CN117777047B - N-substituted azaspirodecanone compounds and preparation methods and applications thereof - Google Patents

Abstract

The present invention provides N-substituted azaspirodecanone compounds and preparation methods and applications thereof, and belongs to the technical field of spirodecanone compounds. N-substituted azaspirodecanone compounds have a structure as shown in Formula I or Formula II; the present invention realizes efficient construction of N-substituted azaspirodecanone compounds through nucleophilic addition reaction. The preparation method of the present invention is simple, the prepared compound has a stable structure, small toxic and side effects, good water solubility and anti-tumor activity, and can be used to prepare anti-tumor drugs.

Description

N-substituted azaspirodecanone compounds and preparation methods and applications thereof

Technical Field

The invention belongs to the technical field of spirodecanone compounds, and specifically relates to N-substituted azaspirodecanone compounds and preparation methods and applications thereof.

Background technique

Malignant tumors are major diseases that endanger human health and life. Therefore, the research and development of new anti-tumor drugs with low toxicity and high efficacy based on different structure-activity relationships and different targets is of great significance. Spirodecenone is a class of derivatives containing a fully substituted quaternary carbon center (spiroatom), which has multiple biological activities, such as anti-tumor, anti-inflammatory, antibacterial, antiviral and anti-degenerative diseases. In addition, spirodecanone also has a unique rigidity and conformational structure. It has attracted widespread attention because of the special structure and multi-faceted biological activities.

Previous studies have found that sulfonamide has good nitrene reactivity and stability. At the same time, sulfonamide compounds can also be selectively enriched in tumor tissues and have the potential of tumor-targeted drug delivery carriers. Using sulfonamide as a nitrogen source to design and synthesize a series of sulfonamide azaspirodecanedione H1 and H2 derivatives have certain anti-tumor activity. However, although this type of compound can show the characteristics of enrichment in tumor sites and good anti-tumor activity, it has strong toxicity, poor water solubility and low in vitro activity. These defects limit its further application research. The structural formula of sulfonamide azaspirodecanedione H1 and H2 derivatives is shown below:

.

Summary of the invention

The problem to be solved by the present invention is to provide N-substituted azaspirodecanone compounds and preparation methods and applications thereof, so as to solve the problems of strong toxicity, poor water solubility and low in vitro activity of sulfonamide azaspirodecanone derivatives.

The technical solution adopted to solve the technical problem is as follows: provide an N-substituted azaspirodecanone compound, which has a structure as shown in Formula I or Formula II:

;

Where:

R is H, C _1-20 straight or branched alkyl, halogen, aliphatic ring, aromatic ring, carboxyl, hydroxyl, amino, thiol, carbonyl derivative, phosphoryl derivative, phosphitylated derivative or sulfonylated derivative;

_R2 is an aromatic sulfonyl or substituted aromatic sulfonyl, an aromatic carbonyl or substituted aromatic carbonyl, an aromatic phosphoryl or substituted aromatic phosphoryl, an aromatic ring or substituted aromatic ring, an aliphatic ring or substituted aliphatic ring, a heterocyclic ring or substituted heterocyclic ring, a benzyl or substituted benzyl;

_R3 is a _C1-20 straight or branched alkyl group, halogen, aliphatic ring, aromatic ring, heterocyclic ring, carboxyl, hydroxyl, amino, thiol, carbonyl derivative, phosphoryl derivative, phosphitylated derivative or sulfonylated derivative;

X is oxygen, sulfur, nitrogen or carbon, and n is 0, 1, 2, 3, 4 or 5.

Preferably, R is H, halogen or aromatic ring; _R2 is aromatic sulfonyl or substituted aromatic sulfonyl, aromatic ring or substituted aromatic ring, heterocycle or substituted heterocycle, benzyl or substituted benzyl; _R3 is heterocycle, carboxyl, thiol or carbonyl derivative; X is oxygen; and n is 1.

Preferably, the specific structural formula of the N-substituted azaspirodecanone compound is as follows:

.

The preparation method of the above-mentioned N-substituted azaspirodecanone compound comprises the following synthetic route:

Synthesis route 1: Compound A and compound B are dissolved in an organic solvent, reacted under the action of a catalyst to obtain an intermediate compound 1, then an oxidant is added to react to obtain an intermediate compound 2, and then a nucleophilic reagent is added to react to obtain an N-substituted azaspirodecanone compound as shown in formula I; the synthesis route is as follows:

;

Synthesis route 2: Compound C and compound D are dissolved in an organic solvent, reacted under the action of a dehydrating agent to obtain an intermediate compound 3, then an oxidant is added to react to obtain an intermediate compound 4, then compound B is added to react under the action of a catalyst to obtain an intermediate compound 5, and then a nucleophilic reagent is added to react to obtain an N-substituted azaspirodecanone compound as shown in formula II; the synthesis route is as follows:

;

Synthesis route 3: Compound E and compound F are added with a dehydrating agent to react to obtain intermediate compound 6, and then an oxidant and a copper salt catalyst are added to react to obtain intermediate compound 5, and then a nucleophilic reagent is added to react to obtain an N-substituted azaspirodecanone compound as shown in formula II; the synthesis route is as follows:

.

More preferably, compound E is The preparation of N-substituted azaspirodecanone compounds comprises the following steps: dissolving 3-chlorobenzaldehyde, cyanamide and methanol in N-bromosuccinimide, reacting under the action of a catalyst to obtain an intermediate compound 7, then adding phenylhydrazine and a catalyst to react to obtain an intermediate compound 8, then adding 2-(4-methoxyphenoxy)acetic acid, carbodiimide and 1-hydroxybenzotriazole to react to obtain an intermediate compound 9, then adding an oxidant and a copper salt catalyst to react to obtain an intermediate compound 10, and then adding a nucleophilic reagent to react to obtain an N-substituted azaspirodecanone compound as shown in formula II; the synthesis route is as follows:

.

The preferred beneficial effect of the present invention is that the intermediate compound before the addition of the nucleophilic reagent is a Michael acceptor molecule, which can react with a variety of nucleophilic biologically active molecules in the body due to its electrophilicity, thereby exhibiting biological effects. This type of molecule can covalently bind to nucleophilic amino acid residues (such as cysteine, arginine, etc.), hydroxyl groups or sulfhydryl groups of a variety of proteins to regulate intracellular signaling pathways such as NF-κB, STAT3, etc. However, proteins containing nucleophilic amino acid residues and other groups may be the target of Michael acceptor molecules, which is easy to produce off-target effects. The toxic and side effects caused by the reaction have great potential risks in clinical applications. By subjecting the intermediate compound to a nucleophilic addition reaction with a nucleophilic reagent, the addition reaction sites of the Michael acceptor molecule in the body can be reduced, that is, the N-substituted azaspirodecanone compounds finally obtained can effectively reduce the toxic and side effects and have good safety. At the same time, the introduction of a three-membered ring, polar or hydrophilic group into the spirodecanone structure can further improve its water solubility. For example, polynitrogen heterocyclic structures such as imidazole, triazole and piperazine can also help to improve the binding force between the compound and the target, improve the lipid-water partition coefficient and thus improve its pharmacological activity.

More preferably, the catalyst is triethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene or potassium tert-butoxide.

More preferably, the oxidant is iodobenzene trifluoroacetate or diacetoxy iodobenzene; and the copper salt catalyst is tetraacetonitrile copper perchlorate.

More preferably, the dehydrating agent in the synthesis route 2 is 1,3-dicyclohexylcarbodiimide; and the dehydrating agent in the synthesis route 3 is a mixture of carbodiimide and 1-hydroxybenzotriazole.

More preferably, the nucleophiles are ethanethiol, dimethyl malonate, 2H -1,2,3-triazole, imidazole or 1-methylpiperazine.

More preferably, the organic solvent is dichloromethane, acetonitrile, methanol, ethanol, diethyl ether, 1,2-dichloroethane, chloroform, toluene or xylene.

The present invention also provides the use of the N-substituted azaspirodecanone compound in the preparation of anti-tumor drugs.

The present invention has the following beneficial effects:

(1) The method for preparing N-substituted azaspirodecanone compounds provided by the present invention has a moderate reaction, and the prepared N-substituted azaspirodecanone compounds are not easy to open the ring and have good biological activity.

(2) The N-substituted azaspirodecanone compound provided by the present invention has good anti-tumor activity, low toxicity, good water solubility and high purity, and the structure is a completely new compound.

(3) The N-substituted azaspirodecanone compounds provided by the present invention are simple to synthesize, the raw materials are readily available, and they can be produced on a large scale.

Detailed ways

The following examples are only used to explain the present invention and are not intended to limit the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

The millimole (mmol) ratios of the feed materials in Examples 1 to 12 are all consistent.

Example 1

10-(Ethylthio)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one (I-6) has the following structure:

.

The preparation process of 10-(ethylthio)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one in this embodiment is as follows:

S1. Dissolve 2-phenoxyethylamine (1.2 mmol) in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, triethylamine (1.5 mmol) and p-toluenesulfonyl chloride (1 mmol) were sequentially dissolved in 2-phenoxyethylamine solution in an ice bath, and stirred at room temperature for 8 h. After the reaction, the reaction solution was washed with saturated brine, and then separated and purified with polyamide to obtain intermediate compound 1;

S3, adding intermediate compound 1 (0.4 mmol), iodobenzene trifluoroacetate (1 mmol), dimerized rhodium acetate (0.04 mmol) and calcium oxide (4 mmol) to 25 mL of dichloromethane, stirring and reacting at 50°C for 6 h, separating and purifying the reaction solution after the reaction with polyamide to obtain intermediate compound 2;

S4. The intermediate compound 2 (0.1 mmol) was dissolved in a 50% volume fraction ethanol solution, ethyl mercaptan (0.2 mmol) was added, and the reaction was stirred at room temperature for 10 h. After the reaction, polyamide was used for separation and purification to obtain 10-(ethylthio)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one (yield 39%).

The NMR data of 10-(ethylthio)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one are as follows:

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 7.79 (d, J = 8.3 Hz, 2H), 7.36 (d, J = 8.3 Hz, 2H), 6.65 (d, J = 10.1 Hz, 1H), 6.05 (d, J = 10.1 Hz, 1H), 4.38 – 4.27 (m,1H), 4.13 – 4.00 (m, 2H), 3.81 – 3.61 (m, 2H), 2.92 – 2.85 (m, 2H), 2.78 –2.53 (m, 2H), 2.46 (s, 3H), 1.24 (t, J = 7.4 Hz, 3H);

¹³ C-NMR (400 MHz, CDCl ₃ ) δ (ppm): 196.9, 146.5, 144.3, 136.7, 130.3,129.9, 127.5, 94.2, 66.4, 49.5, 47.9, 43.9, 25.9, 21.6, 14.9;

HRMS ( _ESI ) m/z ( _% ) for _{C17H21NNaO4S2} (M ₊ Na): Calcd. 390.0810Found. 390.0811.

Example 2

Dimethyl 2-(8-oxo-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dec-9-en-6-yl)malonate (I-7) has the structure shown in the following formula:

.

The preparation process of dimethyl 2-(8-oxo-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dec-9-en-6-yl)malonate in this embodiment is as follows:

S1, dissolving 2-phenoxyethylamine in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, sequentially dissolving triethylamine and p-toluenesulfonyl chloride in a 2-phenoxyethylamine solution in an ice bath, stirring and reacting at room temperature for 8 h, washing the reaction solution with saturated brine, and then separating and purifying with silica gel to obtain intermediate compound 1;

S3, adding the intermediate compound 1, iodobenzene trifluoroacetate, dimerized rhodium acetate and calcium oxide into dichloromethane, stirring and reacting at 50° C. for 8 h, and separating and purifying the reaction solution after the reaction with silica gel to obtain the intermediate compound 2;

S4. The intermediate compound 2 was dissolved in acetonitrile, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) was added, and then dimethyl malonate was added. The mixture was stirred at room temperature for 7 h. After the reaction, it was separated and purified by silica gel to obtain dimethyl 2-(8-oxo-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dec-9-en-6-yl)malonate (yield 67%).

The NMR data of dimethyl 2-(8-oxo-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dec-9-en-6-yl)malonate are as follows:

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 7.74 (d, J = 8.3 Hz, 2H), 7.33 (d, J = 8.3 Hz, 2H), 6.62 (d, J = 10.2 Hz, 1H), 6.03 (d, J = 10.2 Hz, 1H), 4.06 – 3.91 (m,2H), 3.80 (m, 2H), 3.72 (s, 3H), 3.69 (s, 3H), 3.63 (m, 1H), 3.51 (d, J = 3.6 Hz, 1H), 3.02 (m, 1H), 2.82 (m, 1H), 2.45 (s, 3H);

¹³ C NMR (400 MHz, CDCl ₃ ) δ (ppm): 197.2, 168.4, 167.9, 145.3, 144.5,136.6, 130.5, 130.0, 127.4, 94.1, 64.9, 52.8, 52.3, 49.9, 47.3, 42.3, 37.9,21.6;

HRMS ( _ESI ) m/z ₍ %) for _C20H24NO8S (M+H): Calcd. 438.1223Found. 438.1214.

Example 3

4-Toluenesulfonyl-10-( 2H -1,2,3-triazol-2-yl)-1-oxo-4-azaspiro[4.5]dec-6-en-8-one (I-5) has the following structure:

.

The preparation process of 4-toluenesulfonyl-10-( 2H -1,2,3-triazol-2-yl)-1-oxo-4-azaspiro[4.5]dec-6-en-8-one in this embodiment is as follows:

S1, dissolving 2-phenoxyethylamine in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, sequentially dissolving triethylamine and p-toluenesulfonyl chloride in a 2-phenoxyethylamine solution in an ice bath, stirring and reacting at room temperature for 8 h, washing the reaction solution with saturated brine, and then separating and purifying with polyamide to obtain an intermediate compound 1;

S3, adding the intermediate compound 1, iodobenzene trifluoroacetate, dimerized rhodium acetate and calcium oxide into dichloromethane, stirring and reacting at 50° C. for 8 h, and separating and purifying the reaction solution after the reaction with polyamide to obtain the intermediate compound 2;

S4. The intermediate compound 2 was dissolved in acetonitrile, DBU was added, and then 2H -1,2,3-triazole was added. The reaction was stirred at room temperature for 8 h. After the reaction, polyamide was used for separation and purification to obtain 4-toluenesulfonyl-10-( 2H -1,2,3-triazole-2-yl)-1-oxo-4-azaspiro[4.5]dec-6-en-8-one (yield 47%).

The NMR data of 4-toluenesulfonyl-10-( 2H -1,2,3-triazol-2-yl)-1-oxo-4-azaspiro[4.5]dec-6-en-8-one are as follows:

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 7.98 (d, J = 8.4 Hz, 2H), 7.67 (s, 2H),7.36 (d, J = 8.4 Hz, 2H), 6.82 (d, J = 10.2 Hz, 1H), 6.27 (dd, J = 10.2, 1.1 Hz,1H), 6.03 (m, 1H), 3.93 – 3.86 (m, 1H), 3.71 (dd, J = 16.7, 12.6 Hz, 1H), 3.47– 3.39 (m, 1H), 3.06 (m, 1H), 2.96 – 2.87 (m, 2H), 2.46 (s, 3H);

¹³ C NMR (400 MHz, CDCl ₃ ) δ (ppm): 195.2, 144.7, 144.1, 135.9, 134.6,131.3, 129.7, 128.4, 92.7, 65.7, 63.7, 46.6, 40.5, 21.7;

HRMS (ESI) m/z (%) for C ₁₇ H ₁₉ N ₄ O ₄ S (M+H): Calcd. 375.1127 Found. 375.1119.

Example 4

10-( 1H -imidazol-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one (I-3) has the following structure:

.

The preparation process of 10-( 1H -imidazol-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one in this embodiment is as follows:

S1, dissolving 2-phenoxyethylamine in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, sequentially dissolving triethylamine and p-toluenesulfonyl chloride in a 2-phenoxyethylamine solution in an ice bath, stirring and reacting at room temperature for 8 h, washing the reaction solution with saturated brine, and then separating and purifying with silica gel to obtain intermediate compound 1;

S3, adding the intermediate compound 1, iodobenzene trifluoroacetate, dimerized rhodium acetate and calcium oxide into dichloromethane, stirring and reacting at 50° C. for 8 h, and separating and purifying the reaction solution after the reaction with silica gel to obtain the intermediate compound 2;

S4. The intermediate compound 2 was dissolved in acetonitrile, DBU was added, and then imidazole was added. The mixture was stirred at room temperature for 8 h. After the reaction, silica gel was used for separation and purification to obtain 10-( 1H -imidazole-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one (yield 61%).

The NMR data of 10-( 1H -imidazol-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one are as follows:

¹ H NMR (400 MHz, Chloroform-d) δ 7.73 (d, J = 8.1 Hz, 2H), 7.69 (s, 1H),7.38 (d, J = 8.0 Hz, 2H), 7.13 (d, J = 25.3 Hz, 2H), 6.54 (d, J = 10.2 Hz, 1H),6.18 (d, J = 10.2 Hz, 1H), 5.50 (dd, J = 13.6, 4.4 Hz, 1H), 3.88 (td, J = 7.2, 4.2Hz, 1H), 3.42 – 3.32 (m, 1H), 3.32 – 3.19 (m, 2H), 3.13 (ddd, J = 8.5, 5.9, 4.2 Hz, 1H), 2.98 – 2.89 (m, 1H), 2.47 (s, 3H);

¹³ C NMR (101 MHz, Chloroform-d) δ 194.69, 145.07, 144.07, 137.40,135.91, 131.21, 130.17, 129.10, 127.58, 119.27, 92.51, 65.82, 57.47, 47.00,41.46, 21.65

HRMS(ESI) m/z : calculated for C ₁₈ H ₁₉ N ₃ O ₄ S [M+H] ⁺ 374.1173, found374.1173.

Example 5

10-(4-methylpiperazine-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one (I-4) has the following structure:

.

The preparation process of 10-(4-methylpiperazine-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one in this embodiment is as follows:

S1, dissolving 2-phenoxyethylamine in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, sequentially dissolving triethylamine and p-toluenesulfonyl chloride in a 2-phenoxyethylamine solution in an ice bath, stirring and reacting at room temperature for 8 h, washing the reaction solution with saturated brine, and then separating and purifying with polyamide to obtain intermediate compound 1;

S3, adding the intermediate compound 1, iodobenzene trifluoroacetate, dimerized rhodium acetate and calcium oxide into dichloromethane, stirring and reacting at 50° C. for 8 h, and separating and purifying the reaction solution after the reaction with polyamide to obtain the intermediate compound 2;

S4. The intermediate compound 2 was dissolved in acetonitrile, DBU was added, and then 1-methylpiperazine was added, and the reaction was carried out at room temperature for 8 h. After the reaction, polyamide was used for separation and purification to obtain 10-(4-methylpiperazine-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one (yield 36%).

The NMR data of 10-(4-methylpiperazine-1-yl)-4-toluenesulfonyl-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 7.90 – 7.70 (m, 2H), 7.34 (d, J = 8.1 Hz, 2H), 6.64 (d, J = 10.2 Hz, 1H), 6.04 (dd, J = 10.2, 1.3 Hz, 1H), 4.20 – 4.11 (m, 1H), 4.02 (q, J = 7.1 Hz, 1H), 3.90 – 3.81 (m, 1H), 3.75 (dd, J = 13.0, 4.2 Hz, 1H), 3.66 (ddd, J = 8.1, 6.6, 4.6 Hz, 1H), 3.00 – 2.63 (m, 4H), 2.59 – 1.76(m, 12H);

¹³ C NMR (101 MHz, Chloroform- d ) δ 198.67, 146.88, 144.03, 137.43,130.35, 129.83, 127.32, 95.68, 65.80, 63.58, 55.87, 48.06, 45.83, 36.89,21.56;

HRMS(ESI) m/z : calculated for C ₂₀ H ₂₇ N ₃ O ₄ S [M+H] ⁺ 406.1788, found 406.1788.

Example 6

4-((4-fluorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one (I-2) has the following structure:

.

The preparation process of 4-((4-fluorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one in this embodiment is as follows:

S1, dissolving 2-phenoxyethylamine in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, sequentially dissolving triethylamine and p-fluorobenzenesulfonyl chloride in a 2-phenoxyethylamine solution in an ice bath, stirring and reacting at room temperature for 8 h, washing the reaction solution with saturated brine, and then separating and purifying with silica gel to obtain intermediate compound 1;

S3. The intermediate compound 1, iodobenzene trifluoroacetate, dimerized rhodium acetate and calcium oxide were mixed and added into dichloromethane, and the mixture was stirred at 50°C for 8 h. The reaction solution after the reaction was separated and purified by silica gel to obtain the intermediate compound 2. The specific NMR data of the intermediate compound 2 are as follows:

¹ H NMR (400 MHz, Chloroform-d) δ 7.83 (dd, J = 8.6, 5.0 Hz, 2H), 7.22 (t, J = 8.4 Hz, 2H), 6.50 (d, J = 9.8 Hz, 2H), 6.22 (d, J = 9.8 Hz, 2H), 4.19 (t, J =6.1 Hz, 2H), 3.73 (t, J = 6.1 Hz, 2H);

¹³ C NMR (101 MHz, Chloroform-d) δ 184.63, 165.46, 143.03, 135.04,130.50, 129.68, 116.48, 86.29, 65.56, 47.05;

HRMS(ESI) m/z : calculated for C ₁₄ H ₁₂ FNO ₄ S [M+H] ⁺ 310.0545, found310.0545;

S4. Dissolve the intermediate compound 2 in acetonitrile, add DBU, then add imidazole, and stir the mixture at room temperature for 8 h. After the reaction, separate and purify the mixture using silica gel to obtain 4-((4-fluorophenyl)sulfonyl)-10-(1H-imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one (yield 55%).

The NMR data of 4-((4-fluorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 7.95 – 7.80 (m, 2H), 7.73 (s, 1H),7.34 – 7.22 (m, 2H), 7.15 (d, J = 19.7 Hz, 2H), 6.52 (d, J = 10.2 Hz, 1H), 6.20(dd, J = 10.2, 1.1 Hz, 1H), 5.51 (dd, J = 13.6, 4.4 Hz, 1H), 3.91 (ddd, J = 8.0,6.5, 4.4 Hz, 1H), 3.43 – 3.21 (m, 3H), 3.19 – 3.08 (m, 1H), 2.95 (ddd, J = 15.8, 4.4, 1.2 Hz, 1H);

¹³ C NMR (101 MHz, Chloroform- d ) δ 194.43, 165.73, 143.63, 137.31,134.92, 131.52, 130.39, 129.07, 119.26, 116.96, 92.75, 65.81, 57.50, 47.03,41.50;

HRMS(ESI) m/z : calculated for C ₁₇ H ₁₆ FN ₃ O ₄ S [M+H] ⁺ 378.0916, found378.0916.

Example 7

4-((4-chlorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one (I-1) has the following structure:

.

The preparation process of 4-((4-chlorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one in this embodiment is as follows:

S1, dissolving 2-phenoxyethylamine in dichloromethane to obtain a 2-phenoxyethylamine solution;

S2, sequentially dissolving triethylamine and p-chlorobenzenesulfonyl chloride in a 2-phenoxyethylamine solution in an ice bath, stirring and reacting at room temperature for 8 h, washing the reaction solution with saturated brine, and then separating and purifying with polyamide to obtain an intermediate compound 1;

S3. The intermediate compound 1, iodobenzene trifluoroacetate, dimerized rhodium acetate and calcium oxide were mixed and added into dichloromethane, and the mixture was stirred at 50°C for 8 h. The reaction solution after the reaction was separated and purified by polyamide to obtain the intermediate compound 2. The specific NMR data of the intermediate compound 2 are as follows:

¹ H NMR (400 MHz, Chloroform-d) δ 7.80 – 7.66 (m, 2H), 7.57 – 7.43 (m,2H), 6.57 – 6.40 (m, 2H), 6.29 – 6.14 (m, 2H), 4.19 (t, J = 6.1 Hz, 2H), 3.73(t, J = 6.1 Hz, 2H);

¹³ C NMR (101 MHz, Chloroform-d) δ 184.60, 142.91, 140.06, 137.44,129.75, 129.51, 129.11, 86.36, 65.56, 47.08;

HRMS(ESI) m/z : calculated for C ₁₄ H ₁₂ ClNO ₄ S [M+H] ⁺ 326.0246, found326.0246;

S4. The intermediate compound 2 was dissolved in acetonitrile, DBU was added, and then imidazole was added. The mixture was stirred at room temperature for 8 h. After the reaction, polyamide was used for separation and purification to obtain 4-((4-chlorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-8-one (yield 47%).

The NMR data of 4-((4-chlorophenyl)sulfonyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-en-8-one are as follows:

¹ H NMR (400 MHz, Chloroform-d) δ 7.79 (d, J = 8.2 Hz, 2H), 7.73 (s, 1H),7.57 (d, J = 8.2 Hz, 2H), 7.15 (d, J = 19.7 Hz, 2H), 6.50 (d, J = 10.2 Hz, 1H),6.20 (d, J = 10.2 Hz, 1H), 5.50 (dd, J = 13.6, 4.4 Hz, 1H), 3.91 (dt, J = 11.9, 5.4Hz, 1H), 3.45 – 3.04 (m, 4H), 2.95 (dd, J = 15.8, 4.4 Hz, 1H);

¹³ C NMR (151 MHz, Chloroform-d) δ 194.38, 143.50, 140.65, 137.28,137.24, 131.54, 129.90, 129.07, 128.91, 119.23, 92.73, 65.78, 57.50, 47.01,41.44;

HRMS(ESI) m/z : calculated for C ₁₇ H ₁₆ ClN ₃ O ₄ S [M+H] ⁺ 394.0626, found394.0626.

Example 8

Dimethyl-2-(4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-3,8-dioxo-1-oxo-4-azaspiro[4.5]dodec-9-en-6-yl)malonate (II-3) has the structure shown in the following formula:

.

The preparation process of dimethyl-2-(4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-3,8-dioxo-1-oxo-4-azaspiro[4.5]dodec-9-ene-6-yl)malonate in this embodiment is as follows:

S1. Add 3-chlorobenzaldehyde, cyanamide, methanol, potassium tert-butoxide and N-bromosuccinimide into a reaction bottle, stir evenly and react at room temperature for 8 hours. After the reaction, the reaction solution is separated and purified by polyamide to obtain intermediate compound 7.

S2, adding intermediate compound 7, phenylhydrazine and triethylamine into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with polyamide to obtain intermediate compound 8;

S3, adding intermediate compound 8, 2-(4-methoxyphenoxy)acetic acid, carbodiimide and 1-hydroxybenzotriazole into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with polyamide to obtain intermediate compound 9;

S4, adding intermediate compound 9, iodobenzene trifluoroacetate and copper perchlorate tetraacetonitrile into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with polyamide to obtain intermediate compound 10;

S5. The intermediate compound 10 was dissolved in acetonitrile, DBU was added, and then dimethyl malonate was added. The reaction was stirred at 30°C for 8 h. After the reaction, polyamide was used for separation and purification to obtain dimethyl 2-(4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-3,8-dioxo-1-oxo-4-azaspiro[4.5]dodec-9-ene-6-yl)malonate (yield: 59.6%).

The NMR data of dimethyl-2-(4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-3,8-dioxo-1-oxo-4-azaspiro[4.5]dodec-9-en-6-yl)malonate are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 8.05 (d, J = 1.7 Hz, 1H), 7.95 (dt, J =7.0, 1.6 Hz, 1H), 7.63–7.51 (m, 5H), 7.44–7.33 (m, 2H), 6.52 (d, J = 10.2 Hz,1H), 6.00 (d, J = 9.5 Hz, 1H), 4.49–4.36 (m, 2H), 3.78 (d, J = 2.7 Hz, 1H), 3.74(s, 3H), 3.74 (s, 3H), 3.24–3.13 (m, 1H), 3.03 (dd, J = 16.7, 12.8 Hz, 1H),2.77 (dd, J = 16.4, 4.4 Hz, 1H);

¹³ C NMR (100 MHz, Chloroform- d ) δ 196.1, 171.1, 168.1, 167.5, 160.8,143.7, 142.8, 135.9, 134.7, 132.1, 131.6, 130.4, 123.0, 129.7, 126.6, 125.0,124.4, 95.0, 66.3, 53.1, 52.5, 49.1, 41.3,36.3;

HRMS (ESI-TOF) m/z: [M+H] ⁺ calcd for C ₂₇ H ₂₄ ClN ₄ O ₇ , 551.1328; found,551.1324.

Example 9

4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodecan-6-ene-3,8-dione (II-2) has the following structure:

.

The preparation process of 4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione in this embodiment is as follows:

S1. Add 3-chlorobenzaldehyde, cyanamide, methanol, potassium tert-butoxide and N-bromosuccinimide into a reaction bottle, stir evenly and react at room temperature for 8 hours. After the reaction, the reaction solution is separated and purified by silica gel to obtain intermediate compound 7.

S2, adding intermediate compound 7, phenylhydrazine and triethylamine into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 8;

S3, adding intermediate compound 8, 2-(4-methoxyphenoxy)acetic acid, carbodiimide and 1-hydroxybenzotriazole into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 9;

S4, adding intermediate compound 9, iodobenzene trifluoroacetate and copper perchlorate tetraacetonitrile into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 10;

S5. The intermediate compound 10 was dissolved in acetonitrile, DBU was added, and then imidazole was added. The mixture was stirred at 30°C for 8 h. After the reaction, the mixture was separated and purified using silica gel to obtain 4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-( 1H -imidazole-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione (yield: 46.6%).

The NMR data of 4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 8.13 (s, 1H), 8.03 (d, J = 8.9 Hz, 2H),7.58–7.51 (m, 3H), 7.50–7.42 (m, 4H), 7.20 (s, 1H), 6.38 (d, J = 10.2 Hz, 1H),6.25 (d, J = 10.2 Hz, 1H), 5.72 (dd, J = 13.8, 4.3 Hz, 1H), 4.25–4.06 (m, 2H),3.50–3.38 (m, 2H), 3.06 (dd, J = 15.8, 3.8 Hz, 1H);

HRMS (ESI-TOF) m/z: [M+H] ⁺ calcd for C _{2 5} H _{2 0} ClN ₆ O ₃ , 487.1280; found,487.1281.

Example 10

4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodecan-6-ene-3,8-dione (II-5) has the following structure:

.

The preparation process of 4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione in this embodiment is as follows:

S1. Add 3-chlorobenzaldehyde, cyanamide, methanol, potassium tert-butoxide and N-bromosuccinimide into a reaction bottle, stir evenly and react at room temperature for 8 hours. After the reaction, the reaction solution is separated and purified by silica gel to obtain intermediate compound 7.

S2, adding intermediate compound 7, phenylhydrazine and triethylamine into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 8;

S3, adding intermediate compound 8, 2-(4-methoxyphenoxy)acetic acid, carbodiimide and 1-hydroxybenzotriazole into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 9;

S4, adding intermediate compound 9, iodobenzene trifluoroacetate and copper perchlorate tetraacetonitrile into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 10;

S5. The intermediate compound 10 was dissolved in acetonitrile, DBU was added, and then ethyl mercaptan was added. The mixture was stirred at 30°C for 8 h. After the reaction, the mixture was separated and purified using silica gel to obtain 4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione (yield: 42.3%).

The NMR data of 4-(3-(3-chlorophenyl)-1-phenyl- 1H -1,2,4-triazol-5-yl)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodecan-6-ene-3,8-dione are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 8.06 (d, J = 1.8 Hz, 1H), 7.95 (td, J =6.8, 1.7 Hz, 1H), 7.65 (dd, J = 8.2, 1.5 Hz, 2H), 7.59–7.55 (m, 1H), 7.55–7.49(m, 2H), 7.42–7.35 (m, 2H), 6.56 (d, J = 10.1 Hz, 1H), 6.12 (dd, J = 10.1, 1.0Hz, 1H), 4.75 (d, J = 14.4 Hz, 1H), 4.37 (d, J = 14.4 Hz, 1H), 4.23 (dd, J = 13.0,5.0 Hz, 1H), 3.12–2.87 (m, 3H), 2.69 (dd, J = 12.5, 7.5 Hz, 1H), 1.33–1.28 (m,3H);

¹³ C NMR (100 MHz, Chloroform- d ) δ 196.1, 169.7, 160.1, 144.4, 143.3,137.0, 134.8, 133.4, 131.7, 130.0, 130.0, 129.7, 129.7, 126.4, 124.3, 124.3,94.9, 68.0, 49.1, 43.1, 26.2, 15.1.

Embodiment 11

4-(2,6-Dichlorophenyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione

(II-7) has the following structure:

.

The preparation process of 4-(2,6-dichlorobenzene)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione in this embodiment is as follows:

S1, adding p-aminophenol, glycolic acid and dimethyl dicyanamide into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 3;

S2, adding intermediate compound 3, diacetoxyiodobenzene, copper perchlorate tetraacetonitrile and dichloromethane into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 4;

S3, adding intermediate compound 4, DBU, tetrahydrofuran and 2,6-dichlorobenzyl chloride into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 5;

S4. The intermediate compound 5 was dissolved in acetonitrile, DBU was added, and then imidazole was added. The mixture was stirred at 30°C for 8 h. After the reaction, silica gel was used for separation and purification to obtain 4-(2,6-dichlorobenzene)-10-( 1H -imidazole-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione (yield: 53.3%).

The NMR data of 4-(2,6-dichlorophenyl)-10-( 1H -imidazol-1-yl)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 7.53 (s, 1H), 7.41–7.36 (m, 2H), 7.31 (dd, J = 9.1, 6.8 Hz, 1H), 7.15 (s, 1H), 7.00 (s, 1H), 6.06 (d, J = 10.2 Hz, 1H),5.79 (d, J = 10.2 Hz, 1H), 5.36 (d, J = 15.2 Hz, 1H), 5.22 (dd, J = 13.4, 4.3 Hz,1H), 4.59 (d, J = 15.2 Hz, 1H), 4.26 (d, J = 13.8 Hz, 1H), 3.71 (d, J = 1 13.8 Hz,1H), 3.34 (dd, J = 15.6, 13.5 Hz, 1H), 2.91 (dd, J = 15.6, 4.3 Hz, 1H);

¹³ C NMR (100 MHz, Chloroform- d ) δ 193.9, 168.3, 144.4, 136.8, 136.5,131.2, 131.1, 130.3, 130.1, 129.1, 118.3, 92.2, 66.4, 54.8, 40.5, 39.2;

HRMS (ESI-TOF) m/z: [M+H] ⁺ calcd for C ₁₈ H ₁₆ N ₃ O ₃ Cl ₂ , 392.0563, found,392.0577.

Example 12

4-(4-chlorophenyl)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodecan-6-ene-3,8-dione (II-8) has the following structure:

.

The preparation process of 4-(4-chlorobenzene)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione in this embodiment is as follows:

S1, adding p-aminophenol, glycolic acid and dimethyl dicyanamide into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 3;

S2, adding intermediate compound 3, diacetoxyiodobenzene, copper perchlorate tetraacetonitrile and dichloromethane into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 4;

S3, adding intermediate compound 4, DBU, tetrahydrofuran and 2,6-dichlorobenzyl chloride into a reaction bottle, stirring evenly and reacting at room temperature for 8 hours, and separating and purifying the reaction solution after the reaction with silica gel to obtain intermediate compound 5;

S4. The intermediate compound 5 was dissolved in acetonitrile, DBU was added, and then ethyl mercaptan was added. The mixture was stirred at 30°C for 8 h. After the reaction, silica gel was used for separation and purification to obtain 4-(4-chlorobenzene)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione (yield: 41.1%).

The NMR data of 4-(4-chlorophenyl)-10-(ethylthio)-1-oxo-4-azaspiro[4.5]dodec-6-ene-3,8-dione are as follows:

¹ H NMR (400 MHz, Chloroform- d ) δ 7.33–7.27 (m, 4H), 6.38 (d, J = 10.1 Hz,1H), 6.06 (dd, J = 10.1, 1.1 Hz, 1H), 4.57 (d, J = 13.5 Hz, 1H), 4.48–4.37 (m,3H), 3.19 (dd, J = 13.0, 4.4 Hz, 1H), 3.01 (dd, J = 16.2, 13.0 Hz, 1H), 2.81 (td, J = 16.2, 4.4, 1.1 Hz, 1H), 2.34 (td, J = 11.8, 7.4 Hz, 1H), 2.13 (dd, J = 13.5, 7.4 Hz, 1H). = 11.8,7.4 Hz, 1H), 1.13 (t, J = 7.4 Hz, 3H);

¹³ C NMR (100 MHz, Chloroform- d ) δ 196.3, 170.7, 146.1, 134.4, 134.3,132.5, 130.2, 129.0, 93.8, 68.0, 48.8, 43.3, 42.4, 25.2, 14.6;

HRMS (ESI-TOF) m/z: [M+H] ⁺ calcd for C ₁₇ H ₁₉ NO ₃ SCl, 352.0769, found,352.0785.

Example 13 Antitumor Activity Assay

The proliferation of adherent cells was detected by sulforhodamine B (SRB) method. Bendamustine and vorinostat were used as positive control drugs. Human breast cancer MDA-MB-231 cells in logarithmic growth phase were seeded in 96-well plates (200 µL/well) and cultured at 37°C and 5% carbon dioxide for 24 h to adhere to the plate. Then, the cells were treated with different concentrations of the test compound, with 3 replicates for each concentration, and corresponding blank controls and zero wells were set. After incubation for 72 h, the cells were fixed with 10% trichloroacetic acid (TCA) at 4°C for 1 h, the fixative was poured out, the cells were rinsed with distilled water 5 times, and the cells were naturally air-dried. After fixation, the cells were stained with 0.4% (w/v) SRB at room temperature for 20 min, the liquid was poured out, and the cells were washed with 1% glacial acetic acid 5 times to remove the excess unbound SRB dye, and the cells were naturally air-dried. The protein bound to SRB was dissolved with 10 mM tris (hydroxymethylamino) methane (Tris) and shaken on a horizontal shaker for 20 min. An equal amount of DMSO was used as a vehicle control, and bendamustine (Bendamustiune, a bifunctional alkylating agent) and vorinostat (SAHA, an HDAC inhibitor) were used as positive controls. The number of cells was counted using a cell viability analyzer (Beckman-Coulter). The viability was the ratio of the number of cells treated with the compound to the number of cells treated with the vehicle, and the IC ₅₀ value was obtained from the dose-response curve of each compound. The experiment was performed three times, and the data are expressed as the average value.

The compounds of Examples 1 to 12 were subjected to in vitro antitumor activity experiments, and bendamustine (Bendamustiune, a bifunctional alkylating agent) and vorinostat (SAHA, an HDAC inhibitor) were used as positive drug controls to test the in vitro cell growth inhibition ability of the target compounds on human breast cancer cells MDA-MB-231, and the results are shown in Table 1. As can be seen from Table 1, after the nucleophilic site in the intermediate compound 2 is added, the activities of the prepared N-substituted azaspirodecanone compounds are significantly improved compared with those before the addition of the nucleophilic site, except for Examples 4 and 5; at the same time, the N-substituted azaspirodecanone compounds prepared in Examples 1 to 12 also exhibited certain proliferation inhibition activity (IC ₅₀ 0.13~4.66μM) on lung cancer cell line A549 and cervical cancer cell line Hela cells.

Table 1

Example 14 Acute toxicity assay and compound dissolution behavior analysis

The intermediate compound 2 and N-substituted azaspirodecanone compounds prepared in Examples 1 to 7 were subjected to acute toxicity tests, and SPF-grade Km mice were divided into two groups. The two groups of mice were administered with different doses of intermediate compound 2 and N-substituted azaspirodecanone compounds, and the doses were 120 to 1000 mg/kg, and the mice after administration were observed for 14 days. The results showed that there were mice deaths in the intermediate compound 2 group, while there were no deaths in the N-substituted azaspirodecanone compound group, and there was no obvious change in the behavior of the mice, the diet was normal, there was no abnormal secretion, and the weight continued to grow; indicating that the toxicity of intermediate compound 2 is higher than that of N-substituted azaspirodecanone compounds, that is, the toxicity can be reduced after the addition of the nucleophilic site in intermediate compound 2. At the same time, the results of in vivo and in vitro activity studies showed that the activity of some compounds (I-2) in the N-substituted azaspirodecanone compounds synthesized after the unilateral α, β-unsaturated ketone structure of intermediate compound 2 was eliminated or other types of modifications were performed, and the toxicity was greatly reduced; animal toxicity experiments showed that the _LD50 of intermediate compound 2 was 66.1~80 mg/kg, which was in the moderate toxicity range, while the median lethal dose of N-substituted azaspirodecanone compounds for Km mice was greater than 120~1000 mg/kg.

Animal level anti-tumor activity experiment was conducted, 24 BALB/c tumor-bearing mice were randomly divided into 4 groups, 6 mice in each group, namely blank control group, positive drug control-bendamustine control group 10 mg/kg, low-dose N-substituted azaspirodecanone compound group 10 mg/kg, high-dose N-substituted azaspirodecanone compound group 20 mg/kg; four groups of mice were given medication once every two days, for 7 consecutive times, and the physical signs of the mice were observed within 14 days. The results showed that the tumor inhibition rate of N-substituted azaspirodecanone compounds on BALB/c breast cancer model mice reached 61%; the results of tissue pathological sections and staining showed that these compounds did not cause obvious lesions in normal tissues. That is, the N-substituted azaspirodecanone compounds of the present invention reduce the protein toxicity sites on the mother nucleus while retaining the basic structure of spirodecanone, thereby reducing the toxicity of the compounds in vivo.

At the same time, since the nucleophilic site addition reaction is carried out under the conditions of ethanol/water or acetonitrile/organic base, the residual transition metal salt is dissolved in the system, and the final product is further purified after the process, so that the purity of the prepared N-substituted azaspirodecanone compound is improved.

In addition, the lipid-water partition coefficient is a very important indicator that determines the distribution of the compound in the body. Appropriate water solubility helps the drug dissolve in body fluids and reach an effective concentration at the site of action, thereby producing a therapeutic effect. For anti-tumor drugs, good water solubility not only facilitates absorption and distribution, but also allows the drug to be quickly excreted, reducing accumulation and toxic side effects on the kidneys. The introduction of one or more heteroatoms into N-substituted azaspirodecanone compounds increases the possibility of forming hydrogen bonds with water, and its dissolution behavior is improved compared to intermediate compound 2. For example, in the animal level anti-tumor activity exploration experiment, it was clearly observed that 2 mg of N-substituted azaspirodecanone compounds were completely dissolved in 200 μL of DMSO and 1.8 mL of olive oil solution, while 1 mg of intermediate compound 2 was still a suspension in 200 μL of DMSO and 1.8 mL of olive oil solution. This is more helpful to improve the binding force between the compound and the target, improve the lipid-water partition coefficient, and thus improve its pharmacological activity.

The present invention has been described according to the above embodiments. It should be understood that the above embodiments do not limit the present invention in any form. Any technical solutions obtained by equivalent replacement or equivalent transformation shall fall within the scope of the present invention.

Claims (8)

1. An n-substituted azaspiro-decarenone compound characterized by the specific structural formula:

   。
2. 2. The process for the preparation of an N-substituted azaspiro-decarenone compound of claim 1, selected from the following synthetic routes:

   Synthesis path 1: dissolving a compound A and a compound B in an organic solvent, reacting under the action of a catalyst to obtain an intermediate compound 1, adding an oxidant to react to obtain an intermediate compound 2, and adding a nucleophile to react to obtain an N-substituted azaspiro-decamonoalkone compound shown in a formula I; the synthetic route is as follows:

   ；

   synthesis path 2: dissolving a compound C and a compound D in an organic solvent, reacting under the action of a dehydrating agent to obtain an intermediate compound 3, adding an oxidant to react to obtain an intermediate compound 4, adding a compound B, reacting under the action of a catalyst to obtain an intermediate compound 5, and adding a nucleophile to react to obtain the N-substituted azaspiro-decone compound shown in the formula II; the synthetic route is as follows:

   ；

   Synthesis path 3: adding a dehydrating agent into the compound E and the compound F to react to obtain an intermediate compound 6, adding an oxidant and a copper salt catalyst to react to obtain an intermediate compound 5, and adding a nucleophile to react to obtain the N-substituted azaspiro-decamonoalkenone compound shown in the formula II; the synthetic route is as follows:

   ；

   R, R ₂、R₃, X and N in the synthetic routes 1-3 are corresponding and consistent with the structures of the N-substituted azaspiro-decarenone compounds I-1-II-12 in claim 1.
3. 3. The process for the preparation of N-substituted azaspiro-decone compounds as claimed in claim 2, wherein the catalysts are selected from triethylamine, 1, 8-diazabicyclo [5.4.0] undec-7-ene or potassium tert-butoxide.
4. 4. The method of preparing an N-substituted azaspiro-decarenone compound of claim 2, wherein the oxidizing agents are each selected from iodobenzene trifluoroacetate or diacetoxyiodobenzene; the copper salt catalyst is tetraacetonitrile copper perchlorate.
5. 5. The method for preparing an N-substituted azaspiro-decarenone compound according to claim 2, wherein the dehydrating agent in the synthetic pathway 2 is 1, 3-dicyclohexylcarbodiimide; the dehydrating agent in the synthetic route 3 is a mixture of carbodiimide and 1-hydroxybenzotriazole.
6. 6. The method for preparing an N-substituted azaspiro-decarenone compound according to claim 2, wherein the nucleophiles are each selected from the group consisting of ethanethiol, dimethyl malonate, 2H-1,2, 3-triazole, imidazole, and 1-methylpiperazine.
7. 7. The method for preparing an N-substituted azaspiro-decarenone compound according to claim 2, wherein the organic solvents are selected from dichloromethane, acetonitrile, methanol, ethanol, diethyl ether, 1, 2-dichloroethane, chloroform, toluene, and xylene.
8. 8. The use of an N-substituted azaspiro-decarenone compound of claim 1 for the preparation of an antitumor drug.