AU2020103289A4

AU2020103289A4 - Preparation and Application of Alkaloid Derivatives for Reversing Human Tumor Drug Resistance and Anti-canine Breast Cancer

Info

Publication number: AU2020103289A4
Application number: AU2020103289A
Authority: AU
Inventors: Yingying GAO; Changliang He; Xiaoxia Liang; Shangxian Luan; Qiang Wu; Lizi Yin; Li Zhang
Original assignee: Sichuan Agricultural University
Current assignee: Sichuan Agricultural University
Priority date: 2020-11-06
Filing date: 2020-11-06
Publication date: 2021-01-14
Anticipated expiration: 2028-11-06

Abstract

The invention relates to a method for synthesizing and preparing a series of 8-position long-chain fatty acyloxy substituted diterpene alkaloid derivatives by taking natural diterpene alkaloids as raw materials; Experiments have proved that these compounds can be used as anti-tumor preparations for reversing tumor drug resistance and anti canine breast cancer preparations. -1/15 R9RB R 9 sR R1 , e f l R RI - r Ro-0--NHl \ "R X io - R3R R4 Figure 1

Description

-1/15 R9RB R 9sR R1 , e

fl - r R RI0 --NHl Ro- \ "R X io -

R3R R4

Figure 1

Preparation and Application of Alkaloid Derivatives for Reversing Human Tumor Drug Resistance and Anti-canine Breast Cancer

TECHNICAL FIELD

The present invention relates to a process for preparing 8-position long-chain fatty acyloxy substituted diterpene alkaloids from natural diterpene alkaloids;The invention also relates to the use of the compound in preparing preparations for reversing multidrug resistance of human tumors and treating canine breast cancer, belonging to the pharmaceutical field.

BACKGROUND ART

[0002] Breast cancer is one of the recognized public health problems in the world, which seriously threatens the physical and mental health of women all over the world. Drug chemotherapy is one of the main methods to treat breast cancer. However, with the development of breast cancer chemotherapy, the resistance of breast cancer to chemotherapy has become the main obstacle to breast cancer chemotherapy, often leading to the failure of breast cancer chemotherapy. Multidrug resistance (MDR) of tumor cells is one of the main causes of tumor chemotherapy failure. Doxorubicin (ADM) is the most commonly used drug for clinical treatment of breast cancer, but long-term use will lead to multidrug resistance of breast cancer cells, which will lead to failure of chemotherapy. Multidrug resistance of breast cancer will lead to treatment failure and spread of breast cancer,directly affecting the prognosis and overall survival cycle of patients. It is of great significance to explore the mechanism of anti-breast cancer reversing agents that reverse adriamycin resistance and find new drugs to overcome drug resistance of tumor cells, which has attracted more and more pharmaceutical workers' attention.

[0003] In addition, dogs, as companion animals of human beings, live in a very similar environment and contact with carcinogenic factors to human beings. Therefore, dog breast tumors are also the most common malignant tumors in female dogs, with an incidence rate as high as 25% - 42%, which is 3 times the malignant rate of human breast tumors. The same inducement also leads to similarities in the pathogenesis of human and dog breast cancer. Studies have shown that the gene sequences controlling the morphology, biological behavior and clinical development of canine breast tumors have high homology with human relative gene sequences, and both have the same susceptibility genes BRCA1 and BRCA2. Therefore, canine breast tumors are also natural models of human breast tumors. In-depth research on drugs for the treatment of canine breast tumors can not only save human faithful partners, but also undoubtedly open up new research approaches for human tumor medicine, which has dual significance.

[0004] Diterpenoid alkaloids are a kind of natural products with complex structures, which are mainly distributed in Aconitium, Delphinium and Spirea of the family Trifoliaceae, and have interesting chemical properties and extensive biological activities. Diterpenoid alkaloids have long aroused scientists' great interest in phytochemistry, synthesis and pharmaceutical chemistry due to their important pharmacological activities and structural complexity such as anti-inflammatory, analgesic, anti-platelet aggregation, anti-tumor, insecticidal, immune regulation, etc. However, these components often have obvious toxicity to nervous system, circulatory system, digestive system, etc., so their clinical application is very limited. How to improve its anti-tumor activity while reducing toxicity and enhancing its targeting to tumor tissues has very important theoretical value and practical significance.

The invention uses natural diterpenoid alkaloids as raw materials to synthesize and prepare a series of 8-position long-chain fatty acyloxy substituted diterpenoid alkaloid derivatives, from which anti-tumor preparations with low toxicity and high efficiency for reversing tumor drug resistance are screened, and compounds with inhibitory activity for canine breast cancer are found.

SUMMARY

The present invention aims to provide an anti-tumor compound with unique structure, which has good reversal of multidrug resistance of tumor and also has inhibition of canine breast cancer. Its structure is as follows: formula and its salt (see fig. 1)

Characterized in that Ro is hydrogen, methyl and ethyl; R1 , R3, R4, R 8 are hydrogen, hydroxyl,C1-6 alkoxy groups such as methoxy or ethoxy, C1-6 alkanoyloxy such as acetyl; R2 is hydroxyl; R6 , R9, Rio is hydrogen, hydroxyl or C1-6 alkanoyloxy, for example acetyl, R 5 is a saturated or unsaturated or halogen-containing long-chain fatty acyloxy group containing 8 to 24 carbon atoms, such as octanoyl, linoleoyloxy, linolenoyloxy, oleoyloxy, palmitic acyloxy, stearoyloxy, eicosapentaenoyloxy, 10 fluorooctadecyloxy,9,13-difluorooctadecyloxy and 10-bromooctadecyloxy,9,13 dibromooctadecyloxy,R 7 is hydrogen or hydroxyl or benzoyloxy or p-halobenzoyloxy; X is an acid radical commonly used in pharmacy,such as chlorine,bromine, trifluoroformyloxy, etc.

IIn the structure of the preferred compound formula of the present invention, Ro is

preferred hydrogen and ethyl; R1 , R3, R4 , R8 are preferred methoxy, R2 is preferred hydroxy,R 6 ,R9,Rio is preferred hydrogen or hydroxy or acetyl,R 5 is preferred saturated or unsaturated or halogen-containing long-chain fatty acyloxy containing 8 to 18 carbon atoms,such as octanoyl,linoleoyloxy,oleoyloxy,palmitoyloxy,stearoyloxy,R7 is preferred p-methoxybenzoyloxy,and X is preferred chlorine.

I The preferred compounds of formula for the present invention are selected from the

group consisting of the following compounds (see FIG. 2):

The formula of the present invention

I The compound of formula of the invention can change the substituents in the

structure and introduce the 8-position long-chain fatty acid ester by chemical methods by taking the corresponding diterpenoid alkaloid as a raw material.

The following examples of the present invention list examples of the preparation of 8 position long-chain fatty acid ester alkaloid derivatives using diterpene alkaloids as raw materials.

[0010] The following examples of the present invention list the proliferation inhibitory activity of the 8-position long-chain fatty acid ester alkaloid derivative to adriamycin-resistant human breast cancer cells and canine breast cancer cells, and the acute toxicity test examples of representative drugs.

Description of FIGURES

[0011] FIG. 1 is a structural formula of the compound formula.

[0012]FIG. 2 is a structural formula of a preferred compound of formula.

[0013]FIG. 3 is a structural formula for synthesize derivatives 2-33.

[0014]FIG. 4 is a road map for that preparation of synthetic derivative 2-21, 23-31.

[0015]FIG. 5 is a road map for the preparation of synthetic derivatives 22, 32, 33.

13C-NMR

[0016]FIG. 6 spectroscopic data for compound 1-6 in CDCl 3 100 ( 13 C).

[0017]FIG. 7 13 C-NMR spectroscopic data for compound 7-13 in CDCl 3 100 ( 13C).

13C-NM Rspectroscopic data for compound 14-17 in CDCl3 100 or 150

[0018FIG. 8

( 13C).

13C-NMR

[0019]FIG. 9 spectroscopic data for compound 21-25 in CDCl 3 100 ( 13C).

[0020]FIG. 10 13C-NMR spectroscopic data for compound 26-29 in CDCl 3 100 ( 13 C).

13

[00211FIG. 11 C-NMR spectroscopic data for compound 30-33 in CDCl 3 100 ( 13 C).

[0022]FIG. 12 IC 5 0 (M) result of inhibition of compounds 1-33 on drug-resistant human breast cancer cell (ADR-MCF-7) and canine breast cancer cell (CMT1211).

[0023]FIG. 13 organ diagram of each dose group of aconitine linoleate (7).

[0024]FIG. 14 that change of liver structure of mice in different treatment group (SP. Times. 400).

[0025] FIG. 15 Changes in lung structure of mice in different treatment groups (SP. Times.400).

DETAILED DESCRIPTION:

EXAMPLE 1: The structural formulas of compounds 1-6 referred to in the following examples are shown in FIG. 3.

[0027] Aconitine (1) is used as raw material and catalyzed by p-toluenesulfonic acid to react with long-chain fatty acid acyl chloride reagents (octanoyl chloride, myristoyl chloride, palmitoyl chloride, stearoyl chloride, oleoyl chloride and linoleoyl chloride) at 45 °C to obtain compounds 1-6 respectively. The synthesis route is shown in Fig. 4.

[0028] 3-capryloyl-aconitine (1). Light yellow oil, yield 28.44%. IR (KBr): 3457, 2928, 1725, 1638, 1453, 1383, 1278, 1098, 985, 749, 710, 615 cm-'; 'H-NMR (400

MHz, CDC 3 ) 6 8.02 (d, J= 7.2 Hz, 1H, H-2", 6"), 7.57 (t, J= 7.4 Hz, 1H, H-4"),

7.45 (t, J= 7.6 Hz, 2H, H-3", 5"), 4.87 (d, J= 5.0 Hz, 1H, H-14 1 ), 4.46 (dd, J= 5.2,

2.8 Hz, 1H, H-15 ), 4.37 (d, J= 2.8 Hz, 1H, H-31 ), 4.07 (d, J= 6.5 Hz, 1H, H-6 ),

3.89 (s, 1H, H-17), 3.73 (s, 3H, 16'-OCH 3 ), 3.36 (d, J= 8.8 Hz, 1H, H-18 a ), 3.31 (d,

J= 5.3 Hz, 1H, H-16 a), 3.24 (s, 3H, l'-OCH3), 3.18 (s, 3H, 6'-OCH3 ), 3.17 (s, 3H,

18'-OCH 3), 1.10 (t, J= 7.1 Hz, 3H, N-CH 2CH3 ), 0.87 (t, J= 6.8 Hz, 3H, H-8"'). 13 C

NMR Figure 6. HRMS calculated for C 4 1H 62 NO1 2 772.4272, found 772.4277[M+H]+.

3-myristoyl-aconitine (2). Light yellow oil, yield 30.17%. IR (KBr): 3467, 2926, 2854, 1724, 1640, 1452, 1383, 1279, 1110, 1095, 985, 895, 836, 710, 614 cm-1; 'H-NMR

(400 MHz, CDC 3 ) 6 8.05 (d, J= 7.4 Hz, 1H, H-2", 6"), 7.59 (t, J= 7.3 Hz, 1H, H

4"), 7.47 (t, J= 7.6 Hz, 2H, H-3", 5"), 4.90 (d, J= 5.1 Hz, 1H, H-14 1 ), 4.49 (dd, J=

4.9, 2.7 Hz, 1H,H-15 1 ), 4.40 (d, J 2.6 Hz, 1H, H-3 1 ), 4.10 (d, J= 6.7 Hz, 1H, H

6 1 ), 3.91 (s, 1H, H-17), 3.76 (s, 3H, 16'-OCH3), 3.33 (d, J= 5.0 Hz, 1H, H-16 a ),

3.27 (s, 3H, 18'-OCH 3), 3.22 (s, 3H, l'-OCH3), 3.21 (s, 3H,6'-OCH 3), 1.12 (t, J= 7.1 Hz, 3H, N-CH 2 CH3), 0.90 (t, J= 6.7 Hz, 3H, H-14"'). 13C NMR Figure 6. HRMS calculated for C 48 H 74 NO 12 856.5211, found 856.5267[M+H]+.

3-palmityl-aconitine (3). Light yellow oil, yield 35.60%. IR (KBr): 3462, 2924, 2853, 1723, 1642, 1452, 1383, 1278, 1117, 1095, 985, 710, 614 cm-1; IH-NMR (400 MHz,

CDC 3 ) 6 8.05 (d, J = 7.3 Hz, 1H, H-2", 6"), 7.59 (t, J= 7.4 Hz, 1H, H-4"), 7.47 (t, J

= 7.6 Hz, 2H, H-3", 5"), 4.89 (d, J= 5.1 Hz, 1H, H-14 1 ), 4.48 (dd, J= 5.0, 2.7 Hz,

1H, H-15 ), 4.39 (d, J= 2.6 Hz, 1H, H-3 1 ), 4.10 (d, J= 6.5 Hz, 1H, H-61 ), 3.91 (s,

1H, H-17), 3.76 (s, 3H, 16'-OCH 3), 3.33 (d, J= 5.0 Hz, 1H, H-16 a),3.27 (s, 3H,

18'-OCH 3), 3.22 (s, 3H, l'-OCH 3), 3.21 (s, 3H,6'-OCH 3), 1.12 (t, J=7.1 Hz, 3H, N

CH 2 CH3 ), 0.90 (t, J= 6.7 Hz, 3H, H-16"'). 13C NMR Figure 6. HRMS calculated for C 5 H 7 NO 12 884.5524, found 884.5597[M+H]+.

3-stearatyl-aconitine(4). Light yellow oil, yield 28.32%. IR (KBr): 3458, 2922, 2851, 1722, 1635, 1450, 1384, 1277, 1093, 709 cm-1; H-NMR (400 MHz, CDC 3) 6 8.05

(d, J= 7.4 Hz, 1H, H-2", 6"), 7.59 (t, J= 7.4 Hz, 1H, H-4"), 7.47 (t, J= 7.6 Hz, 2H, H 3", 5"), 4.89 (d, J 5.1 Hz, 1H, H-14 1 ), 4.53 - 4.45 (m, 1H, H-15 ), 4.39 (d, J=

2.4 Hz, 1H, H-3 ),4.10 (d, J= 6.5 Hz, 1H, H-6 ), 3.92 (s, 1H, H-17), 3.75 (s, 3H,

16'-OCH 3), 3.33 (d, J= 5.1 Hz, 1H, H-16 a ), 3.27 (s, 3H, 18'-OCH3), 3.22 (s, 3H, '

OCH 3 ), 3.21 (s, 3H,6'-OCH 3), 1.12 (t, J= 7.1 Hz, 3H, N-CH 2 CH3), 0.90 (t, J= 6.6 Hz, 3H, H-18"'). 13C NMR Figure 6. HRMS calculated for C 2 H 2NO1 2 912.5837, found 912.5805[M+H]+.

3-oleoyl-aconitine (5). Light yellow oil, yield 33.70%. IR (KBr): 3454, 2923, 2852, 1638, 1384, 1276, 1093, 450, 709 cm-1; IH-NMR (400 MHz, CDCl 3) 6 8.05 (d, J=

7.3 Hz, 1H, H-2", 6"), 7.59 (t, J= 7.4 Hz, 1H, H-4"), 7.48 (t, J= 7.6 Hz, 2H, H-3", 5"), 5.38 (d, J= 13.5 Hz, 2H,H-9"', 10"'), 4.90 (d, J= 5.0 Hz, 1H, H-14 1 ), 4.49 (dd, J=

5.0, 2.7 Hz, 1H, H-15 ), 4.40 (d, J 2.7 Hz, 1H, H-3 1 ), 4.10 (d, J= 6.4 Hz, 1H, H

3.27 (s, 3H, 18'-OCH3), 3.22 (s, 3H,1'-OCH3 ), 3.21 (s, 3H,6'-OCH3), 1.12 (t, J=

7.1 Hz, 3H, N-CH 2CH 3), 0.90 (t, J= 6.6 Hz, 3H, H-18"'). 13C NMR Figure 6. HRMS calculated for C5 2 H8 oNO 12 910.5681, found 910.5789[M+H]+.

3-linoleoyl-aconitine(6). Light yellow oil, yield 33.70%. IR (KBr): 3494, 2927, 2855, 1725, 1452, 1381, 1316, 1279, 1186, 1098, 1027, 985, 917, 710, 615 cm-1; 'H-NMR

(400 MHz, CDCl 3 ) 6 8.05 (d, J= 7.4 Hz, 1H, H-2", 6"), 7.59 (t, J= 7.4 Hz, 1H, H

4"), 7.47 (t, J= 7.6 Hz, 2H, H-3", 5"), 5.37 (m, 4H, H- 9"', 10"', 12"', 13"'), 4.89 (d, J= 5.0 Hz, 1H, H-14 ), 4.48 (d, J= 2.7 Hz, 1H, H-15 ),4.39 (d, J= 2.7 Hz, 1H, H-3

1 ), 4.09 (d, J= 6.6 Hz, 1H, H-6 ), 3.92 (s, 1H, H-17), 3.75 (s, 3H, 16'-OCH3), 3.33

(d, J= 5.1 Hz, 1H, H-16 a), 3.26 (s, 3H, 18'-OCH3), 3.21 (s, 3H, l'-OCH3), 3.20 (s,

3H, 6'-OCH3), 1.12 (t, J 6.5 Hz, 3H, N-CH 2CH 3), 0.90 (t, J= 6.8 Hz, 3H, H-18"').

13 C NMR Figure6. HRMS calculated for C52 H 7 8NO 12 908.5524, found 908.5520[M+H]+.

EXAMPLE 2: The structural formulas of compounds 7-12 and 14-17 referred to in the following examples are shown in FIG. 3.

0.1 mmol of aconitine was mix with 0.3 mmol of different fatty acids and react in vacuum at 110 °C for 20 to 30 min. The crude product was purified by silica gel rapid chromatography (petroleum ether-acetone ratio 15: 1 to 4: 1) to obtain products 7-12 and 14-17 respectively.

[0030]8-capryloyl-aconitine(7). Orange oil, yield 52.20%. IR (KBr): 3456, 2924, 1639, 1383, 1274, 1094, 749, 709 cm-1; 1 H-NMR (400 MHz, CDC 3) 6 8.02 (d, J

7.3 Hz, 1H, H-2", 6"), 7.55 (t, J= 7.4 Hz, 1H, H-4"), 7.43 (t, J= 7.7 Hz, 2H, H-3", 5"), 4.85 (d, J= 4.9 Hz, 1H, H-14 1 ), 4.48 (d, J= 2.6 Hz, 1H, H-15 ), 4.43 (dd, J= 5.3,

2.6 Hz, 1H, H-3 1 ), 4.02 (d, J= 6.3 Hz, 1H, H-6 ), 3.94 (s, 1H, H-17), 3.75 (s, 3H,

16'-OCH 3), 3.60 (d, J= 8.9 Hz, 1H, H-18 a ), 3.45 (d, J= 8.9 Hz,1H, H-18 ), 3.33

(d, J= 5.4 Hz, 1H, H-16 a ), 3.28 (s, 3H, 18' -OCH 3), 3.25 (s, 3H, l ' -OCH 3), 3.14 (s,

3H, 6'-OCH3), 1.08 (t, J= 7.0 Hz, 3H, N-CH 2CH3), 0.87 (t, J= 6.6 Hz, 3H, H-8"'). 13 C NMRThe data is shown in Figure7. HRMS calculated for C 4aH 6 NO 11 730.4166, found 730.4166[M+H]+.

8-myristoyl-aconitine (8). Light yellow oil, yield 59.89%. IR (KBr): 3492, 2925, 2854, 1719, 1452, 1382, 1277, 1190, 1099, 1030, 984, 919, 710, 601 cm-1; 'H-NMR (400

MHz, CDC 3 ) 6 8.01 (d, J= 7.2 Hz, 1H, H-2", 6"), 7.55 (t, J= 7.4 Hz, 1H, H-4"),

7.43 (t, J= 7.6 Hz, 2H, H-3", 5"), 4.84 (d, J= 4.9 Hz, 1H, H-14 ),4.47 (d, J= 2.6 Hz, 1H, H-15 ), 4.42 (dd, J= 5.2, 2.5 Hz, 1H, H-3 1 ), 4.01 (d, J 6.4 Hz, 1H, H-6

1 ), 3.94 (s, 1H, H-17), 3.74 (s, 3H, 16'-OCH 3), 3.59 (d, J= 8.9 Hz, 1H, 1H, H-18 a ),

3.44 (d, J= 8.8 Hz, 1H, H-18 1 ), 3.32 (d, J= 5.4 Hz, 1H, H-16 a ), 3.28 (s, 3H, 18'

OCH 3 ), 3.24 (s, 3H, l'-OCH 3), 3.14 (s, 3H, 6'-OCH3), 1.08 (t, J= 7.1 Hz, 3H, N

CH 2 CH3 ), 0.87 (t, J= 6.7 Hz, 3H, H-14"'). 13 C NMRThe data is shown in Figure 7.. HRMS calculated for C 4 H 72 NO 1 1 814.5105, found 814.5108[M+H]+.

8-palmityl-aconitine (9). Orange oil, yield 59.82%. IR (KBr): 3473, 2924, 2853, 1718, 1452, 1383, 1277, 1189, 1097, 1030, 984, 709, 601 cm-1; IH-NMR (400 MHz, CDC 3

) 6 8.02 (d, J= 7.3 Hz, 1H, H-2", 6"), 7.55 (t, J= 7.4 Hz, 1H, H-4"), 7.43 (t, J= 7.7 Hz, 2H, H-3", 5"), 4.85 (d, J= 4.9 Hz, 1H, H-14 1 ), 4.48 (d, J= 2.6 Hz, 1H, H-15 3 ),

4.43 (dd, J= 5.3, 2.6 Hz, 1H, H-3 1 ), 4.02 (d, J= 6.3 Hz, 1H, H-6 1 ), 3.94 (s, 1H, H

17), 3.75 (s, 3H, 16'-OCH3), 3.60 (d, J= 8.9 Hz, 1H, H-18 a ), 3.45 (d, J= 8.9 Hz,

1H, H-18 1), 3.33 (d, J= 5.4 Hz, 1H, H-16 a ),3.28 (s, 3H, 18'-OCH3), 3.25 (s, 3H,

1'-OCH 3), 3.14 (s, 3H, 6'-OCH3), 1.08 (t, J= 7.0 Hz, 3H, N-CH 2 CH3), 0.87 (t, J=

6.6 Hz, 3H, H-16"'). 13C NMRThe data is shown in Figure 7. HRMS calculated for C 4 8H 7 6NO 11 842.5418, found 842.5430[M+H]+.

8-stearatyl-aconitine (10). Orange oil, yield 52.82%. IR (KBr): 3452, 2918, 2850, 1635,1384,1094,708 cm-1; IH-NMR(400 MHz, CDC 3) 68.02(d,J=7.2Hz,1H,

H-2", 6"), 7.56 (t, J= 7.4 Hz, 1H, H-4"), 7.44 (t, J= 7.7 Hz, 2H, H-3", 5"), 4.86 (d, J= 5.0Hz,1H,H-143),4.49(d,J= 2.6Hz,1H,H-153),4.44(dd,J= 5.3,2.4Hz,1H,

H-313),4.03(d,J=6.3Hz,1H,H-6),3.96(s,1H,H-17),3.76(s,3H,16'-OCH 3),

3.60(d,J= 8.8Hz,1H,H-18a),3.45(d,J= 8.9Hz,1H,H-1813),3.34(d,J= 5.4

Hz, 1H, H-16 a ), 3.29 (s, 3H, 18' -OCH 3), 3.26 (s, 3H, l'-OCH3), 3.15 (s, 3H, 6' 13 C OCH 3 ), 1.10 (t, J= 7.1 Hz, 3H, N-CH 2CH3 ), 0.87 (t, J= 6.8 Hz, 3H, H-18''').

NMRThe data is shown in Figure 7... HRMS calculated for CsH 8 aNO 1 1 870.5731,

found 870.5759 [M+H]+.

8-oleoyl-aconitine (11). Light yellow oil, yield 55.22%. IR (KBr): 3451, 2924, 2851, 1636, 1384, 693, 620 cm- 1; H-NMR (400 MHz, CDC 3) 68.04 (d, J= 7.2 Hz, 1H,

H-2", 6"), 7.57 (t, J= 7.4 Hz, 1H, H-4"), 7.45 (t, J= 7.6 Hz, 2H, H-3", 5"), 5.46

5.29 (m, 2H, 9"', 10"'), 4.87 (d, J= 4.9 Hz, 1H, H-14 ),4.49 (d, J= 2.4 Hz, 1H, H

), 4.45 (dd, J= 5.2, 2.5 Hz, 1H, H-3 1 ), 4.04 (d, J 6.4 Hz, 1H, H-61 ), 3.96 (s,

1H, OH-13), 3.77 (s, 3H, 16'-OCH3), 3.62 (d, J= 8.9 Hz, 1H, H-18 a ), 3.47 (d, J=

8.8 Hz, 1H,H-18 1 ), 3.35 (d, J= 5.3 Hz, 1H, H-16 a ), 3.30 (s, 3H, 18'-OCH 3), 3.27

(s, 3H, 1'-OCH3), 3.16 (s, 3H, 6'-OCH3), 1.10 (t, J= 7.1 Hz, 3H, N-CH 2CH 3), 0.93

- 0.84 (m, 3H, H-18"'). 13 C NMRThe data is shown in Figure 7... HRMS calculated

for C5 H 7 8NO 1 1 868.5575, found 868.5588[M+H]+.

8-linoleoyl -aconitine (12). Light yellow oil, yield 55.22%. IR (KBr): 3451, 2924, 2851,1636,1384,693,620 cm-1; IH-NMR (400 MHz, CDC 3) H-NMR(CDCl 3 , 400

MHz) 6 : 1.09 (3H , t , J =7.2 Hz, N-CH 2CH3 ), 3.17, 3.29 , 3.30, 3.77 (each 3H , s, 4

XOCH 3 ), 4.87 (1H ,d, J =4.0 Hz, H-141), 4.49 (1H , d, J =2.8 Hz, H-15 3), 3.99

(1H, d, J= 8 Hz, H-6 1 ),3.65 (1H, d, J = 8.0 Hz, H-18 a ), 3.35 (1H, d, J = 8.0 Hz,

H-18 1 ); 5.38 (4H, m, H-9"',6"', 11"', 12"'), 8.05 (2H, d, J =8.0 Hz, H-2',6'), 7.58

(1H,t,J =8.0 Hz,H-4'), 7.46 (2H,t,J =8.0 Hz, H-3',5'). 13C NMRThe data is shown

in Figure7. HRMS calculated for C5oH76NO11866.5418, found 866.5488[M+H]+.

8-(9,13-difluorooctadecanoyl)oxy)-aconitine (14). White solid, yield 12.14%. IR (KBr): 3549, 2926, 2855, 1714, 1636, 1453, 1384, 1277, 1098, 710 cm-1; IH-NMR

(400 MHz, CDCl 3 ) 6 8.02 (d, J= 7.3 Hz, 2H, H-2", 6"), 7.56 (t, J= 7.4 Hz, 1H, H

4"), 7.44 (t, J= 7.6 Hz, 2H, H-3", 5"), 4.85 (d, J= 4.9 Hz, 1H, H-14 ),4.48 (dd, J=

2.9, 1.2 Hz, 1H, OH-15), 4.43 (dd, J= 5.5, 2.7 Hz, 1H, H-15 ), 4.02 (dd, J= 6.5, 1.8

Hz, 1H, H-6 ),3.94 (s, 1H, OH-13), 3.75 (s, 3H, 16'-OCH3), 3.60 and 3.45 (d, J=

8.9 Hz, each 1H, H-18), 3.33 (d, J= 5.4 Hz, 1H, H-16), 3.29 (s, 3H, 18'-OCH3), 3.25

(s, 3H,1'-OCH 3), 3.15 (s, 3H, 6'-OCH3), 1.09 (t, J= 7.1 Hz, 3H, N-CH 2 CH3), 0.88 (t,

J= 6.4 Hz, 3H, H-18"'). 13 C NMRThe data is shown in Figure 8... HRMS calculated for C5 H 7 8F 2NO 11906.5543, found 906.5534 [M+H]+.

8-(1O-fluorooctadecanoyl)oxy)-aconitine (15). White solid, yield 38.26%. IR (KBr): 3490, 2926, 2855, 1721, 1452, 1382, 1277, 1099, 1030, 984, 710 cm-1; IH-NMR (400

MHz, CDCl 3 ) 6 8.01 (d, J= 8.2 Hz, 2H, H-2", 6"), 7.55 (t, J= 7.4 Hz, 1H, H-4"),

7.43 (t, J = 7.6 Hz, 2H, H-3", 5"), 4.84 (d, J= 4.9 Hz, 1H, H-14 ),4.47 (d, J= 2.9 Hz, IH, OH-15), 4.42 (dd, J 5.6, 2.5 Hz, IH, H-15 ), 4.01 (d, J 6.6 Hz, IH, H-6

1 ), 3.94 (s, IH, OH-13), 3.74 (d, J= 1.6 Hz, 3H, 16' -OCH 3), 3.59 and 3.44 (d, J=

8.9 Hz, each IH, H-18), 3.32 (d, J= 5.5 Hz, IH, H-16), 3.28 (d, J= 1.5 Hz, 3H, 18'

OCH 3 ), 3.24 (d, J= 1.6 Hz, 3H, 1'-OCH3), 3.14 (d, J= 1.4 Hz, 3H, 6'-OCH3), 1.08 (t,

J= 7.0 Hz, 3H, N-CH 2 CH3), 0.86 (t, J = 6.6 Hz, 3H, H-18"'). 13C NMRThe data is shown in Figure 8. HRMS calculated for C 5 oH79FNOii 888.5637, found 888.5640

[M+H]+.

8-(9,13-di-bromoctadecanoyl)oxy)-aconitine (16). Light yellow oil, yield 53.15%. 3488, 2928, 2856, 1719, 1451, 1278, 1098, 710, 526 cm-1; IH-NMR (400 MHz,

CDC 3 ) 6 8.01 (d, J= 7.3 Hz, 2H,H-2", 6"), 7.56 (t, J 7.3 Hz, 1H,H-4"), 7.44 (t, J=

7.6 Hz, 2H, H-3",5"), 4.84 (d, J= 5.0 Hz, 1H, H-14 ),4.46 (d, J= 2.3 Hz, 1H, OH

), 4.42 (dd, J= 5.5, 2.7 Hz, 1H, H-15 ), 4.01 (d, J= 6.2 Hz, 1H H-61 ), 3.93 (s,

1H, OH-13), 3.74 (s, 3H, 16'-OCH3), 3.59 and 3.44 (d, J= 9.0 Hz, J= 8.9 Hz, each

1H H-18), 3.32 (d, J= 5.4 Hz, 1H, H-16), 3.28 (s, 3H, 18'-OCH3), 3.24 (s, 3H, 1'

OCH 3 ), 3.14 (s, 3H, 6'-OCH3), 1.08 (t, J= 7.1 Hz, 3H, N-CH 2 CH3), 0.89 - 0.85 (m,

3H, H-18"'). 13 C NMRThe data is shown in Figure 8. HRMS calculated for C 5oH 7 Br 2 NO 11 1028.3921, found 1028.3937 [M+H]+.

8-(1-bromoctadecanoyl)oxy)-aconitine (17). Light yellow oil, yield 49.27%. IR (KBr): 3484, 2926, 2854, 1723, 1451, 1278, 1099, 710, 601 cm-1 cm- 1; IH-NMR

(400 MHz, CDC 3 ) 6 8.01 (d, J= 7.5 Hz, 2H, H-2", 6"), 7.60 (t, 1H, H-4"), 7.43 (t, J

= 8.5, 4.4 Hz, 2H, H-3", 5"), 4.83 (d, J= 4.9 Hz, 1H, H-14 1 ), 4.55 - 4.44 (m, 1H, OH-15), 4.44 - 4.37 (m, 1H, H-15 ),4.01 (d, J= 6.6 Hz, 1H, H-6 ), 3.94 (s, 1H,

OH-13), 3.74 (d, J= 2.1 Hz, 3H, 16'-OCH3), 3.58 and 3.43 (d, J= 8.9 Hz, each 1H,

H-18), 3.32 (d, J= 5.5 Hz, 1H, H-16), 3.27 (s, 3H, 18'-OCH3), 3.23 (s, 3H, 1'

OCH 3 ), 3.13 (s, 3H, 6'-OCH3), 1.07 (t, J= 7.1 Hz, 3H, N-CH 2 CH3), 0.85 (t, J= 6.7

Hz, 3H, H-18"'). 13C NMR The data is shown in Figure 8. HRMS calculated for C 5 oH79BrNOii 950.4816, found 950.4873 [M+H]+.

EXAMPLE 3 that structural formula of compound 13 refer to in the following example is shown in FIG. 3.

0.1 mmol of aconitine linoleate and 0.3 mmol of pyridine were dissolve in 3 mL of DCM (dichloromethane), followed by addition of 0.12 mmol of acetic anhydride, stir at room temperature for 10 h, then adjusting that pH of the reaction solution to above 9 with concentrated ammonia water, and extracting twice with (5mL). The solvent is removed under reduced pressure to obtain a yellow oil. It was purified by silica gel rapid chromatography (petroleum ether-acetone system: 15: 1 to 4: 1) to obtain product 13, and the synthesis route is shown in FIG. 4.

[0032]3-acetyl-8-linoleate-aconitine(13). Light yellow oil, yield 52.07%. IR (KBr): 3452, 2922, 2855, 1638, 1384, 1276, 1095, 749 cm-1; 1H-NMR (400 MHz, CDC 3

) 68.02 (d, J= 7.1 Hz, 2H, H-2", 6"), 7.55 (t, J= 7.4 Hz, 1H, H-4' '), 7.44 (t, J= 7.7

Hz, 2H, H-3", 5"), 5.43-5.26 (m, 4H, 9"', 10"',12"', 13"'), 4.91 (dd, J= 12.7, 5.6 Hz, 1H, H-3), 4.85 (d, J= 5.0 Hz, 1H, H-14 ),4.46 (d, J= 2.5 Hz, 1H, OH-15), 4.42 (dd,

J= 5.4, 2.8 Hz, 1H, H-15 ), 4.06 (d, J= 7.0 Hz, 1H, H-61 ), 3.88 (s, 1H, OH-13),

3.77 (d, J= 8.9 Hz, 1H, H-18 a ) 3.74 (s, 3H, 16'-OCH3), 3.31 (d, J= 5.3 Hz, 1H, H

16 a ), 3.25 (s, 3H, 18'-OCH3), 3.19 (s, 3H, l'-OCH 3), 3.18 (s, 3H, 6' -OCH 3), 2.06

(s, 3H, 3'-COCH3), 1.10 (t, J= 7.1 Hz, 3H, N-CH 2CH3), 0.90 (t, 3H, H-18"'). 13 C

NMR Figure 7. HRMS calculated for C 2H 78 NO1 2 908.5524, found 908.5525[M+H]+.

EXAMPLE 4: The structural formulas of compounds 18-20 referred to in the following examples are shown in FIG. 3.

[0033] 0.1 mmol aconitine and 0.3 mmol linoleic acid were reacted under 110 °C vacuum for 30 min, and the oil was purified by silica gel rapid chromatography to obtain aconitine linoleic acid ester. Then 0.02 mmol aconitine linoleate was dissolved in 1.5 mL acetone, 0.1 mL 48% hydrobromic acid/concentrated hydrochloric acid or 0.2 mL trifluoroacetic acid was added to the solution respectively, and after stirring at room temperature for 30 min, the reaction solution was evaporated under reduced pressure to remove the solvent to obtain three salts 18-20. The synthesis route is shown in Fig. 4.

[0034] Example 5: The structural formula of compound 21 referred to in the following examples is shown in FIG. 3.

0.1 mmol of aconitine and 0.3 mmol of NBS (N-bromosuccinimide) were dissolve in 0.4 mL of acetic acid, Stir for 0.5 h at normal temperature, and the reaction process is monitored by TLC. After the reaction is completed, the pH of the reaction solution is adjusted to greater than 9 with concentrated ammonia water. DCM is dissolved and transferred to a liquid separation funnel. After washing the organic phase with water for 2 times, anhydrous Na 2 SO 4 is dried, the organic phase is concentrated under reduced pressure, and the crude product is purified by silica gel rapid chromatography to obtain denitroethyl aconitine. Then it reacted with 3 eq linoleic acid under 110 °C vacuum for 30 min, and the oil was purified by silica gel rapid chromatography to obtain product 21. The synthesis route is shown in FIG. 4.

[0036]N-deethyl-8-linoleate aconitine (21). IR (KBr): 3450, 2919, 2850, 1637, 1384, 1273, 1099, 708 cm-1; 'H-NMR (400 MHz, CDC 3) 6 8.02 (d, J= 7.1 Hz, 2H, H-2", 6"), 7.55 (d, J= 7.4 Hz,1H,H-4"), 7.43 (t, J= 7.7 Hz, 2H, H-3", 5"), 5.43 - 5.28 (m, 4H, H-9"',10",12"',13"'), 4.86 (d, J= 5.0 Hz, 1H, H-14 1 ), 4.45 (d, J= 5.4 Hz, 1H, H 1 ), 4.05 (d, J= 6.9 Hz, 1H, H-6 1 ), 3.75 (s, 3H, 16'-OCH3), 3.59 and 3.53 (d, J= 9.0 Hz, each 1H, H-18),3.35 (d, J= 5.5 Hz, 1H, H-16), 3.30 (s, 3H, 18'-OCH3),3.28 (s,3H, 1'-OCH3), 3.14 (s,3H, 6'-OCH3), 0.88 (s,3H, H-18"'). 13C NMR Figure 9. HRMS calculated forC 48 H 72 NO 1 1838.5105, found 838.5231[M+H]+.

EXAMPLE 6 The structural formula of compound 22 referred to in the following examples is shown in FIG. 3.

[0037] Reacting 0.1 mmol of aconitine with 0.3 mmol of linoleic acid under 110 °C vacuum for 30 min, the oil was purified by silica gel rapid chromatography to obtain product 22, and the synthesis route is shown in FIG. 5.

[0038] 8-linoleate mesaconitine (22). IR (KBr): 3460, 2926, 2853, 1636, 1453, 1384, 1276, 1191, 1097, 1031, 986, 710 cm-1; 1 H-NMR (400 MHz, CDC 3) 6 8.06 (m, 2H, H-2", 6"), 7.56 (s, 1H, H-4"), 7.44 (s, 2H, H-3", 5"), 5.35 (dd, J= 4.2, 1.6 Hz, 4H, H 9"',10"',12"',13"'),4.85 (d, J= 5.0 Hz, 1H, H-14 ),4.44 (s, 2H, H-15 ), 4.02 (dd, J = 6.7, 1.7 Hz, 1H, H-61 ), 3.74 (s, 3H, 16'-OCH3), 3.62 and 3.52 (d, J= 9.0 Hz, each

1H, H-18), 3.32 (d, J = 5.0 Hz, 1H, H-16), 3.29 (s,3H,18'-OCH 3), 3.28 (s,3H,1' OCH3 ), 3.15 (s, 3H, 6'-OCH3),3.11 (d, J= 2.5 Hz, 1H, H-1), 3.03 (s, 1H, H-17), 2.34 (s, 3H, NCH 3 ), 0.88 (s, 3H, H-18"'). 13C NMR Figure 9. HRMS calculated for C 4 9 H7 4 NO 11852.5262, found 852.5383[M+H]+.

EXAMPLE 7 The structural formulas of compounds 23-24 referred to in the following examples are shown in FIG. 3.

Dissolve 0.1 mmol 21 and 0.3 mmol DMAP (4-dimethylaminopyridine) in 4 mL DCM, then add 0.2 mmol acetic anhydride, the reaction process is monitored by TLC, the pH of the reaction solution is adjusted to greater than 9 with concentrated ammonia water after the reaction is completed, the DCM is dissolved and transferred to a separating funnel, the organic phase is washed with water twice, anhydrous Na2SO4 is dried, the organic phase is concentrated under reduced pressure, and the crude product is purified by silica gel rapid chromatography to obtain 23.

2) 0.1 mmol of 3-TBDMS aconitine derivative and 0.3 mmol of NBS (N bromosuccinimide) are dissolve in 0.4 mL of acetic acid, Stirring at room temperature for 0.5 h, the reaction process was monitored by TLC, after the reaction was completed, the pH of the reaction solution was adjusted to be greater than 9 with concentrated ammonia water, DCM was dissolved and transferred to a separating funnel, the organic phase was washed with water for 2 times, anhydrous Na 2 SO 4 was dried, the organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel rapid chromatography to obtain 3-TBDMS protected denitroethyl aconitine derivatives. It was then reacted with 3 eq linoleic acid under 110 °C vacuum for 30 min, After the oil was purified by rapid chromatography on silica gel, 3 eq DMAP and 2 eq acetic anhydride were dissolved in DCM and stirred at normal temperature. The reaction process was monitored by TLC. After the reaction was completed, the pH of the reaction solution was adjusted to greater than 9 with concentrated ammonia water. After DCM was dissolved, the organic phase was washed with water twice, dried with anhydrous Na2 SO 4 , and the organic phase was concentrated under reduced pressure. The product of 3-TBDMS protected nitroacetyl linoleate was obtained by rapid silica gel chromatography. Finally, it was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF and stirred at 75 °C for 10 hours. The reaction process was detected by TLC. After the reaction is completed, the reaction solution is extracted twice with diethyl ether/ethyl acetate (1: 1), and the solvent is removed by evaporation under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain product 24. The synthesis route is shown in FIG. 4.

[0041] 3-acetyl-N-acetyl-8-linoleateaconitine(23). IR (KBr): 3463, 2930, 2856, 1724, 1635, 1448, 1386, 1365, 1277, 1241, 1095, 989, 711, 607 cm-; H-NMR (400 MHz,

CDC 3 ) 68.02 - 7.95 (m, 2H, H-2", 6"), 7.54 (t, J= 7.4 Hz, 1H, H-4"), 7.42 (t, J= 7.6

Hz, 2H, H-3", 5"), 5.32 (m, 4H, H-9"',10"',12"',13"'),4.84 (d, J= 5.1 Hz, 1H, H-14 0 ),

4.44 (dd, J= 5.5, 2.6 Hz, 1H, H-15 3), 4.13 (d, J= 7.0 Hz,1H, H-6 ), 3.73 (s, 3H,

16'-OCH 3), 3.25 (d, J= 5.4 Hz, 1H, H-16), 3.20 (s, 3H, 18'-OCH3), 3.18 (s, 3H, 1'-

OCH3 ), 3.14 (s, 3H, 6'-OCH3), 2.31 (s, 3H, NCOCH 3), 2.01 (s, 3H, 3-OAc), 0.87 0.83 (m, 3H, H-18"'). 13C NMR Figure 9. HRMS calculated for C2H76NO13 922.5317, found 922.5443[M+H]+.

N-acetyl-8-linoleate aconitine (24). IR (KBr): 3455, 2927, 2855, 1714, 1602, 1454, 1384, 1276, 1099, 711 cm-1; 1 H-NMR (400 MHz, CDCl 3) 6 8.05-7.99 (m, 2H,H-2",

6"), 7.60-7.53 (m, 1H, H-4"), 7.45 (t, J = 7.7 Hz, 2H, H-3",5"), 5.41-5.30 (m, 4H, H 9"',10"',12"',13"'), 4.86 (d, J = 5.0 Hz, 1H, H-14 1 ), 4.49 (d, J=3.1 Hz, 1H, H-15),

4.46 (dd, J = 5.5, 3.2 Hz, 1H, H-15 ), 4.26 (d, J = 14.2 Hz, 1H, H-6 1 ), 4.12 (s, 1H,

OH-15), 3.94 (s, 1H, OH-13), 3.77 (s, 3H, 16'-OCH3), 3.69-3.62 (m, 1H, H-18 a),

3.59 (d, J = 9.1 Hz, 1H, H-18 1 ), 3.30 (s, 3H, 18'-OCH3), 3.26 (s, 3H,1'-OCH3 ), 13 C 3.14 (s, 3H, 6'-OCH3), 2.33 (s, 3H, NCOCH 3 ), 0.88 (d, J = 6.9 Hz, 3H, H-18"').

NMR Figure 9 HRMS calculated for CsH 7 4 NO 1 2 880.5211, found 880.5181 [M+H]+.

EXAMPLE 8 that structural formula of compound 25 refer to in the following example is shown in FIG. 3.

[0042] 0.2 mmol aconitine and 1.2 mmol mCPBA (m-chloroperoxybenzoic acid) were dissolved in 10 mL DCM and stirred at room temperature for 3 h. The reaction process was detected by TLC. After the reaction was finished, the pH of the reaction solution was adjusted to greater than 9 with concentrated ammonia water, and DCM

(10 mL) was used for extraction twice. The solvent was removed by evaporation under reduced pressure, and the crude product was purified by silica gel rapid chromatography to obtain aconitine oxynitroxide, which was then reacted with 3 eq linoleic acid under 110 °C vacuum for 30 min, and the oil was purified by silica gel rapid chromatography to obtain product 25.

[0043] 8-linoleateaconitine oxynitride (25). IR (KBr): 3456, 2924, 2852, 1721, 1637, 1451, 1384,1276,1190,1097,710 cm- 1; IH-NMR(400 MHz, CDC 3) 6 8.06-7.98

(m, 2H, H-2", 6"), 7.58 (m, IH, H-4"), 7.44 (t, J= 7.6 Hz, 2H, H-3", 5"), 5.32 (m, 4H, H-9",10"',12"',13"'), 4.86 (d, J= 5.0 Hz, IH, H-14 1 ), 4.44 (dd, J= 5.5, 2.8 Hz, IH,

H-15 ), 4.03 (m, IH, H-6 1 ), 3.76 (s, 3H, 16'-OCH3), 3.60 and 3.46 (d, J= 8.8 Hz,

1H each, H-18), 3.33 (d, J= 5.4 Hz, 1H, H-16), 3.29 (s, 3H, 18'-OCH3), 3.27 (s,3H,

1'-OCH 3), 3.16 (s, 3H, 6'-OCH3), 1.38 (t, J= 7.1 Hz, 3H, N-CH 2 CH3), 0.88 (m,3H,

H-18"'). 13 C NMR Figure 9. HRMS calculated for CsH 7 NO1 2 882.5368, found 882.5436[M+H]+.

EXAMPLE 9 The structural formulas of compounds 26-29 referred to in the following examples are shown in FIG. 3.

[0044] 1) 1.7 mmol of aconitine and 8.5 mmol of imidazole were dissolved in 8 mL of dry DCM, followed by 2 mmol of TBDMSCl (tert-butyldimethylchlorosilane), and the reaction process was detected by TLC at room temperature for 48 h. After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and the solvent was extracted twice with chloroform (200 mL). The solvent was removed under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain 3-TBDMS-aconitine.

1 mmol of 3-TBDMS-aconitine was dissolve in 5% sodium hydroxide/methanol solution, stirred at 55 °C under reflux for 1h, then that reaction solution was concentrated under reduced pressure to remove methanol, then 50 mL of distilled water was added to the crude product and extracted twice with DCM (200 mL). The solvent was removed under reduced pressure to obtain a white solid crude product. The crude product was purified by silica gel rapid chromatography to obtain 3 TBDMS-8, 13, 14,15-OH aconitine.

[0046] 3) The product obtained in 2) was dissolved with p-halobenzoyl chloride or p methoxybenzoyl chloride (1.2 eq) in dry DCM, followed by addition of DMAP (4 dimethylaminopyridine 3 eq), stirred at room temperature for 24 h, and the reaction process was detected by TLC. After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, extracted twice with DCM,

dried with anhydrous sodium sulfate after the organic phases were combined, concentrated under reduced pressure, and the residue was purified by silica gel rapid chromatography to obtain C14 modified derivatives.

[0047] 4) The benzene ring-introduced product at position 14 obtained in 3) was dissolved with Ac20 (acetic anhydride 3 eq) in dry DCM, followed by TsOH (p toluenesulfonic acid 3 eq), stirred at room temperature for 24 h, and the reaction process was detected by TLC. After the reaction is finished, the pH value of the reaction solution is adjusted to > 9 with concentrated ammonia water, and DCM is used for extraction twice. The solvent was removed by evaporation under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain C8,15 acetylated products or C8,13,15 acetylated products.

The product obtained from 4) was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, stirred at 75 °C of reflux for 10 hours, and the reaction process was detected by TLC. After the reaction was completed, the reaction solution was extracted twice with diethyl ether/ethyl acetate (1: 1), and the solvent was evaporated under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain deprotected aconitine derivatives.

[0049] 6) Each product obtained in 5) was reacted with 3 eq linoleic acid in vacuum at 110 °C for 20 to 30 min. The crude product was purified by silica gel rapid chromatography (petroleum ether-acetone system: 15: 1 to 4: 1) to obtain products 26 to 29 respectively. The synthesis route is shown in FIG. 4.

[005013,15-di-acetyl-14-(4' ' -F)-benzoyl-8-linoleate aconitine (26). Light yellow oil, yield 5.2%. IR (KBr): 3454, 2925, 2852, 1733, 1508, 1265, 1103, 1031, 738 cm-';

'H-NMR (400 MHz, CDCl 3 ) 68.19 (dd, J= 8.7, 5.5 Hz, 2H, H-2", 6"), 7.16 (t, J=

8.6 Hz, 2H, H-3", 5"), 6.06 (d, J= 6.0 Hz, 1H, H-14 1 ), 5.42 (m, 4H, 9"', 10"', 12"',

13"'), 5.11 (d, J= 5.3 Hz, 1H, H-15 3 ), 4.02 (d, 1H, H-6 ),3.58 (s, 1H, OH-13), 3.40

(s, 3H, 16'-OCH 3), 3.29 (s, 3H, 18'-OCH3), 3.26 (s, 3H, l'-OCH3), 3.15 (s, 3H, 6'

OCH 3 ), 2.14 (s, 3H, 13-OAc), 2.05 (s, 3H, 15-OAc), 1.15 (s, 3H, N-CH 2 CH3), 0.88 (d, J = 3.3 Hz, 3H, H-18"'). 13 C NMR Figure 10. HRMS calculated for C 4 H79FNO13 968.5535, found 968.5545 [M+H]+.

13,15-di-acetyl-14-(4"-Br)-benzoyl-8-linoleateaconitine (27). Light yellow oil, yield 6.8%. IR (KBr): 3452,2926,2855,1639,1457, 1384,1273,1103,1032,589 cm - 1; 1 H

NMR (400 MHz, CDC 3) 6 8.03 (d, J= 8.6 Hz, 2H, H-2", 6"), 7.64 (d, J=8.5 Hz, 2H,

H-3", 5"), 6.05 (d, J= 5.9 Hz, 1H, H-14 1 ),5.35 (dd, J= 9.8, 4.5 Hz, 4H, H-9"', 10"',

12"', 13"'), 5.11 (d, J= 5.3 Hz,1H, H-15 ),3.98 (d,J= 6.2 Hz,1H,H-6 0 ),3.84 (d,J=

5.9 Hz,1H,3-OH),3.76 (m, 1H, H-18),3.58 (d, J=2.2 Hz,1H,H-18 1 ),3.39 (s,3H,16'-

OCH 3 ),3.29(s,3H,18'-OCH 3),3.26(s,3H,1'-OCH 3),3.15(s,3H,6'-OCH 3),2.14

(s,3H,13'-OAc ),2.05 (s, 3H, 15'-OAc), 1.15 (s, 3H, N-CH 2 CH3), 0.87 (d, J= 3.2 Hz,

3H, H-18"'). 13C NMR Figure 10. HRMS calculated for C 4 H79BrNO13 1028.4735, found 1028.4727 [M+H]+.

-acetyl-14-(45"-Cl)-benzoyl-8-linoleateaconitine (28). Light yellow oil, yield 4.8%. IR (KBr): 3547, 2924, 2852, 1726, 1638, 1488, 1456, 1247, 1245, 1101, 461, 683 cm I; 1 H-NMR (400 MHz, CDC 3 )6 7.99 (d, J= 8.5 Hz, 2H, H-25", 65"), 7.44 (d, J

= 8.5 Hz, 2H, H-35", 55"), 5.35 (dd, J=6.1, 2.3 Hz, 4H, H-9", 10"', 12"', 13"'), 5.06 (d,

J= 5.1 Hz,1H, H-14 0 ),4.41 (dd,J= 2.8 Hz, 1H, H-15 3 ),4.01 (d, J=6.6 Hz,1H,H-6

),3.85 (d,J= 5.2 Hz,1H,3-OH),3.77 (dd,J= 9.3,4.6 Hz,1H,H-18),3.59 (s, 3H, 16'

OCH 3 ),3.43 (d, J = 8.9 Hz, 1H, H-181 ),3.29 (s, 3H, 18'-OCH3),3.25 (s,3H,1'

OCH 3 ),3.16 (s,3H,6'-OCH 3), 2.04 (s,3H,15'-OAc),1.10 (t,J= 7.1Hz,3H, N-CH 2CH3),

0.89-0.86(m,3H,H-18"'). 13 C NMR Figure 10. HRMS calculated for C2H77ClNO12 942.5134, found 942.511 [M+H]+.

-acetyl-14-(4' '-OCH)-benzoyl-8-inoeate aconitine (29). Light yellow oil, yield

8.2%. IR (KBr): 3447, 2921, 2853, 1637, 1384, 1243, 1102 cm-1; H-NMR (400

MHz, CDC 3 ) 6 8.00 (d, J= 8.9 Hz, 2H, H-2',6'), 6.92 (d, J= 8.9 Hz, 2H, H-3",5"),

5.42-5.28 (m,4H,H-9"',10"',12"',13"'), 5.04 (d, J= 5.1 Hz, 1H, H-14 1 ),4.41 (d, J=

2.8 Hz,1H, H-15 ),4.04-3.98 (m, 1H, H-6 ),3.84 (s,3H,4"-OCH 3), 3.77 (dJ =4.3

Hz,1H each,H-18),3.59 (s,3H,16'-OCH 3), 3.29 (s,3H,18'-OCH 3),3.25 (s,3H,1'

OCH 3 ),3.15 (s,3H,6'-OCH 3),2.04 (s,3H,15'-OAc),1.09 (tJ=7.1 Hz,3H,N-CH 2CH3),

0.89-0.86 (m,3H,H-18"'). 13C NMR Figure 10. HRMS calculated for C 3H 8 0 NO 13 938.5630, found 938.5626 [M+H]+.

EXAMPLE 10 The structural formula of the compound 30 referred to in the following examples is shown in FIG. 3.

[0051] 1) 1.7 mmol of aconitine and 8.5 mmol of imidazole were dissolved in 8 mL of dry DCM, followed by 2 mmol of TBDMSCl (tert-butyldimethylchlorosilane), and the reaction process was detected by TLC at room temperature for 48 h. After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and the solvent was extracted twice with chloroform (200 mL). The solvent was removed under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain 3-TBDMS-aconitine.

1 mmol of 3-TBDMS-aconitine was dissolve in 5% sodium hydroxide/methanol solution, stirred under reflux at 55 °C for 1h, then that reaction solution was concentrated under reduced pressure to remove methanol, then 50 mL of distilled water was added to the crude product and extracted twice with DCM (200 mL). The solvent was removed under reduced pressure to obtain a white solid crude product. The crude product was purified by silica gel rapid chromatography to obtain 3 TBDMS-8, 13, 14,15-OH aconitine.

Dissolve Ac20 (acetic anhydride 4eq) obtained in 2) in dry DCM, then add TsOH (p toluenesulfonic acid 3eq), and the reaction process is detected by TLC. After the reaction, adjust the pH value of the reaction solution to > 9 with concentrated ammonia water, extract twice with DCM, dry anhydrous sodium sulfate after combining organic phases, concentrate under reduced pressure, and purify the crude product by silica gel rapid chromatography to obtain acylated products of C8, 14, 15.

The product obtained from 3) was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, stirred at 75 °C of reflux for 10 hours, and the reaction process was detected by TLC. After the reaction was completed, the reaction solution was extracted twice with diethyl ether/ethyl acetate (1: 1), and the solvent was evaporated under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain deprotected aconitine derivatives.

[0055] 5) Each product obtained in 4) was reacted with 3 eq linoleic acid in vacuum at 110. D egree. C. for 20 to 30 min. The crude product was purified by silica gel rapid chromatography (petroleum ether-acetone system: 15: 1 to 4: 1) to obtain product 30 respectively. The synthesis route is shown in FIG. 4.

[0056] 14,15-di-acetyl-8-linoleate aconitine (30). IR (KBr): 3447,2921,2853,1637,1384,1243,1102 cm- 1; IH-NMR (400 MHz, CDC 3) 6 5.35 (q,

J= 5.2, 4.2 Hz,4H,H-9",10",12",13"), 4.86 (d,J=5.1 Hz,1H,H-15 0 ),4.50 (d,J=2.7 Hz,

1H, H-14 1 ), 4.35 (dd,J =5.8,2.6 Hz,1H,H-16 1 ),4.01 (dd,J =6.5,1.9 Hz,1H,H-6

1 ),3.77 and 3.59 (d, J=8.8 Hz,1H each,H-18),3.54 (s,3H,16'-OCH 3),3.30 (s,3H, 18'-

OCH 3 ),3.23 (s,3H,1K'-OCH 3), 3.22 (s, 3H, 6'-OCH3 ),2.07 (s,3H,14-OAc),2.06 (s, 3H,

-OAc),1.09 (t,J=7.1 Hz,3H,N-CH 2CH 3),0.89 (t,3H, H-18"). 13C NMR see Figure 11. HRMS calculated for C47H76NO2 846.5368, found 846.5366 [M+H]+.

EXAMPLE 11 that structural formula of compound 31 refer to in the following example is shown in FIG. 3.

The 3-TBDMS aconitine derivative (1 eq) was reacted with linoleic acid (3 eq) in vacuum at 110 °C for 20 to 30 min. The crude product was purified by rapid chromatography on silica gel (eluted with petroleum ether-acetone at 40: 1 to 10: 1) to give the 3-TBDMS-8-lipo derivative.

The product obtained in 1) was dissolved with Ac20 (acetic anhydride 2 eq) in dry dichloromethane, then TsOH (p-toluenesulfonic acid 3 eq) was added, stirred at room temperature for 24 h, and the reaction process was detected by TLC. After the reaction was completed, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and the solvent was extracted twice with DCM. The solvent was evaporated under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain 15-acetylated product.

[0059] 3) The product obtained in 2) was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, stirred at 75 °C of reflux for 10 hours, and the reaction process was detected by TLC. After the reaction was completed, the reaction solution was extracted twice with diethyl ether/ethyl acetate (1: 1), and the solvent was evaporated under reduced pressure. The crude product was purified by silica gel rapid chromatography to obtain product 31. The synthesis route is shown in FIG. 4.

[0060] 15-acetyl-8-linoleate aconitine (31). Light yellow oil, yield 12.1%. IR (KBr): 3451,2922,2851,723,1636,1384,1276,1102,747 cm-1; IH-NMR (400 MHz, CDC 3 )

6 8.05 (dd, J= 8.4, 1.4 Hz, 2H, H-2", 6"), 7.57 (t, J 7.4 Hz, 1H, H-4"), 7.45 (t, J=

7.7 Hz, 2H, H-3", 5"), 5.35 (m, 4H, H-9"',10"',12"',13"'),5.08 (d, J= 5.1 Hz, 1H, H-15 ), 4.49 (dd, J= 5.4, 2.8 Hz,1H,H-14 0 ),3.60 (d, J= 3.1 Hz,3H,16'-OCH 3), 3.29

(s,3H,18'-OCH 3),3.26 (s,3H,6'-OCH 3 ), 3.16 (s,3H),2.05 (s, 3H, l'-OCH3), 1.04 (t, 3H, N-CH 2CH3), 0.88 (t, J=3.7 Hz, 3H, H-18"'). 13 C NMRThe data are shown in Figure 11. HRMS calculated for C 2 H 7 8NO 12 908.5524, found 908.5516 [M+H]+.

EXAMPLE 12 that structural formula of compound 32 refer to in the following example is shown in FIG. 3.

[0061] 0.1 mmol of Aconitine a and 0.3 mmol of linoleic acid were reacted under 110 °C vacuum for 30 min, and the oil was purified by silica gel rapid chromatography to obtain product 32. The synthesis route is shown in FIG. 5.

[0062] 8-linoleate vilmorrianine A (32) (light yellow oil, 90.23% yield). IR (KBr): 3448, 2921, 2852, 1652, 1447, 1384, 1265, 1088, 736 cm-1; H-NMR (400 MHz,

CDC 3 ) 6 7.99 (d, J= 8.9 Hz, 2H, H-2", 6"), 6.89 (d,J=8.9 Hz,2H,H-3", 5"), 5.33 (d,

J=5.6 Hz,4H, H-9"',10"',12"',13"'),5.01 (s,1H,H-14 0 ),4.05 (m,1H,H-6 0 ),3.83 (s,3H,

16'-OCH 3), 3.81 -3.77 (m,1H), 3.60 and 3.42 (d, J=8.9 Hz,each 1H,H-18),3.39 (s,3H, 4'-OCH 3), 3.28 (s, 3H, 18'-OCH3), 3.24 (s,3H,1'-OCH 3), 3.16 (s,3H, 6'-OCH3), 1.08 (s, 3H, N-CH 2 CH3 , 0.87 (s,3H, H-18"'). 13C NMR The data are shown in Figure 11. HRMS calculated for C5 1H 7 8NO 1o 864.5626, found 864.5756[M+H]+.

EXAMPLE 13 that structural formula of compound 33 refer to in the following example is shown in FIG. 3.

To 0.2 mL of dichloromethane, 0.05 mmol of lappaconitine and 0.1 mmol of p toluenesulfonic acid were added, followed by 0.15 mmol of acetic anhydride, and stirred at room temperature for 24 h. The reaction process was monitored by TLC. After the reaction, the pH was adjusted to be greater than 9 with ammonia water.

After chloroform was dissolved, the reaction was transferred to a separating funnel. After washing the organic phase with water for 2 times, anhydrous Na2 SO 4 was dried, the organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel rapid chromatography to obtain 8-position acetylated lappaconitine. Then it reacted with 3 eq linoleic acid under 110 °C vacuum for 30 min, and the oil was purified by silica gel rapid chromatography to obtain product 33. The synthesis route is shown in FIG. 5.

[0064] 8-linoleate lappaconitine (33). IR (KBr): 3477, 2925, 2850, 1607, 1384, 1255, 1169, 1099, 691 cm- 1; 1 H-NMR (400 MHz, CDC 3 ) 6 11.07 (s, 1H, Ar-NH), 8.67

(dd, J= 8.5, 1.2 Hz, 1H, H-3"), 7.91 (dd, J= 8.1, 1.7 Hz, 1H, H-6"), 7.49 (ddd, J= 8.7, 7.3, 1.7 Hz, 1H, H-4"), 7.09- 6.98 (m, 1H, H-5"), 5.35 (dt, J= 6.6, 1.6 Hz, 2H, H

9"',10"'), 5.33-5.31 (m, 2H, H-12"',13"'), 3.52 (d, J=11.4 Hz,1H, Ha-19), 3.40 (s,1H,

H-14), 3.38 (s, 3H, 14'-OCH 3), 3.31 (s,3H,16'-OCH 3), 3.29 (s,3H, l'-OCH3), 3.18 (dd, J= 10.3,6.8 Hz, 1H,H-1),2.99 (d,J=3.4Hz, 1H,H-17),2.67(m, 1H, Ha-6),2.59(q, J= 2.1 Hz, 1H, Ha-3), 2.56 (d, J= 7.7 Hz, 1H, Ha-21), 2.52 (dd, J= 5.0, 2.4 Hz, 1H, Hb-19), 2.49 (d, J= 9.9 Hz, 1H, Hb-21), 2.45 (dd, J= 7.7 Hz, 1H, Ha-12), 2.34 (dt, J=

7.8, 3.8 Hz, 1H, Ha-15), 2.32 (m, 1H, Ha-15), 2.27 (d, J= 3.4 Hz, 1H, H-13), 2.26 (s, 1H, Ha-2), 2.22 (s, 3H, NHCOCH 3), 2.17 (d, J= 6.5 Hz, 1H, Hb-2), 2.10 (dd, J= 12.4, 4.5 Hz, 1H, H-7), 1.97 (t, J= 3.3 Hz,1H, Hb-12), 1.88 (s, 1H, Hb-3), 1.57 (s, 1H, Hb 6), 1.11 (t, J= 7.1 Hz, 3H, NCH 2 CH3), 0.88 (t, J= 7.2, 2.1 Hz, 3H, H-18"'). 13C NMR Figure 11. HRMS calculated for CsH 7 N 2 0 9 847.5473, found 847.5605[M+H]+.

EXAMPLE 14:

Test of Anti-tumor Activity in Vitro:

1. Experimental Samples and Methods

Activity experiments were carried out with each of the compounds prepared in Examples 1-33.

The IC50 values of adriamycin-resistant human breast cancer cells (ADR-MCF-7) of compounds 1-33 were determined by MTS method with adriamycin and etoposide as positive controls.

[0066] Principle of MTS method for detecting cell activity: MTS is a brand-new MTT analog, all called 3-(4, 5-dimethylthiazol-2-yl)-5 (3-carboxy-methoxyphenyl)-2-(4 sulfopheny)-2H-tetrazolium, which is a yellow dye. Succinate dehydrogenase in living cell mitochondria can metabolize and reduce MTS to generate soluble Formazan compound, and the content of Formazan can be determined at 490nm by

enzyme labeling instrument. Under normal circumstances, the amount of formazan

produced is proportional to the number of living cells, so the number of living cells can be inferred from the OD value of optical density.

[0067] Experimental method:

1) Sample configuration: 1 mg of each compound prepared in Examples 1-33 was dissolved in 1 mL of weakly acidic buffer solution (pH 5.5-7) with a certain pH value, and stored in cold storage as a stock solution, with blank buffer solution with corresponding pH as diluent and blank control.

[0068] 2). Inoculating cells: A single cell suspension was prepared with a culture solution containing 10% fetal bovine serum (DMEM or RMPI1640), and 3000-15000 cells per well were inoculated into a 96-well plate with a volume of 100ul per well, and adherent cells were inoculated and cultured 12-24 hours in advance.

[0069] 3). Add the compound solution to be tested, the final volume of each well is 200ul, and 3 multiple pores are set up for each treatment.

[0070] 4). Color development: After 48 hours of culture at 37 °C, adherent cells discard the culture solution in the pores and add 20ul of MTS solution and 100ul of culture solution per well; Suspension cells discard 100ul of culture supernatant and add 20ul of MTS solution per well; Set up 3 blank multiple pores (mixture of 20ul of MTS solution and 100ul of culture solution) and continue incubating for 2-4 hours to determine the light absorption value after the reaction is fully carried out.

Colorimetry: The wavelength of 492nm was selected, the light absorption value of each well was read by multi-functional enzyme labeling instrument (MULTISKAN FC), and the results were recorded. After data processing, the cell growth curve was drawn with concentration as abscissa and cell survival rate as ordinate, and the IC50 value of the compound was calculated by two-point method (Reed and Muench method). 6) Positive control compounds: Adriamycin and etoposide were used as positive compounds in each experiment. Cell growth curves were drawn with concentration as abscissa and cell survival rate as ordinate. IC50 values of compounds were calculated by two-point method (Reed and Muench method).

[0072] 2. Experimental results (see Fig. 6)

3. Conclusions

Compounds 7-12, 14, 16-22, 26, 28-29, 31-32 have obvious effects of reversing multidrug resistance of tumor cells and anti-canine breast cancer cells, and can be used as anti-tumor agents for human drug resistance reversal and canine breast cancer

inhibitors.

EXAMPLE 15:

Acute Toxicity Test of Compound 7 and Aconitine

1.1 Experimental animals

Kunming mice, weighing 18 ~ 22g, half male and half female, were raised under strict conditions, in which the temperature was 22 °C and the humidity was 55 5%, which satisfied the free intake of food and water by mice.

1.2 Test Operation

One hundred and ten healthy mice (half male and half female) were randomly divided into 11 groups. The experimental group had 10 mice in each group and the blank group had 10 mice. The control group was aconitine. Compound 7 (180, 190, 200, 210, 220mg/kg respectively) and aconitine (0.8, 0.4, 0.2, 0.1, 0.05 mg/kg respectively) were administered intraperitoneally. The mice were fasted for 8 ~ 10 hours before intragastric administration, but could not help drinking water. The blank group was injected with the same dose of solvent. Before the test, the drug was prepared into a certain concentration so that the dosage was 0.02 mL/g according to the body weight. After administration, the general health status, poisoning manifestations and death process of mice were observed and recorded for 14 days. At 14 days, the mice were killed and the data were collected by necropsy.

[0075] 1.3 Experimental Results

After calculation, the LD50 of compound 7 for intraperitoneal administration to mice is 192.944 mg/kg, and its 95% confidence limit is 179.692 ~ 201.337 mg/kg. The LD50 of aconitine in mice was 0.2 mg/kg in acute toxicity test. After dissection, the viscera of the undead mice were observed, and it was found that there was no obvious abnormality in the organ of aconitine linoleate (7) (see Fig. 13). Aconitine (0.2 mg/kg) group and aconitine linoleate (7) (192.944 mg/kg) were taken for slice observation. It was found that except for slight changes in liver and lung, other organs such as heart, spleen, kidney and thymus had no obvious changes compared with the blank group. The H-E staining results in the two groups are shown in the appendix.

[0076] As shown in FIG. 14, compared with the blank group, the liver tissue in the aconitine medium dose group is obviously damaged, with moderate diffuse vacuolar degeneration and more punctate necrosis foci. In the high dose group of aconitine linoleate (7), punctate necrosis foci were occasionally found.

[0077] As can be seen from FIG. 15, the pathological section results of lung tissue show that compared with the blank group, the alveolar septum in the aconitine medium dose group is significantly thickened [66], and the number of alveoli is reduced; The high dose of aconitine linoleate (7) also has slight alveolar septum thickening, but it is significantly improved compared with the medium dose group of aconitine.

[00781

1.4 Conclusion

After the introduction of long-chain fatty acid esters into the 8-position, the toxicity of aconitine derivatives is greatly reduced, and its safe dose is about 900 times higher than aconitine, which is more promising for application and clinical use.

Claims

Claims 1. A compound of formula I and its salt

R? 3 R?

Characterized in that Rois hydrogen, methyl and ethyl; R1 , R3 , R 4 , Rsare hydrogen, hydroxyl, C1-6 alkoxy groups such as methoxy or ethoxy, C1-6 alkanoyloxy such as acetyl;R2 is hydroxyl;R6, R9 , Rio is hydrogen,hydroxylor C-6 alkanoyloxy,for example acetyl, Rsis asaturated or unsaturated or halogen-containing long-chain fatty acyloxy group containing 8to 24 carbon atoms,such as octanoyl,linoleoyloxy, linolenoyloxy,oleoyloxy,palmitic,acyloxy,stearoyloxy,eicosapentaenoyloxy,10 fluorooctadecyloxy,9,13-difluorooctadecyloxy and 10-bromooctadecyloxy,9,13 dibromooctadecyloxy,R7 is hydrogen or hydroxyl or benzoyloxy or p-halobenzoyloxy; X is an acid radical commonly used in pharmacy,such as chlorine,bromine, trifluoroformiyloxy, etc.
2. The use according to claim 1, wherein the Cl-6 alkoxy group is selected from the group consisting of methoxy or ethoxy; Cl-6 alkanoyloxy is selected from that group consisting of acetyl; the long-chain fatty acyloxy group is selected from the group consisting of octanoyl,linoleoyloxy,oleoyloxy,palmitoyloxy,stearoyloxy,9,13 difluorooctadecyloxy, 10-bromooctadecyloxy, 9, 13-dibromooctadecyloxy.
3. Use according to claim 1, wherein Rois preferred hydrogen and ethyl; R 1 , R 3 , R 4 ,

Rs are preferred methoxy, R2 is preferred hydroxy, R6 , R9 , Rio is preferred hydrogen or hydroxy or acetyl, Rsis preferred saturated orunsaturated orhalogen-containing long-chain fatty acyloxy containing 8 to 18 carbon atoms,such as octanoyl, linoleoyloxy,oleoyloxy,palmitoyloxy,stearoyloxy, R7 is preferred p methoxybenzoyloxy, and Xis preferred chlorine.
4. Use according to claim 1, characterized in that it is selected from the group consisting of:

g 414

3 41ri1n Hi"

1.............
5. That compound according to claim 1, wherein the compound is prepared from a natural diterpenoid alkaloid by chemically changing of the substituents in the structure and introducing a long-chain fatty acid ester at position 8, wherein the

preparation method is as follows:
6. That compound of claim 1, characterize by the use in the preparation of reversal agents for tumor multidrug resistance and anti-canine breast cancer.

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Figure 15