ ,ppm):δ170.86,159.80,155.56,151.99,148.69,147.10,143.40,137.14,130.94,126.51,125.90,123.33,121.24,110.77,110.26,43.34,33.45 , 22.24, 13.36... Yield 61%.

Compound 14 of Example 7, R ₁ =propyl-S-, preparation of

Using isopropylamine instead of ethanolamine, the same method as in Example 7 was used to prepare ^14.1 HNMR (400MHz, CDCl ₃ , ppm): δ8.10-8.12 (d, J=8.4Hz, 1H), 7.96-7.98 (d, J =8.0Hz,1H),7.53-7.57(m,1H),7.47-7.50(m,1H),5.17-5.23(m,1H),3.01-3.05(t,J=7.2Hz,2H),1.74- 1.76(d, J=6.8Hz, 6H), 1.61-1.70(m, 2H), 0.90-0.94(t, J=7.4Hz, 3H). ¹³ CNMR(100MHz, CDCl ₃ , ppm): δ169.85, 159.59, 155.77, 151.99, 148.24, 137.14, 131.41, 126.50, 125.87, 123.31, 121.24, 51.52, 33.43, 22.28, 21.94, 13.35. Yield 60%.

Example 8 Compound 15, R ₁ =propyl-S-, preparation of

Using mercaptobenzoxazole instead of mercaptobenzothiazole, the same method as in Example ⁷ was used to prepare 15.1 HNMR (400MHz, DMSO-d ₆ , ppm): δ7.92-7.94 (d, J=8.0Hz, 1H) ,7.86-7.88(d,J=8.0Hz,1H),7.56-7.60(m,1H),7.50-7.54(m,1H),4.95(br,1H),4.61-4.63(t,J=5.4Hz ,2H),3.89-3.93(m,2H),2.76-2.79(t,J=7.2Hz,2H),1.33-1.42(m,2H),0.67-0.71(t,J=7.4Hz,3H). ¹³ CNMR (100MHz, DMSO-d ₆ , ppm): δ169.51, 160.03, 153.67, 152.79, 149.94, 141.77, 131.46, 127.52, 125.78, 120.93, 111.76, 59.23, 50.35, 33.13, 132.220. Yield 66%.

Compound 16 of Example 9, R ₁ =propyl-S-, preparation of

Using mercaptopyrimidine instead of mercaptobenzothiazole, the same method as in Example ⁷ was used to prepare 16.1 HNMR (400MHz, DMSO-d ₆ , ppm): δ8.85-8.86 (d, J=4.8Hz, 2H), 7.58- 7.60(t,J=4.8Hz,1H),4.98(br,1H),4.62-4.65(t,J=5.4Hz,2H),3.93-3.94(m,2H),3.02-3.04(t,J= 7.0Hz, 2H), 1.58-1.67(m, 2H), 0.91-0.95(t, J＝7.2Hz, 3H). ¹³ CNMR(100MHz, DMSO-d ₆ , ppm): δ169.52, 165.21, 160.78, 159.51, 149.96, 132.73, 121.32, 59.19, 50.28, 33.14, 22.38, 13.63... Yield 71%.

Compound 17 of Example 10, R ₁ =propyl-S-, preparation of

Using 2-mercapto-1,3,4-thiadiazole instead of mercaptobenzothiazole, the same method as in Example 7 was used to prepare 17. ¹ HNMR (400MHz, DMSO-d ₆ , ppm): δ9.96(s, 1H ),4.94-4.97(t,J=6.0Hz,1H),4.65-4.67(t,J=5.4Hz,2H),3.92-3.96(m,2H),3.03-3.07(t,J=7.2Hz, 2H), 1.58-1.67(m, 2H), 0.93-0.97(t, J=7.4Hz, 3H). ¹³ CNMR(100MHz, DMSO-d ₆ , ppm): δ169.48, 159.38, 158.04, 155.12, 149.59, 131.18 , 59.25, 50.38, 33.12, 22.36, 13.63... Yield 70%.

Compound 18 of Example 11, R ₁ =propyl-S-, preparation of

Using 2-amino-5-mercapto-1,3,4-thiadiazole instead of mercaptobenzothiazole, the same method as in Example 7 was used to prepare 18. ¹ H NMR (400MHz, DMSO-d ₆ , ppm): δ7. 79(s,1H),4.90-4.93(t,J=5.8Hz,1H),4.62-4.64(t,J=5.4Hz,2H),3.90-3.94(m,2H),3.03-3.07(t, J＝7.2Hz, 2H), 1.58-1.67(m, 2H), 0.94-0.97(t, J＝7.4Hz, 3H). ¹³ C NMR (100MHz, DMSO-d ₆ , ppm): δ173.62, 168.96, 160.34 , 149.11, 138.42, 130.82, 58.77, 49.78, 32.78, 22.13, 13.09... Yield 65%.

Compound 19 of Example 12, R ₁ =propyl-S-, preparation of

Using 2-mercapto-5-methyl-1,3,4-thiadiazole instead of mercaptobenzothiazole, the same method as in Example 7 was used to prepare 19. ¹ H NMR (400MHz, CDCl ₃ ) δ7.29(d, J=1.2Hz,1H),7.25(d,J=1.3Hz,1H),4.71-4.73(m,2H),4.17(m,2H),3.73(s,3H),2.89-2.93(m,2H ), 1.62-1.65 (m, 2H), 0.97-1.00 (t, J=7.4Hz, 3H). ¹³ C NMR (100MHz, CDCl ₃ ) δ174.95, 167.39, 154.15, 136.25, 136.12, 135.73, 129.97, 64.56, 54.53, 38.84, 38.15, 27.01, 18.12... Yield 69%.

Compound 20 of Example 13, R ₁ =propyl-S-, preparation of

Using 2-mercaptothiazole instead of mercaptobenzothiazole, the same method as in Example 7 was used to prepare 20. ¹ H NMR (400MHz, CDCl ₃ ) δ8.02(d, J=3.4Hz, 1H), 7.68(d, J= 3.4Hz, 1H), 4.74-4.77(m, 2H), 4.17-4.21(m, 2H), 2.97-3.00(t, J＝6.7Hz, 2H), 1.64-1.69(m, 2H), 0.98-1.02 (t, J=7.3Hz, 3H). ¹³ C NMR (100MHz, CDCl ₃ ) δ170.27, 160.66, 152.04, 149.15, 143.59, 131.03, 125.00, 59.98, 49.91, 33.32, 22.23, 13.37. Yield 62%.

Compound 21 of Example 14, R ₁ =propyl-S-, preparation of

Using methimazole instead of mercaptobenzothiazole, the same method as in Example 7 was used to prepare 21, ¹ H NMR (400MHz, CDCl ₃ ) δ7.29(d, J=1.2Hz, 1H), 7.25(d, J=1.3 Hz,1H),4.71-4.73(m,2H),4.17(m,2H),3.73(s,3H),2.89-2.93(d,J＝7.2Hz,2H),1.62-1.65(m,2H) ,0.97-1.00(t,J=7.4Hz,3H). ¹³ C NMR(100MHz,CDCl ₃ )δ174.95,167.39,154.15,136.25,136.12,135.73,129.97,64.56,54.53,38.84,38.15,27.02.1,18. . Yield 68%.

The LSD1 inhibitory activity assay of the above-mentioned compound of embodiment 15:

1. Experimental method:

The sample is obtained by purification of the above compound synthesized in the example; sample stock solution: weigh 3-5mg sample and place it in a 1.5mL EP tube, then prepare a solution with a concentration of 20mM with DMSO, store it at 4°C, and store it in the experiment Dilute with DMSO according to the desired concentration. After incubating the sample to be tested with the LSD1 protein at room temperature, add the LSD1 reaction substrate H3K4me2 and incubate the reaction, and finally add the fluorescent dye Amplex and horseradish peroxidase HRP to incubate at room temperature, and detect on a microplate reader with an excitation light of 530nm and an emission light of 590nm Fluorescence value:

The test results were calculated by using SPSS software to calculate the IC ₅₀ value.

The experimental results are shown below.

Table 1. The inhibitory activity data of the above compounds 8-21 on LSD1:

Claims (5)

1. A pyrimidotriazole-containing LSD1 inhibitor, characterized in that it has the structure described in general formula I, In general formula I, R ₂ is C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, Any one of them, wherein, R is halogen, H, -OH, -NO ₂ , -OCH ₃ , C1-4 acyl, n=1~6; MsO- is methylsulfonic acid group, TsO- is toluenesulfonic acid base; R ₁ is any one of the following groups: C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, hydrogen atom, wherein, X is nitrogen, oxygen or sulfur atom, R is halogen, H, -OH, -NO ₂ , -OCH ₃ , C1-4 acyl, n=1~6; R ₃ is any one of the following groups: Wherein, R is halogen, H, -OH, -NO ₂ , -OCH ₃ , CF ₃ , C1-4 alkyl or C1-4 acyl. 2. containing pyrimidotriazole class LSD1 inhibitor as claimed in claim 1, it is characterized in that, Preferred in general formula I: R ₂ is C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, Any one of them; n=1～3; R ₁ is any one of the following groups: C1-5 straight chain alkyl, C1-5 branched chain alkyl or C3-5 cycloalkyl, hydrogen atom, wherein, X is a sulfur atom, n=1~3; _R3 is any one of the following groups: Wherein, R is C1-4 alkyl, chlorine, bromine, H. 3. The pyrimidotriazole-containing LSD1 inhibitor as claimed in claim 1, characterized in that: preferably one of the following compounds: 8:R ¹ =propyl-S-, 9:R ¹ =propyl-S-, 10:R ¹ =propyl-S-, 11:R ¹ =propyl-S-, 12:R ¹ =propyl-S-, 13:R ¹ =propyl-S-, 14:R ¹ =propyl-S-, 15:R ¹ =propyl-S-, 16:R ¹ =propyl-S-, 17:R ¹ =propyl-S-, 18:R ¹ =propyl-S-, 19:R ¹ =propyl-S-, 20:R ¹ =propyl-S-, 21:R ¹ =propyl-S-, 4. The method for preparing the pyrimidotriazole-containing LSD1 inhibitor as claimed in claim 1 is characterized in that: it is synthesized by the following steps: (1) The preparation method of general formula 2: In a solvent, compound 1 and a brominated hydrocarbon are reacted under the action of an alkaline substance and stirred at room temperature. After the reaction is completed, filter, wash, and dry to obtain a series of compound 2; the solvent used is selected from acetone, methanol, ethanol, propanol, Isopropanol, tetrahydrofuran, acetonitrile, water, DMF, dichloromethane, chloroform, dioxane, or a mixture of two or more; the basic substances used are triethylamine, diisopropylethylamine, pyridine, One of sodium hydroxide and potassium hydroxide; the brominated hydrocarbon corresponds to the R1 substituent in the general formula; (2) The preparation method of general formula 3: In acetic acid, add nitrating reagent, then add serial compound 2 in batches, stir and react, after the reaction, pour into water, suction filter, wash, dry to obtain serial compound 3; said nitrating reagent is fuming nitric acid or concentrated nitric acid ; (3) The preparation method of general formula 4: In the solvent, add a chlorinating reagent, then add the series of compound 3 in batches, add an organic base dropwise, and reflux the reaction. After the reaction, cool to room temperature, hydrolyze, extract with an organic solvent, wash with water, neutralize, and dry the organic phase to obtain the series of compounds 4; the chlorination reagent is selected from phosphorus oxychloride and phosphorus pentachloride; the solvent is selected from toluene, dioxane, THF, ethyl acetate; the organic base is selected from triethylamine, N,N-dimethylaniline , N, N-diethylaniline, dimethylacetamide or pyridine; (4) The preparation method of general formula 5: Dissolve series compound 4 in a mixed solvent of ethanol and acetic acid, then add reduced iron powder in batches, raise the temperature and reflux for reaction, filter with suction, spin dry the solvent, extract with organic solvent, wash with water, and dry the organic phase to obtain series compound 5; (5) The preparation method of general formula 6: Dissolve the series compound 5 and the amine compound in the solvent, then add an organic base, reflux reaction, after the reaction, evaporate the solvent to dryness, add ethyl acetate, wash with water, and dry the organic phase to obtain the series compound 6; the solvent used is methanol, Ethanol, isopropanol, DMF, dioxane, THF, acetonitrile; the organic base is selected from triethylamine, pyridine or diisopropylethylamine; the amine compounds correspond to the R substituent in the general _formula ; (6) The preparation method of general formula 7: The series compound 6 was dissolved in the mixed solution of acetic acid and water, and the aqueous solution of sodium nitrite was added dropwise under ice bath for reaction. After the reaction was completed, ethyl acetate and water were added, the layers were separated, washed with water, neutralized, and the organic phase was dried to obtain Series compound 7; (7) The preparation method of compound 8～21 in general formula: Dissolve the series compound 7 and the mercapto aromatic ring compound in the solvent, add an acid-binding agent, react at room temperature or under reflux, after the reaction is completed, evaporate the solvent to dryness, dissolve in ethyl acetate, wash with water, dry, and pass through the column; the solvent For methanol, ethanol, isopropanol, THF, acetonitrile, DMF, dioxane; acid-binding agent for triethylamine, pyridine, isopropylethylamine. 5. The application of the pyrimidotriazole-containing LSD1 inhibitor according to any one of claims 1-3 in the preparation of medicines, characterized in that: it is used as an anti-tumor lead compound based on targeting LSD1.