Preparation method and application of tanshinone IIA and cryptotanshinone amide derivative

Abstract

The invention discloses a preparation method and application of tanshinone IIA and cryptotanshinone amide derivatives. The tanshinone IIA and cryptotanshinone amide derivatives are compounds shown in a general formula (I) or stereoisomers, racemes, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs of the compounds. The tanshinone IIA and cryptotanshinone amide derivative can be used for preparing anti-inflammatory drugs, antibacterial drugs, tumor drugs, drugs for treating cardiovascular and cerebrovascular diseases, drugs for treating burns and scalds and drugs for treating infectious diseases.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to tanshinone IIA, cryptotanshinone amide derivatives, and preparation methods and applications thereof. Background Art

[0002] Danshen, a traditional Chinese medicinal herb, is the dried root or rhizome of the Lamiaceae plant Salvia miltiorrhiza Bunge. It is widely used clinically to treat inflammation, viral hepatitis, liver fibrosis, and cardiovascular disease. The main active ingredients of Danshen are fat-soluble diterpenoid quinone compounds and water-soluble phenolic acid compounds. Both have anti-myocardial ischemia, increased coronary flow, improved cardiac function, anti-thrombotic and lipid-regulating effects, and anti-inflammatory and antioxidant properties, but each has its own specific focus. The water-soluble components are more prominent in antioxidant effects, while the fat-soluble components of Danshen are more prominent in antibacterial, anti-inflammatory, and anti-cancer effects. This diversity of effects is the basis for Danshen's wide range of indications.

[0003] Tanshinones are fat-soluble phenanthrenequinone compounds extracted from Danshen (Salvia miltiorrhiza). The main components are dihydrotanshinone I (DHT), cryptotanshinone (CPT), tanshinone I (TI), and tanshinone IIA (TIIA). Tanshinones possess a wide range of biological activities, including but not limited to anti-inflammatory, antibacterial, and anticancer properties.

[0004] However, there is currently a lack of research and design of novel tanshinone derivatives that enhance the therapeutic efficacy of various diseases, including anti-inflammatory, antibacterial, anti-infective, cancer, cardiovascular, and respiratory diseases. In light of this, the present invention conducts a derivatization reaction at the active methyl groups of cryptotanshinone and tanshinone IIA. Through a multi-step reaction, novel amide derivatives of tanshinone IIA and cryptotanshinone are obtained, increasing their water solubility and improving their drug-like properties, resulting in excellent drugability. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for preparing novel amide derivatives of tanshinone IIA and cryptotanshinone and their application. The present invention obtains novel amide derivatives of tanshinone IIA and cryptotanshinone through multi-step reactions, significantly improving the pharmacological activity of such compounds in the treatment of inflammatory system diseases, bacterial and other infectious diseases, cancer, immune system diseases, cardiovascular system diseases, and respiratory system diseases.

[0006] In order to achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0007] A tanshinone IIA or cryptotanshinone amide derivative, which is a compound represented by general formula (I) or a stereoisomer, racemate, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof;

[0008]

[0009] in, express:

[0010]

[0011] wherein R1 and R2 are independently selected from H, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted ester, substituted or unsubstituted aromatic heterocyclic group, substituted or unsubstituted aryl, or R1, R2 and the nitrogen atom to which they are attached together form a substituted or unsubstituted 5-6 membered heterocyclic group;

[0012] The 5- to 6-membered heterocyclic group is a group formed by replacing a 5- to 6-membered cycloalkyl group with one or more NH, S or O groups;

[0013] The substituent on the C1-C3 alkyl group is selected from halogen, hydroxyl, C1-C4 alkoxycarbonyl, di-C1-C4 alkylamino or C1-C4 alkylamino;

[0014] The substituent on the aromatic heterocyclic group or aryl group is halogen, C1-C4 alkoxy or C1-C4 alkyl, the aromatic heterocyclic group is further preferably thiazolyl or benzothiazolyl, and the aryl group is further preferably phenyl;

[0015] The substituent on the 5- to 6-membered heterocyclic group is a C1-C4 alkyl group.

[0016] Preferably, each R1 and R2 are independently H, methyl, chloroethyl, substituted or unsubstituted hydroxyethyl, substituted or unsubstituted C2 ester, ethyl substituted by methoxycarbonyl, N,N-dimethylethyl, phenyl, p-methoxyphenyl, p-fluorophenyl, thiazolyl or benzothiazolyl; or, R1, R2 and the nitrogen atom to which they are attached together form a substituted or unsubstituted morpholinyl, N-isopropylpiperazinyl, tetrahydropyrrolyl or piperidinyl;

[0017] Wherein, the substituent of the morpholinyl group is an optically active methyl group.

[0018] Furthermore, the compound represented by formula (1) is selected from:

[0019]

[0020]

[0021] On the other hand, the present invention provides a method for preparing the above-mentioned compound, comprising the following synthetic route:

[0022]

[0023] Formula 1 Reaction route I, reagents and reaction conditions are: (a) Pd / C, DMA, H2; K2CO3, BnCl; (b) SeO2, Dioxane, 110°C; (c) NH2SO3H, NaClO2, t-BuOH, THF; (d) HATU, amines, DIPEA, DCM; (e) H2, Pd / C, EA / MeOH

[0024]

[0025] Formula 2, Reaction Scheme II, reagents and reaction conditions are: (f) K2CO3, CH3I, Acetone, 50°C; (g) Mg, NH4Cl, MeOH, THF; (h) NaOH, THF, MeOH, H2O; 2N HCl; (i) i-PrOH, (S)-(-)-α-methylbenzylamine, 110°C; (j) 4N HCl; (d) HATU, amines, DIPEA, DCM; (e) H2, Pd / C, EA / MeOH; (k) i-PrOH, (R)-(+)-α-methylbenzylamine, 110°C

[0026] The present invention also provides a pharmaceutical composition comprising an effective amount of the above-mentioned tanshinone IIA, cryptotanshinone amide derivative, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0027] The present invention also provides the use of the above-mentioned tanshinone IIA, cryptotanshinone amide derivatives, or the above-mentioned pharmaceutical compositions in the preparation of anti-inflammatory drugs, antibacterial drugs, tumor drugs, drugs for treating cardiovascular and cerebrovascular diseases, drugs for treating burns and scalds, and drugs for treating infectious diseases.

[0028] Preferably, the use is to prepare antibacterial drugs or anti-inflammatory drugs.

[0029] Preferably, the antibacterial drug is used to inhibit Bacillus subtilis and Staphylococcus aureus;

[0030] The anti-inflammatory drug is used for treating keratitis.

[0031] Compared with the prior art, the beneficial effects of the present invention are embodied in:

[0032] (1) The present invention starts with commercially available tanshinone IIA of relatively low purity. After protecting the keto carbonyl group of tanshinone IIA with a benzyl group, the active methyl group is oxidized to an acid, and then debenzylated by acid-amine condensation to obtain tanshinone IIA amide derivative 6. Furthermore, 4 is subjected to esterification and double bond reduction to obtain ester 9 of the cryptotanshinone core structure. The ester is then hydrolyzed and resolved with a chiral amine to obtain chiral acids 11 and 15. Finally, high-purity chiral cryptotanshinone core amide derivatives 13 and 17 are obtained by acid-amine condensation and debenzylation.

[0033] (2) The present invention uses benzyl to protect the two ketone carbonyl groups of tanshinone IIA and cryptotanshinone, which greatly improves the yield of each subsequent reaction step.

[0034] (3) The present invention oxidizes the active methyl group of tanshinone IIA to carboxylic acid, which not only allows the double bonds at positions 1 and 2 to be efficiently reduced after esterification, but also utilizes its acidity to form salts with chiral amines for efficient chiral resolution. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 is the HPLC chromatogram of compound 11;

[0036] Figure 2 is the HPLC chromatogram of compound 15;

[0037] Figure 3 is the IL-1β inhibitory activity of the compound of the present invention at a concentration of 10 μM;

[0038] Figure 4 is the TNF-α inhibitory activity of the compound of the present invention at a concentration of 10 μM;

[0039] Figure 5 is the IL-6 inhibitory activity of the compound of the present invention at a concentration of 10 μM;

[0040] Figure 6 The following are photos of the healing model in Test Example 5: (a) Photos of mouse wounds on days 0, 3, 7, 11, and 14 after treatment with saline, blank gel, 6D gel, and 6K gel. (b) Statistics of the wound healing rates of mice in different treatment groups on days 3, 7, 11, and 14. (c) Statistics of the bacterial colony load in mouse wound tissue on day 4 of treatment. (d) Photos of the bacterial colonies formed on the MH plate after the wound tissue homogenate was applied on day 4 of treatment. (Dilution factor: 10 4 )

[0041] Figure 7 This is an H&E staining image of the wound skin tissue of mice on the 14th day of treatment in the wound healing model of Test Example 5;

[0042] Figure 8These are H&E staining images of the main organ tissues of mice on the 14th day of treatment in the wound healing model of Test Example 5;

[0043] Figure 9 The following are photos of the keratitis model in Test Example 5: (a) Photos of corneal infection in different groups before and after treatment. (b) Statistics of corneal colony loads in different treatment groups. (c) Photos of colonies formed by corneal homogenates from different treatment groups applied on MH plates. (Dilution factor: 10 4 );

[0044] Figure 10 This is an H&E staining image of the mouse corneal tissue after treatment in the keratitis model of Test Example 5;

[0045] Figure 11 These are H&E staining images of the main organ tissues of mice after treatment in the keratitis model of Test Example 5. DETAILED DESCRIPTION

[0046] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of the present invention, but are not intended to limit the present invention.

[0047] Example 1 Preparation of Compound 2

[0048]

[0049] Commercially available tanshinone IIA (29.4 g, 100 mmol) and 10% palladium on carbon (600 mg, 2 wt%) were placed in a 250 mL two-necked flask equipped with a magnetic stirrer. 150 mL of N,N-dimethylformamide was added, and the atmosphere was replaced with hydrogen several times. The mixture was allowed to react at room temperature for 3 hours. After the reaction solution turned from red to black, it was transferred via syringe to a 250 mL two-necked flask under argon, containing potassium carbonate (56.3 g, 408 mmol) and a magnetic stirrer. After stirring at room temperature for 10 minutes, benzyl chloride (21.9 mL, 255 mmol) was added, and the temperature was raised to 55°C for 6 hours. After the reaction was complete, the solid was removed by filtration through celite, and the residue was rinsed with ethyl acetate. 125 mL of water and 50 mL of ethyl acetate were added to the filtrate. After separation, the aqueous layer was extracted with ethyl acetate (50 mL x 3). The organic layers were combined, washed several times with water and once with saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated to remove ethyl acetate to obtain a crude product. Appropriate amounts of petroleum ether and methanol were added to the crude product, stirred for 10-15 minutes, and filtered to obtain a white or light yellow solid 2 (31.0 g, 65%).

[0050] 1H NMR(400MHz, CDCl3)δ8.11(dd,J=8.7,2.9Hz,1H),7.64–7.49(m,4H),7.46–7.32(m,9H),5.30(d,J=2.5Hz,2H ),5.10(d,J=2.7Hz,2H),3.59(dd,J=7.8,4.6Hz,2H),2.33(s,2H),1.98–1.62(m,4H),1.42(d,J=2.7Hz,6H). 13 C NMR (101MHz, CDCl3) δ149.25,145.47,144.73,143.35,140.93,137.98,137.63,132.79,128.95,128.56,128.51,128.08, 127.95,127.76,126.39,125.39,119.10,118.43,118.12,116.42,76.42,76.36,39.00,35.10,32.34,30.95,20.44,9.80.

[0051] Example 2 Preparation of Compound 3

[0052]

[0053] Compound 2 (4.8 g, 10 mmol) and selenium dioxide (2.2 g, 20 mmol) were weighed and placed in a 100 mL single-necked flask equipped with a magnetic stirrer. 30 mL of dioxane was added and the mixture was refluxed for 24 hours. After the reaction was complete, 50 mL of water and 30 mL of ethyl acetate were added. After separation, the aqueous layer was extracted with ethyl acetate (15 mL x 3). The organic layers were combined, washed several times with water and once with saturated brine. The organic layer was dried over anhydrous sodium sulfate and rotary evaporated to remove the ethyl acetate to obtain a crude product. An appropriate amount of petroleum ether and a small amount of ethyl acetate were added to the crude product, stirred for 10-15 minutes, and filtered to obtain a white or light yellow solid 3 (3.4 g, 70%).

[0054] 1 H NMR (400MHz, CDCl3) δ10.32(s,1H),8.30(s,1H),8.12(d,J=8.7Hz,1H),7.62(d,J=8.7Hz,1H),7.55(d,J=6.7Hz,2H),7. 50–7.26(m,8H),5.35(s,2H),5.15(s,2H),3.61(t,J=5.8Hz,2H),1.91–1.82(m,2H),1.78(d,J=5.5Hz,2H),1.44(s,6H). 13C NMR (101MHz, CDCl3) δ186.26,186.23,149.61,149.54,146.58,144.58,144.15,137.46,136.88,133.13,128.93,128.6 0,128.54,128.44,127.92,127.21,126.20,123.83,118.16,117.91,115.66,76.39,38.78,35.14,32.23,30.95,20.28.

[0055] Example 3 Preparation of Compound 4

[0056]

[0057] Compound 3 (4.9 g, 10 mmol) and aminosulfonic acid (2.9 g, 30 mmol) were weighed and placed in a 250 mL single-necked flask equipped with a magnetic stirrer. 90 mL of tetrahydrofuran and 10 mL of tert-butanol were added. After stirring at -10°C for 20 minutes, 30 mL of an aqueous solution of sodium chlorite (2.7 g, 30 mmol, 80 wt%) was added dropwise. After the addition was complete, the reaction was maintained at -10°C for 20-30 minutes. After the reaction was complete, an aqueous solution of sodium sulfite (4.0 g, 31.8 mmol) was added dropwise. After the addition was complete, 50 mL of ethyl acetate was added. After separation, the aqueous layer was extracted with ethyl acetate (15 mL x 3). The organic layers were combined, washed several times with water and once with saturated brine. The organic layer was dried over anhydrous sodium sulfate and the ethyl acetate was removed by rotary evaporation to obtain the crude product. An appropriate amount of petroleum ether and a small amount of ethyl acetate were added to the crude product, stirred for 10-15 minutes, and filtered to obtain 4 (4.6 g, 91%) as a white or light yellow solid.

[0058] 1 H NMR (400MHz, CDCl3) δ8.44(d,J=1.2Hz,1H),8.12(d,J=8.6Hz,1H),7.66(d,J=8.6Hz,1H),7.59–7.19 (m,10H),5.52(s,2H),5.14(s,2H),3.59(t,J=5.7Hz,2H),1.83(dd,J=36.5,5.2Hz,4H),1.44(s,6H). 13CNMR(101MHz, CDCl3)δ162.55,152.38,149.34,145.60,145.08,140.89,136.91,134.26,133.15,129.91,129.35,128.7 8,128.69,128.22,127.85,127.05,126.79,118.61,118.05,115.54,113.84,77.57,38.73,35.19,32.21,30.95,20.25.

[0059] Example 4 Preparation of Compound 5

[0060]

[0061] Compound 4 (101.2 mg, 0.2 mmol) and HATU (150 mg, 0.4 mmol) were weighed and placed in a Schlenk tube equipped with a magnetic stirrer. The atmosphere was replaced with argon. 4 mL of dichloromethane, DIPEA (0.1 mL, 0.6 mmol), and various amines (0.4 mmol) were added and reacted at room temperature for 8-10 hours. After the reaction was complete, 15 mL of water and 10 mL of dichloromethane were added, along with a small amount of 1N HCl. After separation, the aqueous layer was extracted with dichloromethane (15 mL x 3). The organic layers were combined, washed several times with water and once with saturated brine. The organic layer was dried over anhydrous sodium sulfate and rotary evaporated to remove dichloromethane to yield the crude product. The crude product was purified by silica gel column chromatography to yield the colloidal products 5A-5T.

[0062] Example 6 Preparation of Compound 6A

[0063]

[0064] Compound 5A (101 mg, 0.2 mmol) and 10% palladium on carbon (10 mg, 10 wt%) were weighed and placed in a 50 mL single-necked flask equipped with a magnetic stirrer. 4 mL of ethyl acetate and 4 mL of methanol were added, and the atmosphere was replaced with hydrogen several times. The reaction was allowed to proceed at 45°C for 2 hours. After the reaction was complete, the palladium on carbon was removed by filtration through diatomaceous earth, the filter residue was rinsed with ethyl acetate, and the filtrate was collected and rotary evaporated to remove the ethyl acetate to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain 6A (57.5 mg, 89%) as a red solid.

[0065] 1 H NMR(400MHz, CDCl3)δ9.02(s,1H),8.17(s,1H),7.75–7.63(m,2H),5.70(s,1 H), 3.20 (t, J = 6.4Hz, 2H), 1.88–1.77 (m, 2H), 1.72–1.64 (m, 2H), 1.33 (s, 6H).13 C NMR (101MHz, CDCl3) δ181.41,175.56,164.19,151.97,150.59,150.42,145.39,13 4.10,126.12,125.92,121.37,121.23,115.30,37.42,34.64,31.41,29.85,18.73.

[0066] Example 7 Preparation of Compound 6B

[0067]

[0068] The amine in Example 5 was converted to 5B using methylamine hydrochloride, and compound 6B was synthesized using the same method as compound 6A in Example 6.

[0069] 1 H NMR (400MHz, CDCl3) δ9.08 (s, 1H), 8.13 (s, 1H), 7.73–7.59 (m, 2H), 3.18 (t, J = 6.4 Hz,2H),2.98(d,J=4.8Hz,3H),1.86–1.77(m,2H),1.69–1.63(m,2H),1.32(s,6H). 13 CNMR(101MHz,CDCl3)δ181.70,175.93,163.93,160.88,151.88,149.80,145.38,134.0 2,126.48,126.17,122.53,121.25,115.75,37.69,34.87,31.81,30.01,26.24,18.99.

[0070] Example 8 Preparation of Compound 6C

[0071]

[0072] The amine in Example 5 was converted to 5C using dimethylamine hydrochloride, and compound 6C was synthesized using the same method as compound 6A in Example 6.

[0073] 1 H NMR (400MHz, CDCl3) δ7.65 (ddd, J=23.4, 8.2, 1.5Hz, 3H), 3.19 (t, J=6.4Hz, 2H), 3.12 (s, 3H), 2.96(d,J=1.5Hz,3H),1.80(h,J=5.3Hz,2H),1.67(dt,J=10.8,6.2Hz,2H),1.34–1.29(m,6H). 13C NMR (101MHz, CDCl3) δ182.39,174.07,162.45,161.59,151.36,145.22,144.24,133.84,1 26.75,126.61,121.41,120.93,117.70,38.31,37.85,35.26,34.91,31.94,30.09,19.15.

[0074] Example 9 Preparation of Compound 6D

[0075]

[0076] The amine in Example 5 was reacted with morpholine to obtain 5D, and compound 6D was synthesized using the same method as compound 6A in Example 6.

[0077] 1 H NMR (400MHz, CDCl3) δ7.71 (s, 1H), 7.69–7.60 (m, 2H), 3.80 (s, 4H), 3.66 (t, J = 4.8Hz, 2H), 3.3 8(t,J=4.8Hz,2H),3.19(t,J=6.4Hz,2H),1.84–1.77(m,2H),1.70–1.63(m,2H),1.32(s,6H). 13 C NMR (151MHz, CDCl3) δ182.27,174.08,161.82,161.29,151.60,145.37,144.69,133.89,126.60,126.59,12 1.00,120.62,117.34,66.65,66.58,47.46,42.66,37.83,34.94,31.94,31.71,30.09,22.78,19.14,14.24.

[0078] Example 10 Preparation of Compound 6E

[0079]

[0080] The amine in Example 5 was reacted with 3-(S)-3-methylmorpholine to obtain 5E, and compound 6E was synthesized using the same method as compound 6A in Example 6.

[0081] 1H NMR(400MHz, CDCl3)δ7.68(d,J=8.1Hz,2H),7.62(d,J=8.2Hz,1H),4.02–3.42(m,6H),3.23–3.10 (m,3H),1.80(qd,J=7.9,4.3Hz,2H),1.70–1.63(m,2H),1.51–1.38(m,3H),1.32(d,J=1.4Hz,6H). 13 CNMR(101MHz, CDCl3)δ182.29,174.09,164.46,161.77,151.57,145.38,144.28,133.89,130.64,126 .61,126.58,120.98,117.39,70.78,66.77,55.31,45.66,37.82,34.93,31.93,30.10,19.14,14.78.

[0082] Example 11 Preparation of Compound 6F

[0083]

[0084] The amine in Example 5 was treated with isopropylpiperazine to obtain 5F, and compound 6F was synthesized using the same method as compound 6A in Example 6.

[0085] 1 H NMR (400MHz, DMSO) δ8.19(s,1H),7.86(d,J=8.2Hz,1H),7.66(d,J=8.1Hz,1H),3.34(s,5H),3.19–2 .91(m,5H),2.08(s,1H),1.79–1.67(m,2H),1.66–1.59(m,2H),1.29(s,6H),1.24(d,J=6.6Hz,6H). 13 C NMR (101MHz, DMSO) δ181.83,174.09,161.20,160.68,150.57,145.58,143.46,133.92,127.29,1 26.69,120.89,119.50,117.90,58.11,47.98,47.65,37.88,34.93,31.99,30.08,19.24,16.97.

[0086] Example 12 Preparation of Compound 6G

[0087]

[0088] The amine in Example 5 was converted to aniline to obtain 5G, and compound 6G was synthesized using the same synthesis method as compound 6A in Example 6.

[0089] 1 H NMR (400MHz, CDCl3) δ11.13(s,1H),8.22(s,1H),7.87–7.79(m,2H),7.75–7.62(m,2H),7.40–7.31(m,2H),7 .13(ddt,J=8.6,7.3,1.2Hz,1H),3.22(t,J=6.4Hz,2H),1.89–1.78(m,2H),1.72–1.65(m,2H),1.34(s,6H). 13 C NMR (101MHz, CDCl3) δ181.50,176.36,164.42,158.22,152.28,150.72,145.75,138.36,134.21,12 9.13,126.41,126.24,124.58,123.56,121.48,120.22,115.50,37.77,35.02,31.91,30.16,19.09.

[0090] Example 13 Preparation of Compound 6H

[0091]

[0092] The amine in Example 5 was converted to 5H using p-methoxyaniline, and compound 6H was synthesized using the same synthesis method as compound 6A in Example 6.

[0093] 1 H NMR(400MHz, CDCl3)δ11.03(s,1H),8.20(s,1H),7.76–7.72(m,2H),7.72–7.64(m,2H),6.92–6.8 4(m,2H),3.80(s,3H),3.21(t,J=6.4Hz,2H),1.89–1.78(m,2H),1.72–1.64(m,2H),1.33(s,6H). 13 C NMR (101MHz, CDCl3) δ181.52,176.30,164.34,157.84,156.56,152.20,150.47,145.69,134.17,131.6 2,126.44,126.22,123.59,121.61,121.44,115.54,114.27,55.60,37.76,35.00,31.90,30.14,19.08.

[0094] Example 14 Preparation of Compound 6I

[0095]

[0096] The amine in Example 5 was converted to 5I using p-fluoroaniline, and compound 6I was synthesized using the same method as compound 6A in Example 6.

[0097] 1 H NMR (400MHz, CDCl3) δ11.15(s,1H),8.21(s,1H),7.83–7.74(m,2H),7.75–7.62(m,2H),7. 09–6.99(m,2H),3.21(t,J=6.4Hz,2H),1.89–1.79(m,2H),1.73–1.65(m,2H),1.34(s,6H). 13 C NMR (101MHz, CDCl3) δ181.44,176.40,164.51,160.78,158.36,158.12,152.40,150.69,145.81,134.45,134.24 ,126.34,126.23,123.37,121.79,121.71,121.50,115.88,115.66,115.45,37.76,35.03,31.91,30.17,19.08.

[0098] Example 15 Preparation of Compound 6J

[0099]

[0100] The amine in Example 5 was converted to 5J using chloroethylamine hydrochloride, and compound 6J was synthesized using the same method as compound 6A in Example 6.

[0101] 1 H NMR (400MHz, CDCl3) δ9.50 (s, 1H), 8.15 (s, 1H), 7.71 (d, J = 8.2Hz, 1H), 7.65 (d, J = 8.2Hz, 1H), 3.84–3.67(m,4H),3.20(t,J=6.4Hz,2H),1.87–1.77(m,2H),1.71–1.64(m,2H),1.33(s,6H). 13C NMR (101MHz, CDCl3) δ181.62,175.97,164.13,160.67,152.12,150.24,145.60,134.14,1 26.46,126.29,122.33,121.36,115.78,43.12,41.62,37.77,34.99,31.90,30.11,19.08.

[0102] Example 16 Preparation of Compound 6K

[0103]

[0104] The amine in Example 5 was converted to 5K using ethanolamine, and compound 6K was synthesized using the same method as compound 6A in Example 6.

[0105] 1 H NMR (400MHz, CDCl3) δ9.46(s,1H),8.14(s,1H),7.70(d,J=8.2Hz,1H),7.64(d,J=8.1Hz,1H),3.86(dd,J=5.6 ,4.4Hz,2H),3.62(q,J=5.3Hz,2H),3.19(t,J=6.4Hz,2H),1.87–1.77(m,2H),1.71–1.64(m,2H),1.33(s,6H). 13 C NMR (151MHz, CDCl3) δ181.62,176.03,164.22,161.80,152.18,150.20,145.64,134.17,1 26.44,126.26,122.39,121.40,115.79,62.79,43.05,37.77,35.00,31.91,30.13,19.08.

[0106] Example 17 Preparation of Compound 6L

[0107]

[0108] The amine in Example 5 was prepared using glycine methyl ester hydrochloride to obtain 5L, and compound 6L was synthesized using the same method as compound 6A in Example 6.

[0109] 1H NMR (400MHz, CDCl3) δ9.59(t,J=5.5Hz,1H),8.18–8.13(m,1H),7.70(d,J=8.2Hz,1H),7.64(d,J=8.1Hz,1H),4.2 2(d,J=5.5Hz,2H),3.80–3.76(m,3H),3.19(t,J=6.4Hz,2H),1.87–1.76(m,2H),1.71–1.63(m,2H),1.32(s,6H). 13 C NMR (101MHz, CDCl3) δ181.57,175.89,169.85,164.03,160.71,152.07,150.35,145.53,134.1 0,126.43,126.30,121.98,121.33,115.84,52.48,41.54,37.77,34.98,31.90,30.11,19.08.

[0110] Example 18 Preparation of Compound 6M

[0111]

[0112] The amine in Example 5 was converted to 5M using L-aminopropanol, and compound 6M was synthesized using the same method as compound 6A in Example 6.

[0113] 1 H NMR (400MHz, CDCl3) δ9.28(d,J=6.6Hz,1H),8.13(d,J=1.3Hz,1H),7.70(d,J=8.1Hz,1H),7.64(d,J=8.1Hz,1H),4.24(tt,J=7 .8,3.5Hz,1H),3.80–3.62(m,2H),3.18(t,J=6.4Hz,2H),1.82(h,J=5.8Hz,2H),1.67(t,J=5.9Hz,2H),1.33(d,J=5.7Hz,9H). 13 C NMR (101MHz, CDCl3) δ181.59,175.98,164.21,161.26,152.16,150.19,145.67,134.18,126. 44,126.20,122.63,121.40,115.75,68.04,48.93,37.76,34.99,31.90,30.13,19.08,16.85.

[0114] Example 19 Preparation of Compound 6N

[0115]

[0116] 6N

[0117] The amine in Example 5 was converted to 5N using alanine methyl ester hydrochloride, and compound 6N was synthesized using the same synthesis method as compound 6A in Example 6.

[0118] 1 H NMR(400MHz, CDCl3)δ9.55(d,J=6.9Hz,1H),8.13(d,J=1.4Hz,1H),7.70(dd,J=8.2,1.3Hz,1H),7.64(dd,J=8.1,1.3Hz,1H),4.74–4.62(m,1H), 3.76(d,J=1.4Hz,3H), 3.19(t,J=6.4Hz,2H), 1.82(p,J=6.1Hz,2H), 1.68(dd,J=8.6,3.8Hz,2H), 1.57(dd,J=7.3,1.4Hz,3H), 1.35–1.30(m,6H). 13 C NMR (101MHz, CDCl3) δ181.58,175.85,172.97,164.00,160.03,152.05,150.28,145.57,134.13,1 26.45,126.23,122.20,121.33,115.83,52.53,48.59,37.76,34.97,31.88,30.11,19.07,17.76.

[0119] Example 20 Preparation of Compound 6O

[0120]

[0121] The amine in Example 5 was converted to 2-aminothiazole to give 5O, and compound 6O was synthesized using the same method as compound 6A in Example 6.

[0122] 1 H NMR (400MHz, CDCl3) δ12.60 (s, 1H), 8.31 (s, 1H), 7.77–7.66 (m, 2H), 7.57 (d, J = 3.5Hz, 1H), 7. 04(d,J=3.5Hz,1H),3.23(t,J=6.4Hz,2H),1.89–1.78(m,2H),1.73–1.65(m,2H),1.34(s,6H).

[0123] Example 21 Preparation of Compound 6P

[0124]

[0125] The amine in Example 5 was converted to 5P using 2-aminobenzothiazole, and compound 6P was synthesized using the same method as compound 6A in Example 6.

[0126] 1 H NMR (400MHz, CDCl3) δ12.72(s,1H),8.35(s,1H),7.91(d,J=8.2Hz,1H),7.84(d,J=7.8Hz,1H),7.72(q,J=8.2Hz,2H),7. 45(d,J=8.2Hz,1H),7.33(d,J=7.3Hz,1H),3.24(t,J=6.3Hz,2H),1.85(d,J=8.1Hz,2H),1.77–1.67(m,2H),1.34(s,6H).

[0127] Example 22 Preparation of Compound 6Q

[0128] The amine in Example 5 was converted to 5Q using tetrahydropyrrole, and compound 6Q was synthesized using the same method as compound 6A in Example 6.

[0129] 1 H NMR (400MHz, CDCl3) δ7.70(s,1H),7.68(d,J=8.2Hz,1H),7.62(d,J=8.1Hz,1H),3.65(t,J =6.9Hz,2H),3.33(t,J=6.6Hz,2H),3.19(t,J=6.3Hz,2H),2.05–1.75(m,8H),1.32(s,6H).

[0130] Example 23 Preparation of Compound 6R

[0131]

[0132] The amine in Example 5 was substituted with piperidine to give 5R, and compound 6R was synthesized using the same method as compound 6A in Example 6.

[0133] 1 H NMR (400MHz, CDCl3) δ7.64–2.94(m,3H),3.24–2.94(m,6H),2.05–1.75(m,10H),1.68(s,6H).

[0134] Example 24 Preparation of Compound 6S

[0135]

[0136] The amine in Example 5 was converted to 5S using N,N-dimethylethylenediamine, and compound 6S was synthesized using the same synthesis method as compound 6A in Example 6.

[0137] 1 H NMR (600MHz, Acetone) δ9.52(s,1H),8.34(d,J=2.7Hz,1H),7.92(d,J=8.1Hz,1H),7.65(d,J=8.1Hz,1H),4.03(q,J=5 .6Hz,2H),3.79–3.74(m,2H),3.31(s,6H),3.14(t,J=6.4Hz,2H),1.86–1.79(m,2H),1.74–1.69(m,2H),1.36(s,6H). 13 C NMR(151MHz,Acetone)δ182.08,176.63,165.00,163.23,152.80,151.72,145.57,134 .74,127.05,122.23,116.47,60.16,44.59,38.47,36.37,35.61,32.01,30.74,20.34.

[0138] Example 25 Preparation of Compound 6T

[0139]

[0140] The amine in Example 5 was converted to 5T using 3-aminopropylmorpholine, and compound 6T was synthesized using the same method as compound 6A in Example 6.

[0141] 1 H NMR(600MHz,MeOD)δ8.29(s,1H),7.86(d,J=8.2Hz,1H),7.72(d,J=8.1Hz,1H),3.95(s,4H),3.59(t,J=6.5Hz, 2H),3.32(s,6H),3.20(t,J=6.4Hz,2H),2.19–2.11(m,2H),1.90–1.83(m,2H),1.75–1.70(m,2H),1.37(s,6H). 13C NMR(151MHz,MeOD)δ181.04,175.77,163.95,161.94,151.58,149.85,144.53,133.83,126.25,126 .12,121.41,120.94,115.35,63.82,54.72,51.94,37.51,35.86,34.44,30.67,29.74,23.68,18.71

[0142] Example 26 Preparation of Compound 7

[0143]

[0144] Compound 4 (5.06 g, 10 mmol) and potassium carbonate (4.12 g, 30 mmol) were weighed and placed in a 100 mL single-necked flask equipped with a magnetic stirrer. 40 mL of acetone was added and stirred at room temperature for 10 minutes. Iodomethane (3.7 mL, 60 mmol) was then added dropwise, and the temperature was raised to 50°C for 5 hours. After the reaction was complete, the solid was removed by filtration, and the residue was rinsed with ethyl acetate. The filtrate was collected and the solvent removed by rotary evaporation to obtain the crude product. An appropriate amount of petroleum ether and a small amount of ethyl acetate were added to the crude product, stirred for 10-15 minutes, and filtered to obtain 7 (4.94 g, 95%) as a white or light yellow solid.

[0145] 1 H NMR(500MHz, CDCl3)δ8.31(s,1H),8.10(d,J=8.7Hz,1H),7.58(d,J=8.7Hz,1H),7.53–7.46(m,2H),7.42(dd,J=7.5,1.8Hz,2H),7.39–7 .27(m,6H),5.26(s,2H),5.10(s,2H),3.79(s,3H),3.53(t,J=6.1Hz,2H),1.79(dt,J=6.4,3.8Hz,2H),1.74–1.67(m,2H),1.38(s,6H). 13 C NMR (101MHz, CDCl3) δ163.25,150.70,149.82,147.30,144.39,137.80,137.73,133.06,131.05,128.86,128.44,128.30,127.98, 127.86,127.73,127.03,126.10,118.19,117.99,115.74,115.68,76.65,76.34,51.98,38.86,35.17,32.28,30.95,29.86,20.32.

[0146] Example 27 Preparation of Compound 8

[0147]

[0148] Compound 7 (2.6 g, 5.0 mmol), magnesium turnings (2.4 g, 100 mmol), and ammonium chloride (1.3 g, 25 mmol) were weighed into a 250 mL two-necked flask equipped with a magnetic stirrer. 100 mL of anhydrous tetrahydrofuran and 25 mL of anhydrous methanol were added. The atmosphere was replaced with argon several times and the mixture was stirred vigorously at room temperature for 2-3 hours. After the reaction was complete, aqueous ammonium chloride was added at 0°C until the solution clarified. 50 mL of ethyl acetate was added, and after separation, the aqueous layer was extracted with ethyl acetate (15 mL x 3). The organic layers were combined, washed several times with water and once with saturated brine. The organic layers were dried over anhydrous sodium sulfate and the solvent was removed by rotary evaporation to obtain the crude product. An appropriate amount of petroleum ether and a small amount of ethyl acetate were added to the crude product, stirred for 10-15 minutes, and filtered to obtain 8 (2.5 g, 96%) as a white solid.

[0149] 1 H NMR (400MHz, CDCl3) δ7.72(d,J=8.8Hz,1H),7.45(d,J=6.5Hz,2H),7.42–7.28(m,9H),5.37(d,J=11.1Hz,1H),5.04(d,J=11.1Hz,1H),4.96(q,J=11. 1Hz,2H),4.90–4.77(m,2H),4.37(dd,J=9.6,6.2Hz,1H),3.62(s,3H),3.5 2–3.42(m,2H),1.74(ddd,J=26.1,11.1,5.6Hz,4H),1.35(d,J=7.2Hz,6H). 13 C NMR (101MHz, CDCl3) δ172.95,153.82,147.28,144.64,143.04,137.84,137.76,132.23,129.86,128.52,128.44,128.33, 128.09,128.01,127.83,124.79,119.59,117.04,113.41,76.37,75.07,52.51,47.57,38.95,35.11,32.12,30.83,20.34.

[0150] Example 28 Preparation of Compound 9

[0151]

[0152] Compound 8 (10.5 g, 20 mmol) was weighed and placed in a 250 mL single-necked flask equipped with a magnetic stirrer. 75 mL of tetrahydrofuran and 25 mL of methanol were added dropwise, and 20 mL of 2M sodium hydroxide solution was added dropwise. The reaction was allowed to proceed at 45°C for 2-3 hours. After completion of the reaction, the tetrahydrofuran and methanol were removed by rotary evaporation. The concentrate was acidified with 1N hydrochloric acid to a pH of 1-2. 100 mL of dichloromethane was added. After separation, the aqueous layer was extracted with dichloromethane (50 mL x 3). The organic layers were combined and washed once with saturated brine. After drying over anhydrous sodium sulfate, the dichloromethane was removed by rotary evaporation to yield crude yellow oil, 9 (9.9 g, 97%). Because 9 was unstable and easily deteriorated, it was resolved without further purification.

[0153] 1 H NMR (600MHz, CDCl3) δ7.59 (d, J=9.0Hz, 1H), 7.33–7.21 (m, 6H), 7.14–7.08 (m, 2H), 7.07 –7.01(m,3H),5.09(d,J=11.4Hz,1H),4.99(d,J=11.4Hz,1H),4.84(d,J=11.4Hz,1H),4. 64(d,J=11.4Hz,1H),4.61–4.54(m,1H),4.34(t,J=9.0Hz,1H),3.83–3.68(m,1H),3.50 (dt,J=17.4,4.8Hz,1H),3.30–3.16(m,1H),1.81–1.56(m,4H),1.32(s,3H),1.31(s,3H)

[0154] Example 29 Preparation of Compound 11

[0155]

[0156] Weigh compound 9 (1 g, 2 mmol) and place it in a 500 mL single-necked flask equipped with a magnetic stirrer. Add 150 mL of dichloromethane and (S)-(-)-α-methylbenzylamine (1.21 g, 10 mmol) and stir at room temperature for 15 to 25 minutes. After both are fully salted, remove the dichloromethane by rotary evaporation to obtain a yellow solid. Add 350 mL of isopropyl alcohol and reflux with stirring to completely dissolve the yellow solid. If some salt still does not dissolve, add methanol or isopropyl alcohol as needed until the solution is completely clear. After it is completely clear, keep the solution at 30 to 40°C and let the solid precipitate for 4 hours. Filter, and the filtrate is rotary evaporated to remove the isopropyl alcohol and retain the white filter cake. Place it in a 250 mL single-necked flask equipped with a magnetic stirrer. Add 200 mL of isopropyl alcohol and reflux with stirring to completely dissolve the white solid. If some salt still does not dissolve, add methanol or isopropyl alcohol as needed until the solution is completely clear. After the solution was completely clarified, the solution was kept at 40-50°C for 2 hours to precipitate solids. The solution was filtered, and the filtrate was rotary evaporated to remove isopropanol and retained to collect the white filter cake, i.e., compound 10.

[0157] Ethyl acetate was added to compound 10, acidified with 1N hydrochloric acid, and the aqueous layer was extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated to remove ethyl acetate to obtain a yellow oil 11 (ee value >95%).

[0158] Example 30 Preparation of Compound 15

[0159]

[0160] The (S)-(-)-α-methylbenzylamine in Example 29 was replaced with (R)-(+)-α-methylbenzylamine, and compound 15 (ee value>90%) was obtained by the same synthesis method as compound 11 in Example 29.

[0161] Figure 1 and Figure 2 The HPLC spectra of compound 11 and compound 15 are shown respectively, wherein the chromatographic conditions and chromatographic data are as follows:

[0162] Compound 11:

[0163] Daicel Chiralcel AD-H (0.46 cm × 25 cm), Hexane:i-PrOH = 95:5; v = 1.0 mL / min; λ = 254 nm. Data are as follows:

[0164]

[0165] Compound 15:

[0166] Daicel Chiralcel OD-H (0.46 cm × 25 cm), Hexane:i-PrOH = 95:5; v = 1.0 mL / min; λ = 254 nm. The data are as follows:

[0167]

[0168] Example 31 Preparation of Compound 12 and Compound 16

[0169]

[0170] Compound 11 or 15 (101.6 mg, 0.2 mmol) and HATU (150 mg, 0.4 mmol) were weighed and placed in a Schlenk tube equipped with a magnetic stirrer. The atmosphere was replaced with argon. 4 mL of dichloromethane, DIPEA (0.1 mL, 0.6 mmol), and various amines (0.4 mmol) were added and reacted at room temperature for 8-10 hours. After the reaction was complete, 15 mL of water and 10 mL of dichloromethane were added, along with a small amount of 1N HCl. After separation, the aqueous layer was extracted with dichloromethane (15 mL x 3). The organic layers were combined, washed several times with water and once with saturated brine. The organic layer was dried over anhydrous sodium sulfate and rotary evaporated to remove dichloromethane to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain the colloidal products 12A-12R and 16A-16R.

[0171] Example 32 Preparation of Compounds 13A and 17A

[0172]

[0173] 13Aor17A

[0174] Compound 12A or 16A (101 mg, 0.2 mmol) and 10% palladium on carbon (10 mg, 10 wt%) were weighed and placed in a 50 mL single-necked flask equipped with a magnetic stirrer. 4 mL of ethyl acetate and 4 mL of methanol were added, and the atmosphere was replaced with hydrogen several times. The reaction was allowed to proceed at 45°C for 1-2 hours. After the reaction was complete, the palladium on carbon was removed by filtration through celite, and the residue was rinsed with ethyl acetate. The filtrate was collected and the ethyl acetate was removed by rotary evaporation to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain 13A or 17A as a red solid.

[0175] 1H NMR (400MHz, CDCl3) δ7.66(dd,J=8.1,2.0Hz,1H),7.60–7.51(m,2H),5.57(ddd,J=9.9,5.9,2.0Hz,1H),5.51(s,1H),4.87(td,J=10.3,2.0Hz,1 H),4.30(ddd,J=10.5,5.9,2.0Hz,1H),3.20(dt,J=6.7,3.8Hz,2H),1.79(qt,J=6.5,3.7Hz,2H),1.72–1.62(m,2H),1.30(dd,J=3.7,2.0Hz,6H). 13 C NMR (101MHz, CDCl3) δ183.22,175.99,173.43,171.03,153.84,144.30,132.99,128.22,125.35 ,123.38,111.56,77.40,77.08,76.77,76.00,44.73,37.68,35.05,31.92,31.88,29.79,19.00.

[0176] Example 33 Preparation of Compounds 13B and 17B

[0177]

[0178] The amine in Example 31 was replaced with methylamine hydrochloride to obtain 12B or 16B, and compounds 13B and 17B were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0179] 1 H NMR (500MHz, CDCl3) δ7.66(d,J=8.1Hz,1H),7.55(d,J=8.0Hz,2H),5.63(dd,J=9.9,5.9Hz,1H),4.87(t,J=10.2Hz,1H),4.25(dd,J=1 0.5, 5.9Hz, 1H), 3.20 (t, J = 6.5Hz, 2H), 2.80 (d, J = 4.8Hz, 3H), 1.79 (td, J = 7.3, 4.7Hz, 2H), 1.69–1.65 (m, 2H), 1.31 (d, J = 4.8Hz, 6H). 13C NMR (126MHz, CDCl3) δ183.37,176.20,173.53,169.77,153.87,144.36,133.06,128. 28,125.54,123.45,112.13,44.99,37.78,35.13,32.00,31.96,29.87,26.69,19.09.

[0180] Example 34 Preparation of Compounds 13C and 17C

[0181]

[0182] The amine in Example 31 was replaced with dimethylamine hydrochloride to obtain 12C or 16C, and compounds 13C and 17C were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0183] 1 H NMR (400MHz, CDCl3) δ7.57(d,J=8.1Hz,1H),7.44(d,J=8.1Hz,1H),5.22(dd,J=9.2,5.6Hz,1H),4.84(t,J=9.4Hz,1H),4.56(dd, J=9.7,5.6Hz,1H),3.36(s,3H),3.16(t,J=6.4Hz,2H),2.96(s,3H),1.81–1.71(m,2H),1.65–1.59(m,2H),1.27(d,J=7.9Hz,6H). 13 C NMR (101MHz, CDCl3) δ183.52,174.55,172.30,170.37,152.84,143.78,132.67,128.27,125.92,12 3.03,113.98,78.33,77.44,77.12,76.80,41.74,38.32,37.75,36.44,34.91,31.92,29.72,19.05.

[0184] Example 35 Preparation of Compounds 13D and 17D

[0185]

[0186] The amine in Example 31 was replaced with morpholine to obtain 12D or 16D, and compounds 13D and 17D were synthesized using the same method as compounds 13A and 17A in Example 32.

[0187] 1H NMR (400MHz, CDCl3) δ7.63(d,J=8.1Hz,1H),7.53(d,J=8.1Hz,1H),5.34(dd,J=9.2,5.1Hz,1H),4.85(t,J=9. 4Hz,1H),4.51(dd,J=9.5,5.1Hz,1H),4.23(dt,J=13.1,3.6Hz,1H),4.06–3.93(m,2H),3.85(dt,J=11.9,3.8 Hz,1H),3.76(dt,J=11.7,3.9 Hz,1H),3.65–3.51(m,2H),3.28(ddd,J=13.1,9.1,3.6 Hz,1H),3.19(t,J=6.4 Hz,2H),1.79(p,J=6.3 Hz,2H),1.69–1.61(m,2H),1.30(d,J=5.8 Hz,6H). 13 C NMR(101 MHz, CDCl3)δ183.51,174.63,172.62,169.11,153.17,143.99,132.76,128.32,125.79,123.18,113.43,7 8.30,77.39,77.08,76.76,66.99,66.65,46.91,42.84,41.44,37.75,34.97,31.95,31.91,29.74,19.06.

[0188] Example 36 Preparation of Compounds 13E and 17E

[0189]

[0190] The amine in Example 31 was replaced with 3-(S)-3-methylmorpholine to obtain 12E or 16E, and compounds 13E and 17E were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0191] 1H NMR (400 MHz, CDCl3) δ7.63 (d, J = 8.1 Hz, 1H), 7.54 (d, J = 8.1 Hz, 1H), 5.37 (dd, J = 9.3, 4.9 Hz, 1H), 4.83 (t, J = 9.3 Hz, 1H), 4.56 (dt, J = 8.8, 4.4 Hz,1H),4.44(dd,J=9.4,4.9 Hz,1H),4.29(d,J=12.3 Hz,1H),4.03–3.87(m,2H),3.70–3.44(m,3H),3.19(t,J=6.5 Hz,2H),1.79(p,J=6.4Hz,2H),1.64(s,2H),1.28(dd,J=11.5,6.6 Hz,9H). 13 CNMR(101 MHz, CDCl3)δ183.64,174.77,172.72,168.86,153.25,144.04,132.86,128.41,125.92,123.32 ,113.42,78.43,70.84,67.29,45.61,41.87,37.84,35.05,32.04,31.98,29.81,19.15,15.19.

[0192] Example 37 Preparation of Compounds 13F and 17F

[0193]

[0194] The amine in Example 31 was replaced with isopropylpiperazine to obtain 12F or 16F, and compounds 13F and 17F were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0195] 1 H NMR (400MHz, MeOD) δ7.84(t,J=8.0Hz,1H),7.65(t,J=7.9Hz,1H),5.20(dq,J=8.8,4.9Hz,1H),5.01(t,J=9.0Hz,1H),4.81(dt,J=8.9,4.3Hz,1H),3. 66–3.59(m,3H),3.32(d,J=6.7Hz,6H),3.20(q,J=6.9Hz,2H),1.84(q,J=6 .6Hz,2H),1.71(q,J=5.4Hz,2H),1.43(d,J=6.9Hz,6H),1.38–1.32(m,6H). 13C NMR(101MHz,MeOD)δ184.40,178.89,175.74,174.77,154.60,144.97,134.15,128.99,126.45, 124.23,114.30,79.69,59.80,44.36,42.09,40.17,38.48,35.72,32.06,30.69,19.78,17.04.

[0196] Example 38 Preparation of Compounds 13G and 17G

[0197]

[0198] The amine in Example 31 was replaced with aniline to obtain 12G or 16G, and compounds 13G and 17G were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0199] 1 H NMR(400MHz, CDCl3)δ9.80(s,1H),7.66(d,J=8.1Hz,1H),7.61–7.55(m,3H),7.33–7.24(m,3H),7.07(t,J=7.4Hz,1H),5.71(dd,J=10.0,6.0Hz,1 H),4.95(t,J=10.3Hz,1H),4.48(dd,J=10.5,6.0Hz,1H),3.30–3.11(m,2H),1.80(h,J=6.7Hz,2H),1.66(t,J=5.9Hz,2H),1.31(d,J=8.1Hz,6H). 13 C NMR (101MHz, CDCl3) δ183.11,176.47,173.88,166.78,154.05,144.39,138.15,133.01,128.94,128.16,125.24 ,124.23,123.45,119.83,111.41,77.40,77.08,76.76,75.88,46.11,37.66,35.07,31.90,31.86,29.77,18.98.

[0200] Example 39 Preparation of Compounds 13H and 17H

[0201]

[0202] The amine in Example 31 was replaced with p-methoxyaniline to obtain 12H or 16H, and compounds 13H and 17H were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0203] 1 H NMR (400MHz, CDCl3) δ9.66(s,1H),7.67(d,J=8.2Hz,1H),7.57(d,J=8.1Hz,1 H),7.52–7.46(m,2H),6.87–6.79(m,2H),5.70(dd,J=10.0,6.0Hz,1H),4.94 (t,J=10.3Hz,1H),4.46(dd,J=10.5,5.9Hz,1H),3.77(s,3H),3.21(q,J=6.7 Hz, 2H), 1.80 (h, J = 6.7Hz, 2H), 1.66 (t, J = 5.9Hz, 2H), 1.31 (d, J = 7.9Hz, 6H). 13 C NMR (101MHz, CDCl3) δ182.94,176.24,173.61,166.30,156.05,153.78,144.15,132.78,131.15,127.95, 125.06,123.22,121.18,113.83,111.32,75.73,55.26,45.68,37.44,34.84,31.69,31.64,29.55,18.76.

[0204] Example 40 Preparation of Compounds 13I and 17I

[0205]

[0206] The amine in Example 31 was replaced with p-fluoroaniline to obtain 12I or 16I, and compounds 13I and 17I were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0207] 1 H NMR(400MHz, CDCl3)δ9.83(s,1H),7.68(d,J=8.1Hz,1H),7.61–7.49(m,3H),7.04–6.93(m,2H),5.70(dd,J=10.1,6.0Hz,1H),4.95(t,J= 10.3Hz,1H),4.47(dd,J=10.5,6.0Hz,1H),3.31–3.12(m,2H),1.80(dt,J=12.6,6.2Hz,2H),1.67(t,J=5.9Hz,2H),1.31(d,J=8.7Hz,6H). 13CNMR(101MHz, CDCl3)δ182.88,176.32,173.82,166.54,153.95,144.25,133.97,132.82,127.96,124.97, 123.27,121.29,121.21,115.45,115.23,111.08,75.64,45.77,37.43,34.86,31.69,31.63,29.56,18.76.

[0208] Example 41 Preparation of Compounds 13J and 17J

[0209]

[0210] The amine in Example 31 was replaced with chloroethylamine hydrochloride to obtain 12J or 16J, and compounds 13J and 17J were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0211] 1 H NMR (400MHz, CDCl3) δ7.97(t,J=5.6Hz,1H),7.66(d,J=8.1Hz,1H),7.55(d,J=8.1Hz,1H),5.60(dd,J=10.0,5.9Hz,1H),4.89(t,J=10.3Hz,1H), 4.30(dd,J=10.5,5.9Hz,1H),3.73–3.45(m,4H),3.25–3.17(m,2H),1.8 0(qdd,J=6.5,3.7,1.6Hz,2H),1.68–1.64(m,2H),1.31(d,J=5.4Hz,6H). 13 C NMR (101MHz, CDCl3) δ182.92,175.82,173.20,169.22,153.63,144.09,132.74,127.95,125 .11,123.14,111.42,75.77,44.77,42.69,41.50,37.44,34.82,31.70,31.63,29.55,18.76.

[0212] Example 42 Preparation of Compounds 13K and 17K

[0213]

[0214] The amine in Example 31 was replaced with ethanolamine to obtain 12K or 16K, and compounds 13K and 17K were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0215] 1 H NMR (400MHz, CDCl3) δ7.86(t,J=5.7Hz,1H),7.64(d,J=8.1Hz,1H),7.52(d,J=8.1Hz,1H),5.55(dd,J=9.9,6.1Hz,1H),4.89(t,J=10.2Hz,1H),4.30( dd,J=10.4,6.0Hz,1H),3.70(t,J=5.1Hz,2H),3.51–3.34(m,2H),3.16(q, J=6.0Hz,2H),1.85–1.71(m,2H),1.70–1.60(m,2H),1.29(d,J=6.1Hz,6H). 13 C NMR (101MHz, CDCl3) δ183.21,175.92,173.58,170.21,153.82,144.28,132.97,128.12,125.35,123.3 8,112.00,77.41,77.09,76.78,76.40,62.10,45.15,42.98,37.65,35.03,31.90,31.86,29.77,18.97.

[0216] Example 43 Preparation of Compounds 13L and 17L

[0217]

[0218] The amine in Example 31 was replaced with glycine methyl ester hydrochloride to obtain 12L or 16L, and compounds 13L and 17L were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0219] 1 H NMR (400MHz, CDCl3) δ8.07(t,J=5.8Hz,1H),7.65(d,J=8.1Hz,1H),7.53(d,J=8.1H z,1H),5.59(dd,J=10.0,5.9Hz,1H),4.89(t,J=10.3Hz,1H),4.36(dd,J=10.5,5.9 Hz,1H),4.07(dd,J=17.9,5.9Hz,1H),3.93(dd,J=17.9,5.5Hz,1H),3.71(s,3H),3 .19(t,J=6.4Hz,2H),1.86–1.72(m,2H),1.70–1.61(m,2H),1.30(d,J=4.4Hz,6H). 13C NMR (101MHz, CDCl3) δ183.17,176.01,173.41,169.83,169.68,153.82,144.27,132.96,128.22,125.37,12 3.37,111.61,77.43,77.11,76.79,75.98,52.33,44.89,41.62,37.68,35.05,31.93,31.87,29.79,19.00.

[0220] Example 44 Preparation of Compounds 13M and 17M

[0221]

[0222] The amine in Example 31 was replaced with L-aminopropanol to obtain 12M or 16M, and compounds 13M and 17M were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0223] 1 H NMR (400MHz, CDCl3) δ7.64(t,J=7.0Hz,2H),7.53(d,J=8.1Hz,1H),5.54(dd,J=9.9,6. 2Hz,1H),4.89(t,J=10.2Hz,1H),4.27(dd,J=10.5,6.2Hz,1H),4.09–3.99(m,1H),3.6 1(dd,J=11.4,3.7Hz,1H),3.47(dd,J=11.2,6.3Hz,1H),3.17(q,J=6.9Hz,2H),1.79(h ept,J=6.4Hz,2H),1.65(q,J=7.0Hz,2H),1.30(d,J=6.5Hz,6H),1.21(d,J=6.8Hz,3H). 13 CNMR(101MHz, CDCl3)δ183.24,176.06,173.67,169.51,153.86,144.29,132.96,128.14,125.34,123.40,1 11.93,77.41,77.09,76.77,76.32,66.93,48.64,45.37,37.66,35.04,31.92,31.86,29.76,18.98,16.93.

[0224] Example 45 Preparation of Compounds 13N and 17N

[0225]

[0226] The amine in Example 31 was replaced with alanine methyl ester hydrochloride to obtain 12N or 16N, and compounds 13N and 17N were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0227] 1 H NMR (400MHz, CDCl3) δ7.99 (d, J = 6.9 Hz, 1H), 7.65 (d, J = 8.1 Hz, 1H), 7.53 (d, J = 8. 1Hz,1H),5.57(dd,J=9.9,6.0Hz,1H),4.87(t,J=10.2Hz,1H),4.46(p,J=7.2Hz, 1H),4.34(dd,J=10.5,6.0Hz,1H),3.74(s,3H),3.20(q,J=6.0Hz,2H),1.79(h,J =5.9Hz,2H),1.65(t,J=5.8Hz,2H),1.39(d,J=7.2Hz,3H),1.30(d,J=7.6Hz,6H). 13 C NMR (101MHz, CDCl3) δ183.26,175.96,173.20,172.95,168.68,153.75,144.25,132.94,128.20,125.41,123.3 3,111.61,77.41,77.09,76.77,75.92,52.44,48.77,45.02,37.69,35.03,31.94,31.86,29.76,19.01,17.71.

[0228] Example 46 Preparation of Compounds 13O and 17O

[0229]

[0230] The amine in Example 31 was replaced with 2-aminothiazole to obtain 12O or 16O, and compounds 13O and 17O were synthesized using the same method as compounds 13A and 17A in Example 32.

[0231] 1H NMR (600MHz, CDCl3) δ7.66(d,J=8.1Hz,1H),7.55(d,J=8.1Hz,1H),7.46(d,J=3.5Hz,1H),6.94(d,J=3.5Hz,1H),5.65(dd,J=10.0,6.0Hz,1H),5.0 1(t,J=10.2Hz,1H),4.58(dd,J=10.4,6.0Hz,1H),3.25–3.11(m,2H),1.8 4–1.74(m,2H),1.65(ddt,J=7.2,5.4,2.2Hz,2H),1.31(d,J=8.6Hz,6H). 13 C NMR (151MHz, CDCl3) δ182.84,176.31,173.93,167.02,157.56,154.34,144.53,138.23,133.03,1 28.36,125.06,123.53,113.81,110.64,75.66,45.55,37.73,35.18,31.98,31.95,29.85,19.04.

[0232] Example 47 Preparation of Compounds 13P and 17P

[0233]

[0234] The amine in Example 31 was replaced with 2-aminobenzothiazole to obtain 12P or 16P, and compounds 13P and 17P were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0235] 1 H NMR (400MHz, CDCl3) δ11.37(s,1H),7.80(dd,J=15.7,8.1Hz,2H),7.67(d,J=8.1 Hz,1H),7.56(d,J=8.1Hz,1H),7.41(t,J=7.7Hz,1H),7.33–7.25(m,1H),5.70(dd ,J=10.2,6.0Hz,1H),5.03(t,J=10.3Hz,1H),4.62(dd,J=10.5,6.0Hz,1H),3.30– 3.10(m,2H),1.79(h,J=7.0Hz,2H),1.66(t,J=5.9Hz,2H),1.31(d,J=9.3Hz,6H). 13C NMR (101MHz, CDCl3) δ182.76,176.47,174.12,167.67,156.96,154.48,148.98,144.64,133.05,132.46,128.39,1 26.29,124.98,124.04,123.56,121.77,121.29,110.26,75.44,45.83,37.72,35.21,32.00,31.95,29.88,19.05.

[0236] Example 48 Preparation of Compounds 13Q and 17Q

[0237]

[0238] The amine in Example 31 was replaced with tetrahydropyrrole to obtain 12Q or 16Q, and compounds 13Q and 17Q were synthesized using the same synthesis method as compounds 13A and 17A in Example 32.

[0239] 1 H NMR (400MHz, CDCl3) δ7.60(d,J=8.1Hz,1H),7.49(d,J=8.1Hz,1H),5.25(dd,J=9.2,6.0Hz,1H),4.87(t,J=9.5Hz,1H),4.46–4.33(m,2 H),3.46(qd,J=7.3,2.6Hz,3H),3.17(t,J=6.4Hz,2H),2.14–1.82(m,4H),1.81–1.72(m,2H),1.69–1.58(m,2H),1.29(d,J=5.2Hz,6H).

[0240] Example 49 Preparation of Compounds 13R and 17R

[0241]

[0242] The amine in Example 49 was replaced with piperidine to obtain 12R or 16R, and compounds 13R and 17R were synthesized using the same method as that for compounds 13A and 17A in Example 32.

[0243] 1H NMR (400MHz, CDCl3) δ7.58(dd,J=8.1,5.8Hz,1H),7.45(d,J=7.9Hz,1H),5.24(dt,J =8.9,5.5Hz,1H),4.84(td,J=9.3,5.8Hz,1H),4.57(dd,J=9.8,5.3Hz,1H),3.64(ddd d,J=18.8,11.1,7.6,4.8Hz,2H),3.17(q,J=6.2Hz,2H),2.95(d,J=5.8Hz,2H),1.81 –1.72(m,2H),1.66–1.61(m,6H),1.55(d,J=7.0Hz,2H),1.28(dd,J=8.9,5.7Hz,6H).

[0244] The following test methods, unless otherwise specified, are conventional test methods in this field.

[0245] Test Example 1 In vitro antibacterial activity test

[0246] The minimum inhibitory concentrations of 6A-6T, 13A-13R, and 17A-17R against Gram-positive bacteria such as Bacillus subtilis and Staphylococcus aureus were determined by the MIC method.

[0247] Table 1 MICs of Tanshinone IIA and Cryptotanshinone amide derivatives (μg / mL)

[0248]

[0249]

[0250] Test Example 2 In vitro NLRP3 inflammasome activation experiment

[0251] The inhibitory activities of 6A-6T, 13A-13R and 16A-16R at a concentration of 10 μM on LPS-induced IL-1β, TNF-α and IL-6 release in RAW264.7 macrophages were as follows: Figures 3-5 As shown (the performance not listed in the figure is to promote the release of inflammatory factors). The content of these three inflammatory factors was determined by ELISA method. It can be seen from the figure that it has good inhibitory activity against IL-1β, but poor inhibitory activity against TNF-α and IL-6.

[0252] Test Example 3 IL-1βIC 50

[0253] Next, we selected 16 compounds with good anti-IL-1β activity at a concentration of 10 μM from the test compounds and performed IC 50 Determination of.

[0254] Table 2 IC values ​​of preferred compounds for inhibition of IL-1β 50 value

[0255]

[0256]

[0257] Test Example 4: Toxicity of mouse peritoneal macrophages

[0258] In addition, cytotoxicity tests on mouse peritoneal macrophages (MPMs) were also performed, and the results are shown in Table 3.

[0259] Table 3 Cytotoxicity of tanshinone IIA and cryptotanshinone amide derivatives on MPMs

[0260]

[0261] Test Example 5 Animal Model

[0262] Based on the above activity test results, compounds 6D and 6K with good antibacterial and anti-inflammatory activities and high stability were selected to prepare hydrogels with a concentration of 0.2%, and were tested in wound healing models and keratitis models.

[0263] 5.1 Wound healing model

[0264] Forty male Kunming mice aged 6-8 weeks, weighing 20±2 g, were selected and fed adaptively for one week before modeling. The mice were anesthetized by intraperitoneal injection of sodium pentobarbital (60 mg / kg). After depilation of the back of the mice, full-thickness skin excision was performed to create a circular wound with a diameter of 7 mm. Then, 50 μL of 1×10 7 The MRSA bacterial suspension was inoculated on the wound. The mice were then randomly divided into 4 groups: model group, blank hydrogel group, 6D hydrogel group, and 6K hydrogel group, with 10 mice in each group. The model group was given normal saline every day, and the hydrogel group was given hydrogel treatment. The wounds were photographed on the 3rd, 7th, 11th, and 14th days, and the wound healing rate was calculated. On the 4th day of treatment, 5 mice in each group were killed, and the skin tissue was homogenized. After continuous dilution with normal saline, it was spread on the MH bacterial culture plate, cultured at 37°C for 18 hours, and the colony count was performed. All mice were killed on the 14th day, and the skin wound tissue and major organs (heart, liver, spleen, lung, and kidney) were fixed with 4% formaldehyde solution, paraffin sliced, and histopathological examination was performed with H&E staining. The results are shown in the table. Figures 6-8 .

[0265] Result analysis: Figure 6As shown in Figure a, on the 3rd and 7th day of treatment, the wounds in the 6D and 6K gel groups healed significantly compared to the model group and the blank gel group, which is consistent with the trend of wound healing rate in Figure b. On the 3rd and 7th day, the wound healing rates in the 6D and 6K gel groups were statistically different from those in the model group (P<0.05). Figure 6 c and Figure 6 As shown in d, on the fourth day of treatment, the wound tissue colony load in the 6D and 6K gel groups was significantly lower than that in the model group. Figure 7 As shown in the figure, after 14 days of treatment, the epidermis of the wounds of mice in each group had been completely reconstructed. Compared with the model group and the blank gel group, more hair follicles, sweat glands and new blood vessels were observed in the wound tissues of the 6D and 6K gel groups. In addition, there were no significant changes in the major organs (heart, liver, spleen, lungs, and kidneys). Figure 8 ), indicating that the hydrogel has excellent biosafety in vivo.

[0266] 5.2 Keratitis model

[0267] Thirty-two male Kunming mice aged 6-8 weeks, weighing 20±2 g, were selected and fed adaptively for one week before modeling. The mice were anesthetized by intraperitoneal injection of sodium pentobarbital (60 mg / kg), and then 5 μL of (0.3%) procaine hydrochloride was dripped into the right eyeball for local anesthesia. The corneal epithelium of the mice was scratched with a 30G sterile needle. The scratched area was a circle with a diameter of 3 mm centered on the pupil. 5 μL of 1×10 8 CFU / mL of MRSA bacterial liquid was used to suture the eyelids with an 8-0 suture needle. The sutures were removed after 12 hours, and the cornea was observed to have an abscess and turbid area, indicating that the keratitis model was successfully established. The mice with successful modeling were randomly divided into 4 groups and given normal saline, 6D (2 mg / mL), 6K (2 mg / mL), and linezolid (1 mg / mL) respectively. The drugs were given once every 5 minutes within the first hour, and once every half an hour for the next 7 hours. Half an hour after the last dose, all mice were killed, and the eyeballs of the mice were removed. Tissue homogenates of 6 mice in each group were continuously diluted with normal saline and spread on MH bacterial culture plates. They were cultured at 37°C for 18 hours and the colonies were counted. The eyeballs and major organs (heart, liver, spleen, lungs, and kidneys) of 2 mice in each group were removed and fixed with 4% formaldehyde solution, paraffin sliced, and histopathological examination was performed with H&E staining. The results are shown in the figure. Figures 9-11 .

[0268] Result analysis: Figure 9 As shown in Figure a, before treatment, obvious abscesses and turbidity were observed in mice in each group, indicating that the MRSA-infected keratitis model was successfully established. After treatment, the corneal abscesses and turbidity of the model group mice were significantly deeper than before treatment, while the corneal abscesses and turbidity of the mice in the 6D, 6K and linezolid treatment groups showed varying degrees of relief. Figure 9 b and Figure 9 As shown in c, the colony load of corneal tissue in the 6D, 6K and linezolid treatment groups was significantly lower than that in the model group. Figure 10 As shown in the figure, after treatment, the corneal epithelium of the model group showed obvious damage, while after treatment with 6D, 6K and linezolid, the corneal tissue was intact and the edema was reduced. In addition, there were no significant changes in the major organs (heart, liver, spleen, lung, and kidney) ( Figure 11 ), indicating that the drug has good biosafety in the body.

Claims

1. A tanshinone IIA or cryptotanshinone amide derivative, characterized in that: The compound represented by the general formula (I) or its stereoisomer, racemate, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug; wherein R1 and R2 are independently selected from H, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted ester, substituted or unsubstituted aromatic heterocyclic group, substituted or unsubstituted aryl, or R1, R2 and the nitrogen atom to which they are attached together form a substituted or unsubstituted 5-6 membered heterocyclic group; The substituent on the C1-C3 alkyl group is selected from halogen, hydroxyl, C1-C4 alkoxycarbonyl, di-C1-C4 alkylamino or C1-C4 alkylamino; The substituent on the aromatic heterocyclic group or aryl group is halogen, C1-C4 alkoxy or C1-C4 alkyl; The substituent on the 5- to 6-membered heterocyclic group is a C1-C4 alkyl group.

2. The tanshinone IIA or cryptotanshinone amide derivative according to claim 1, characterized in that: wherein each R1 and R2 are independently H, methyl, chloroethyl, substituted or unsubstituted hydroxyethyl, substituted or unsubstituted C2 ester, ethyl substituted by methoxycarbonyl, N,N-dimethylethyl, phenyl, p-methoxyphenyl, p-fluorophenyl, thiazolyl or benzothiazolyl; or, R1, R2 and the nitrogen atom to which they are attached together form a substituted or unsubstituted morpholinyl, N-isopropylpiperazinyl, tetrahydropyrrolyl or piperidinyl; Wherein, the substituent of the morpholinyl group is an optically active methyl group.

3. The tanshinone IIA or cryptotanshinone amide derivative according to claim 2, characterized in that: The following compound or its stereoisomer, racemate, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug:

4. The method for preparing tanshinone IIA and cryptotanshinone amide derivatives according to claim 3, characterized in that: Including the preparation of tanshinone IIA amide derivatives or cryptotanshinone amide derivatives; The preparation of the tanshinone IIA amide derivative is as follows: The following processes are included: The two ketone carbonyl groups of compound 1 were protected with benzyl groups to give compound 2, which was then oxidized with selenium dioxide to give compound 3. This was followed by Pinnick oxidation to give compound 4, which then underwent acid-amine condensation with the corresponding amine in the presence of HATU to form compound 5. Finally, it was hydrogenated and debenzylated in the presence of palladium on carbon to give the target compound 6. The preparation of the cryptotanshinone amide derivative is as follows: The following processes are included: Compound 4 was esterified with iodomethane under alkaline conditions to produce compound 7, followed by magnesium-methanol double bond reduction to produce compound 8, which was then hydrolyzed to produce compound 9. Compound 9 was then reacted with two configurations of α-methylbenzylamine for chiral resolution to produce compounds 10 and 14, which were then subjected to acid stripping, acid-amine condensation, and palladium-carbon hydrogenation to produce the R-configuration target compound 13 and the S-configuration target compound 17. Wherein, R1 and R2 are defined as in any one of claims 1 to 3.

5. A pharmaceutical composition comprising an effective amount of the tanshinone IIA or cryptotanshinone amide derivative according to any one of claims 1 to 3, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

6. Use of the tanshinone IIA or cryptotanshinone amide derivative according to any one of claims 1 to 3, or the pharmaceutical composition according to claim 5, in the preparation of anti-inflammatory drugs, antibacterial drugs, anti-tumor drugs, drugs for treating cardiovascular and cerebrovascular diseases, drugs for treating burns and scalds, and drugs for treating infectious diseases.

7. The use according to claim 6, characterized in that The application is to prepare antibacterial drugs or anti-inflammatory drugs.

8. The use according to claim 7, characterized in that The antibacterial drug is used to inhibit Bacillus subtilis and Staphylococcus aureus; The anti-inflammatory drug is used for treating keratitis.

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