CN104003939B - Diaryl substitution thiohydantoin class compound and preparation method and application - Google Patents

Abstract

The invention discloses a diaryl-substituted thiohydantoin compound, the structure of which is shown in the general formula (I): Wherein, R ₁ is hydrogen, halogen or trifluoromethyl; R ₂ is hydrogen, halogen, trifluoromethyl, nitrile or alkoxy; R ₃ is hydrogen, halogen or nitrile; R ₄ is hydrogen or halogen; R is hydrogen, halogen, methyl, N _- methylcarbamoyl, N-ethylcarbamoyl, N-isopropylcarbamoyl, N-cyclopropylcarbamoyl. The diaryl-substituted thiohydantoin compounds of the present invention, especially 7b-7g, 7i-7k, 7m-7p, 7r-7t have strong growth inhibitory activity on hormone-dependent prostate cancer cell LNCaP, and can be used for the preparation of antineoplastic drugs.

Description

Diaryl substituted thiohydantoin compound and its preparation method and application

technical field

The invention relates to the field of organic compound synthesis and medical application, in particular to thiohydantoin compounds, their preparation method and pharmaceutical application.

Background technique

Prostate cancer (PCa) is a common tumor that threatens men's health. In the past decade, the incidence of PCa in my country has shown an obvious upward trend, and has become one of the fastest-growing tumors. The vast majority of prostate cancer cells need to grow and proliferate under the stimulation of androgen (testosterone or dihydrotestosterone DHT), so androgen deprivation therapy (androgen deprivation therapy, ADT, also known as castration therapy) is currently clinically used for prostate cancer First-line endocrine therapy for treatment. Androgen ablation therapy works by blocking androgen production (eg, radical prostatectomy or the gonadotropin-releasing hormone analog GnRH) and blocking endogenous androgen interaction with the androgen receptor (androgen receptor, AR) binding (AR antagonists) to block androgen action on cancer cells to inhibit their growth. However, almost all prostate cancer patients who initially respond to ADT will relapse after 12 to 18 months of treatment. Even if the serum testosterone is at the castration level, the patient's tumor will still progress to castration resistance prostate cancer (CRPC) .

Studies on the pathogenesis of CRPC have shown that the transformation of prostate cancer from hormone dependence to CRPC may have the following reasons: 1) AR overexpression makes it sensitive to low concentrations of androgen; 2) AR gene mutation makes AR hypersensitive or ligand The specificity of the binding domain is reduced so that it is activated by other steroid hormones or even AR antagonists; 3) the expression level of key enzymes involved in androgen synthesis in the tumor is up-regulated, resulting in excessive local androgen concentration; 4) growth factors or cytokines have an effect on AR signaling 5) Abnormal expression of AR auxiliary activators (such as SRC-1). Taken together, the reactivation of the AR signaling pathway is the main reason for the progression of prostate cancer to CRPC. Therefore, AR is not only a therapeutic target in androgen-dependent prostate cancer, but also a key target in the treatment of CRPC.

AR antagonists play an important role in the treatment of prostate cancer. The traditional non-steroidal AR antagonists flutamide, nilutamide, and bicalutamide can compete with DHT for binding to the ligand binding domain (LBD) of AR, resulting in the failure of helix 12 to translocate correctly to form AF-2 Functional domain, the receptor-ligand complex forms a dimer and translocates to the nucleus, which can still weakly bind to androgen response elements (AREs), but cannot promote the synthesis of the co-activator SRC, so that the repressor NCoR AR and SMRT content increased, and AR combined with the repressor could not initiate the transcription of target genes, thereby inhibiting the growth of prostate cancer cells. See: "Cancer" 2009,15(6):461, Shen et al., (Shen HC,BalkPC.Development of Androgen Receptor Antagonists with Promising Activity in Cartration-Resistant Prostate Cancer[J].2009,15(6):461) . However, AR mutations allow it to bind to co-activators after combining with traditional AR antagonists, resulting in AR antagonists becoming agonists, inducing target gene transcription, and leading to treatment failure, which is also one of the main reasons for CRPC.

Due to the limitations of traditional AR antagonists in the treatment of CRPC, the research and development of new AR antagonists has become a hot issue for researchers. MDV3100 is a new type of non-steroidal AR antagonist jointly developed by Medivation and Astellas, which was approved by the FDA for the treatment of CRPC in August 2012. The affinity between MDV3100 and AR is much higher than that of bicalutamide, and it can block the biological function of mutant AR, and has no agonistic activity on AR. The aryl-substituted thiohydantoin derivatives have good application prospects as anti-prostate cancer drugs.

Contents of the invention

Aiming at the prior art above, the present invention provides a thiohydantoin androgen receptor antagonist with anti-prostate cancer activity——diaryl-substituted thiohydantoin compounds, and the present invention also provides such Process for the preparation of compounds and their use in pharmacy.

The present invention is achieved through the following technical solutions:

Diaryl substituted thiohydantoin compound, the structure is as shown in general formula (I):

Wherein, R ₁ is hydrogen, halogen or trifluoromethyl; R ₂ is hydrogen, halogen, trifluoromethyl, nitrile or alkoxy; R ₃ is hydrogen, halogen or nitrile; R ₄ is hydrogen or halogen; R ₅ is hydrogen, halogen, methyl, N-ethylcarbamoyl, N-isopropylcarbamoyl, N-cyclopropylcarbamoyl.

Preferably, R ₁ is hydrogen, fluorine or bromine; R ₂ is hydrogen, fluorine, chlorine, bromine, trifluoromethyl, nitrile or methoxy; R ₃ is hydrogen, fluorine, nitrile; R ₄ is hydrogen or Fluorine, R ₅ is fluorine, methyl, N-ethylcarbamoyl or N-isopropylcarbamoyl.

Further preferably, the diaryl-substituted thiohydantoin compounds of the present invention include but are not limited to one of the following compounds:

4-(8-oxo-6-thio-5-(4-fluorophenyl)-5,7 diazaspiro[3.4]oct-7 base)-2-trifluoromethylbenzonitrile (7a );

5-(4-fluorophenyl)-6-thioxo-7-(2,3,4-trifluorophenyl)-5,7-diazaspiro[3.4]oct-8-one (7b);

4-(5-(4-fluorophenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-7-yl)phthalonitrile (7c);

7-(3-Chloro-4-fluorophenyl)-5-(4-fluorophenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7d);

2-Chloro-4-(5-(4fluorophenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-7-yl)benzonitrile (7e);

4-(5-(4-fluorophenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-7-yl)-2-methoxybenzonitrile (7f );

7-(3-Chloro-2-fluorophenyl)-5-(4-fluorophenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7g);

4-(8-oxo-6thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-7-yl)-2-trifluoromethylbenzonitrile (7h);

4-(8-oxo-6-thioxo-5-p-tolyl-5,7-diazaspiro[3.4]oct-7yl)phthalonitrile (7i);

2-Chloro-4-(8-oxo-6-thioxo-5-p-tolyl-5,7-diazaspiro[3.4]oct-7yl)benzonitrile (7j);

2-methoxy-4-(8-oxo-8thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-7yl)benzonitrile (7k);

7-(4-fluorophenyl)-6-thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-8-one (7l);

7-(3-Chloro-4-fluorophenyl)-6-thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-8-one (7m);

7-(2-bromophenyl)-6-thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-8-one (7n);

6-thio-5-p-tolyl-7-(2,3,4-trifluoromethyl)-5,7-diazaspiro[3.4]oct-8-one (7o);

7-(3-Chloro-2-fluorophenyl)-6-thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-8-one (7p);

7-(3-Bromophenyl)-6-thio-5-p-tolyl-5,7-diazaspiro[3.4]oct-8-one (7q);

4-(7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-5-yl)-N - isopropylbenzamide (7r);

4-(7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-5-yl)-N -Ethyl-2-fluorobenzamide (7s);

4-(7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-5-yl)-N - Isopropyl-2-fluorobenzamide (7t).

The brackets behind the names of the above-mentioned preferred 20 compounds are their corresponding codes. For the convenience of description, the codes in the above brackets will be directly used in the following content of this specification.

The preparation method of the above-mentioned diaryl substituted thiohydantoin compounds,

The preparation method of diaryl substituted thiohydantoin compound of the present invention comprises the following steps (synthetic route is as follows):

Synthetic route I:

Wherein, R' is fluorine or hydrogen; R is methyl, ethyl, isopropyl or cyclopropyl.

Synthetic route II:

Wherein, R ₁ is hydrogen, halogen or trifluoromethyl; R ₂ is hydrogen, halogen, trifluoromethyl, nitrile or alkoxy; R ₃ is hydrogen, halogen or nitrile; R ₄ is hydrogen or halogen; R ₅ is hydrogen, halogen, methyl, N-ethylcarbamoyl, N-isopropylcarbamoyl, N-cyclopropylcarbamoyl. Preferably, R ₁ is hydrogen, fluorine or bromine; R ₂ is hydrogen, fluorine, chlorine, bromine, trifluoromethyl, nitrile or methoxy; R ₃ is hydrogen, fluorine, nitrile; R ₄ is hydrogen or Fluorine, R ₅ is fluorine, methyl, N-ethylcarbamoyl or N-isopropylcarbamoyl.

Reagents and reaction conditions: (i) thionyl chloride, DMF, THF, alkylamine; (ii) Pd/C, H ₂ ; (iii) sodium cyanide, cyclobutanone, 90% acetic acid; (iv) sulfur Phosgene, water; (v)a: DMF, room temperature; b: methanol, hydrochloric acid.

Further, the specific steps are as follows:

(i) Dissolve the raw material 2-substituted-4-nitrobenzoic acid (compound 1) in anhydrous THF, add 3 drops of DMF, slowly add thionyl chloride under ice-bath conditions, drop it, and react at room temperature for 1 hour, Remove the solvent and excess thionyl chloride under reduced pressure, add anhydrous tetrahydrofuran to dissolve, and slowly drop into the tetrahydrofuran solution of alkylamine. After the reaction is complete, remove tetrahydrofuran and excess alkylamine under reduced pressure; the remaining liquid is washed with ethyl acetate Ester extraction, the organic layer was washed with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to remove the solvent under reduced pressure to obtain intermediate compound 2; wherein, 2-substituted 4-nitrobenzoic acid (compound 1 ), the molar ratio of thionyl chloride and alkylamine is 1:(1～1.5):(2～3).

(ii) Dissolve the above-mentioned intermediate N-alkyl-2-substituted-4-nitrobenzamide (compound 2) in anhydrous methanol, add Pd/C, and react overnight at room temperature under a hydrogen atmosphere (6~12h) , filtered, and the solvent was distilled off under reduced pressure to obtain intermediate 3;

(iii) Put substituted aniline A (compound 3), cyclobutanone, and 90% acetic acid solution (v/v) in a two-necked flask, stir evenly, add sodium cyanide, heat up to 70-80°C, and react for 24 hours ; Cool to room temperature, dilute with distilled water, extract with ethyl acetate, wash the organic phase with saturated sodium chloride solution, dry over anhydrous magnesium sulfate, filter, evaporate the solvent under reduced pressure, and obtain intermediate 4 by silica gel column chromatography, eluting system Petroleum ether: ethyl acetate volume ratio = 3:1; wherein, the molar ratio of substituted aniline A, cyclobutanone, and sodium cyanide is 1:2:2; every 5 mmol of substituted aniline A uses 20 ml of 90% acetic acid solution;

(iv) Add thiophosgene to water, stir well below 21°C to form a heterogeneous system, add substituted aniline B (compound 5) into the above heterogeneous system, and continue to stir fully for 1-2.5h; after the reaction is completed, put The reaction solution was extracted with dichloromethane, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered, evaporated under reduced pressure to remove the solvent, and silica gel column chromatography to obtain the intermediate substituted phenyl isothiocyanate (compound 6), the elution system is petroleum ether: ethyl acetate volume ratio = 6:1; wherein, the molar ratio of thiophosgene and substituted aniline B (compound 5) is (1-1.5):1.

(v) Dissolve the substituted phenyl isothiocyanate (compound 6), the intermediate (compound 4) in DMF, and react at room temperature for 24 hours; after the reaction, add methanol to the reaction solution, slowly drop 2mol/L of hydrochloric acid, heated to 70-80°C, reacted for 6 hours, cooled to room temperature, poured into ice water, extracted with ethyl acetate, dried the organic phase with anhydrous magnesium sulfate, filtered, evaporated the solvent under reduced pressure, 95% ethanol (v/ v) recrystallization to obtain the final product diaryl substituted thiohydantoin (compound); wherein, the substituted phenyl isothiocyanate, intermediate (compound 4), and the molar ratio of hydrochloric acid are 2:(1.5 -2): 6; the amount of methanol used is: 15ml of methanol for every 2mmol of substituted phenyl isothiocyanate.

Preferably, in the above step (i), the alkylamine is methylamine, ethylamine or isopropylamine.

Preferably, in the above step (ii), N-alkyl-2-substituted-4-nitrobenzamide (compound 2) is N-ethyl-2-fluoro-4-nitrobenzamide or N- Isopropyl-2-fluoro-4-nitrobenzamide, N-isopropyl-4-nitrobenzamide.

Preferably, in the above step (iii), the substituted aniline A (compound 3) is p-toluidine, p-fluoroaniline, N-ethyl-2-fluoro-4-aminobenzamide, N-isopropyl-2- Fluoro-4-aminobenzamide or N-isopropyl-4-aminobenzamide.

Preferably, in the above step (iv), the substituted aniline B (compound 5) is 3-methoxy-4-cyanoaniline, 3-chloro-4-cyanoaniline, 3,4-dicyanoaniline, 2 ,3,4-Trifluoroaniline, 3-chloro-4-fluoroaniline, 3-bromoaniline, 2-fluoro-3-chloroaniline, 3-trifluoromethyl-4-cyanoaniline, 2-bromoaniline or 4-fluoroaniline.

Preferably, in the above step (v), the substituted phenyl isothiocyanate (compound 6) is 3-trifluoromethyl-4-cyanophenyl isothiocyanate, 2,3,4-trifluoro Phenyl isothiocyanate, 3,4-dicyanophenyl isothiocyanate, 3-chloro-4-fluorophenyl isothiocyanate, 3-chloro-4-cyanophenyl isothiocyanate Cyanate, 3-methoxy-4-cyanophenylisothiocyanate, 2-fluoro-3-chlorophenylisothiocyanate, 4-fluorophenylisothiocyanate, 2- Bromophenylisothiocyanate, 3-bromophenylisothiocyanate; intermediate 4 is N-ethyl-4-((1-cyanocyclobutyl)amino)-2-fluorobenzamide , N-isopropyl-4-((1-cyanocyclobutyl)amino)-2-fluorobenzamide, N-isopropyl-4-((1-cyanocyclobutyl)amino)benzene Formamide, 1-p-tolylamino-1-cyanocyclobutane or 1-p-fluorophenylamino-1-cyanocyclobutane.

The diaryl-substituted thiohydantoin compounds of the present invention, especially 7b-7g, 7i-7k, 7m-7p, 7r-7t have strong growth inhibitory activity on hormone-dependent prostate cancer cell LNCaP, and can be used for the preparation of antineoplastic drugs.

Compared with the prior art, the excellent effect of the present invention is: the present invention has synthesized compounds based on MDV3100 with different structures, and the innovative point is to use substituents on the benzene ring in MDV3100 with different sizes or different electron-donating or absorbing compounds. Electron-effect groups are substituted to obtain new compounds that have growth-inhibiting effects on prostate cells. The method for evaluating the growth inhibitory activity of prostate cancer cells adopts the conventional tetramethylazolazolium blue colorimetric method (MTT method), and the results are shown in Table 1.

The results of activity experiments show that the diphenyl substituted thiohydantoin compounds of the present invention have obvious cell growth inhibitory activity on LNCaP cells (AR positive), and are hormone-independent except for 7c, 7e, 7i, and 7j. The prostate cancer cell line PC-3 (AR negative) had no cytostatic activity. Compounds 7b-7g, 7i-7k, 7m-7p, 7r-7t have growth inhibitory activity on hormone-dependent prostate cancer cells LNCaP similar to or superior to the lead compound. Among them, the growth inhibitory activity of 7i, 7j and 7t on LNCaP was significantly better than that of the lead compound, with IC ₅₀ of 5.46 μM, 5.76 μM and 1.78 μM, respectively. The independent prostate cancer cell PC-3 has obvious cytostatic activity, which may suggest that 7i and 7j exert cytostatic activity not only through AR signaling pathway, but also through other pathways.

Table 1 Numbering, structure and activity test results of thiohydantoin compounds

detailed description

The present invention will be further described below in conjunction with the examples for a deeper understanding of the present invention and its advantages and effects, but the examples are only used to illustrate the present invention rather than limit the present invention.

Methods and reagents not described in detail in the examples are conventional methods and reagents in the field.

The synthesis of embodiment 1 substituted aniline 3a-3c

(1) Preparation of N-alkyl-2-substituted-4-nitrobenzamide

Put 2-substituted-4-nitrobenzoic acid (10mmol) in a 100mL reaction flask, add 30mL of anhydrous tetrahydrofuran, add 3 drops of DMF, stir well at room temperature, until the solid is completely dissolved, put the reaction flask in an ice alcohol bath , slowly add thionyl chloride (12mmol) dropwise, react at room temperature for 1h, evaporate the solvent under reduced pressure, dissolve the remaining solid with anhydrous DMF, and slowly add dropwise to 15ml of tetrahydrofuran solution of methylamine, ethylamine or isopropylamine (2mol/L), evaporate the organic solvent under reduced pressure, extract with ethyl acetate, combine the organic layers, wash with water, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, remove anhydrous sodium sulfate, evaporate under reduced pressure Solvent, to obtain intermediate N-alkyl-2-fluoro-4-nitrobenzamide (compound 2, including three kinds, as follows):

N-ethyl-2-fluoro-4-nitrobenzamide (2a): yellow solid, yield 84%, ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.66 (s, 1H), 8.21(dd, J ₁ =1.8Hz, J ₂ =9.6Hz, 1H), 8.13(dd, J ₁ =1.8Hz, J ₂ =7.8Hz, 1H), 7.83(t, J=7.8Hz, 1H), 3.29 (m, 2H), 1.12 (t, J=7.8Hz, 3H), MS (calcd/found) [M+H] ⁺ : 213.1/213.3.

N-isopropyl-2-fluoro-4-nitrobenzamide (2b): yellow solid, yield 97%, ¹ H-NMR(DMSO-d ₆ )δ(ppm): 8.56(d, J= 7.8Hz, 1H), 8.18(dd, J ₁ =1.8Hz, J ₁ =9.0Hz, 1H), 8.12(J ₁ =1.8Hz, J ₂ =9.0Hz, 1H), 7.77(dd, J ₁ =6.6 Hz, J ₂ =8.4 Hz, 2H) MS (calcd/found) [M+H] ⁺ : 227.1/227.3.

N-isopropyl-4-nitrobenzamide (2c): white solid, yield 99%, ¹ H-NMR (DMSO-d ₆ )δ (ppm): 8.58 (d, J=7.2Hz, 1H ), 8.30(d, J=8.4Hz, 2H), 8.07(d, J=9.0Hz, 2H), 4.10(q, J=6.6Hz, 1H), 1.18(d, J=6.6Hz, 6H), MS(calcd/found)[M+H] ⁺ :209.1/209.4.

(2) Preparation of N-alkyl-2-substituted-4-aminobenzamide 3

Dissolve the N-alkyl-2-substituted-4-nitrobenzamide (compound 2a, 2b, 2c) (8mmol) obtained above in 15mL of anhydrous methanol, add 430mg of 10% Pd/C, and react under 1atm hydrogen atmosphere Overnight (10 h), filtered, and solvent intermediates (3a-3c) evaporated under reduced pressure.

The obtained N-alkyl-2-substituted-4-nitrobenzamide (3a-3c) has three kinds, namely N-ethyl-2-fluoro-4-nitrobenzamide, N-isopropyl-2 -Fluoro-4-nitrobenzamide, N-ethyl-4-nitrobenzamide, specifically as follows:

N-ethyl-2-fluoro-4-aminobenzamide (3a): white solid, yield 97%, ¹ H-NMR(DMSO-d ₆ )δ(ppm): 7.59(d, J=5.4Hz ,1H),7.42(t,J=8.4Hz,1H),7.37(dd,J ₁ =1.8Hz,J ₂ =8.4Hz,1H),6.27(J ₁ =1.8Hz,J ₂ =8.4Hz,1H ), 5.89 (s, 1H), 3.22 (m, 2H), 1.07 (t, J=6.6Hz, 3H), MS (calcd/found) [M+H] ⁺ : 183.1/183.4.

N-isopropyl-2-fluoro-4-aminobenzamide (3b): white solid, yield 95%, ¹ H-NMR(DMSO-d ₆ )δ(ppm): 7.37(t, J=9.0 Hz, 1H), 7.31 (t, J = 6.6Hz, 1H), 6.37 (dd, J ₁ = 1.8Hz, J ₂ = 8.4Hz, 1H), 5.26 (dd, J ₁ = 1.8Hz, J ₂ = 8.4 Hz,1H),5.86(s,2H),4.01(q,J=6.6Hz,1H),1.12(d,J=6.6Hz,6H), MS(calcd/found)[M+H] ⁺ :, 197.1/197.4.

N-isopropyl-4-aminobenzamide (3c): white solid, yield 90%, ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.70 (d, J=7.8Hz, 1H) ,7.56(d,J=8.4Hz,2H),6.51(d,J=8.4Hz,2H),6.55(s,1H),4.04(q,J=7.2Hz,1H),1.12(d,J= 6.0Hz,6H), MS(calcd/found)[M+H] ⁺ :179.1/179.3.

The preparation of embodiment 2 intermediate compound 4

Put the intermediate compound 3 (5mmol) and cyclobutanone (10mmol) in a 100mL two-neck flask, add 20mL of 90% glacial acetic acid to dissolve it completely, carefully add sodium cyanide (10mmol), quickly seal it, and reflux at 80°C for 24h. Pour the reaction mixture into 100 mL of ice water, adjust the pH value to neutral, extract with ethyl acetate, combine the organic phases, wash with water, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain The crude product was separated and purified by silica gel column chromatography to obtain intermediate 4, and the elution system was petroleum ether:ethyl acetate=3:1.

The intermediate compound 3 used was p-toluidine, p-fluoroaniline, N-isopropyl-4-aminobenzamide, N-ethyl-2-fluoro-4-aminobenzamide or N-isopropyl-2 -Fluoro-4-aminobenzamide, correspondingly, the obtained intermediate compound 4 is one of the following 5 types, specifically as follows:

N-ethyl-4-((1-cyanocyclobutyl)amino)-2-fluorobenzamide (4a): white solid, 82% yield, ¹ H-NMR(DMSO-d ₆ )δ( ppm): 7.85 (d, J = 4.2Hz, 1H), 7.54 (t, J = 9.0Hz, 1H), 7.33 (s, 1H), 6.45 (dd, J ₁ = 1.8Hz, J ₂ = 9.0Hz, 1H), 6.29(J ₁ ＝1.8Hz, J ₂ ＝12.0Hz), 3.23(m, 2H), 2.75(m, 2H), 2.35(m, 2H), 2.01-2.11(m, 2H), 1.08( t, J=7.2Hz, 6H), MS (calcd/found) [M+H] ⁺ : 262.1/262.3.

N-isopropyl-4-((1-cyanocyclobutyl)amino)-2-fluorobenzamide (4b): white solid, yield 79%, ¹ H-NMR (DMSO-d ₆ )δ (ppm): 7.64 (d, J = 7.2Hz, 1H), 7.48 (t, J = 8.4Hz, 1H), 7.31 (s, 1H), 7.45 (dd, J ₁ = 2.4Hz, J ₂ = 9.0Hz ,1H),6.30(dd,J ₁ =2.4Hz,J ₂ =13.8Hz,1H),7.02(m,1H),4.02(m,1H),2.75(m,2H),2.35(m,2H) , 2.03-2.10 (m, 2H), 1.12 (t, J = 7.2Hz, 6H), MS (calcd/found) [M+H] ⁺ : 276.1/276.5.

N-isopropyl-4-((1-cyanocyclobutyl)amino)benzamide (4c): white solid, yield 86%, ¹ H-NMR(DMSO-d ₆ )δ(ppm): 7.87(d, J=7.8Hz, 1H), 7.71(d, J=8.4Hz, 2H), 7.05(s, 1H), 6.58(d, J=8.4Hz, 2H), 4.06(m, 1H), 2.74(m,2H),2.36(m,2H),2.03-2.12(m,2H),1.13(d,J＝7.2Hz,6H), MS(calcd/found)[M+H] ⁺ :258.2/ 258.3.

1-p-Tolylamino-1-cyanocyclobutane (4d): brown solid, yield 88.7%, mp: 66.0-68.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 6.99(d, J=7.8Hz, 2H), 6.50(d, J=8.4Hz, 2H), 6.38(s, 1H), 2.67(m, 2H), 2.32(m, 2H), 2.17(s, 3H), 2.05(m, 2H). MS(calcd/found)[M+H] ⁺ :197.1/197.4.

1-p-Fluorophenylamino-1-cyanocyclobutane (4e): white solid, yield 76.1%, mp: 53.5-55.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.03 (t, J = 8.4 Hz, 2H), 6.58 (m, 3H), 2.68 (m, 2H), 2.32 (m, 2H), 2.05 ( m,2H).MS(calcd/found)[M+H] ⁺ :191.1/191.4.

The preparation of embodiment 3 intermediate 6 substituted phenyl isothiocyanates

Add C(S)Cl ₂ (6.5mmol) and water (11mL) into a 25mL round bottom flask, stir well at room temperature to form a heterogeneous system, and add substituted aniline (compound 5) (6mmol) into the reaction solution , react at room temperature for 1h to 2.5h, extract the reaction solution with dichloromethane (15mL×3), combine the organic phases and wash with saturated NaCl (30mL), dry the organic phases with anhydrous MgSO ₄ , filter, and evaporate the solvent under reduced pressure , the intermediate compound 6 was separated by silica gel column chromatography, and the elution system was petroleum ether: ethyl acetate volume ratio 6:1.

The substituted aniline (compound 5) used above is 3-methoxy-4-cyanoaniline, 3-chloro-4-cyanoaniline, 3,4-dicyanoaniline, 2,3,4-trifluoroaniline, 3-Chloro-4-fluoroaniline, 3-bromoaniline, 2-fluoro-3-chloroaniline, 3-trifluoromethyl-4-cyanoaniline, 2-bromoaniline, 4-fluoroaniline.

3-Methoxy-4-cyanophenylisothiocyanate (6a): white solid, yield 64.8%, mp: 150.5-153.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 7.81(d, J=8.4Hz, 1H), 7.38(s, 1H), 7.15(d, J=8.4Hz, 1H), 3.93(s, 3H).

3-Chloro-4-cyanophenylisothiocyanate (6b): yellow-white solid, yield 87.9%, mp: 73.0-74.5°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.06 (d, J = 8.4Hz, 1H), 7.94 (d, J = 1.8Hz, 1H), 7.61 (dd, J ₁ = 1.8Hz, J ₂ = 8.4Hz, 1H).

3,4-Dicyanophenylisothiocyanate (6c): yellow solid, yield 91.9%, mp: 91.0.0-93.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.30 (d, J = 2.4Hz, 1H), 8.21 (d, J = 7.8Hz, 1H), 7.95 (dd, J ₁ = 2.4Hz, J ₂ =8.4Hz,1H).

2,3,4-Trifluorophenylisothiocyanate (6d): yellow oil, yield 70.%. ¹ H-NMR(DMSO-d ₆ )δ(ppm):6.95(m,2H).

3-Chloro-4-fluorophenylisothiocyanate (6e): colorless oil, yield 47.9%. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.50 (m, 1H), 7.39 (m, 2H).

3-Bromophenylisothiocyanate (6f): yellow oil, yield 79.5%. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 7.71(s, 1H), 7.58(d, J=7.8Hz, 1H), 7.45(d, J=7.8Hz, 1H), 7.40(t, J＝7.8Hz,1H).

2-fluoro-3-chlorophenylisothiocyanate (6g): yellow oil, yield 75.5%. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.59 (dt, J ₁ =1.2Hz, J ₂ =7.8Hz, 1H), 7.47 (dt, J ₁ =1.2Hz, J ₂ =7.8Hz, 1H),7.28(dt,J ₁ =1.2Hz,J ₂ =7.8Hz,1H).

3-Trifluoromethyl-4-cyanophenylisothiocyanate (6h): Brown solid, 46.5% yield, mp: 38.0-39.5°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.25 (d, J = 8.4Hz, 1H), 8.11 (d, J = 1.8Hz, 1H), 7.94 (dd, J ₁ = 1.8Hz, J ₂ = 8.4Hz, 1H).

2-Bromophenylisothiocyanate (6i): colorless oil, yield 45.0%. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 7.67(d, J=7.8Hz, 1H), 7.62(d, J=7.2Hz, 1H), 7.39(t, J=7.2Hz, 1H) ,7.19(m,1H).

4-Fluorophenylisothiocyanate (6j): yellow oil, yield 64.0%. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.52 (m, 2H), 7.30 (t, J = 9.0 Hz, 2H).

The preparation of embodiment 4 diphenylthiohydantoin 7

Substituted phenyl isothiocyanate (compound 6) (2mmol), intermediate compound 4 (1.5mmol), DMF (1mL) were added into a 50mL round bottom flask, and reacted at room temperature for 24h. Add methanol (15mL) to the reaction solution, and slowly add 2mol/L hydrochloric acid (3mL) dropwise, heat up to 70-80°C, react for 6h, cool to room temperature, pour into ice water (25mL), and extract with ethyl acetate (30mL×3), the organic phase was dried with anhydrous magnesium sulfate, filtered, the solvent was evaporated under reduced pressure, and the solid was recrystallized with 95% ethanol to obtain the target compound 7.

The used substituted phenyl isothiocyanate (compound 6) and intermediate compound 4 are all selected from the compounds involved in Examples 2 and 3. After the combination of different raw materials, the following 20 kinds of target compounds can be obtained:

4-(8-oxo-6-thio-5-(4-fluorophenyl)-5,7 diazaspiro[3.4]oct-7-yl)-2-trifluoromethylbenzonitrile ( 7a): white solid powder, yield 51.6%, mp: 177.0-179.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 8.38(d, J=8.4Hz, 1H), 8.25(s, 1H), 8.06(d, J=8.4Hz, 1H), 7.47(m, 4H), 2.62(m, 2H), 2.41(m, 2H), 1.95(m, 1H), 1.53(m, 1H). IR(KBr,cm ^-1 ):υ _Ar-H : 3064.21,3001.07;υ _CH :2962.22;υ _CN :2233.70;υ _C=O :1764.53;υ _C=C : _{1612.80,1511.76} ; ; δ _Ar-H : 1182.83, 1139.57, 1055.32, 832.19, 805.73. HRMS (ESI) m/z calcd for C ₂₀ H ₁₃ F ₄ N ₃ OS [M+H] ⁺ : 420.0788, found 420.0787. HPLC purity = 98.4%.

7-(2,3,4-trifluorophenyl)-5-(4-fluorophenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7b): White solid powder, yield 28.0%, mp: 143.0-144.5°C. ¹ H-NMR(DMSO-d ₆ )δ(ppm):7.55(m,4H),7.44(m,2H),2.62(m,1H),2.55(m,1H),2.48(m,1H), 2.41(m,1H),1.97(m,1H),1.54(m,1H). IR(KBr,cm ^-1 ):υAr _-H : 3083.60; _υCH :2945.02;υC _=O :1762.14;υC _=C :1621.48,1509.58; _δCH :1424.51,1319.94;δAr _-H :1163.92 , 1107.45, 1011.38, 837.65, 801.20. HRMS (ESI) m/z calcd for C ₁₈ H ₁₂ F ₄ N ₂ OS [M+H] ⁺ : 381.0679, found 381.0683. HPLC purity = 99.3%.

4-(8-oxo-6-thioxo-5-(4-fluorophenyl)-5,7diazaspiro[3.4]oct-7-yl)phthalonitrile (7c): white solid Powder, yield 45.1%, mp: 252.0-254.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.33 (dd, J ₁ =3.0Hz, J ₂ =5.4Hz, 2H), 8.09 (dd, J ₁ =1.8Hz, J ₂ =8.4Hz, 1H), 7.47(m, 4H), 2.62(m, 2H), 2.41(m, 2H), 1.95(m, 1H), 1.53(m, 1H). IR(KBr,cm ^-1 ):υAr _-H :3079.60,3002.98; _υCH :2947.33; _υCN :2235.43;υC _=O :1759.19;υC _=C :1601.14,1511.31; _δCH :1431.67,1314.18 ; δ _Ar-H : 1119.92, 834.35, 805.85. HRMS (ESI) m/z calcd for C ₂₀ H ₁₃ FN ₄ OS[M+H] ⁺ : 377.0867, found 377.0866. HPLC purity = 98.6%.

7-(3-Chloro-4-fluorophenyl)-5-(4-fluorophenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7d): white Solid powder, yield 47.4%, mp: 154.0-156.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.78 (dd, J ₁ = 1.8Hz, J ₂ = 7.2Hz, 1H), 7.59 (t, J = 8.4Hz, 1H), 7.50 (m, 3H),7.47(t,J＝8.4Hz,2H),2.62(m,1H),2.55(m,1H),2.58(m,2H),2.39(m,2H),1.95(m,1H), 1.53(m,1H). IR(KBr,cm ^-1 ):υAr _-H :3075.43,3001.55; _υCH :2959.07;υC _=O :1744.93;υC _=C :1601.16,1501.23; _δCH :1428.34,1314.07;δAr _-H : 1185.83, 1109.55, 837.91, 820.45, 805.65. HRMS (ESI) m/z calcd for C ₁₈ H ₁₃ ClF ₂ N ₂ OS [M+H] ⁺ : 379.0478, found 379.0483. HPLC purity = 95.9%.

4-(8-oxo-6-thioxo-5-(4-fluorophenyl)-5,7diazaspiro[3.4]oct-7-yl)-2-chlorobenzonitrile (7e): White solid powder, yield 31.0%, mp: 224.0-226.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.16 (d, J = 8.4Hz, 1H), 7.97 (d, J = 1.2Hz, 1H), 7.70 (dd, J ₁ = 1.8Hz, J ₂ =8.4Hz,1H),7.47(m,4H),2.81(d,J=4.2Hz,3H),2.60(m,2H),2.40(m,2H),1.95(m,1H),1.53( m, 1H). IR (KBr, cm ^-1 ): υAr-H: 3075.12, 3001.35; υ _CH : 2955.66; υ _CN : 2232.49; υ _{C =O :} 1758.40; δ _Ar-H : 1185.47, 1106.54, 826.51. HRMS (ESI) m/z calcd for C ₁₉ H ₁₃ ClFN ₃ OS [M+H] ⁺ : 386.0525, found 386.0528. HPLC purity = 98.8%.

4-(8-oxo-6-thioxo-5-(4-fluorophenyl)-5,7 diazaspiro[3.4]oct-7-yl)-2-methoxybenzonitrile (7f ): white solid powder, yield 32.4%, mp: 290.0-292.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.90 (d, J = 7.8Hz, 1H), 7.49 (m, 2H), 7.44 (d, J = 8.42H), 7.41 (d, J = 1.8Hz,1H),7.21(dd,J ₁ =1.8Hz,J ₂ =8.4Hz,1H),3.92(s,3H),2.60(m,2H),2.41(m,2H),1.95(m, 1H),1.53(m,1H).IR(KBr,cm ^-1 ):υAr _-H :3070.75; _υCH :2991.32,2954.09; _υCN :2229.48;υC _=O :1749.33;υC _=C : 1605.78,1509.47; δ _CH :1428.34,1316.80; υ _CO :1254.47; δ _Ar-H :1168.22,1106.59,837.47,805.30.HRMS(ESI)m/z calcd for C ₂₀ H ₁₆ FN ₃ O ₂ S H] ⁺ :382.1020, found 382.1024. HPLC purity = 97.8%.

7-(3-Chloro-2-fluorophenyl)-5-(4-fluorophenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7g): white Solid powder, yield 31.6%, mp: 135.0-137.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 7.77(t, J=7.2Hz, 1H), 7.56(m, 3H), 7.44(m, 3H), 2.61(m, 1H), 2.54( m,1H), 2.42(m,1H), 1.97(m,1H), 1.55(m,1H). IR(KBr, cm ^-1 ): υ _Ar-H : 3086.18, 3058.39; υ _CH : 2990.49, 2945.03; υ _C=O : 1760.18; υ _{C=C :} 1601.95, 1513.71, 1485.07; δ _Ar-H : 1178.96, 1151.84, 1151.09, 820.97, 797.05. HRMS (ESI) m/z calcd for C ₁₈ H ₁₃ ClF ₂ N ₂ OS [M+H] ⁺ : 379.0478, found 379.0484. HPLC purity = 98.3%.

4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7 diazaspiro[3.4]oct-7-yl)-2-trifluoromethylbenzonitrile (7h): white solid powder, yield 64.5%, mp: 166.0-168.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 8.37(d, J=7.8Hz, 1H), 8.26(s, 1H), 8.06(d, J=8.4Hz, 1H), 7.41(d, J=7.8Hz, 2H), 7.29(d, J=8.4Hz, 2H), 2.60(m, 2H), 2.42(m, 5H), 1.95(m, 1H), 1.52(m, 1H). IR(KBr,cm ^-1 ):υAr _-H :3093.99,3036.43; _υCH :2985.35,2943.25; _υCN :2235.53;υC _=O :1767.22;υC _=C :1631.46,1612.02,1578.78,1514.33, 1504.26; δ _CH : 1436.21, 1417.88, 1377.68, 1316.67; δ _Ar-H : 1184.92, 1150.74, 1056.14, 838.06, 819.42, 790.33. HRMS (ESI) m/z calcd for C ₂₁ H ₁₆ F ₃ N ₃ OS [M+H] ⁺ : 416.1039, found 416.1039. HPLC purity = 99.0%.

4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7 diazaspiro[3.4]oct-7-yl)phthalonitrile (7i): white Solid powder, yield 39.4%, mp: 247.5-248.6°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.34 (m, 2H), 8.10 (dd, J ₁ = 1.8Hz, J ₂ = 8.4Hz, 1H), 7.41 (d, J = 7.8Hz, 2H), 7.29 (d, J=7.8Hz, 2H), 2.59 (m, 2H), 2.43 (m, 5H), 1.95 (m, 1H), 1.53 (m, 1H). IR(KBr,cm ^-1 ):υAr _-H :3039.18,3004.37; _υCH :2958.37; _υCN :2235.73;υC _=O :1749.38;υC _=C :1600.55,1514.75,1493.33; _δCH :1434.49 , 1380.40, 1313.35; δ _Ar-H : 1192.04, 1163.76, 1119.99, 833.09, 819.56, 789.26. HRMS (ESI) m/z calcd for C ₂₁ H ₁₆ N ₄ OS [M+H] ⁺ : 373.1118, found 373.1121. HPLC purity = 98.2%.

4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7 diazaspiro[3.4]oct-7-yl)-2-chlorobenzonitrile (7j) : White solid powder, yield 66.4%, mp: 189.0-190.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.17 (d, J = 8.4Hz, 1H), 7.98 (d, J = 1.8Hz, 1H), 7.71 (dd, J ₁ = 1.8Hz, J ₂ =8.4Hz,1H),7.40(d,J=8.4Hz,2H),7.29(d,J=7.8Hz,2H),2.59(m,2H),2.41(m,5H),1.95(m, 1H), 1.51(m, 1H). IR(KBr,cm ^-1 ):υAr _-H :3088.41,3037.21; _υCH :2956.11; _υCN :2231.32;υC _=O :1756.83;υC _=C :1598.32,1514.25,1492.81; _δCH :1423.66 , 1377.29, 1310.33; δ _Ar-H : 1187.05, 1174.32, 1109.24, 1054.94, 816.40, 788.27. HRMS (ESI) m/z calcd for C ₂₀ H ₁₆ CN ₃ OS [M+H] ⁺ : 382.0775, found 382.0783. HPLC purity = 98.6%.

4-(8-oxo-6-thio-5-(4-methylphenyl)-5,7 diazaspiro[3.4]oct-7-yl)-2-methoxybenzonitrile ( 7k): white solid powder, yield 58.3%, mp: 233.5-235.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.89 (d, J = 8.4Hz, 1H), 7.42 (d, J = 1.8Hz, 1H), 7.40 (d, J = 8.4Hz, 2H) ,7.30(d,J=8.4Hz,2H),7.21(dd,J ₁ ＝1.8Hz,J ₂ ＝8.4Hz,1H),2.59(m,2H),2.43(m,5H),1.95(m, 1H), 1.52(m, 1H). IR(KBr, cm ^-1 ):υAr _-H :3095.04,3036.53; _υCH :2985.44,2943.38; _υCN :2226.00;υC _=O :1759.76;υC _=C :1605.57,1578.50,1506.37; _δCH : 1415.87, 1316.64; υ _CO : 1282.98, 1257.18; δ _Ar-H : 1187.73, 1165.31, 1107.42, 853.07, 816.92, 790.79. HRMS (ESI) m/z calcd for C ₂₁ H ₁₆ N ₄ OS [M+H] ⁺ : 378.1271, found 378.1273. HPLC purity = 98.4%.

7-(4-fluorophenyl)-5-(4-methylphenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7l): white solid powder, Yield 20.1%, mp: 218.0-220.1°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.47 (m, 2H), 7.39 (d, J = 7.8Hz, 2H), 7.34 (m, 2H), 7.30 (d, J = 8.4Hz, 2H), 2.57(m, 2H), 2.41(m, 5H), 1.95(m, 1H), 1.51(m, 1H). IR(KBr, cm ^-1 ): υ _Ar-H : 3085.56, 3045.25, 3010.57; υ _CH : 2983.05, 2940.68; υ _C=O : 1751.97; υ _{C=C :} 1598.85, 1507.38; δ _Ar-H : 1187.84, 1116.55, 837.74, 815.36, 791.45. HRMS (ESI) m/z calcd for C ₁₉ H ₁₇ FN ₂ OS [M+H] ⁺ : 341.1118, found 341.1120. HPLC purity = 98.8%.

7-(3-Chloro-4-fluorophenyl)-5-(4-methylphenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7m): White needle-like crystals, yield 51.2%, mp: 157.1-158.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.79 (dd, J ₁ =2.4Hz, J ₂ =6.6Hz, 1H), 7.59 (t, J = 9.0Hz, 1H), 7.50 (m, 1H), 7.39(d, J=7.8Hz, 2H), 7.29(d, J=8.4Hz, 2H), 2.57(m, 2H), 2.41(m, 5H), 1.95(m, 1H), 1.51( m, 1H). IR(KBr,cm ^-1 ):υAr _-H :3071.00,3051.68; _υCH :2984.84,2938.03;υC _=O :1744.40;υC _=C :1595.99,1501.90; _δCH :1421.34,1314.55; _{δAr -H} : 1186.34, 1128.25, 1059.18, 816.16, 792.05. HRMS (ESI) m/z calcd for C ₁₉ H ₁₆ ClFN ₂ OS [M+H] ⁺ : 375.0729, found 375.0734. HPLC purity = 99.6%.

7-(2-Bromophenyl)-5-(4-methylphenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7n): white flaky crystal , yield 31.7%, mp: 200.0-203.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 7.82(d, J=7.8Hz, 1H), 7.56(d, J=3.6Hz, 2H), 7.45(m, 1H), 7.41(d, J=8.4Hz, 2H), 7.29(d, J=7.8Hz, 2H), 2.59(m, 1H), 2.45(m, 6H), 1.99(m, 1H), 1.56(m, 1H). IR(KBr, cm ^-1 ): υ _Ar-H : 3040.10, 3045.25, 3003.00; υ _CH : 2942.56, 2853.41; υ _C=O : 1745.54; υ _{C=C :} 1515.29; δ _Ar-H : 1195.66, 1115.32, 821.40, 793.60, 763.83. HRMS (ESI) m/z calcd for C ₂₁ H ₁₆ N ₄ OS [M+H] ⁺ : 401.0318, found 401.0314. HPLC purity = 99.4%.

7-(2,3,4-trifluorophenyl)-5-(4-methylphenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7o) : White solid, yield 53.6%, mp: 158.0-160.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 7.54(m, 2H), 7.40(d, J=7.8Hz, 2H), 7.33(d, J=8.4Hz, 2H), 2.61(m, 1H), 2.52(m,1H), 2.47(m,1H), 1.97(m,1H), 2.40(m,4H), 1.96(m,1H), 1.53(m,1H). IR(KBr,cm ^-1 ):υAr _-H :3083.07; _υCH :2986.29,2939.77;υC _=O :1757.20;υC _=C :1625.52,1512.17; _δCH :1421.21,1321.43;δAr _-H : 1161.96, 1055.32, 1012.02, 817.48, 790.50. HRMS (ESI) m/z calcd for C ₁₉ H ₁₅ F ₃ N ₂ OS [M+H] ⁺ : 377.0930, found 377.0933. HPLC purity = 98.7%.

7-(3-Chloro-2-fluorophenyl)-5-(4-methylphenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7p): White solid powder, yield 16.1%, mp: 140.0-142.0°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.77 (m, 1H), 7.58 (m, 1H), 7.42 (m, 3H), 7.33 (d, J=8.4Hz, 2H), 2.61 ( m,1H), 2.53(m,1H), 2.48(m,1H), 2.42(m,4H), 1.97(m,1H), 1.54(m,1H). IR(KBr, cm ^-1 ): υ _Ar-H : 3077.99, 3044.10; υ _CH : 2991.79, 2959.82; υ _C=O : 1758.49; υ _{C=C :} 1601.94, 1515.47, 1487.96; δ _Ar-H : 1180.15, 1151.84, 1116.64, 820.44, 793.44. HRMS (ESI) m/z calcd for C ₁₉ H ₁₆ ClFN ₂ OS [M+H] ⁺ : 375.0729, found 375.0731. HPLC purity = 97.6%.

7-(3-bromophenyl)-5-(4-methylphenyl)-6-thio-5,7-diazaspiro[3.4]oct-8-one (7q): white solid powder, Yield 11.7%, mp: 125.0-128.5°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 7.73 (t, J = 1.8Hz, 1H), 7.67 (dt, J ₁ = 1.8Hz, J ₂ = 7.8Hz, 1H), 7.47 (m, 2H), 7.39(d, J=8.4Hz, 2H), 7.29(d, J=8.4Hz, 2H), 2.57(m, 2H), 2.41(m, 5H), 1.95(m, 1H), 1.51( m, 1H). IR(KBr, cm ^-1 ): υ _Ar-H : 3067.63, 3044.10; υ _CH : 2992.73, 2955.64; υ _C=O : 1749.47; υ _{C=C :} 1575.22, 1513.07, 1480.75; δ _Ar-H : 1189.62, 1112.09, 851.59, 792.45. HRMS (ESI) m/z calcd for C ₁₉ H ₁₇ BrN ₂ OS [M+H] ⁺ : 401.0318, found 401.0317. HPLC purity = 97.0%.

4-(7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-5-yl)-N - Isopropylbenzamide (7r): white solid powder, yield 80.0%, mp: 221.0-223.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 8.40(s, 1H), 8.39(d, J=7.80Hz, 1H), 8.26(d, J=1.20Hz, 1H), 8.06(dd, J ₁ ＝7.80Hz, J ₂ ＝1.80Hz, 1H), 8.03(d, J＝8.40Hz, 1H), 7.52(d, J＝8.40Hz, 1H), 4.12(m, 1H), 2.63(m, 2H), 2.43(m, 2H), 1.95(m, 1H), 1.53(m, 1H), 1.19(d, J=6.00Hz, 6H). HRMS(ESI) m/zcalcd for C ₂₄ H ₂₁ F ₃ N ₄ O ₂ S[M+H] ⁺ : 487.1410, found 487.1405. HPLC purity = 97.9%.

4-(7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-5-yl)-N -Ethyl-2-fluorobenzamide (7s): white solid powder, yield 48.4%, mp: 127.0-128.0°C. ¹ H-NMR (DMSO-d ₆ )δ(ppm): 8.55(s, 1H), 8.40(d, J=8.40Hz, 1H), 8.25(s, 1H), 8.06(d, J=8.40Hz, 1H), 7.80(t, J=8.10Hz, 1H), 7.47(t, J=10.20Hz, 1H), 7.38(dd, J ₁ ＝8.40Hz, J ₂ ＝6.60Hz, 1H), 3.29(m, 2H), 2.64(m, 2H), 1.97(m, 1H), 1.57(m, 1H), 1.13(t, J=7.20Hz, 3H). HRMS(ESI) m/zcalcd for C ₂₃ H ₁₈ F ₄ N ₄ O ₂ S[M+H] ⁺ : 491.1159, found 491.1162. HPLC purity = 99.3%.

4-(7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thio-5,7-diazaspiro[3.4]oct-5-yl)-N -Isopropyl-2-fluorobenzamide (7t): white solid powder, yield 70.0%, mp: 190.0-192.5°C. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.43 (d, J = 7.8Hz, 1H), 8.39 (d, J = 8.4Hz, 1H), 8.24 (d, J = 1.2Hz, 1H) ,8.06(dd,J ₁ =7.8Hz,J ₂ =6.60Hz,1H),7.75(t,J=7.8Hz,1H),7.44(dd,J ₁ =10.8Hz,J ₂ =1.2Hz,1H) ,7.37(d,J ₁ =7.8Hz,J2=6.00Hz,1H),4.08(q,J=7.2Hz,1H),2.63(m,2H),2.45(m,2H),1.97(m,1H ),1.56(m,1H),1.17(d,J＝7.2Hz,6H).HRMS(ESI)m/zcalcdforC ₂₄ H ₂₀ F ₄ N ₄ O ₂ S[M+H] ⁺ :505.1316,found505.1307 . HPLC purity = 99.4%.

Claims (4)

1. A diaryl-substituted thiohydantoin compound, characterized in that, The chemical structural formula of 7j is as follows: The chemical structural formula of 7t is as follows: . 2. the preparation method of the diaryl substituted thiohydantoin compound according to claim 1, is characterized in that: the synthetic route of described compound 7j is as follows: Wherein, R1 is hydrogen; R2 is chlorine; R3 is cyano; R4 is hydrogen; R5 is methyl; The synthetic route of described compound 7t is as follows: Synthetic route I: Wherein, R' is fluorine; R is isopropyl; Synthetic route II: Wherein, R1 is hydrogen; R2 is trifluoromethyl; R3 is cyano; R4 is fluorine; R5 is N-isopropylcarbamoyl; Reagents and reaction conditions: (i) thionyl chloride, DMF, THF, alkylamine; (ii) Pd/C, H2; (iii) sodium cyanide, cyclobutanone, 90% acetic acid; (iv) sulfur Gas, water; (v)a: DMF, room temperature; b: methanol, hydrochloric acid; Specific steps are as follows: (i) Dissolve the raw material 2-substituted-4-nitrobenzoic acid in anhydrous THF, add DMF, add thionyl chloride under ice-bath conditions, drop it, react at room temperature for 1 hour, evaporate the solvent and excess under reduced pressure Thionyl chloride was dissolved in anhydrous tetrahydrofuran, and dropped into the tetrahydrofuran solution of alkylamine. After the reaction was complete, tetrahydrofuran and excess alkylamine were evaporated under reduced pressure; the remaining liquid was extracted with ethyl acetate, and the organic layer was washed with water. Washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to remove the solvent under reduced pressure to obtain intermediate compound 2; wherein, 2-substituted 4-nitrobenzoic acid, thionyl chloride, and alkylamine The molar ratio is 1:(1～1.5):(2～3); (ii) Dissolve the above obtained intermediate N-alkyl-2-substituted-4-nitrobenzamide in anhydrous methanol, add Pd/C, react overnight at room temperature under a hydrogen atmosphere, filter, and evaporate the solvent under reduced pressure to obtain Intermediate 3; (iii) Put substituted aniline A, cyclobutanone, and 90% acetic acid solution in a two-necked flask, stir evenly, add sodium cyanide, heat up to 70-80°C, and react for 24 hours; cool to room temperature, dilute with distilled water, and dilute with acetic acid Ethyl extraction, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered, the solvent was evaporated under reduced pressure, and intermediate 4 was obtained by silica gel column chromatography. The elution system was petroleum ether: ethyl acetate volume ratio =3:1; Wherein, the mol ratio of substituted aniline A, cyclobutanone and sodium cyanide is 1:2:2; (iv) Add thiophosgene to water, stir fully below 21°C to form a heterogeneous system, add substituted aniline B to the above heterogeneous system, and continue to stir fully for 1-2.5h; Extract with methyl chloride, combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous magnesium sulfate, filter, evaporate the solvent under reduced pressure, and obtain the intermediate substituted phenyl isothiocyanate by silica gel column chromatography, and the elution system is petroleum Ether: ethyl acetate volume ratio=6:1; Wherein, the molar ratio of thiophosgene and substituted aniline B is (1-1.5):1; (v) Dissolve the substituted phenyl isothiocyanate and intermediate 4 in DMF, and react at room temperature for 24 hours; ℃, reacted for 6 hours, cooled to room temperature, poured into ice water, extracted with ethyl acetate, dried the organic phase with anhydrous magnesium sulfate, filtered, evaporated the solvent under reduced pressure, and recrystallized from 95% ethanol to obtain the final product diaryl substituted Thiohydantoin; Wherein, substituted phenyl isothiocyanate, intermediate 4, the molar ratio of hydrochloric acid three is 2:(1.5-2):6; In the preparation steps of the compound 7j: in the step (iii), the substituted aniline A is p-toluidine, and in the step (iv), the substituted aniline B is 3-chloro-4-cyanoaniline; In the preparation steps of the compound 7t: in the step (i), the alkylamine is isopropylamine, and in the step (ii), the N-alkyl-2-substituted-4-nitrobenzamide is N-isopropyl- 2-fluoro-4-nitrobenzamide, in step (iii), substituted aniline A is N-isopropyl-2-fluoro-4-aminobenzamide, in step (iv), substituted aniline B is 3 -Trifluoromethyl-4-cyanoaniline. 3. The application of the diaryl substituted thiohydantoin compound according to claim 1 in the preparation of antitumor drugs. 4. The application according to claim 3, characterized in that: the anti-tumor drug is an anti-prostate cancer drug.