CN111763218B - A kind of thienopyrimidinone thioacetic acid derivatives and preparation method and application thereof - Google Patents

Abstract

The invention relates to a thienopyrimidinone thioacetic acid derivative and a preparation method and application thereof. The compound has the structure shown in formula I or II. The present invention also relates to a preparation method and a pharmaceutical composition containing the compound of formula I or II. The present invention also provides the application of the above-mentioned compounds in the preparation of anti-gout medicines.

Description

A kind of thienopyrimidinone thioacetic acid derivatives and preparation method and application thereof

technical field

The invention belongs to the technical field of organic compound synthesis and medical application, and in particular relates to a thienopyrimidinone thioacetic acid derivative and a preparation method and application thereof.

Background technique

Gout is a metabolic disease caused by the increased production or decreased excretion of uric acid due to the disorder of purine metabolism, and the formation of monosodium urate crystals deposited in joint tissues. Hyperuricemia is its main clinical manifestation. The end product of purine metabolism in the body is uric acid, and hyperuricemia is a metabolic disease caused by abnormal purine metabolism in the body. In recent decades, the number of patients with gout and hyperuricemia in the world has been increasing, which seriously affects the quality of human life and brings a heavy burden to the society. my country is a country with a high incidence of gout, the number of gout patients is about 1%-3%, and the number of hyperuricemia patients is 5.0%-23.4%. At present, the clinical drugs used to treat gout and hyperuricemia mainly include anti-inflammatory drugs, uric acid production inhibitors mainly based on xanthine oxidase inhibitors, and uricosuric drugs. There are serious toxic and side effects, which greatly limit the clinical use. Urate transporter 1 (URAT1) is a novel target of drugs that promote uric acid excretion. The increased URATI activity or gene expression caused by its gene mutation is one of the important pathogenesis of hyperuricemia. Lesinurad is a newly marketed oral uric acid excretion-increasing drug for the treatment of gout that inhibits the renal proximal tubule uric acid transporter URAT1. However, the compound has serious liver toxicity, so the modification of this chemical structure is of great significance for the discovery of new anti-gout drugs with high efficiency, low toxicity and independent intellectual property rights.

SUMMARY OF THE INVENTION

In view of the deficiencies of the prior art, the present invention provides a thienopyrimidinone thioacetic acid derivative and a preparation method thereof. The present invention also provides the activity screening results and application of the above compounds as anti-gout drugs.

The technical scheme of the present invention is as follows:

1. Thienopyrimidinone Thioacetic Acid Derivatives

The thienopyrimidinone thioacetic acid derivatives of the present invention have the structure shown in the following general formula I or II:

in,

R is -H, -CH ₃ or

R ₁ is -CH ₂ -,- ^* CH(CH ₃ )- or -C(CH ₃ ) ₂ -; * represents a chiral carbon atom;

R ₂ is -OH or -OCH ₃ .

Preferably according to the present invention, the thienopyrimidinone thioacetic acid derivatives are one of the following:

Table 1. Structural formulas of compounds M1～T8

Second, the preparation method of thienopyrimidinone thioacetic acid derivatives

The preparation method of thienopyrimidinone thioacetic acid derivatives of the present invention is one of the following:

(1) Preparation method of 4-oxo-3,4-dihydrothieno[3,2-d]pyrimidine target compound (I)

The preparation method of 4-oxo-3,4-dihydrothieno[3,2-d]pyrimidine target compound, the steps are as follows:

Using 4-bromo-1-naphthylamine as the starting material, the Suzuki coupling reaction with cyclopropylboronic acid produces the intermediate 4-cyclopropyl-1-naphthylamine a, which is then reacted with 1,1'-thiocarbonyldiimidazole Obtain intermediate b, then react with methyl 3-aminothiophene-2-carboxylate containing different substituents in absolute ethanol or pyridine solution to obtain intermediate I-1(ac), and then in DMF solution K ₂ CO ₃ Under the catalysis of substituted methyl bromoacetate, a nucleophilic substitution reaction occurs to obtain the target product I-2(ai), and finally in the mixed solution of tetrahydrofuran and methanol, the target product I-3(ai) is obtained by hydrolysis with lithium hydroxide or sodium hydroxide. ).

The synthetic route one is as follows:

Reagents and conditions: (i) K ₃ PO ₄ , Pd(PPh ₃ ) ₄ , cyclopropylboronic acid, toluene: water=25:2 v/v, N ₂ protection, 100°C, 12h; (ii) CH ₂ Cl ₂ , 1,1'-thiocarbonyldiimidazole, room temperature, 12h; (iii) methyl 3-aminothiophene-2-carboxylate or methyl 3-amino-5-methylthiophene-2-carboxylate, absolute ethanol, 90 ℃, 4h, methyl 3-amino-4-methylthiophene-2-carboxylate, pyridine, 50℃, 12h, NaOH, 90℃, 12h; (iv) K ₂ CO ₃ , DMF, 2-bromoisobutyric acid Methyl ester or methyl 2-bromopropionate or methyl bromoacetate, 100°C, 3h; (v) methanol, tetrahydrofuran, lithium hydroxide/sodium hydroxide, room temperature, 12h.

(2) Preparation method of 4-oxo-3,4-dihydrothieno[2,3-d]pyrimidine target compound (II)

The preparation method of 4-oxo-3,4-dihydrothieno[2,3-d]pyrimidine target compound, the steps are as follows:

Using 4-bromo-1-naphthylamine as the starting material, the Suzuki coupling reaction with cyclopropylboronic acid produces the intermediate 4-cyclopropyl-1-naphthylamine a, which is then reacted with 1,1'-thiocarbonyldiimidazole Obtain intermediate b, and then react with methyl 2-aminothiophene-3-carboxylate containing different substituents in pyridine or absolute ethanol to generate intermediate II-1(ae), then in DMF solution, in K ₂ CO ₃ Under the catalysis, nucleophilic substitution reaction with various esters to obtain the corresponding target product II-2(ao), and finally in the mixed solution of tetrahydrofuran and methanol, the target product II-3(ao) is obtained by hydrolysis with lithium hydroxide or sodium hydroxide. ).

The second synthetic route is as follows:

Reagents and conditions: (i) K ₃ PO ₄ , Pd(PPh ₃ ) ₄ , cyclopropylboronic acid, toluene: water=25:2 v/v, N ₂ protection, 100°C, 12h; (ii) CH ₂ Cl ₂ , 1,1'-thiocarbonyldiimidazole, room temperature, 12h; (iii) various substituted methyl 2-aminothiophene-3-carboxylate, absolute ethanol, 90℃, 4h, pyridine, 50℃, 12h, NaOH, 90°C, 12h; (iv) K ₂ CO ₃ , DMF, methyl 2-bromoisobutyrate or methyl 2-bromopropionate or methyl bromoacetate, 100°C, 3h; (v) methanol, tetrahydrofuran, hydrogen Lithium oxide or sodium hydroxide, room temperature, 12h.

The room temperature in the present invention refers to 20-30°C.

3. Application of Thienopyrimidinone Thioacetic Acid Derivatives

The invention discloses the screening results of thienopyrimidinone thioacetic acid derivatives for lowering blood uric acid and their first application as anti-gout drugs. It is proved by experiments that the thienopyrimidinone thioacetic acid derivatives of the present invention can be used as blood uric acid lowering drugs. Specifically, it is used as a compound for lowering blood uric acid in the preparation of anti-gout drugs. The present invention also provides the application of the above-mentioned compounds in the preparation of anti-gout medicines.

Antigout activity of target compounds

For the 48 compounds synthesized according to the above method (see Table 1 for the structural formula of the compounds), the carboxylic acid compounds were screened for their in vitro target inhibitory activity, and their target inhibitory activity data were listed in Table 2, with Lesinurad as the positive control. . It can be seen from Table 2 that compounds I-3a, I-3b, I-3e, II-3a, II-3b and II-3h all showed better URAT1 inhibitory activity, and they were all stronger than the positive control drug.

Therefore, the thienopyrimidinone thioacetic acid derivatives of the present invention are a series of URAT1 inhibitors with novel structures, which can be used as leading compounds against gout.

The thienopyrimidinone thioacetic acid derivatives of the present invention can be used as URAT1 inhibitors. Specifically, it is used as a URAT1 inhibitor for the preparation of anti-gout drugs.

An anti-gout pharmaceutical composition, comprising the thienopyrimidinone thioacetic acid derivatives of the present invention and one or more pharmaceutically acceptable carriers or excipients.

Description of drawings

Figure 1 is the mean plasma concentration-time curve of compound II-3a by oral administration and injection.

Detailed ways

The following examples are helpful to understand the present invention, but do not limit the content of the present invention. In the following examples, the numbers of all target compounds are the same as those in Table 1.

synthetic route:

Example 1. Preparation of intermediate 1-cyclopropyl-4-isothiocyanatonaphthalene (b)

Preparation of intermediate 4-cyclopropyl-1-naphthylamine (a):

The raw material 4-bromo-1-naphthylamine (3.0g, 13.50mmol) and cyclopropylboronic acid (1.5g, 17.55mmol) were mixed and placed in a 250mL double-neck flask, and K ₃ PO ₄ (10.2g) was added to the flask. , 47.10 mmol), tetrakistriphenylphosphine palladium (1.5 g, 1.35 mmol), after mixing, add about 60 mL of toluene and 5 mL of water as a solvent, seal, carry out N ₂ protection, and heat under reflux at 100 ° C for 12h reaction; TLC monitoring, to be reacted After completion, the reaction solution was cooled to room temperature, filtered, washed with ethyl acetate (20 mL×3), the filtrate was collected, and then the filtrate was dried to obtain a dark brown residue, and about 30 mL of water was added to the residue, followed by extraction with ethyl acetate (40mL×3), combine the organic phases, add an appropriate amount of anhydrous magnesium sulfate for drying, filter and dry after 2 hours to obtain crude product a, which is purified by column chromatography (petroleum ether:ethyl acetate=6:1) to obtain the corresponding intermediate The product was a brown-red oil, yield: 60%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.34-8.21 (m, 1H), 8.02 (d, J=8.3Hz, 1H), 7.48 (ddd, J=8.2, 6.6, 1.3Hz, 1H), 7.42(ddd,J＝8.3,6.7,1.4Hz,1H),7.00(d,J＝7.7Hz,1H),6.57(d,J＝7.7Hz,1H),5.58(s,2H),2.16(ddd , J=13.5, 8.4, 5.3Hz, 1H), 1.00–0.85 (m, 2H), 0.63–0.49 (m, 2H).

Preparation of Intermediate 1-Cyclopropyl-4-isothiocyanatonaphthalene (b)

Intermediate a (0.62 g, 3.36 mmol) was added to a 250 mL round-bottomed flask, then about 40 mL of dichloromethane was added to dissolve, and then 1,1'-thiocarbonyldiimidazole (1.0 g, 5.6 mmol) was added and stirred at room temperature for 12 h , monitored by TLC, after the reaction was over, the solvent was evaporated to dryness, eluted (pure petroleum ether) and purified to obtain intermediate b as a colorless oil with a yield of 80.0%. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.54-8.42 (m, 1H), 8.02 (dd, J=7.9, 2.0Hz, 1H), 7.74 (td, J=7.4, 6.7, 3.5Hz, 2H) ),7.55(d,J＝7.7Hz,1H),7.24(d,J＝7.7Hz,1H),2.42(td,J＝8.4,4.3Hz,1H),1.13–1.02(m,2H),0.73 (dd,J=5.5,1.8Hz,2H).

Example 2. Preparation of intermediate 3-(4-cyclopropylnaphthalen-1-yl)-2-mercaptothieno[3,2-d]pyrimidin-4(3H)-one (I-1a)

Intermediate b (0.507 g, 2.25 mmol) and methyl 3-aminothiophene-2-carboxylate (0.307 g, 1.95 mmol) were dissolved in 20 mL of absolute ethanol, heated under reflux at 90 °C for 4 h; TLC monitoring showed that the reaction was complete After cooling to room temperature, crystals were precipitated, filtered, the filter cake was taken, a small amount of ethanol was added for recrystallization, the solid was taken out by filtration, and 30 mL of 2mol/L KOH solution was added, heated to reflux at 90 °C until the compound was dissolved, stopped heating, cooled to room temperature, At this time, a small amount of flocs were precipitated, and then the pH was adjusted to about 3 with 3mol/L HCl, and a large amount of white solids were precipitated. At this time, slowly filtered, washed with clean water (10 mL×2), and dried to obtain intermediate I-1a, which is White solid, 18.3% yield; melting point: >250°C; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.63 (s, 1H), 8.48 (d, J=8.4 Hz, 1H), 8.26 (d , J=5.3Hz, 1H), 7.72–7.57 (m, 2H), 7.54–7.46 (m, 1H), 7.44–7.30 (m, 2H), 7.15 (d, J=5.3Hz, 1H), 2.47 ( dd, J=8.3, 5.5Hz, 1H), 1.12 (d, J=8.4Hz, 2H), 0.90–0.76 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ176.91, 157.34, 146.57, 140.18, 138.29, 134.64, 134.07, 129.88, 127.22, 127.03, 126.53, 125.17, 123.51, 123.29, 117.98, 115.74, 13.34, 7.51, 7.23.ESI-MS: m/z _{34 H 14 N} [MH] ^- , ₂ OS ₂ [350.05]

Example 3. Intermediate 3-(4-Cyclopropylnaphthalen-1-yl)-2-mercapto-7-methylthieno[3,2-d]pyrimidin-4(3H)-one (I-1b ) preparation

Intermediate b (0.224g, 0.99mmol) and reactant methyl 3-amino-5-methylthiophene-2-carboxylate (0.135g, 0.79mmol) were dissolved in about 5mL of pyridine solution, heated at 45°C for 12h, TLC monitoring, after the reaction was completed, cooled to room temperature, added about 40 mL of ice water, extracted with dichloromethane (40 mL × 3), combined the organic phases, dried over anhydrous magnesium sulfate, filtered, took the filtrate, evaporated to dryness, and reused 30mL of 1% NaOH solution was heated to reflux at 90°C for 12h, cooled to room temperature, filtered, the filtrate was taken, and then the pH was adjusted to about 3 with 1mol/L hydrochloric acid, a large amount of white solid was precipitated, filtered, and the filter cake was washed with water, Intermediate I-1b was obtained as white solid, yield 53.0%; melting point: >250°C; ¹ H NMR (400MHz, DMSO-d ₆ )δ13.40(s, 1H), 8.48(d, J=8.5Hz ,1H),7.90(s,1H),7.64–7.54(m,2H),7.52–7.46(m,1H),7.36(q,J=7.6Hz,2H),2.47(dd,J=8.3,5.5 Hz, 1H), 2.39(s, 3H), 1.18–1.09(m, 2H), 0.83(q, J=8.0, 5.4Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ177.48,157.36, ^{145.70,140.15,134.74,134.12,133.23,129.87,127.66,127.23,127.01,126.51,125.19,123.53,123.25,116.12,13.78,13.36,7.52,7.26.3} ₂₀ H ₁₆ N ₂ OS ₂ [364.07].

Example 4. Intermediate 3-(4-Cyclopropylnaphthalen-1-yl)-2-mercapto-6-methylthieno[3,2-d]pyrimidin-4(3H)-one (I-1c ) preparation

The preparation method of this intermediate is the same as that of Example 3, and it is a white solid with a yield of 50.0%; melting point: 209-211°C; ¹ HNMR (400MHz, DMSO-d ₆ )δ13.53(s, 1H), 8.47(d, J=8.4Hz, 1H), 7.65–7.53 (m, 2H), 7.51–7.45 (m, 1H), 7.34 (q, J=7.6Hz, 2H), 6.91 (s, 1H), 2.61 (s, 3H) ), 2.48–2.42 (m, 1H), 1.11 (d, J=8.6Hz, 2H), 0.81 (tt, J=12.1, 7.6Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ176. 88,156.81,152.88,146.72,140.14,134.69,134.09,129.91,127.24,127.00,126.49,125.17,123.47,123.25,116.53,113.54,16.66,13.33,7.50,7.24.ESI-MS:m/z 363.3[MH] ^- , C ₂₀ H ₁₆ N ₂ OS ₂ [364.07].

Example 5. Preparation of compound I-2a

Intermediate I-1a (0.17 mmol) and K ₂ CO ₃ (0.61 mmol) were mixed in a 25 mL round-bottomed flask, dissolved in about 8 mL of DMF, heated at 90° C. for 5 min, and then added dropwise methyl bromoacetate (0.30 mmol), continued to heat to 100 °C, refluxed for 3 h, monitored by TLC, after the reaction was complete, the system was cooled to room temperature, and ice water was slowly added to the reaction solution, the solution first became clear and then turbid, and added dropwise until there was no new solid Generate, filter, wash the filter cake with water, take the filter cake and dry to obtain the target product I-2a; white solid, yield: 85.2%, melting point: 157-159 ° C.

Spectral data of compound I-2a: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.4 Hz, 1H), 8.28 (d, J=5.2 Hz, 1H), 7.69 (t, J= 7.6Hz, 1H), 7.66–7.52 (m, 2H), 7.43 (d, J=7.9Hz, 2H), 7.38 (d, J=5.3Hz, 1H), 4.03–3.85 (m, 2H), 3.64 ( s,3H),2.56(dq,J=8.2,4.2,2.7Hz,1H),1.15(dq,J=8.5,3.2,2.8Hz,2H),0.86(dq,J=14.0,9.2,7.4Hz, 2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ169.09,159.78,157.66,156.64,142.80,136.96,134.13,130.47,129.67,128.48,128.05,127.26,125.48,125.14,123.22,122.60,119.45,52.85, 34.61, 13.42, 7.83, 7.54. ESI-MS: m/z 423.3 [M+H] ⁺ , C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 6. Preparation of compound 1-2b

The procedure is the same as in Example 5, except that the ester used is methyl 2-bromopropionate, white solid, yield: 75.5%, melting point: 165-167°C.

Spectral data of compound I-2b: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.4 Hz, 1H), 8.29 (d, J=5.2 Hz, 1H), 7.69 (t, J= 7.5Hz, 1H), 7.60 (dq, J=14.0, 7.0, 6.3Hz, 2H), 7.50–7.31 (m, 3H), 4.44 (dq, J=14.7, 7.3Hz, 1H), 3.64 (d, J = 18.9Hz, 3H), 2.63–2.54 (m, 1H), 1.37 (dd, J=13.4, 7.3Hz, 3H), 1.22–1.08 (m, 2H), 0.87 (ddd, J=29.7, 9.9, 5.3 Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ172.06,159.13,157.65,156.60,142.82,137.01,134.10,130.40,129.64,128.46,128.05,127.27,125.49,125.07,12 53.01, 44.38, 17.25, 13.40, 7.94, 7.49. ESI-MS: m/z 437.4[M+H] ⁺ , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 7. Preparation of compound I-2c

The operation is the same as in Example 5, except that the ester used is methyl 2-bromoisopropionate, white solid, yield 68.7%, melting point: 198-201°C.

Spectral data of compound I-2c: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.4 Hz, 1H), 8.27 (d, J=5.3 Hz, 1H), 7.69 (t, J= 7.6Hz, 1H), 7.59(dd, J=7.6, 4.1Hz, 2H), 7.39(dd, J=7.9, 4.5Hz, 2H), 7.27(d, J=5.2Hz, 1H), 3.66(s, 3H), 2.56(dd, J=9.6, 4.3Hz, 1H), 1.52(s, 3H), 1.46(s, 3H), 1.22–1.08(m, 2H), 0.97–0.75(m, 2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ173.86,158.99,157.58,156.48,142.70,137.08,134.10,130.37,129.56,128.36,128.08,127.24,125.49,124.74,123.15,122.42,119.34,53.28,53.07,26.03, 25.80, 13.39, 8.04, 7.42. ESI-MS: m/z 451.3 [M+H] ⁺ , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 8. Preparation of compound I-2d

The operation is the same as in Example 5, except that the intermediate I-1b is used, white solid, yield: 81.7%, melting point: 96-98°C.

Spectral data of compound I-2d: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.5 Hz, 1H), 7.93 (s, 1H), 7.69 (t, J=7.6 Hz, 1H) ,7.64–7.54(m,2H),7.42(t,J=7.8Hz,2H),3.90(d,J=3.7Hz,2H),3.65(s,3H),2.56(dq,J=8.2,4.2 , 2.8Hz, 1H), 2.33(s, 3H), 1.23-1.10(m, 2H), 0.87(h, J=9.7, 8.4Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169. 29,159.67,157.81,155.45,142.79,134.15,133.49,131.65,130.51,129.66,128.42,128.04,127.25,125.47,123.21,122.59,119.17,52.78,34.86,13.42,12.56,7.82,7.55.ESI-MS:m/ z 437.4[M+H] ⁺ , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 9. Preparation of compound I-2e

The procedure is the same as in Example 8, except that the ester used is methyl 2-bromopropionate, white solid, yield: 80.0%, melting point: 152-154°C.

Spectral data of compound I-2e: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.5 Hz, 1H), 7.93 (s, 1H), 7.69 (t, J=7.5 Hz, 1H) ,7.64–7.52(m,2H),7.47–7.32(m,2H),4.35(dq,J=14.8,7.3Hz,1H),3.64(d,J=11.5Hz,3H),2.62–2.53(m , 1H), 2.34(s, 3H), 1.39(t, J=6.8Hz, 3H), 1.21–1.09(m, 2H), 0.96–0.77(m, 2H). ¹³ C NMR (100MHz, DMSO-d) ₆ )δ172.34,158.99,157.77,155.48,142.77,134.12,133.56,131.63,130.47,129.63,128.41,128.07,127.25,125.49,123.18,122.47,119.25,52.95,44.41,16.47,13.40,12.62,7.94,7.50. ESI-MS: m/z 451.4 [M+H] ⁺ , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 10. Preparation of compound 1-2f

The operation was the same as that of Example 8, except that the ester used was methyl 2-bromoisopropionate, white solid, yield: 83.0%, melting point: 130-132°C.

Spectral data of compound I-2f: ¹ H NMR (400MHz, Chloroform-d) δ 8.52 (d, J=8.5Hz, 1H), 7.60 (t, J=7.5Hz, 1H), 7.54-7.47 (m, 1H) ), 7.42(d, J=7.8Hz, 3H), 7.37(d, J=7.6Hz, 1H), 3.71(s, 3H), 2.45(td, J=8.3, 4.0Hz, 1H), 2.39(s , 3H), 1.59(s, 3H), 1.56(s, 3H), 1.14(dd, J=8.4, 4.3Hz, 2H), 0.88(dq, J=22.5, 5.6Hz, 2H). ¹³ C NMR( 100MHz,DMSO-d ₆ )δ173.91,158.74,157.74,155.55,142.68,134.11,133.75,131.53,130.43,129.57,128.35,128.06,127.23,125.49,123.14,122.41,119.12,53.13,53.06,26.00,25.73,13.38 , 12.74, 8.04, 7.41. ESI-MS: m/z 465.3 [M+H] ⁺ , C ₂₅ H ₂₄ N ₂ O ₃ S ₂ [464.12].

Example 11. Preparation of compound I-2g

The operation is the same as in Example 5, except that the intermediate used is I-1c, white solid, yield: 86.4%, melting point: 179-181°C.

Spectral data of compound I-2g: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.4 Hz, 1H), 7.69 (q, J=7.5 Hz, 1H), 7.63-7.54 (m, 2H), 7.40(t, J=7.4Hz, 2H), 7.12(s, 1H), 3.98-3.84(m, 2H), 3.63(s, 3H), 2.63(s, 3H), 2.59-2.52(m ,1H),1.15(d,J=8.3Hz,2H),0.90-0.81(m,2H). ¹³ CNMR(100MHz,DMSO-d ₆ )δ169.07,159.74,157.08,156.89,151.24,142.75,134.14,130.54 ,129.70,128.44,128.02,127.23,125.46,123.56,123.18,122.57,117.74,52.81,34.59,16.76,13.41,7.80,7.55.ESI-MS:m/z 437.3[ _M +H] ⁺ ,0 ₂₃ H N ₂ O ₃ S ₂ [436.09].

Example 12. Preparation of compound I-2h

The procedure was the same as in Example 11, except that the ester used was methyl 2-bromopropionate, white solid, yield: 62.7%, melting point: 178-180°C.

Spectral data of compound I-2h: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.4 Hz, 1H), 7.63-7.53 (m, 2H), 7.38(dd, J＝18.1, 7.5Hz, 2H), 7.12(s, 1H), 4.38(dt, J＝14.6, 7.5Hz, 1H), 3.63(d, J＝18.4Hz, 3H), ¹³ C NMR(100MHz,DMSO-d ₆ )δ172.14,159.03,157.07,156.85,151.30,142.77,138.45,134.11,129.67,128.43,128.07,127.24,125.48,123.49,123.17,122.53,117.83,52.98,44.37,17.18, 16.75, 13.39, 7.92, 7.50. ESI-MS: m/z 451.4 [M+H] ⁺ , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 13 Preparation of compound I-2i

The procedure was the same as in Example 11, except that the ester used was methyl 2-bromoisopropionate, white solid, yield: 66.5%, melting point: 176-178°C.

Spectral data of compound I-2i: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.53 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H), 7.57 (dd, J= 14.0, 7.4Hz, 2H), 7.43–7.32(m, 2H), 7.04(s, 1H), 3.66(s, 3H), 2.62(s, 3H), 2.58–2.52(m, 1H), 1.50(s , 3H), 1.44(s, 3H), 1.15(d, J=7.4Hz, 2H), 0.97–0.75(m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ173.84, 158.96, 157.02, 156.74 ,151.42,142.65,134.10,130.44,129.59,128.33,128.05,127.22,125.48,123.21,123.12,122.39,117.6353.24,53.05,26.03,25.79,16.71,13.2-MS:m/z 465.3[M+H] ⁺ ,C ₂₅ H ₂₄ N ₂ O ₃ S ₂ [464.12].

Example 14. Preparation of compound 1-3a

Compound I-2a (40 mg) was dissolved in a mixed solution of 5 mL of methanol and 2.5 mL of THF, and after stirring and dissolving, a newly prepared LiOH solution (210 mg of LiOH was dissolved in 5 mL of water to prepare a 2M lithium hydroxide solution) was added for about 1- 2mL, stirred and reacted at room temperature for 12h, monitored by TLC, after the reaction was complete, add 5mL of water, evaporate to dryness methanol and THF in the system, and then use 1mol/L HCl to adjust the pH of the solution to about 3, precipitate solids, filter, and wash with water , and dried to obtain the target compound as a white solid, yield: 88.3%, melting point: 134-136°C.

Spectral data of compound I-3a: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.86 (s, 1H), 8.55 (d, J=8.4 Hz, 1H), 8.27 (d, J=5.3 Hz, 1H) ,7.68(t,J＝7.5Hz,1H),7.62(d,J＝7.6Hz,1H),7.59–7.54(m,1H),7.42(dt,J＝10.0,4.5Hz,3H),3.88( ^13C NMR (100MHz,DMSO-d ₆ )δ169.69,160.07,157.71,156.74,142.72,136.88,134.13,130.57,129.72,128.47,128.01,127.22,125.47,125.16,123.24,122.64,119.38,35.22,13.43,7.81,7.53. ESI-MS: m/z 407.4 [MH] ^- , C ₂₁ H ₁₆ N ₂ O ₃ S ₂ [408.06].

Example 15. Preparation of compound 1-3b

The procedure was the same as in Example 14, except that the hydrolyzed compound was I-2b, white solid, yield: 91.8%, melting point: 152-155°C.

Spectral data of compound I-3b: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.92 (s, 1H), 8.55 (d, J=8.5 Hz, 1H), 8.28 (d, J=5.2 Hz, 1H) ,7.68(t,J=7.5Hz,1H),7.58(dt,J=13.4,7.3Hz,2H),7.49–7.31(m,3H),4.41(p,J=7.0Hz,1H),2.64– 2.53(m, 1H), 1.40(dd, J=24.6, 7.2Hz, 3H), 1.15(d, J=8.4Hz, 2H), 0.98–0.74(m, 2H). ¹³ C NMR (100MHz, DMSO- d ₆ )δ172.83,159.33,157.72,156.72,142.71,136.87,134.10,130.50,129.69,128.46,128.05,127.22,125.48,125.13,123.23,122.57,119.50,45.01,17.87,13.41,7.89,7.48.ESI-MS : m/z 421.3 [MH] ^- , C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 16. Preparation of compound 1-3c

The procedure is the same as in Example 14, except that the hydrolyzed compound is I-2c. White solid, yield: 86.6%, melting point: 158-161°C.

Spectral data of compound I-3c: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.65 (s, 1H), 8.54 (d, J=8.5 Hz, 1H), 8.26 (d, J=5.3 Hz, 1H) ,7.68(t,J＝7.6Hz,1H),7.57(dd,J＝7.5,4.7Hz,2H),7.39(t,J＝7.7Hz,2H),7.28(d,J＝5.2Hz,1H) ,2.55(dq,J＝8.3,4.2,2.8Hz,1H),1.52(s,3H),1.48(s,3H),1.13(dd,J＝15.0,8.6Hz,2H),0.98–0.76(m ,2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ174.73,159.38,157.65,156.58,142.57,136.88,134.10,130.51,129.63,128.35,127.98,127.18,125.48,124.81,123.17,122.44,119.28,53.53 , 26.14, 25.89, 13.38, 8.02, 7.41. ESI-MS: m/z 435.3 [MH] ^- , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 17. Preparation of Compound I-3d

The operation is the same as in Example 14, except that the hydrolyzed compound is I-2d, white solid, yield: 91.4%, melting point: 139-141°C.

Spectral data of compound I-3d: ¹ H NMR (400MHz, DMSO-d ₆ )δ12.75(s, 1H), 8.48(d, J=8.5Hz, 1H), 7.86(s, 1H), 7.61(t, J=7.6Hz, 1H), 7.56–7.45 (m, 2H), 7.35 (dd, J=7.7, 5.6Hz, 2H), 3.77 (s, 2H), 2.53–2.46 (m, 1H), 2.29 (s , 3H), 1.13–1.03(m, 2H), 0.79(tt, J=9.8, 6.0Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169.87,159.87,157.89,155.55,142.69,134.13, _{133.58,131.57,130.60,129.69,128.39,128.00,127.22,125.47,123.23,122.63,119.04,35.39,13.42,12.73,7.81,7.54.ESI} -MS:m/z 421.3 H ₁₈ N [MH] ^- , ₂ O ₃ S ₂ [422.08].

Example 18. Preparation of compound I-3e

The operation is the same as that of Example 14, except that the hydrolyzed compound is I-3e, white solid, yield: 95.7%, melting point: 210-213°C.

Spectral data of compound I-3e: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.79 (s, 1H), 8.48 (d, J=8.5 Hz, 1H), 7.86 (s, 1H), 7.61 (t, J=7.6Hz, 1H), 7.51 (q, J=9.0, 8.2Hz, 2H), 7.33 (dd, J=13.1, 8.7Hz, 2H), 4.24 (dq, J=14.7, 7.3Hz, 1H), 2.49(dq,J=8.2,4.2,2.7Hz,1H),2.30(s,3H),1.33(dd,J=18.0,7.3Hz,3H),1.07(dd,J=13.4,8.7Hz,2H) ,0.88–0.70(m,2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ173.12,159.25,157.86,155.59,142.68,134.11,133.64,131.53,130.56,129.67,128.39,127.98,127. ,122.55,119.13,45.36,17.19,13.41,12.78,7.90,7.49.ESI-MS: m/z435.4[MH] ^- ,C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 19. Preparation of compound 1-3f

The operation is the same as that of Example 14, except that the hydrolyzed compound is I-2f. White solid, yield: 93.4%, melting point: 242-245°C.

Spectral data of compound I-3f: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.54(s, 1H), 8.54(d, J=8.4Hz, 1H), 7.90(s, 1H), 7.68(t, J=7.6Hz, 1H), 7.56(t, J=8.0Hz, 2H), 7.38(dd, J=14.4, 8.0Hz, 2H), 2.56(d, J=5.1Hz, 1H), 2.35(s, 3H), 1.51 (d, J=19.9Hz, 6H), 1.19–1.11 (m, 2H), 0.95–0.77 (m, 2H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 174.76, 159.05, 157.83 155.70 ,142.57,134.10,133.94,131.39,130.53,129.60,128.31,127.99,127.20,125.49,123.18,122.44,118.91,53.50,26.10,25.81,13.38,12.7.4 MH] ^- ,C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 20. Preparation of compound 1-3g

The operation is the same as in Example 14, except that the hydrolyzed compound is I-2g, white solid, yield: 51.7%, melting point: 218-221°C.

Spectral data of compound I-3g: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.84 (s, 1H), 8.55 (d, J=8.4 Hz, 1H), 7.73-7.65 (m, 1H), 7.58 ( dd,J=14.2,7.3Hz,2H),7.41(dd,J=7.9,3.0Hz,2H),7.15(s,1H),3.87(s,2H),2.63(s,3H),2.56(dt , J=10.8, 5.8Hz, 1H), 1.16 (d, J=8.6Hz, 2H), 0.95–0.79 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169.67, 159.99, 157.13, 156.98 ,151.15,142.67,134.13,130.62,129.74,128.44,127.98,127.20,125.45,123.59,123.20,122.60,117.69,35.13,16.76,13.41,7.79,7.54.ESIMH ^- MS:m/z 42.3 C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 21. Preparation of compound I-3h

The operation is the same as in Example 14, except that the hydrolyzed compound is I-2h, white solid, yield: 89.9%, melting point: 230-233°C.

Spectral data of compound I-3h: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.97 (s, 1H), 8.54 (d, J=8.5Hz, 1H), 7.68 (t, J=7.6Hz, 1H) ,7.57(q,J＝6.7Hz,2H),7.39(t,J＝8.4Hz,2H),7.15(s,1H),4.37(p,J＝6.8Hz,1H),2.63(s,3H) , 2.59–2.53 (m, 1H), 1.38 (dd, J=24.3, 7.2Hz, 3H), 1.14 (d, J=8.4Hz, 2H), 0.95–0.78 (m, 2H). ¹³ C NMR (100MHz) ,DMSO-d ₆ )δ172.92,159.28,157.14,156.97,151.22,142.66,134.10,130.55,129.70,128.44,127.98,127.21,125.49,123.57,123.18,122.53,117.75,44.96,17.79,16.77,13.41,7.87, 7.49. ESI-MS: m/z 435.3 [MH] ^- , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 22. Preparation of Compound I-3i

The operation is the same as in Example 14, except that the hydrolyzed compound is I-2i, white solid, yield: 95.2%, melting point: 241-244°C.

Spectral data of compound I-3i: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.65 (s, 1H), 8.53 (d, J=8.4 Hz, 1H) ,7.67(t,J＝7.3Hz,1H),7.55(dd,J＝14.2,7.7Hz,2H),7.36(dd,J＝14.0,8.0Hz,2H),7.04(s,1H),2.62( ^13C NMR (100MHz,DMSO-d ₆ )δ174.75,159.73,157.11,156.87,151.16,142.47,134.10,130.60,129.65,128.32,127.96,127.16,125.48,123.34,123.17,122.42,117.53,67.77,26.11,23.29,16.71, 13.38, 7.99, 7.41. ESI-MS: m/z 449.4 [MH] ^- , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 23. Preparation of intermediate 3-(4-cyclopropylnaphthalen-1-yl)-2-mercaptothieno[2,3-d]pyrimidin-4(3H)-one (II-1a)

The operation is the same as in Example 2, except that methyl 2-aminothiophene-3-carboxylate is used, white solid, yield: 15.1%, melting point: >250°C; ¹ H NMR (400MHz, DMSO-d ₆ )δ13. 92(s, 1H), 8.47(d, J=8.3Hz, 1H), 7.67–7.54 (m, 2H), 7.52–7.44 (m, 1H), 7.35 (q, J=7.7Hz, 3H), 7.26 (d, J=5.6Hz, 1H), 2.46 (dd, J=8.3, 5.5Hz, 1H), 1.11 (d, J=8.4Hz, 2H), 0.91–0.70 (m, 2H). ¹³ C NMR( 100MHz,DMSO-d ₆ )δ175.81,157.46,151.92,140.10,134.75,134.09,129.86,127.12,126.97,126.51,125.15,123.51,123.33,122.89,120.50,118.79,13.34,7.50,7.23.ESI-MS:m /z 349.4[MH] ^- ,C ₁₉ H ₁₄ N ₂ OS ₂ [350.05].

Example 24. Intermediate 3-(4-Cyclopropylnaphthalen-1-yl)-2-mercapto-6-methylthieno[2,3-d]pyrimidin-4(3H)-one (II-1b ) preparation

The operation is the same as in Example 2, except that methyl 2-amino-5-methylthiophene-3-carboxylate is used, white solid, yield: 16.5%, melting point: >250°C; ¹ H NMR (400MHz, DMSO -d ₆ )δ13.89(s,1H),8.47(d,J=8.5Hz,1H),7.64-7.53(m,2H),7.51-7.45(m,1H),7.34(s,2H), 6.98(s, 1H), 2.48(s, 1H), 2.46(s, 3H), 1.12(d, J=8.6Hz, 2H), 0.87–0.76(m, 2H). ¹³ C NMR (100MHz, DMSO- d ₆ )δ175.34,157.07,150.47,140.05,134.78,134.09,133.87,129.83,127.12,126.97,126.49,125.15,123.50,123.28,120.13,118.51,15.31,13.34,7.49,7.22.ESI-MS:m/z 363.3[MH] ^- ,C ₂₀ H ₁₆ N ₂ OS ₂ [364.07].

Example 25. Intermediate 3-(4-Cyclopropylnaphthalen-1-yl)-2-mercapto-5-methylthieno[2,3-d]pyrimidin-4(3H)-one (II-1c ) preparation

The operation is the same as in Example 2, except that methyl 2-amino-4-methylthiophene-3-carboxylate is used, white solid, yield: 16.3%, melting point: >250°C; ¹ H NMR (400MHz, DMSO -d ₆ )δ13.85(s,1H),8.47(d,J=8.6Hz,1H),7.65-7.57(m,2H),7.52-7.46(m,1H),7.39-7.31(m,2H) ),6.95(s,1H),2.47(dd,J=8.4,5.4Hz,1H),2.32(s,3H),1.12(d,J=8.5Hz,2H),0.80(dq,J=24.7, 5.8Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ175.68, 158.06, 152.86, 139.99, 134.78, 134.69, 134.08, 129.96, 127.13, 126.94, 126.46, 125.11, 123.45. , 15.96, 13.33, 7.19. ESI-MS: m/z 363.3 [MH] ^- , C ₂₀ H ₁₆ N ₂ OS ₂ [364.07].

Example 26. Intermediate 3-(4-Cyclopropylnaphthalen-1-yl)-2-mercapto-5,6-dimethylthieno[2,3-d]pyrimidin-4(3H)-one ( Preparation of II-1d)

The operation is the same as in Example 2, except that 2-amino,4,5-dimethylthiophene-3-carboxylate methyl ester is used as a white solid, yield: 14.5%, melting point: >250°C; ¹ H NMR (400MHz) ,DMSO-d ₆ )δ13.77(s,1H),8.46(d,J=8.5Hz,1H),7.65–7.57(m,1H),7.55(d,J=8.3Hz,1H),7.52– 7.42(m, 1H), 7.32(s, 2H), 2.45(dd, J=8.3, 5.5Hz, 1H), 2.32(s, 3H), 2.23(s, 3H), 1.09(dd, J=18.8, 7.7Hz, 2H), 0.89–0.71 (m, 2H), ESI-MS: m/z 377.4 [MH] ^- , C ₂₁ H ₁₈ N ₂ OS ₂ [378.09].

Example 27. Intermediate 3-(4-Cyclopropylnaphthalen-1-yl)-2-mercapto-5,6,7,8 Tetrahydrobenzo[4,5]thieno[2,3-d] Preparation of Pyrimidine-4(3H)-one (II-1e)

The operation is the same as in Example 3, except that methyl 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate is used, white solid, yield: 35.9%, melting point: >250℃; ¹ H NMR (400MHz, DMSO-d ₆ ) δ 13.84 (s, 1H), 8.47 (d, J=8.5Hz, 1H), 7.59 (dd, J=18.0, 8.3Hz, 2H), 7.52–7.45 (m, 1H), 7.33 (s, 2H), 2.71 (s, 4H), 2.47 (d, J=7.0Hz, 1H), 1.88–1.67 (m, 4H), 1.13 (t, J= 9.7Hz, 2H), 0.91–0.72 (m, 2H). ¹³ CNMR (100MHz, DMSO-d ₆ )δ175.09, 157.59, 139.96, 134.79, 134.08, 131.56, 119.93, 128.98, 127.12, 126.94, 126.46, 125. ,123.35,116.46,25.27,24.47,22.94,22.03,13.32,7.50,7.18.ESI-MS: m/z 403.5[MH] ^- ,C ₂₃ H ₂₀ N ₂ OS ₂ [404.10].

Example 28. Preparation method of target compound II-2a

The operation is the same as in Example 5, the difference is that this example selects Intermediate II-1a, white solid, yield: 88.2%, melting point: 142-144°C.

Spectral data of compound II-2a: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.5 Hz, 1H), 7.69 (t, J=7.5 Hz, 1H), 7.65-7.52 (m, 3H), 7.49–7.32 (m, 3H), 4.01–3.84 (m, 2H), 3.64 (s, 3H), 2.62–2.52 (m, 1H), 1.23–1.08 (m, 2H), 0.87 (dt, J=9.6, 5.0Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ 169.03, 163.61, 158.93, 157.79, 142.76, 134.14, 130.58, 129.56, 128.14, 128.03, 127.26, 125.47, 123.259, 125.47 ,122.57,120.98,52.89,34.60,13.41,7.82,7.55.ESI-MS: m/z 423.3[M+H] ⁺ ,C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 29. Preparation of Compound II-2b

The operation is the same as in Example 28, the difference is that this example is selected from methyl 2-bromopropionate. White solid, yield: 67.0%, melting point: 200-203°C.

Spectral data of compound II-2b: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.4 Hz, 1H), 7.69 (t, J=7.3 Hz, 1H), 7.61 (d, J= 8.1Hz, 1H), 7.57 (d, J=5.7Hz, 2H), 7.46 (d, J=8.4Hz, 1H), 7.40 (d, J=5.7Hz, 2H), 4.40 (dq, J=14.6, 7.2Hz, 1H), 3.64 (d, J=17.0Hz, 3H), 2.59–2.52 (m, 1H), 1.37 (dd, J=10.8, 7.3Hz, 3H), 1.15 (d, J=8.2Hz, 2H),0.96–0.78(m,2H). ¹³ C NMR (100MHz, Chloroform-d)δ172.58,172.18,158.24,142.74,134.59,127.50,127.44,127.28,126.70,125.41,123.61,122.16,12 121.98, 121.48, 121.07, 52.72, 44.26, 16.82, 13.57, 7.03, 6.65. ESI-MS: m/z 437.4[M+H] ⁺ , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 30. Preparation of Compound II-2c

The operation is the same as in Example 28, except that this example is selected from methyl 2-bromoisopropionate. White solid, yield: 76.8%, melting point: 204-206°C.

Spectral data of compound II-2c: ¹ H NMR (400 MHz, Chloroform-d) δ 8.52 (d, J=8.4 Hz, 1H), 7.63-7.57 (m, 1H), 7.54-7.49 (m, 1H), 7.46 –7.41(m,2H),7.41–7.35(m,2H),7.13(d,J=5.7Hz,1H),3.77(s,3H),2.44(p,J=8.4Hz,1H),1.55( d, J=10.9Hz, 6H), 1.19–1.10 (m, 2H), 0.95–0.82 (m, 2H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 173.78, 163.32, 158.11, 157.73, 142.66, 134.11 ,130.45,129.46,128.25,128.06,127.24,125.48,123.47,123.16,122.57,122.43,120.99,53.41,53.14,26.06,25.84,13.38,8.03,7.42.ESI-HMS:m/z ⁺ ,C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 31. Preparation of compound II-2d

The operation is the same as in Example 5, except that this example uses intermediate II-1b, white solid, yield: 68.5%, melting point: 194-196°C.

Spectral data of compound II-2d: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.52 (d, J=8.5 Hz, 1H), 7.66 (t, J=7.6 Hz, 1H), 7.56 (t, J= 8.8Hz, 2H), 7.39(t, J=7.2Hz, 2H), 7.08(s, 1H), 3.97–3.81(m, 2H), 3.61(s, 3H), 2.54(dd, J=8.4, 5.4 Hz, 1H), 2.49–2.45 (m, 3H), 1.24–1.03 (m, 2H), 0.83 (tt, J=9.4, 5.7Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169. 04,162.52,158.08,157.32,142.70,136.81,134.14,130.64,129.54,128.29,128.01,127.24,125.47,123.21,122.56,121.18,119.95,52.86,34.56,15.96,13.41,7.80,7.54.ESI-MS:m/ z 437.4[M+H] ⁺ , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 32. Preparation of Compound II-2e

The operation is the same as in Example 31, except that this example selects methyl 2-bromopropionate, white solid, yield: 70.1%, melting point: 100-102°C.

Spectral data of compound II-2e: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.52 (d, J=8.5 Hz, 1H), 7.66 (t, J=7.6 Hz, 1H), 7.55 (dd, J= 11.7, 5.7Hz, 2H), 7.45–7.30 (m, 2H), 7.08 (s, 1H), 4.42–4.29 (m, 1H), 3.61 (d, J=17.2Hz, 3H), 2.54 (dd, J =8.5,5.2Hz,1H),2.48(s,3H),1.33(dd,J=10.0,7.4Hz,3H),1.13(q,J=5.1Hz,2H),0.93–0.75(m,2H) . ¹³ C NMR(100MHz,DMSO-d ₆ )δ172.10,172.00,162.41,157.31,142.69,136.91,134.11,130.51,129.52,128.28,128.01,127.25,125.48,123.20,122.45,121.29,119.96,53.02,44.34, 17.13, 15.97, 13.39, 7.93, 7.48. ESI-MS: m/z 451.2 [M+H] ⁺ , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 33. Preparation of Compound II-2f

The operation is the same as that of Example 31, the difference is that this example selects methyl 2-bromoisopropionate, white solid, yield: 73.22%, melting point: 182-184°C.

Spectral data of compound II-2f: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.53 (d, J=8.5 Hz, 1H), 7.68 (t, J=7.6 Hz, 1H), 7.57 (dd, J= 15.5, 7.7Hz, 2H), 7.44–7.33(m, 2H), 7.09(s, 1H), 3.65(s, 3H), 2.56(dd, J=8.3, 5.5Hz, 1H), 2.51(s, 3H) ), 1.47(d, J=26.2Hz, 6H), 1.15(q, J=6.6Hz, 2H), 0.97–0.74(m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ173.77,162.22, 157.27,142.61,136.84,134.12,130.53,129.45,128.19,128.03,127.22,125.48,123.13,122.39,121.19,119.93,53.36,53.09,26.07,25.85,15.95,13.37,8.02,7.41.ESI-MS:m/ z 465.3[M+H] ⁺ ,C ₂₅ H ₂₄ N ₂ O ₃ S ₂ [464.12].

Example 34. Preparation of Compound II-2g

The operation is the same as that of Example 5, except that this example selects intermediate II-1g, white solid, yield: 59.1%, melting point: 108-110°C.

Spectral data of compound II-2g: ¹ H NMR (400 MHz, Chloroform-d) δ 8.51 (d, J=8.4 Hz, 1H), 7.60 (t, J=7.4 Hz, 1H), 7.52 (q, J=8.3 Hz, 2H), 7.46–7.35 (m, 2H), 6.73 (s, 1H), 3.96–3.78 (m, 2H), 3.73 (s, 3H), 2.51 (s, 3H), 2.42 (td, J= 8.3, 4.3Hz, 1H), 1.14 (dt, J=8.1, 3.8Hz, 2H), 0.97–0.74 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169.01, 164.16, 158.67, 158.41, 142.66,134.42,134.14,130.56,129.69,128.37,128.00,127.21,125.43,123.19,122.65,119.26,117.78,52.87,34.51,16.49,13.40,7.80,437.3 SI-MS ] ⁺ ,C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 35. Preparation of Compound II-2h

The operation is the same as in Example 34, the difference is that this example is selected from methyl 2-bromopropionate. White solid, yield: 75.2%, melting point: 101-103°C.

Spectral data of compound II-2h: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.5 Hz, 1H), 7.68 (t, J=7.6 Hz, 1H), 7.58 (q, J= 8.7, 7.5Hz, 2H), 7.51–7.37 (m, 2H), 7.14 (s, 1H), 4.39 (dq, J=14.6, 7.2Hz, 1H), 3.63 (d, J=16.6Hz, 3H), 2.55(dq,J=8.3,4.2,2.9Hz,1H),2.40(s,3H),1.36(dd,J=10.1,7.4Hz,3H),1.15(q,J=5.1Hz,2H),0.97 –0.74(m,2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ172.07,164.07,158.41,158.02,142.65,134.44,134.11,130.50,128.35,129.66,128.01,127.22,125.85,123 ,117.86,52.99,44.12,17.16,16.49,13.38,7.92,7.51.ESI-MS: m/z 451.4[M+H] ⁺ ,C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 36. Preparation of Compound II-2i

The operation is the same as in Example 34, except that this example is selected from methyl 2-bromoisopropionate. White solid, yield: 73.0%, melting point: 183-185°C.

Spectral data of compound II-2i: ¹ H NMR (400MHz, Chloroform-d) δ 8.51 (d, J=8.4Hz, 1H), 7.60 (t, J=7.5Hz, 1H), 7.57-7.44 (m, 2H) ), 7.43–7.34(m, 2H), 6.71(s, 1H), 3.77(s, 3H), 2.49(s, 3H), 2.43(td, J=8.4, 4.3Hz, 1H), 1.56(s, 6H), 1.22–1.05 (m, 2H), 0.97–0.77 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ173.78, 163.87, 158.37, 157.90, 142.58, 134.46, 134.11, 130.44, 129.58, 128.27,128.04,127.20,125.45,123.12,122.48,119.29,117.74,53.34,53.12,26.09,25.87,16.47,13.36,8.02,7.43ESI-MS:m/z _465.2 [ _M +H] ⁺ ,C N ₂ O ₃ S ₂ [464.12].

Example 37. Preparation of compound II-2j

The operation is the same as in Example 5, except that the used intermediate is ethyl 2-chloropropionate. White solid, yield: 84.7%, melting point: 99-102°C.

Spectral data of compound II-2j: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.3 Hz, 1H), 7.79-7.63 (m, 1H), 7.63-7.50 (m, 2H), 7.41(d, J=7.6Hz, 2H), 4.00–3.77(m, 2H), 3.62(s, 3H), 2.55(s, 1H), 2.38(s, 3H), 2.32(s, 3H), 1.15 (d, J=7.4Hz, 2H), 0.94–0.68 (m, 2H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ 169.05, 161.23, 158.16, 157.61, 142.61, 134.14, 130.68, 129.67, 129.33, 128.99 ,128.32,127.99,127.20,125.44,123.18,122.61,119.76,52.85,34.46,13.40,13.22,13.07,7.79,7.55.ESI-MS:m/z 451.4[M+H] ⁺ ,C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 38. Preparation of Compound II-2k

The operation is the same as in Example 37, except that the ester used in this example is methyl 2-bromopropionate. White solid, yield: 95.8%, melting point: 108-110°C.

Spectral data of compound II-2k: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.4 Hz, 1H), 7.57 (dt, J= 16.9, 7.7Hz, 2H), 7.46–7.36 (m, 2H), 4.42–4.31 (m, 1H), 3.62 (d, J=17.0Hz, 3H), 2.56 (dd, J=8.7, 5.0Hz, 1H) ), 2.39(s, 3H), 2.31(s, 3H), 1.44–1.29(m, 3H), 1.21–1.10(m, 2H), 0.92–0.78(m, 2H). ¹³ C NMR (100MHz, DMSO) -d ₆ )δ172.02,161.13,158.15,155.89,142.62,134.11,130.63,129.65,129.36,129.09,128.31,128.00,127.21,125.46,123.17,122.58,119.88,53.00,44.04,17.13,13.38,13.22,13.08, 7.90, 7.49. ESI-MS: m/z 465.3 [M+H] ⁺ , C ₂₅ H ₂₄ N ₂ O ₃ S ₂ [464.12].

Example 39. Preparation of compound II-21

The operation is the same as in Example 37, except that the ester used in this example is methyl 2-bromoisopropionate. White solid, yield: 61.9%, melting point: 175-178°C.

Spectral data of compound II-21: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.53 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.6 Hz, 1H), 7.59 (t, J= 7.6Hz, 1H), 7.53(d, J=7.6Hz, 1H), 7.38(d, J=7.8Hz, 2H), 3.65(s, 3H), 2.60–2.52(m, 1H), 2.37(s, 3H), 2.30(s, 3H), 1.49(s, 3H), 1.43(s, 3H), 1.15(q, J=6.6Hz, 2H), 0.95–0.73(m, 2H). ¹³ C NMR(100MHz) ,DMSO-d ₆ )δ173.80,160.93,158.11,156.84,142.52,134.10,130.55,129.55,129.35,129.00,128.22,128.03,127.19,125.46,123.11,122.43,119.79,53.27,53.09,26.10,25.87,13.36, 13.21, 13.07, 8.00, 7.42. ESI-MS: m/z 479.3 [M+H] ⁺ , C ₂₆ H ₂₆ N ₂ O ₃ S ₂ [478.14].

Example 40. Preparation of Compound II-2m

The operation is the same as that in Example 5, except that the intermediate II-1e is used in this example. White solid, yield: 74.1%, melting point: 114-116°C.

Spectral data of compound II-2m: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.6 Hz, 1H), 7.58 (dd, J= 11.9, 7.8Hz, 2H), 7.41 (d, J=7.2Hz, 2H), 3.97–3.82 (m, 2H), 3.62 (s, 3H), 2.82–2.74 (m, 4H), 2.55 (d, J =5.0Hz,1H),1.81(s,2H),1.74(s,2H),1.15(d,J=8.3Hz,2H),0.83(dd,J=16.1,6.8Hz,2H) ^.13C NMR (100MHz,DMSO-d ₆ )δ169.03,161.94,157.87,157.77,142.62,134.14,131.93,131.44,130.63,129.66,128.32,127.99,127.21,125.44,123.19,122.63,118.95,52.84,34.49,25.60,24.93, 22.93, 22.19, 13.40, 7.80, 7.53. ESI-MS: m/z 477.2 [M+H] ⁺ , C ₂₆ H ₂₄ N ₂ O ₃ S ₂ [476.12].

Example 41. Preparation of compound II-2n

The operation is the same as in Example 40, except that methyl 2-bromopropionate is selected in this example. White solid, yield: 81.4%, melting point: 108-110°C.

Spectral data of compound II-2n: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.53 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.3 Hz, 1H), 7.56 (dt, J= 17.0, 7.8Hz, 2H), 7.47–7.35 (m, 2H), 4.37 (dq, J=14.7, 7.1Hz, 1H), 3.62 (d, J=16.6Hz, 3H), 2.83–2.71 (m, 4H) ), 2.59–2.53(m, 1H), 1.80(s, 2H), 1.74(s, 2H), 1.35(t, J=8.0Hz, 3H), 1.14(d, J=8.2Hz, 2H), 0.94 –0.76(m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ172.08,161.84,157.86,157.11,142.61,134.11,132.03,131.46,130.58,129.64,128.30,128.04,123.15,125.3 ,119.07,53.00,44.26,25.61,24.94,22.93,22.19,17.13,13.39,7.89,7.48.ESI-MS: m/z 491.4[M+H] ⁺ ,C ₂₇ H ₂₆ N ₂ O ₃ S ₂ [490.14 ].

Example 42. Preparation of compound II-2o

The operation is the same as in Example 40, except that methyl 2-bromoisopropionate is selected in this example. White solid, yield: 86.2%, melting point: 188-192°C.

Spectral data of compound II-2o: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H), 7.59 (t, J= 7.3Hz, 1H), 7.51(d, J=7.6Hz, 1H), 7.39(d, J=7.6Hz, 2H), 3.65(s, 3H), 2.83–2.72(m, 4H), 2.55(d, J=4.8Hz, 1H), 1.78(d, J=25.9Hz, 4H), 1.47(d, J=23.9Hz, 6H), 1.16(d, J=7.8Hz, 2H), 0.95–0.75(m, 2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ173.76,161.65,157.82,156.99,142.52,134.12,131.92,131.45,130.52,129.57,128.15,127.92,127.10,125.38,123.05,122.42,118.96,53.26, 52.94, 25.98, 25.76, 25.57, 24.89, 22.90, 22.16, 13.29, 7.84, 7.27. ESI-MS: m/z 505.4[M+H] ⁺ ,C ₂₈ H ₂₈ N ₂ O ₃ S ₂ [504.15].

Example 43. Preparation of Compound II-3a

The operation is the same as in Example 14, except that the hydrolyzed compound in this example is II-2a, white solid, yield: 25.5%, melting point: 142-144°C.

Spectral data of compound II-3a: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.80 (s, 1H), 8.55 (d, J=8.5Hz, 1H), 7.68 (t, J=7.6Hz, 1H) ,7.57(dd,J＝18.6,6.8Hz,3H),7.49–7.32(m,3H),3.86(s,2H),2.55(dq,J＝8.2,4.3,2.8Hz,1H),1.14(t , J=8.9Hz, 2H), 0.96–0.73 (m, 2H). ¹³ CNMR (100MHz, DMSO-d ₆ )δ169.58, 163.72, 159.72, 157.83, 142.67, 134.15, 130.68, 129.63, 128.33, 127.96, 127.21, 125.45,123.34,123.24,122.63,122.54,120.94,35.16,13.41,7.80,7.54.ESI-MS:m/z407.4[MH] ^- ,C ₂₁ H ₁₆ N ₂ O ₃ S ₂ [408.06].

Example 44. Preparation of Compound II-3b

The operation is the same as in Example 14, except that the hydrolyzed compound in this example is II-2b. White solid, yield: 33.6%, melting point: 152-154°C.

Spectral data of compound II-3b: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.91 (s, 1H), 8.54 (d, J=8.5Hz, 1H), 7.68 (t, J=7.6Hz, 1H) ,7.58(q,J＝6.8,5.3Hz,3H),7.42(dt,J＝14.2,7.2Hz,3H),4.35(p,J＝7.2Hz,1H),2.55(dd,J＝10.9,5.7 Hz, 1H), 1.39 (dd, J=22.1, 7.3 Hz, 3H), 1.21–1.09 (m, 2H), 0.96–0.78 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ172. 83,163.66,158.57,158.46,157.85,142.67,134.11,130.61,129.59,128.34,128.03,127.22,125.47,123.44,123.22,122.55,121.04,45.00,17.68,13.40,7.88,7.48.ESI-MS:m/z 421.3 [MH] ^- ,C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 45. Preparation of Compound II-3c

The procedure is the same as that of Example 14, except that the hydrolyzed compound is II-2c. White solid, yield: 88.5%, melting point: 155-157°C.

Spectral data of compound II-3c: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.72 (s, 1H), 8.54 (d, J=8.5 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H) ,7.56(q,J＝6.6,5.6Hz,3H),7.48–7.30(m,3H),2.55(dq,J＝8.4,4.2,3.0Hz,1H),1.50(s,3H),1.46(s ,3H),1.22–1.09(m,2H),0.97–0.77(m,2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ174.68,163.39,158.39,157.80,142.55,134.11,130.56,129.50,128.25 ,128.00,127.20,125.49,123.36,123.19,122.50,122.42,120.93,53.44,26.15,25.88,13.38,8.03,7.41.ESI-MS:m/z 435.4[MH] ^- ,C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 46. Preparation of Compound II-3d

The operation is the same as in Example 14, except that the hydrolyzed compound is II-2d. White solid, yield: 94.7%, melting point: 139-142°C.

Spectral data of compound II-3d: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.89 (s, 1H), 8.54 (d, J=8.5Hz, 1H), 7.68 (t, J=7.6Hz, 1H) ,7.56(t,J=9.0Hz,2H),7.41(d,J=7.7Hz,2H),7.10(s,1H),3.84(s,2H),2.54(s,1H),2.52(s, 3H), 1.15(d, J=8.5Hz, 2H), 0.90–0.80(m, 2H). ¹³ CNMR (100MHz, DMSO-d ₆ )δ169.62, 162.67, 158.43, 157.38, 142.60, 136.64, 134.14, 130.75, 129.60,128.28,127.96,127.20,125.46,123.24,122.61,121.13,119.93,35.28,15.97,13.41,7.79,7.52.ESI-MS:m/z 421.3[MH] ^- ,C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 47. Preparation of compound II-3e

The procedure is the same as in Example 14, except that the hydrolyzed compound is II-2e. White solid, yield: 91.3%, melting point: 162-165°C.

Spectral data of compound II-3e: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.94 (s, 1H), 8.54 (d, J=8.5 Hz, 1H), 7.68 (t, J=7.6 Hz, 1H) ,7.56(t,J＝7.8Hz,2H),7.46–7.34(m,2H),7.10(s,1H),4.32(p,J＝7.2Hz,1H),2.56(dd,J＝8.6,5.5 Hz, 1H), 2.52–2.46 (m, 3H), 1.38 (dd, J=23.6, 7.3Hz, 3H), 1.15 (p, J=4.7Hz, 2H), 0.96–0.77 (m, 2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ172.83,162.60,157.67,157.38,142.60,136.72,134.12,130.65,129.58,128.27,127.94,127.19,125.48,123.20,122.54,121.24,119.94,45.08,17.79,15.97, 13.40, 7.86, 7.47. ESI-MS: ESI-MS: m/z 435.4 [MH] ^- , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 48. Preparation of Compound II-3f

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2f. White solid, yield: 94.1%, melting point: 158-161°C.

Spectral data of compound II-3f: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.69 (s, 1H), 8.53 (d, J=8.5 Hz, 1H), 7.67 (t, J=7.6 Hz, 1H) ,7.56(dd,J＝14.9,7.6Hz,2H),7.37(dd,J＝14.1,8.0Hz,2H),7.08(s,1H),2.56(dd,J＝8.4,5.4Hz,1H), 2.50(s, 3H), 1.46(d, J=14.7Hz, 6H), 1.15(q, J=6.4Hz, 2H), 0.96–0.76(m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ174.65,162.32,157.52,157.33,142.49,136.70,134.11,130.65,129.50,128.23,127.96,127.18,125.48,123.16,122.40,121.17,119.86,53.35,26.17,25.89,15.98,13.37,8.01,7.40.ESI - MS: m/z 449.4 [MH] ^- , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 49. Preparation of Compound II-3g

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2g. White solid, yield: 75.14%, melting point: 132-134°C.

Spectral data of compound II-3g: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.86 (s, 1H), 8.54 (d, J=8.5Hz, 1H), 7.68 (t, J=7.6Hz, 1H) ,7.57(t,J＝6.5Hz,2H),7.43(dd,J＝15.6,8.0Hz,2H),7.13(s,1H),3.84(s,2H),2.55(dq,J＝8.1,4.2 , 2.6Hz, 1H), 2.40(s, 3H), 1.13(dd, J=17.6, 5.9Hz, 2H), 0.84(dq, J=11.8, 6.7, 6.1Hz, 2H). ¹³ C NMR(100MHz, DMSO-d ₆ )δ169.60,164.30,159.03,158.47,142.57,134.39,134.14,130.68,129.74,128.36,127.96,127.17,125.43,123.21,122.69,119.20,117.66,35.23,16.51,13.41,7.79,7.55.ESI - MS: m/z 421.3 [MH] ^- , C ₂₂ H ₁₈ N ₂ O ₃ S ₂ [422.08].

Example 50. Preparation of Compound II-3h

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2h. White solid, yield: 85.7%, melting point: 221-224°C.

Spectral data of compound II-3h: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 13.00 (s, 1H), 8.54 (d, J=8.5Hz, 1H), 7.68 (t, J=7.6Hz, 1H) ,7.62–7.52(m,2H),7.50–7.34(m,2H),7.14(s,1H),4.32(p,J=7.0Hz,1H),2.55(dq,J=8.2,4.2,2.9Hz , 1H), 2.40(s, 3H), 1.38(dd, J=23.7, 7.2Hz, 3H), 1.15(d, J=8.4Hz, 2H), 0.95–0.76(m, 2H). ¹³ C NMR( 100MHz,DMSO-d ₆ )δ172.85,164.25,158.49,158.35,142.54,134.39,134.11,130.62,129.71,128.34,127.94,127.17,125.44,123.21,122.62,119.29,117.70,45.02,17.75,16.51,13.40,7.86 , 7.50. ESI-MS: m/z 435.4 [MH] ^- , C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 51. Preparation of compound II-3i

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2i. White solid, yield: 93.7%, melting point: 238-240°C.

Spectral data of compound II-3i: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.62 (s, 1H), 8.47 (d, J=8.4 Hz, 1H), 7.61 (t, J=7.6 Hz, 1H) ,7.56–7.42(m,2H),7.33(t,J=7.2Hz,2H),7.05(s,1H),2.55–2.45(m,1H),2.32(s,3H),1.40(d,J = 12.8Hz, 6H), 1.14–1.01 (m, 2H), 0.88–0.67 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ ) δ174.66, 163.93, 158.43, 158.16, 142.46, 134.36, 134.10, _{130.55,129.64,128.27,127.96,127.15,125.45,123.14,122.48,119.24,117.65,53.35,26.17,25.91,16.50,13.36,8.00,7.42.ESI} ^- MS:m/z MH 449.4[ ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 52. Preparation of compound II-3j

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2j, white solid, yield: 87.8%, melting point: 162-165°C.

Spectral data of compound II-3j: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.95 (s, 1H), 8.54 (d, J=8.5 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H) ,7.56(t,J=7.7Hz,2H),7.41(dd,J=7.6,5.3Hz,2H),3.82(s,2H),2.60–2.52(m,1H),2.37(d,J=7.0 Hz, 3H), 2.31 (d, J=6.6 Hz, 3H), 1.21–1.07 (m, 2H), 0.85 (tt, J=11.7, 6.3 Hz, 2H). ¹³ C NMR (100 MHz, DMSO-d ₆ )δ169.61,161.44,158.23,158.19,142.45,134.14,130.85,129.74,129.28,128.74,128.29,127.92,127.14,125.42,123.22,122.67,119.67,35.77,13.40,13.23,13.07,7.76,7.52.ESI-MS : m/z 435.5[MH] ^- ,C ₂₃ H ₂₀ N ₂ O ₃ S ₂ [436.09].

Example 53. Preparation of Compound II-3k

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2k, white solid, yield: 95.18%, melting point: 150-153°C.

Spectral data of compound II-3k: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.00 (s, 1H), 8.53 (d, J=8.5 Hz, 1H), 7.67 (t, J=7.6 Hz, 1H) ,7.61–7.49(m,2H),7.46–7.34(m,2H),4.31(dt,J=12.8,7.1Hz,1H),2.55(dt,J=10.8,5.6Hz,1H),2.38(s , 3H), 2.31(s, 3H), 1.37(dd, J=25.5, 7.2Hz, 3H), 1.20–1.09(m, 2H), 0.95–0.76(m, 2H). ¹³ C NMR (100MHz, DMSO) -d ₆ )δ172.99,161.34,158.23,157.30,142.46,134.11,130.74,129.70,129.32,128.92,128.30,127.98,127.15,125.45,123.18,122.58,119.83,45.12,17.89,13.39,13.23,13.08,7.84, 7.48. ESI-MS: m/z 449.4 [MH] ^- , C ₂₄ H ₂₂ N ₂ O ₃ S ₂ [450.11].

Example 54. Preparation of compound II-31

The operation is the same as that of Example 14, except that the hydrolyzed compound in this example is II-2l, white solid, yield: 86.2%, melting point: 163-165°C.

Spectral data of compound II-31: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.68 (s, 1H), 8.53 (d, J=8.4Hz, 1H), 7.67 (t, J=7.4Hz, 1H) ,7.63–7.48(m,2H),7.46–7.29(m,2H),2.55(dd,J=9.2,4.1Hz,1H),2.38(s,3H),2.31(s,3H),1.48(d , J=8.7Hz, 3H), 1.44 (d, J=5.3Hz, 3H), 1.22–1.07 (m, 2H), 0.97–0.72 (m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ ) δ174.70,161.03,158.18,157.06,142.38,134.10,130.69,129.62,129.25,128.86,128.23,127.95,127.14,125.46,123.14,122.44,119.76,53.24,26.10,25.87,13.37,13.23,13.10,7.99,7.40. ESI-MS: m/z 463.4 [MH] ^- , C ₂₅ H ₂₄ N ₂ O ₃ S ₂ [464.12].

Example 55. Preparation of Compound II-3m

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2m, white solid, yield: 85.8%, melting point: 158-162°C.

Spectral data of compound II-3m: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.82 (s, 1H), 8.47 (d, J=8.5 Hz, 1H), 7.61 (t, J=7.6 Hz, 1H) ,7.49(t,J=8.6Hz,2H),7.40-7.31(m,2H),3.77(s,2H),2.80-2.65(m,4H),2.56-2.47(m,1H),1.75(s , 2H), 1.68(s, 2H), 1.08(d, J=8.5Hz, 2H), 0.87–0.70(m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169.63,162.09,158.12,157.92 ,142.52,134.13,131.78,131.40,130.73,129.71,128.31,127.95,127.17,125.46,123.22,122.67,118.90,35.21,25.61,24.93,22.94,22.20,13.4 461.4[MH] ^- ,C ₂₅ H ₂₂ N ₂ O ₃ S ₂ [462.58].

Example 56. Preparation of compound II-3n

The procedure is the same as in Example 14, except that the hydrolyzed compound in this example is II-2n, white solid, yield: 92.9%, melting point: 155-157°C.

Spectral data of compound II-3n: ¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.90 (s, 1H), 8.47 (d, J=8.4Hz, 1H), 7.60 (t, J=7.5Hz, 1H) ,7.54–7.43(m,2H),7.34(dd,J=13.1,8.3Hz,2H),4.26(p,J=6.9Hz,1H),2.75–2.65(m,4H),2.52–2.46(m ,1H),1.74(s,2H),1.67(s,2H),1.30(dd,J=23.3,7.2Hz,3H),1.14–1.02(m,2H),0.87–0.69(m,2H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ172.85,162.01,157.93,157.42,142.53,134.10,131.91,131.43,130.62,129.68,128.32,128.01,127.17,125.45,123.21,122.60,119.06,44.91,25.62,24.94 , 22.94, 22.20, 17.62, 13.39, 7.86, 7.47. ESI-MS: m/z 475.4 [MH] ^- , C ₂₆ H ₂₄ N ₂ O ₃ S ₂ [476.12].

Example 57. Preparation of compound II-3o

The operation is the same as that of Example 14, except that the hydrolyzed compound in this example is II-2o, white solid, yield: 80.7%, melting point: 172-174°C.

Spectral data of compound II-3o: ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.71 (s, 1H), 8.52 (d, J=8.5Hz, 1H), 7.67 (t, J=7.5Hz, 1H) ,7.60–7.54(m,1H),7.50(d,J=7.5Hz,1H),7.37(d,J=7.2Hz,2H),2.84–2.70(m,4H),2.53(s,1H), 1.77(d, J=28.1Hz, 4H), 1.47(s, 3H), 1.44(s, 3H), 1.19–1.09(m, 2H), 0.95–0.74(m, 2H). ¹³ C NMR(100MHz, DMSO-d ₆ )δ174.68,161.72,157.88,157.23,142.40,134.10,131.81,131.37,130.65,129.61,128.21,127.94,127.14,125.45,123.14,118.95,53.30,26.18,25.90,25.61,24.96,22.95,22.21 , 13.36, 8.00, 7.38. ESI-MS: m/z 489.4 [MH] ^- , C ₂₇ H ₂₆ N ₂ O ₃ S ₂ [490.14].

Example 58. In vitro target inhibition activity test of target compounds (in two batches)

Test principle

In HEK293T cells stably expressing hURAT1 protein, the ¹⁴ C-labeled substrate uric acid was used to detect the effect of compounds at different concentrations and the positive control drug lesinard on the uptake of the substrate uric acid mediated by hURAT1. The inhibitory effect (IC ₅₀ ) of each compound on this protein was calculated by measuring the radioactivity of uric acid taken up by cells.

Experimental Materials

pcDNA3.1(+)-hURAT1-T2A-eGFP plasmid (Shenzhen Qianhai Dongze Biotechnology Co., Ltd.); Plasmid Extraction Kit (OMEGA Biotechnology Co., Ltd.); Agar (OXOID, USA); Yeast Extract ( U.S. OXOID Company); Peptone Tryptone (U.S. OXOID Company); Ampicillin (U.S. Sigma Company); Purified Water; ); DMSO (Sigma, USA); 96-well microplate (Corning, USA); PBS (Corning, USA); HEPES (Sigma, USA); ¹⁴ C-Uric acid (American Radiolabeled Chemicals, USA).

testing method

The 96-well plate was pre-incubated with poly-D-lysine solution (0.1 mg/mL) for 12 hours for better cell adhesion. Then inoculate the cells into the plate, when the cells reach 90% confluence, mix opti and lip 3000 at 5 μL/well and 0.15 μL/well, respectively, and let stand for 5 min; at the same time, add opti, P3000 and plasmid DNA at 5 μL Mix well with 0.2 μL/well and 500 ng/well, let stand for 5 min; mix the liquid phase mixed in the previous two steps, let stand for 15 min at room temperature; add to 96-well plate with complete medium replaced. Incubate at 37°C in a 5% CO2 incubator for 16-20h, observe the expression of green fluorescent protein EGFP with a fluorescent inverted microscope to verify the success of the transfection, remove the medium after success, and wash the cells twice with PBS. In order to evaluate the inhibitory effect of the drug on URAT1, we took lesinadide as the positive drug, and conducted a preliminary screening of the drug (20uM). CPM). And the _IC50 was determined. Before the absorption experiment, the liquid in the wells was aspirated, and 50 μl of compounds containing various specific concentrations (20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM) were added to each well. The model group and blank group did not add drugs. After 15 minutes of incubation, the cells were aspirated and discarded, and then uric acid absorption buffer containing 50 μM ¹⁴ C-uric acid was added to start uric acid absorption, and incubated at 37° C. for 15 minutes. The liquid in the well was aspirated and washed three times with 100 μL of ice-cold DPBS, and 40 μL of 0.1M NaOH was added to each well to lyse the cells. After lysis at room temperature for 30 minutes, 0.2 mL of scintillation fluid was added to each well, and the plate was placed on a plate shaker and shaken at 260 rpm/min for 15 minutes. The emission value (CPM) of ¹⁴ C-uric acid was measured using a liquid scintillation counter, and the measurement was repeated three times, and the average value was obtained.

Table 2. In vitro target inhibitory activities of I series compounds and II series compounds

Conclusion: It can be seen from Table 2 that in the I series, compounds I-3a, I-3b, I-3e and II series compounds II-3a, II-3b, II-3h all showed significant in vitro target inhibitory activity, Both are better than or equivalent to the positive control drug lesinurad; in the II series, the activities of II-3a and II-3b are far superior to that of the drug lesinurad, and have achieved unexpected results. It can be further developed as a drug lead with a new structure.

Example 59. In vivo anti-gout activity test of the target compound.

Test Materials and Methods

(1) Experimental animals: male Kunming mice, provided by the Experimental Animal Center of Shandong University.

(2) Sample treatment: Select the compounds to be tested and their corresponding esters whose target inhibitory activity is significantly better than lesinurad, and mix them with CMC-Na to an appropriate concentration before use.

(3) Modeling drugs: xanthine, potassium oxonate.

(4) Positive control drug: Lesinurad.

(5) Test method: 20g male Kunming mice were adaptively fed for 1 week and randomly divided into blank group and model group. 0.2 mL of hypoxanthine suspension and 0.2 mL of 400 mg/kg potassium oxonate suspension were subcutaneously injected. After 4 hours, the eyeballs were removed and blood was collected, the supernatant was separated, and the blood uric acid concentration was detected.

Table 3 Activity results in animals

Conclusion: It can be seen from Table 3 that the in vivo uric acid-lowering activity of most compounds is significantly better than that of lesinurad, especially the target inhibitory activity of compound II-3a reached 3.27 μM in the cell experiment screening, and the blood uric acid reduction rate in the animal in vivo activity screening experiment. It reached 90.2%, showing significantly higher activity than lesinard (target inhibitory activity IC ₅₀ = 15.34 μM, blood uric acid reduction rate was 31.4%), which deserves further study.

Example 60. Pharmacokinetic experiment of target compound II-3a

Test principle:

Compound II-3a was administered to SD rats by single intravenous injection (iv) and oral administration (po) respectively, blood was collected at different time points, the plasma drug concentration of II-3a in rats was determined, and the calculated relevant pharmacokinetic parameters.

testing method:

After the health assessment, the rats were adaptively cultured for at least three days under the conditions of a temperature of 20-26°C, a relative humidity of 30%-70%, and 12 hours of alternating light and dark. The rats in the gavage group were fasted for 12 hours before administration, and resumed their diet 4 hours after administration. The rats in the intravenous injection group were allowed to eat freely during the experiment. The test drug II-3a was dissolved in a mixed solution of DMSO:PEG400:physiological saline (3:60:37), and ultrasonically mixed to obtain a clear solution or a uniform suspension. According to the body weight of the rats, 20 mg/kg was given by gavage (n=3) and 1 mg/kg by intravenous injection (n=3). After administration, 0.083, 0.25, 0.5, 1, 2, 4, 250 μL of venous blood was collected at 6, 10 and 24 h. Blood samples were collected into heparin-treated centrifuge tubes and kept on ice until plasma processing. Blood samples will be processed for plasma immediately within half an hour of collection by centrifugation at about 4°C, 3200 g for 10 minutes. Plasma samples will be stored in polypropylene tubes, snap frozen on dry ice, and kept at around -70 ± 10°C until LC/MS/MS analysis. At the end of the experiment, all surviving animals were sacrificed in accordance with the regulations of the Animal Experiment Professional Ethics of the Medical Science Committee. Pharmacokinetics software Phoenix 8.0 was used to calculate parameters such as AUC _0-t , AUC _0-∞ , MRT _0-t and T _1/2 .

Test Results:

Table 4 Pharmacokinetic study of compound II-3a

Conclusion: From the results in Table 4 and Figure 1, after oral administration (20mg/kg), the peak time is 2.33h, II-3a has a medium plasma half-life (t _1/2 = 3.67h), and has a longer plasma half-life (t 1/2 = 3.67h). High bioavailability (II-3a: 95.6%). Meanwhile, the maximum concentration (C _max ) of II-3a in plasma after oral administration was 18938 ng/mL, and the average residence time was 5.98 h.

After II-3a injection (1mg/kg), the maximum plasma concentration was 11213ng/mL, the half-life was 3.88h, the mean residence time was 2.72h, and the apparent volume of distribution and clearance were 0.425L/kg and 2.64, respectively mL/min/kg. II-3a has better pharmacokinetic parameters, and the experimental results preliminarily prove the rationality of the design, which deserves further study.

Claims (6)

1. a thienopyrimidinone thioacetic acid derivatives, is characterized in that, has the structure shown in following general formula I or II:

in, R is -H, -CH ₃ or

R ₁ is -CH ₂ -,- ^* CH(CH ₃ )- or -C(CH ₃ ) ₂ -; * represents a chiral carbon atom; R ₂ is -OH or -OCH ₃ . 2. a thienopyrimidinone thioacetic acid class derivative, it is characterized in that being one of the compounds of following structure:

3. the preparation method of thienopyrimidinone thioacetic acid derivatives as claimed in claim 2 is characterized in that the preparation method of I-3 (a-i) is as follows: Using 4-bromo-1-naphthylamine as the starting material, the Suzuki coupling reaction with cyclopropylboronic acid produces the intermediate 4-cyclopropyl-1-naphthylamine a, which is then reacted with 1,1'-thiocarbonyldiimidazole Obtain intermediate b, then react with methyl 3-aminothiophene-2-carboxylate containing different substituents in absolute ethanol or pyridine solution to obtain intermediate I-1(ac), and then in DMF solution K ₂ CO ₃ Under the catalysis of substituted methyl bromoacetate, a nucleophilic substitution reaction occurs to obtain the target product I-2(ai), and finally in the mixed solution of tetrahydrofuran and methanol, the target product I-3(ai) is obtained by hydrolysis with lithium hydroxide or sodium hydroxide. ); The synthetic route one is as follows:

Reagents and conditions: (i) K ₃ PO ₄ , Pd(PPh ₃ ) ₄ , cyclopropylboronic acid, toluene: water=25:2 v/v, N ₂ protection, 100°C, 12h; (ii) CH ₂ Cl ₂ , 1,1'-thiocarbonyldiimidazole, room temperature, 12h; (iii) methyl 3-aminothiophene-2-carboxylate or methyl 3-amino-5-methylthiophene-2-carboxylate, absolute ethanol, 90 ℃, 4h, NaOH, 90℃, 12h or methyl 3-amino-4-methylthiophene-2-carboxylate, pyridine, 50℃, 12h, NaOH, 90℃, 12h; (iv) K ₂ CO ₃ , DMF , methyl 2-bromoisobutyrate or methyl 2-bromopropionate/methyl bromoacetate, 100°C, 3h; (v) methanol, tetrahydrofuran, lithium hydroxide/sodium hydroxide, room temperature, 12h. 4. the preparation method of thienopyrimidinone thioacetic acid derivatives as claimed in claim 2 is characterized in that the preparation method of II-3 (a-o) is as follows: Using 4-bromo-1-naphthylamine as the starting material, the Suzuki coupling reaction with cyclopropylboronic acid produces the intermediate 4-cyclopropyl-1-naphthylamine a, which is then reacted with 1,1'-thiocarbonyldiimidazole Obtain intermediate b, and then react with methyl 2-aminothiophene-3-carboxylate containing different substituents in pyridine or absolute ethanol to generate intermediate II-1(ae), then in DMF solution, in K ₂ CO ₃ Under the catalysis, nucleophilic substitution reaction with various esters to obtain the corresponding target product II-2(ao), and finally in the mixed solution of tetrahydrofuran and methanol, the target product II-3(ao) is obtained by hydrolysis with lithium hydroxide or sodium hydroxide. ); The second synthetic route is as follows:

Reagents and conditions: (i) K ₃ PO ₄ , Pd(PPh ₃ ) ₄ , cyclopropylboronic acid, toluene: water=25:2 v/v, N ₂ protection, 100°C, 12h; (ii) CH ₂ Cl ₂ , 1,1'-thiocarbonyldiimidazole, room temperature, 12h; (iii) the substituted methyl 2-aminothiophene-3-carboxylate, absolute ethanol, 90°C, 4h, NaOH, 90°C, 12h or the described Substituted methyl 2-aminothiophene-3-carboxylate, pyridine, 50°C, 12h, NaOH, 90°C, 12h; (iv) K ₂ CO ₃ , DMF, methyl 2-bromoisobutyrate or 2-bromopropionic acid Methyl ester or methyl bromoacetate, 100°C, 3h; (v) methanol, tetrahydrofuran, lithium hydroxide or sodium hydroxide, room temperature, 12h. 5. The application of the thienopyrimidinone thioacetic acid derivatives according to claim 1 or 2 in the preparation of an anti-gout medicine. 6. An anti-gout pharmaceutical composition, comprising the thienopyrimidinone thioacetic acid derivative according to claim 1 or 2 and one or more pharmaceutically acceptable carriers or excipients.

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