CN111574449A - Preparation and use of alkaloid derivatives for reversing human tumor resistance and canine breast cancer - Google Patents

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 using natural diterpene alkaloids as raw materials. Antineoplastic preparations, and as a preparation against canine breast cancer.

Description

Preparation and use of alkaloid derivatives for reversing human tumor resistance and canine breast cancer

technical field

The invention relates to a method for preparing 8-position long-chain fatty acyloxy-substituted diterpene alkaloid compounds with natural diterpene alkaloids; the invention also relates to the preparation of such compounds in reversing the multidrug resistance of human tumors and resisting canine breast cancer The use in the preparation belongs to the field of pharmacy.

Background technique

Breast cancer is one of the world's recognized public health problems, which seriously threatens the physical and mental health of women around the world. Drug chemotherapy is one of the main methods for the treatment of breast cancer. However, with the development of breast cancer chemotherapy, breast cancer resistance to chemotherapy has become a problem. The main obstacle to breast cancer chemotherapy often leads to the failure of breast cancer chemotherapy. Multidrug resistance (MDR) of tumor cells is one of the main reasons for the failure of tumor chemotherapy. Doxorubicin (ADM) is the most commonly used drug in the clinical treatment of breast cancer, but long-term use can lead to multidrug resistance in breast cancer cells, resulting in the failure of chemotherapy. Multidrug resistance of breast cancer will lead to treatment failure and spread of breast cancer, which directly affects the prognosis and overall survival of patients. It is of great significance to explore the mechanism of anti-breast cancer reversal agents to reverse doxorubicin resistance and to find new drugs to overcome the resistance of tumor cells, which has attracted more and more attention of pharmaceutical workers.

In addition, dogs, as companion animals of humans, live in a living environment and are exposed to carcinogenic factors that are very similar to those of humans. Therefore, canine mammary tumors are also the most common malignant tumors in female dogs, with an incidence rate of up to 25% to 42%, which are human breast tumors. 3 times the malignancy rate. The same incentives also lead to similarities in the pathogenesis of human and canine breast cancer. Studies have shown that the gene sequences that control the morphology, biological behavior, and clinical development of canine mammary tumors have a high degree of homology with the human relative gene sequences. Both have the same susceptibility genes BRCA1 and BRCA2, therefore, canine mammary tumor is also a natural model of human mammary tumor. In-depth study of drugs for the treatment of canine mammary gland tumors can not only save the faithful companions of human beings, but also will undoubtedly open up new research avenues for human oncology medicine, which has dual significance.

Diterpene alkaloids are a class of natural products with complex structures, mainly distributed in the genus Aconitium, Delphinium and Spirea, with interesting chemical properties and a wide range of biological activity. Diterpenoid alkaloids, with their important pharmacological activities and structural complexity such as anti-inflammatory, analgesic, anti-platelet aggregation, anti-tumor, insecticidal, immune regulation, etc., have long inspired scientists to focus on their phytochemistry, synthesis and medicinal chemistry. interest. However, such components often have obvious toxicity to the nervous system, circulatory system, digestive system, etc., so the clinical application is very limited. How to improve its anti-tumor activity while reducing toxicity and enhance targeting to tumor tissue has very important theoretical value and practical significance.

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

SUMMARY OF THE INVENTION

及其盐(见图1）The present invention aims to provide an anti-tumor compound with a unique structure, which can well reverse the multidrug resistance of tumors and also inhibit canine breast cancer. Its structure is

and its salts (see Figure 1)

It is characterized in that: R ₀ is hydrogen, methyl and ethyl; R ₁ , R ₃ , R ₄ , R ₈ are hydrogen, hydroxyl, C1-6 alkoxy such as methoxy or ethoxy, C1-6 alkoxy Acyloxy such as acetyl; R ₂ is hydroxyl; R ₆ , R ₉ , R ₁₀ are hydrogen, hydroxyl or C1-6 alkanoyloxy such as acetyl, and R ₅ is saturated or unsaturated containing 8-24 carbon atoms. Saturated halogen-containing long-chain fatty acyloxy groups such as octanoyl, linoleoyloxy, linoleyloxy, oleoyloxy, palmitic acid acyloxy, stearic acyloxy, eicosapentaenoyloxy , 10-fluorooctadecanoyloxy, 9,13-difluorooctadecanoyloxy, 10-bromooctadecanoyloxy, 9,13-dibromooctadecanoyloxy, R ₇ is Hydrogen or hydroxyl or benzoyloxy or p-halogenated benzoyloxy; X is an acid group commonly used in pharmacy, such as chlorine, bromine, trifluoroformyloxy and the like.

化合物结构中R₀优选氢和乙基；R₁，R₃，R₄，R₈ 优选为甲氧基，R₂优选羟基，R₆，R₉，R₁₀优选为氢或羟基或乙酰基，R₅优选为含有8-18个碳原子个碳原子的饱和或不饱或含卤素的长链脂肪酰氧基例如辛酰基、亚油酰氧基、油酰氧基、棕榈酸酰氧基、硬酰氧基，R₇优选为为对甲氧基苯甲酰氧基，X优选氯。Preferred formulas of the present invention

In the compound structure, R ₀ is preferably hydrogen and ethyl; R ₁ , R ₃ , R ₄ , R ₈ are preferably methoxy, R ₂ is preferably hydroxyl, R ₆ , R ₉ , R ₁₀ are preferably hydrogen or hydroxyl or acetyl, R ₅ is preferably a saturated or unsaturated or halogen-containing long-chain fatty acyloxy group containing 8-18 carbon atoms such as octanoyl, linoleoyloxy, oleoyloxy, palmitic acid acyloxy, Hard acyloxy, R ₇ is preferably p-methoxybenzoyloxy, X is preferably chlorine.

化合物选自下述化合物 （见图2）：Preferred formulas of the present invention

The compounds were selected from the following compounds (see Figure 2):

Formula of the present invention

化合物可通过以相应二萜生物碱为原料，用化学的方法，将结构中的取代基发生变化，并引入8位长链脂肪酸酯。Formula of the present invention

The compound can use the corresponding diterpene alkaloids as raw materials to change the substituents in the structure by chemical methods, and introduce the 8-position long-chain fatty acid ester.

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

The following examples of the present invention enumerate the proliferation inhibitory activity of the 8-position long-chain fatty acid ester alkaloid derivatives on human breast cancer cells and canine breast cancer cells resistant to doxorubicin, and examples of acute toxicity tests of representative drugs.

Description of drawings

的结构式。Figure 1 Compound formula

's structural formula.

化合物的结构式。Figure 2 Preferred formula of the present invention

The structural formula of the compound.

Figure 3 Structural formulas of synthetic derivatives 2-33.

Figure 4 Schematic diagram of the synthesis of derivatives 2-21 and 23-31.

Figure 5 Schematic diagram of the synthesis of derivatives 22, 32 and 33.

Figure 6 ¹³ C-NMR spectroscopic data for compound 1-6 in CDCl ₃ 100 ( ¹³ C).

Figure 7 ¹³ C-NMR spectroscopic data for compound 7-13 in CDCl ₃ 100 ( ¹³ C).

Figure 8 ¹³ C-NMR spectroscopic data for compound 14-17 in CDCl ₃ 100 or 150 ( ¹³ C).

Figure 9 ¹³ C-NMR spectroscopic data for compound 21-25 in CDCl ₃ 100 ( ¹³ C).

Figure 10 ¹³ C-NMR spectroscopic data for compound 26-29 in CDCl ₃ 100 ( ¹³ C).

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

Figure 12 IC ₅₀ (μM) results of inhibition of drug-resistant human breast cancer cells (ADR-MCF-7) and canine breast cancer cells (CMT1211) by compounds 1-33.

Figure 13 Organogram of aconitine linoleate (7) in each dose group.

Fig. 14 Changes in liver structure of mice in different treatment groups (SP×400).

Fig. 15 Changes in lung structure of mice in different treatment groups (SP×400).

Specific implementation plan:

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

Using aconitine (1) as raw material, under the catalysis of p-toluenesulfonic acid, it was reacted with long-chain fatty acid chloride reagents (octanoyl chloride, myristoyl chloride, palmitoyl chloride, stearoyl chloride, oleoyl chloride, linoleoyl chloride) at 45 ℃, respectively. , to obtain compounds 1-6, respectively, and the synthetic route is shown in Figure 4.

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 ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.02 (d, J = 7.2 Hz, 1H, H-2′′, 6′′), 7.57 (t, J = 7.4Hz, 1H, H-4′′) , 7.45 (t, J = 7.6 Hz, 2H, H-3′′, 5′′), 4.87 (d, J = 5.0 Hz, 1H, H-14β), 4.46 (dd, J = 5.2, 2.8 Hz, 1H, H-15β), 4.37 (d, J = 2.8 Hz, 1H, H-3β), 4.07 (d, J = 6.5 Hz, 1H, H-6β), 3.89 (s, 1H, H-17), 3.73 (s, 3H, 16′-OCH ₃ ), 3.36 (d, J = 8.8 Hz, 1H, H-18α), 3.31 (d, J = 5.3 Hz, 1H, H-16α), 3.24(s, 3H , 1′-OCH ₃ ), 3.18 (s, 3H, 6′-OCH ₃ ), 3.17 (s, 3H, 18′-OCH ₃ ), 1.10 (t, J =7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.87 (t, J = 6.8 Hz, 3H, H-8′′′). The ¹³ C NMR data are shown in Figure 6. HRMS calculated for C ₄₁ H ₆₂ NO ₁₂ 772.4272, 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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 4.49 (dd, J = 4.9 , 2.7 Hz, 1H, H-15β), 4.40 (d, J = 2.6 Hz, 1H, H-3β), 4.10 (d, J = 6.7 Hz, 1H, H-6β), 3.91 (s, 1H, H -17), 3.76(s, 3H, 16′-OCH ₃ ), 3.33 (d, J = 5.0 Hz, 1H, H-16α), 3.27 (s, 3H, 18′-OCH ₃ ), 3.22 (s, 3H, 1′-OCH ₃ ), 3.21 (s, 3H,6′-OCH ₃ ), 1.12 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.90 (t, J = 6.7 Hz, 3H, H-14′′′). The ¹³ C NMR data are shown in 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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.05 (d, J = 7.3 Hz, 1H, H-2′′, 6′′), 7.59 (t, J = 7.4Hz, 1H, H-4′ ′), 7.47 (t, J = 7.6 Hz, 2H, H-3′′, 5′′), 4.89 (d, J = 5.1 Hz, 1H, H-14β), 4.48 (dd, J = 5.0, 2.7 Hz, 1H, H-15β), 4.39 (d, J = 2.6 Hz, 1H, H-3β), 4.10 (d, J = 6.5 Hz, 1H, H-6β), 3.91 (s, 1H, H-17 ), 3.76 (s, 3H, 16′-OCH ₃ ), 3.33 (d, J = 5.0 Hz, 1H, H-16α), 3.27 (s, 3H, 18′-OCH ₃ ), 3.22 (s, 3H, 1′-OCH ₃ ), 3.21 (s, 3H,6′-OCH ₃ ), 1.12 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.90 (t, J = 6.7 Hz, 3H, H-16′′′). The ¹³ C NMR data is shown in Figure 6. HRMS calculated for C ₅₀ H ₇₈ NO ₁₂ 884.5524, found 884.5597[M+H] ⁺ .

3-staratyl-aconitine (4). Light yellow oil, yield 28.32%. IR (KBr):3458, 2922, 2851, 1722, 1635, 1450, 1384, 1277, 1093, 709 cm ^–1 ; ¹ H-NMR ( 400MHz, CDCl ₃ ) δ 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-14β), 4.53 – 4.45 (m, 1H, H-15β), 4.39 (d , J = 2.4 Hz, 1H, H-3β), 4.10 (d, J =6.5 Hz, 1H, H-6β), 3.92 (s, 1H, H-17), 3.75 (s, 3H, 16′-OCH ₃ ), 3.33 (d, J =5.1 Hz, 1H, H-16α), 3.27 (s, 3H, 18′-OCH ₃ ), 3.22 (s, 3H, 1′-OCH ₃ ), 3.21 (s, 3H ,6′-OCH ₃ ), 1.12 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.90 (t, J = 6.6 Hz, 3H, H-18′′′). ¹³ C NMR data See Figure 6. HRMS calculated for C ₅₂ H ₈₂ NO ₁₂ 912.5837, found912.5805[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 ; ¹ H-NMR (400 MHz) , CDCl ₃ ) δ 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-14β), 4.49 (dd, J = 5.0, 2.7 Hz, 1H, H-15β), 4.40(d, J = 2.7 Hz, 1H, H-3β), 4.10 (d, J = 6.4 Hz, 1H, H-6β), 3.91 (s, 1H, H-17), 3.76 (s, 3H, 16′-OCH ₃ ), 3.33 (d, J = 5.0 Hz, 1H, H-16α), 3.27 (s , 3H,18′-OCH ₃ ), 3.22 (s, 3H, 1′-OCH ₃ ), 3.21 (s, 3H,6′-OCH ₃ ), 1.12 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.90 (t, J = 6.6 Hz, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 6. HRMS calculated for C ₅₂ H ₈₀ NO 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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 4.48 (d, J = 2.7 Hz, 1H, H-15β), 4.39 (d, J = 2.7 Hz, 1H, H -3β), 4.09 (d, J = 6.6Hz, 1H, H-6β), 3.92 (s, 1H, H-17), 3.75 (s, 3H, 16′-OCH ₃ ), 3.33 (d, J = 5.1Hz, 1H, H-16α), 3.26 (s, 3H, 18′-OCH ₃ ), 3.21 (s, 3H, 1′-OCH ₃ ), 3.20 (s, 3H, 6′-OCH ₃ ), 1.12 (t, J = 6.5 Hz, 3H, N-CH ₂ CH ₃ ), 0.90 (t, J = 6.8 Hz, 3H, H-18′′′). ¹³ C NMR data are shown in Figure 6. HRMS calculated for C ₅₂ H ₇₈ NO ₁₂ 908.5524, found908.5520[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 mixed with 0.3 mmol of different fatty acids and reacted under vacuum at 110 °C for 20 to 30 min. The crude product was purified by flash chromatography on silica gel (petroleum ether-acetone 15:1 to 4:1) to give products 7-12 and 14-17, respectively.

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, CDCl ₃ ) 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-14β), 4.48 (d, J = 2.6Hz, 1H, H-15β), 4.43 (dd, J = 5.3, 2.6 Hz, 1H, H-3β), 4.02 (d, J = 6.3 Hz, 1H, H-6β), 3.94 (s, 1H, H-17), 3.75 (s, 3H, 16′- OCH ₃ ), 3.60 (d, J = 8.9 Hz, 1H, H-18α), 3.45 (d, J = 8.9 Hz, 1H, H-18β), 3.33 (d, J = 5.4 Hz, 1H, H-16α) ), 3.28 (s, 3H, 18′-OCH ₃ ), 3.25 (s, 3H, 1′-OCH ₃ ), 3.14 (s, 3H, 6′-OCH ₃ ), 1.08 (t, J = 7.0 Hz, 3H, N-CH ₂ CH ₃ ), 0.87 (t, J = 6.6 Hz, 3H, H-8'''). ¹³ C NMR data are shown in Figure 7. HRMS calculated for C ₄₀ H ₆₀ 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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 4.47 (d, J = 2.6 Hz, 1H, H-15β), 4.42 (dd, J = 5.2, 2.5Hz, 1H, H-3β), 4.01 (d, J = 6.4 Hz, 1H, H-6β), 3.94 (s, 1H, H-17 ), 3.74 (s, 3H, 16′-OCH ₃ ), 3.59 (d, J = 8.9 Hz, 1H, 1H, H-18α), 3.44 (d, J = 8.8 Hz, 1H, H-18β), 3.32 (d, J = 5.4 Hz, 1H, H-16α), 3.28 (s, 3H, 18′-OCH ₃ ), 3.24 (s, 3H, 1′-OCH ₃ ), 3.14 (s, 3H, 6′-OCH 3 ) OCH ₃ ), 1.08 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.87 (t, J = 6.7 Hz, 3H, H-14′′′). ^{The 13} C NMR data are shown in Figure 7. HRMS calculated for C ₄₆ H ₇₂ NO ₁₁ 814.5105, 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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 4.48 (d, J = 2.6 Hz, 1H , H-15β), 4.43 (dd, J = 5.3, 2.6 Hz, 1H, H-3β), 4.02 (d, J = 6.3 Hz, 1H, H-6β), 3.94 (s, 1H, H-17) , 3.75 (s, 3H, 16′-OCH ₃ ), 3.60 (d, J = 8.9 Hz, 1H, H-18α), 3.45 (d, J = 8.9 Hz, 1H, H-18β), 3.33 (d, J = 5.4 Hz, 1H, H-16α), 3.28 (s, 3H, 18′-OCH ₃ ), 3.25 (s, 3H, 1′-OCH ₃ ), 3.14 (s, 3H, 6′-OCH ₃ ) , 1.08 (t, J = 7.0 Hz, 3H, N-CH ₂ CH ₃ ), 0.87 (t, J = 6.6 Hz, 3H, H-16′′′). ^{The 13} C NMR data are shown in Figure 7. HRMS calculated for C ₄₈ H ₇₆ NO ₁₁ 842.5418, found842.5430[M+H] ⁺ .

8-staratyl-aconitine (10). Orange oil, yield 52.82%. IR (KBr): 3452, 2918, 2850, 1635, 1384, 1094, 708 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.02 (d, J =7.2 Hz, 1H, H-2′′, 6′′), 7.56 (t, J = 7.4 Hz, 1H, H-4′′), 7.44 (t, J = 7.7Hz, 2H , H-3′′, 5′′), 4.86 (d, J = 5.0 Hz, 1H, H-14β), 4.49 (d, J = 2.6 Hz, 1H, H-15β), 4.44 (dd, J = 5.3, 2.4 Hz, 1H, H-3β), 4.03 (d, J = 6.3 Hz, 1H, H-6β), 3.96 (s, 1H, H-17), 3.76 (s, 3H, 16′-OCH ₃ ), 3.60 (d, J = 8.8 Hz, 1H, H-18α), 3.45 (d, J = 8.9 Hz, 1H, H-18β), 3.34 (d, J = 5.4 Hz, 1H, H-16α), 3.29(s, 3H, 18′-OCH ₃ ), 3.26 (s, 3H, 1′-OCH ₃ ), 3.15 (s, 3H, 6′-OCH ₃ ), 1.10 (t, J =7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.87 (t, J = 6.8 Hz, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 7. HRMS calculated for C ₅₀ H ₈₀ NO ₁₁ 870.5731, 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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.04 (d, J =7.2 Hz, 1H, H-2′′, 6′′), 7.57 (t, J = 7.4 Hz, 1H, H-4′′), 7.45 (t, J = 7.6Hz, 2H, H-3′′, 5′′), 5.46 – 5.29 (m, 2H, 9′′′, 10′′′), 4.87 (d, J = 4.9 Hz, 1H, H-14β), 4.49 ( d, J = 2.4 Hz, 1H, H-15β), 4.45 (dd, J = 5.2, 2.5 Hz, 1H, H-3β), 4.04 (d, J = 6.4 Hz, 1H, H-6β), 3.96 ( s, 1H, OH-13), 3.77 (s, 3H, 16′-OCH ₃ ), 3.62 (d, J = 8.9 Hz, 1H, H-18α), 3.47 (d, J = 8.8 Hz, 1H, H -18β), 3.35(d, J = 5.3 Hz, 1H, H-16α), 3.30 (s, 3H, 18′-OCH ₃ ), 3.27 (s, 3H, 1′-OCH ₃ ), 3.16 (s, 3H, 6′-OCH ₃ ), 1.10 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.93 – 0.84 (m, 3H,H-18′′′). ¹³ C NMR data are shown in Fig. 7.. HRMS calculated for C ₅₀ H ₇₈ NO ₁₁ 868.5575, found868.5588[M+H] ⁺ .

8-linoleoyl-aconitine (12). Light yellow oil, yield 55.22%. IR (KBr):3451, 2924, 2851, 1636, 1384, 693, 620 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) ¹ H-NMR (CDCl ₃ , 400 MHz) δ: 1.09 (3H , t , J =7.2 Hz, N-CH ₂ CH ₃ ), 3.17, 3.29 , 3.30, 3.77 (each 3H , s, 4 ×OCH ₃ ) , 4.87 (1H, d, J = 4.0 Hz, H-14β), 4.49 (1H, d, J=2.8 Hz, H-15β), 3.99 (1H, d, J = 8 Hz, H-6β), 3.65 (1H, d, J = 8.0 Hz, H-18α), 3.35 (1H, d, J = 8.0 Hz, H-18β); 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′). The ¹³ C NMR data are shown in Figure 7. HRMS calculated for C ₅₀ H ₇₆ NO ₁₁ 866.5418, found 866.5488[M+H] ⁺ .

8-(9,13-difluorooctadecanoyl)oxy)-aconitine (14). White solid, yield12.14%. IR (KBr): 3549, 2926, 2855, 1714, 1636, 1453, 1384, 1277, 1098, 710cm ^{– 1} ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 4.48 (dd, J = 2.9, 1.2 Hz, 1H, OH-15), 4.43 (dd, J = 5.5, 2.7 Hz, 1H, H-15β), 4.02 (dd, J = 6.5, 1.8 Hz, 1H, H-6β), 3.94 (s, 1H, OH-13), 3.75 (s, 3H, 16′-OCH ₃ ), 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′-OCH ₃ ), 3.25 (s, 3H, 1′-OCH ₃ ), 3.15 (s, 3H, 6′-OCH ₃ ), 1.09 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.88 (t, J = 6.4 Hz, 3H, H-18′′′). ¹³ C NMR data are shown in Figure 8. HRMS calculated for C ₅₀ H ₇₈ F ₂ NO ₁₁ 906.5543,found 906.5534 [M+H] ⁺ .

8-(10-fluorooctadecanoyl)oxy)-aconitine (15). White solid, yield 38.26%. IR (KBr): 3490, 2926, 2855, 1721, 1452, 1382, 1277, 1099, 1030, 984, 710 cm ^{– 1} ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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 = 4.9Hz, 1H, H-14β), 4.47 (d, J = 2.9 Hz , 1H, OH-15), 4.42 (dd, J = 5.6, 2.5 Hz, 1H, H-15β), 4.01 (d, J = 6.6 Hz, 1H, H-6β), 3.94 (s, 1H, OH- 13), 3.74 (d, J =1.6 Hz, 3H, 16′-OCH ₃ ), 3.59 and 3.44 (d, J = 8.9 Hz, each 1H, H-18), 3.32 (d, J = 5.5 Hz, 1H , H-16), 3.28 (d, J = 1.5 Hz, 3H, 18′-OCH ₃ ), 3.24 (d, J = 1.6 Hz, 3H, 1′-OCH ₃ ), 3.14 (d, J = 1.4 Hz , 3H, 6′-OCH ₃ ), 1.08 (t, J = 7.0 Hz, 3H, N-CH ₂ CH ₃ ), 0.86 (t, J = 6.6 Hz, 3H, H-18′′′). ¹³ C The NMR data are shown in Figure 8. HRMS calculated for C ₅₀ H ₇₉ FNO ₁₁ 888.5637, found 888.5640 [M+H] ⁺ .

8-(9,13-di-bromoctadecanoyl)oxy)-aconitine (16) . Light yellow oil, yield53.15%. 3488, 2928, 2856, 1719, 1451, 1278, 1098, 710, 526 cm ^–1 ; ¹ H-NMR (400MHz, CDCl ₃ ) δ 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-14β), 4.46 (d, J = 2.3 Hz, 1H, OH -15), 4.42 (dd, J = 5.5, 2.7 Hz, 1H, H-15β), 4.01(d, J = 6.2 Hz, 1H H-6β), 3.93 (s, 1H, OH-13), 3.74 ( s, 3H, 16′-OCH ₃ ), 3.59and 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′-OCH ₃ ), 3.24 (s, 3H, 1′-OCH ₃ ), 3.14 (s, 3H, 6′-OCH ₃ ), 1.08 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.89 – 0.85 (m, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 8. HRMS calculated for C ₅₀ H ₇₈ Br ₂ NO ₁₁ 1028.3921, found 1028.3937 [M +H] ⁺ .

8-(10-bromoctadecanoyl)oxy)-aconitine (17) . Light yellow oil, yield49.27%. IR (KBr): 3484, 2926, 2854, 1723, 1451, 1278, 1099, 710, 601 cm ^–1 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 4.55 – 4.44 (m, 1H, OH- 15), 4.44 – 4.37 (m, 1H, H-15β), 4.01 (d, J = 6.6 Hz, 1H, H-6β), 3.94 (s, 1H, OH-13), 3.74 (d, J = 2.1 Hz, 3H, 16′-OCH ₃ ), 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′-OCH ₃ ), 3.23 (s, 3H, 1′-OCH ₃ ), 3.13 (s, 3H, 6′-OCH ₃ ), 1.07 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.85 (t, J = 6.7 Hz, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 8. HRMS calculated for C ₅₀ H ₇₉ BrNO ₁₁ 950.4816, found 950.4873 [M+H] ⁺ .

Example 3: The structural formula of compound 13 referred to in the following examples is shown in FIG. 3 .

0.1 mmol aconitine linoleate and 0.3 mmol pyridine were dissolved in 3 mL DCM (dichloromethane), then 0.12 mmol acetic anhydride was added, stirred at room temperature for 10 h, and then the pH of the reaction solution was adjusted with concentrated ammonia water Above 9, extract twice with (5 mL). The solvent was removed under reduced pressure to give a yellow oil. And it was purified by flash chromatography on silica gel (petroleum ether-acetone system: 15:1 to 4:1) to obtain product 13. The synthetic route is shown in Figure 4.

3-acetyl-8-linoleate-aconitine (13). Light yellow oil, yield 52.07%. IR (KBr): 3452, 2922, 2855, 1638, 1384, 1276, 1095, 749 cm ^–1 ; ¹ H-NMR ( 400 MHz, CDCl ₃ ) δ 8.02 (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-14β), 4.46 (d, J = 2.5 Hz, 1H, OH-15), 4.42 ( dd, J = 5.4, 2.8 Hz, 1H, H-15β), 4.06 (d, J = 7.0 Hz, 1H, H-6β), 3.88 (s, 1H, OH-13), 3.77 (d, J = 8.9 Hz, 1H, H-18α) 3.74 (s, 3H, 16'-OCH ₃ ), 3.31 (d, J = 5.3 Hz, 1H, H-16α), 3.25 (s, 3H, 18′-OCH ₃ ), 3.19 (s, 3H, 1′-OCH ₃ ), 3.18 (s, 3H, 6′-OCH ₃ ), 2.06 (s, 3H, 3′-COCH ₃ ), 1.10 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.90 (t, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 7. HRMS calculated for C ₅₂ H ₇₈ NO ₁₂ 908.5524, found 908.5525[M+H] ⁺ .

Example 4: Structural formulae of compounds 18-20 referred to in the following examples, see Figure 3 .

0.1 mmol of aconitine and 0.3 mmol of linoleic acid were reacted under vacuum at 110 degrees for 30 min, and the oily substance was purified by silica gel flash chromatography to obtain aconitine linoleate. Subsequently, 0.02 mmol of aconitine linoleate was dissolved in 1.5 mL of acetone, 0.1 mL of 48% hydrobromic acid/concentrated hydrochloric acid or 0.2 mL of trifluoroacetic acid were added to the solution, and the reaction solution was stirred at room temperature for 30 min. The solvent was removed by evaporation under reduced pressure to obtain three salts 18-20. The synthetic route is shown in Figure 4.

Example 5: Structural formula of compound 21 referred to in the following examples, see Figure 3 .

0.1 mmol of aconitine and 0.3 mmol of NBS (N-bromosuccinimide) were dissolved in 0.4 mL of acetic acid, and stirred at room temperature for 0.5 h. The reaction process was monitored by TLC. The pH was adjusted to greater than 9, and the DCM was dissolved and transferred to a separatory funnel. The organic phase was washed twice with water, dried over anhydrous Na ₂ SO ₄ , and the organic phase was concentrated under reduced pressure. The crude product was purified by silica gel flash chromatography to obtain denitrogenated ethyl acetate. Chiaconitine. Subsequently, it was reacted with 3 eq of linoleic acid under vacuum at 110 degrees for 30 min, and the oily substance was purified by silica gel flash chromatography to obtain product 21. The synthetic route is shown in Figure 4.

N-deethyl-8-linoleate aconitine (21). IR (KBr): 3450, 2919, 2850, 1637, 1384, 1273, 1099, 708 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.02 ( d, J = 7.1Hz, 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-14β), 4.45 (d, J = 5.4 Hz, 1H, H-15β), 4.05 (d, J = 6.9 Hz, 1H, H-6β), 3.75 (s, 3H, 16′-OCH ₃ ), 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′-OCH ₃ ), 3.28 (s, 3H, 1′-OCH ₃ ), 3.14 (s, 3H, 6′-OCH ₃ ), 0.88 (s, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 9. HRMScalculated for C ₄₈ H ₇₂ NO ₁₁ 838.5105, found 838.5231[M+H] ⁺ .

Example 6: Structural formula of compound 22 referred to in the following examples, see Figure 3 .

The reaction of 0.1 mmol of Zhongwu base with 0.3 mmol of linoleic acid was carried out under vacuum of 110 degrees for 30 min, and the oily substance was purified by silica gel flash chromatography to obtain the product 22. The synthetic route is shown in Figure 5.

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, CDCl ₃ ) 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-14β) , 4.44 (s, 2H, H-15β), 4.02 (dd, J = 6.7, 1.7 Hz, 1H, H-6β), 3.74 (s, 3H, 16′-OCH ₃ ), 3.62 and 3.52 (d, J = 9.0 Hz, each 1H, H-18), 3.32 (d, J = 5.0Hz, 1H, H-16), 3.29 (s, 3H, 18′-OCH ₃ ), 3.28 (s, 3H, 1′- OCH ₃ ), 3.15 (s, 3H, 6′-OCH ₃ ), 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′′′). The ¹³ C NMR data are shown in Figure 9. HRMS calculated for C ₄₉ H ₇₄ NO ₁₁ 852.5262, found 852.5383[M+H] ⁺ .

Example 7: Structural formulas of compounds 23-24 referred to in the examples below, see Figure 3 .

1) Dissolve 0.1 mmol 21 and 0.3 mmol DMAP (4-dimethylaminopyridine) in 4 mL DCM, and then add 0.2 mmol acetic anhydride. The reaction process is monitored by TLC. After the reaction, the pH of the reaction solution is adjusted with concentrated ammonia water. Adjusted to greater than 9, dissolved in DCM and transferred to a separatory funnel, washed the organic phase twice with water, dried over anhydrous Na ₂ SO ₄ , the organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel flash chromatography to obtain 23.

2) Dissolve 0.1 mmol of 3-TBDMS aconitine derivative and 0.3 mmol of NBS (N-bromosuccinimide) in 0.4 mL of acetic acid, and stir at room temperature for 0.5 h. The reaction process is monitored by TLC. The pH of the reaction solution was adjusted to be greater than 9 with concentrated ammonia water, the DCM was dissolved and then transferred to a separatory funnel, the organic phase was washed twice with water, dried over anhydrous Na ₂ SO ₄ , the organic phase was concentrated under reduced pressure, and the crude product was passed through silica gel Purification by flash chromatography gave the 3-TBDMS protected deazoethylaconitine derivative. Then it was reacted with 3 eq linoleic acid under vacuum at 110 degrees for 30 min, the oil was purified by silica gel flash chromatography, then it was dissolved in DCM with 3 eq DMAP and 2 eq acetic anhydride and stirred at room temperature. The reaction process was determined by TLC Monitoring, after the reaction was completed, the pH of the reaction solution was adjusted to greater than 9 with concentrated ammonia water, the DCM was dissolved and then transferred to a separatory funnel, the organic phase was washed twice with water, dried over anhydrous Na ₂ SO ₄ , and the organic phase was decompressed. Concentration and purification by silica gel flash chromatography gave the 3-TBDMS protected azaacetyl linoleate product. Finally, it was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, refluxed at 75 degrees and stirred for 10 h, and the reaction progress was detected by TLC. After the reaction, the reaction solution was extracted twice with ether/ethyl acetate (1:1), and the solvent was evaporated under reduced pressure. The crude product was purified by flash chromatography on silica gel to give product 24. The synthetic route is shown in Figure 4.

3-acetyl-N-acetyl-8-linoleateaconitine (23). IR (KBr): 3463, 2930,2856, 1724, 1635, 1448, 1386, 1365, 1277, 1241, 1095, 989, 711, 607 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.02–7.95 (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-14β), 4.44 (dd, J = 5.5, 2.6 Hz, 1H, H-15β), 4.13 (d, J = 7.0 Hz, 1H, H-6β) , 3.73 (s, 3H, 16′-OCH ₃ ), 3.25 (d, J =5.4 Hz, 1H, H-16), 3.20 (s, 3H, 18′-OCH ₃ ), 3.18 (s, 3H, 1 '-OCH ₃ ), 3.14 (s, 3H,6'-OCH ₃ ), 2.31 (s, 3H, NCOCH ₃ ), 2.01 (s, 3 H, 3-OAc), 0.87 – 0.83 (m, 3H, H -18′′′). The ¹³ C NMR data is shown in Figure 9. HRMS calculated for C ₅₂ H ₇₆ NO ₁₃ 922.5317, found922.5443[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 ) δ 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-14β), 4.49 (d, J = 3.1 Hz, 1H, H-15), 4.46 (dd, J = 5.5, 3.2 Hz, 1H, H-15β), 4.26 (d, J = 14.2 Hz, 1H, H-6β), 4.12 (s, 1H, OH-15), 3.94 (s, 1H, OH-13), 3.77 (s, 3H, 16′-OCH ₃ ), 3.69 – 3.62 (m, 1H, H- 18α), 3.59 (d, J = 9.1 Hz, 1H, H-18β), 3.30 (s, 3H, 18′-OCH ₃ ), 3.26 (s, 3H, 1′-OCH ₃ ), 3.14 (s, 3H , 6′-OCH ₃ ), 2.33 (s, 3H, NCOCH ₃ ), 0.88 (d, J = 6.9 Hz, 3H, H-18′′′). The ¹³ C NMR data are shown in Figure 9. HRMS calculated for C ₅₀ H ₇₄ NO ₁₂ 880.5211, found 880.5181 [M+H] ⁺ .

Example 8: Structural formula of compound 25 referred to in the following examples, see FIG. 3 .

0.2 mmol aconitine and 1.2 mmol m CPBA (m-chloroperoxybenzoic acid) were dissolved in 10 mL of DCM and stirred at room temperature for 3 h. The reaction process was detected by TLC. After the reaction, the pH of the reaction solution was adjusted to more than 9 with concentrated ammonia water, and extracted twice with DCM (10 mL). The solvent was evaporated under reduced pressure, and the crude product was purified by silica gel flash chromatography to obtain aconitine oxynitride, which was then reacted with 3 eq of linoleic acid under vacuum at 110 degrees for 30 min, and the oil was purified by silica gel flash chromatography to obtain product 25.

8-linoleate aconitine oxynitride (25). IR (KBr): 3456, 2924, 2852, 1721, 1637, 1451, 1384, 1276, 1190, 1097, 710 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.06–7.98 (m, 2H, H-2′′, 6′′), 7.58 (m, 1H, 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, 1H,H-14β), 4.44 (dd, J = 5.5, 2.8 Hz, 1H, H-15β), 4.03 (m, 1H, H-6β), 3.76 (s, 3H, 16′-OCH ₃ ), 3.60 and 3.46 (d, J = 8.8 Hz, 1H each, H-18), 3.33 (d, J = 5.4Hz, 1H, H-16), 3.29 (s, 3H, 18′-OCH ₃ ), 3.27 (s, 3H, 1′-OCH ₃ ), 3.16 (s, 3H, 6′-OCH ₃ ), 1.38 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.88 (m, 3H, H-18′′′). ¹³ The C NMR data are shown in Figure 9. HRMS calculated for C ₅₀ H ₇₆ NO ₁₂ 882.5368, found 882.5436[M+H] ⁺ .

Example 9: Structural formulae of compounds 26-29 referred to in the examples below, see Figure 3 .

1) Dissolve 1.7 mmol of aconitine and 8.5 mmol of imidazole in 8 mL of dry DCM, then add 2 mmol of TBDMSCl (tert-butyldimethylsilyl chloride), and react at room temperature for 48 h. The reaction progress is detected by TLC. After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and extracted twice with chloroform (200 mL). The solvent was removed under reduced pressure. The crude product was purified by flash chromatography on silica gel to give 3-TBDMS-aconitine.

2) Dissolve 1 mmol 3-TBDMS-aconitine in 5 % sodium hydroxide/methanol solution, stir under reflux at 55 degrees for 1 h, then concentrate the reaction solution under reduced pressure to remove methanol, and add 50 mL of distilled water to the crude product , and extracted twice with DCM (200 mL). The solvent was removed under reduced pressure to give the crude product as a white solid. The crude product was purified by silica gel flash chromatography to give 3-TBDMS-8,13,14,15-OH aconitine.

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) at room temperature After stirring for 24 h, 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, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel flash chromatography to obtain C14 modified derivatives.

4) Dissolve the product obtained in 3) into the benzene ring at the 14-position and Ac ₂ O (acetic anhydride 3 eq) in dry DCM, then add TsOH (p-toluenesulfonic acid 3 eq), stir at room temperature for 24 h, and react The process is detected with TLC. After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and extracted twice with DCM. The solvent was removed by evaporation under reduced pressure. The crude product was purified by flash chromatography on silica gel to yield either the C8,15 acetylated product or the C8,13,15 acetylated product.

5) The product obtained in 4) was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, and stirred under reflux at 75 degrees for 10h, and the reaction progress was detected by TLC. After the reaction, the reaction solution was extracted twice with ether/ethyl acetate (1:1), and the solvent was evaporated under reduced pressure. The crude product was purified by silica gel flash chromatography to obtain the deprotected aconitine derivative.

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

13,15-di-acetyl-14-(4′′-F)-benzoyl-8-linoleate aconitine (26). Lightyellow oil, yield 5.2%. IR (KBr): 3454, 2925, 2852, 1733, 1508, 1265, 1103, 1031, 738 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.19 (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-14β), 5.42 (m, 4H, 9′′′, 10′′′ , 12′′′, 13′′′), 5.11 (d, J = 5.3 Hz, 1H, H-15β), 4.02 (d, 1H, H-6β), 3.58 (s, 1H, OH-13), 3.40 (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 CH ₃ ), 0.88 (d, J = 3.3 Hz, 3H, H -18′′′). The ¹³ C NMR data are shown in Figure 10. HRMS calculated for C ₅₄ H ₇₉ FNO ₁₃ 968.5535, found968.5545 [M+H] ⁺ .

13,15-di-acetyl-14-(4 ′′ -Br)-benzoyl-8-linoleate aconitine (27). Lightyellow oil, yield 6.8%. IR (KBr): 3452, 2926, 2855, 1639, 1457, 1384, 1273, 1103, 1032, 589 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-14β), 5.35 (dd, J = 9.8, 4.5 Hz, 4H, H- 9′′′, 10′′′, 12′′′, 13′′′), 5.11 (d, J = 5.3 Hz, 1H, H-15β), 3.98 (d, J = 6.2 Hz, 1H, H- 6β), 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-18β ), 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 CH ₃ ), 0.87 (d, J = 3.2 Hz, 3H, H-18 '''). The ¹³ C NMR data is shown in Figure 10. HRMS calculated for C ₅₄ H ₇₉ BrNO ₁₃ 1028.4735, found 1028.4727 [M+H] ⁺ .

15-acetyl-14-(4 ′′ -Cl)-benzoyl-8-linoleate aconitine (28). Light yellowoil, yield 4.8%. IR (KBr): 3547, 2924, 2852, 1726, 1638, 1488, 1456, 1247, 1245, 1101, 461, 683 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.99 (d, J = 8.5 Hz, 2H,H-2′′, 6′′), 7.44 (d , J = 8.5 Hz, 2H, H-3′′, 5′′), 5.35 (dd, J = 6.1, 2.3 Hz, 4H, H-9′′′, 10′′′, 12′′′, 13 ′′′), 5.06 (d, J = 5.1 Hz, 1H, H-14β), 4.41 (dd, J = 2.8 Hz, 1H, H-15β), 4.01 (d, J = 6.6 Hz, 1H, H- 6β), 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-18β), 3.29 (s, 3H, 18′-OCH ₃ ), 3.25 (s, 3H, 1′-OCH ₃ ), 3.16 (s, 3H, 6′ -OCH ₃ ), 2.04 (s, 3 H, 15′-OAc), 1.10 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.89–0.86 (m, 3H, H-18′′ ′). The ¹³ C NMR data are shown in Figure 10. HRMS calculated for C ₅₂ H ₇₇ ClNO ₁₂ 942.5134, found 942.511 [M+H] ⁺ .

15-acetyl-14-(4 ′′ -OCH ₃ )-benzoyl-8-linoleate aconitine (29). Light yellowoil, yield 8.2%. IR (KBr): 3447, 2921, 2853, 1637, 1384, 1243, 1102 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.00 (d, J = 8.9 Hz, 2H, H-2′,6′), 6.92 (d, J = 8.9Hz, 2H, H-3 ′′,5′′), 5.42–5.28 (m, 4H, H-9′′′, 10′′′, 12′′′, 13′′′), 5.04 (d, J = 5.1 Hz, 1H, H-14β), 4.41 (d, J = 2.8 Hz, 1H, H-15β), 4.04 –3.98 (m, 1H, H-6β), 3.84 (s, 3H, 4′′-OCH ₃ ), 3.77 ( d, J = 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 ) OCH ₃ ), 3.15 (s, 3H, 6′-OCH ₃ ), 2.04 (s, 3 H, 15′-OAc), 1.09 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.89 –0.86 (m,3H, H-18′′′). The ¹³ C NMR data are shown in Figure 10. HRMS calculated for C ₅₃ H ₈₀ NO ₁₃ 938.5630, found938.5626 [M+H] ⁺ .

Example 10: Structural formula of compound 30 referred to in the following examples, see FIG. 3 .

1) Dissolve 1.7 mmol of aconitine and 8.5 mmol of imidazole in 8 mL of dry DCM, then add 2 mmol of TBDMSCl (tert-butyldimethylsilyl chloride), and react at room temperature for 48 h. The reaction progress is detected by TLC. After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and extracted twice with chloroform (200 mL). The solvent was removed under reduced pressure. The crude product was purified by flash chromatography on silica gel to give 3-TBDMS-aconitine.

2) Dissolve 1 mmol 3-TBDMS-aconitine in 5 % sodium hydroxide/methanol solution, stir under reflux at 55 degrees for 1 h, then concentrate the reaction solution under reduced pressure to remove methanol, and add 50 mL of distilled water to the crude product , and extracted twice with DCM (200 mL). The solvent was removed under reduced pressure to give the crude product as a white solid. The crude product was purified by silica gel flash chromatography to give 3-TBDMS-8,13,14,15-OH aconitine.

3) The product Ac ₂ O (acetic anhydride 4 eq) obtained in 2) was dissolved in dry DCM, then TsOH (p-toluenesulfonic acid 3 eq) was added, 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, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel flash chromatography to obtain C8,14, The acylated product of 15.

4) The product obtained in 3) was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, and stirred at 75 degrees under reflux for 10h, and the reaction progress was detected by TLC. After the reaction, the reaction solution was extracted twice with ether/ethyl acetate (1:1), and the solvent was evaporated under reduced pressure. The crude product was purified by silica gel flash chromatography to obtain the deprotected aconitine derivative.

5) React each product obtained in 4) with 3 eq of linoleic acid under vacuum at 110 °C for 20 to 30 min. The crude product was purified by silica gel flash chromatography (petroleum ether-acetone system: 15:1 to 4:1) to obtain product 30, respectively. The synthetic route is shown in Figure 4.

14,15-di-acetyl-8-linoleate aconitine (30). IR (KBr): 3447, 2921, 2853, 1637, 1384, 1243, 1102 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 5.35 (q, J = 5.2, 4.2 Hz, 4H, H-9′′, 10′′, 12′′, 13′′), 4.86 (d, J = 5.1 Hz, 1H, H-15β), 4.50 ( d, J = 2.7 Hz, 1H, H-14β), 4.35 (dd, J = 5.8, 2.6 Hz, 1H, H-16β), 4.01 (dd, J =6.5, 1.9 Hz, 1H, H-6β), 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 , 1′-OCH ₃ ), 3.22 (s, 3H,6′-OCH ₃ ), 2.07 (s, 3H, 14-OAc), 2.06 (s, 3H, 15-OAc), 1.09 (t, J = 7.1 Hz, 3H, N-CH ₂ CH ₃ ), 0.89 (t, 3H, H-18′′). The ¹³ C NMR data are shown in Figure 11. HRMS calculated for C ₄₇ H ₇₆ NO ₁₂ 846.5368, found 846.5366 [M+H ] ⁺ .

Example 11: Structural formula of compound 31 referred to in the following examples, see FIG. 3 .

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

2) Dissolve the product obtained in 1) and Ac ₂ O (acetic anhydride 2 eq) in dry dichloromethane, then add TsOH (p-toluenesulfonic acid 3 eq), stir at room temperature for 24 h, and check the reaction progress by TLC . After the reaction, the pH value of the reaction solution was adjusted to > 9 with concentrated ammonia water, and extracted twice with DCM. The solvent was removed by evaporation under reduced pressure. The crude product was purified by silica gel flash chromatography to give 15 acetylated product.

3) The product obtained in 2) was mixed with TBAF (tetrabutylammonium fluoride 2eq) in THF, and stirred at 75 degrees under reflux for 10h, and the reaction progress was detected by TLC. After the reaction, the reaction solution was extracted twice with ether/ethyl acetate (1:1), and the solvent was evaporated under reduced pressure. The crude product was purified by flash chromatography on silica gel to give product 31. The synthetic route is shown in Figure 4.

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 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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-15β), 4.49 (dd, J = 5.4, 2.8 Hz, 1H, H-14β), 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, 1'-OCH ₃ ), 1.04 (t, 3H, N-CH ₂ CH ₃ ), 0.88 (t, J = 3.7 Hz, 3H, H-18'''). ¹³ C NMR data are shown in Figure 11. HRMS calculated for C ₅₂ H ₇₈ NO ₁₂ 908.5524, found 908.5516 [M+H] ⁺ .

Example 12: Structural formula of compound 32 referred to in the following examples, see Figure 3 .

The product 32 was obtained by reacting 0.1 mmol of Coconut base A with 0.3 mmol of linoleic acid at 110 degrees vacuum for 30 min, and the oily substance was purified by silica gel flash chromatography. The synthetic route is shown in Figure 5.

8-linoleate vilmorrianine A (32) (light yellow oil, 90.23% yield). IR(KBr): 3448, 2921, 2852, 1652, 1447, 1384, 1265, 1088, 736 cm ^–1 ; ¹ H-NMR (400MHz) , CDCl ₃ ) δ 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-14β), 4.05 (m, 1H, H-6β), 3.83 (s,3H, 16′-OCH ₃ ), 3.81 – 3.77 (m, 1H), 3.60and 3.42 (d, J = 8.9 Hz, each 1H, H-18), 3.39 (s, 3H, 4′-OCH ₃ ), 3.28 (s, 3H, 18′-OCH ₃ ), 3.24 (s, 3H, 1′-OCH ₃ ), 3.16 (s, 3H, 6′-OCH ₃ ), 1.08 ( s, 3H, N-CH ₂ CH ₃ , 0.87 (s, 3H, H-18'''). ¹³ C NMR data are shown in Figure 11. HRMS calculated for C ₅₁ H ₇₈ NO ₁₀ 864.5626, found 864.5756[M+H ] ⁺ .

Example 13: Structural formula of compound 33 referred to in the examples below, see Figure 3 .

To 0.2 mL of dichloromethane were added 0.05 mmol of homocontamine and 0.1 mmol of p-toluenesulfonic acid, 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. The chloroform was dissolved and then transferred to a separatory funnel. After washing the organic phase twice with water, it was dried over anhydrous Na ₂ SO ₄ , and the organic phase was concentrated under reduced pressure. The product was purified by silica gel flash chromatography to obtain the 8-position acetylated homoconglanine. It was then reacted with 3 eq of linoleic acid under vacuum at 110 degrees for 30 min, and the oily substance was purified by silica gel flash chromatography to obtain product 33. The synthetic route is shown in Figure 5.

8-linoleate lappaconitine (33). IR (KBr): 3477, 2925, 2850, 1607, 1384, 1255, 1169, 1099, 691 cm ^–1 ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 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, H _a -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, 1′-OCH ₃ ), 3.18 ( dd, J =10.3, 6.8 Hz, 1H, H-1), 2.99 (d, J = 3.4 Hz, 1H, H-17), 2.67 (m, 1H, H _a -6), 2.59 (q, J = 2.1 Hz, 1H, H _a -3), 2.56 (d, J = 7.7 Hz, 1H, H _a -21), 2.52 (dd, J =5.0, 2.4 Hz, 1H, H _b -19), 2.49 (d , J = 9.9 Hz, 1H, H _b -21), 2.45 (dd, J = 7.7 Hz, 1H, H _a -12), 2.34 (dt, J = 7.8, 3.8 Hz, 1H, H _a -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, _NHCOC H ₃ ) ,2.17 (d, J = 6.5 Hz, 1H, H _b -2), 2.10 (dd, J = 12.4, 4.5 Hz, 1H, H-7), 1 .97(t, J = 3.3 Hz, 1H, H _b -12), 1.88 (s, 1H, H _b -3), 1.57 (s, 1H, H _b -6), 1.11 (t, J = 7.1 Hz , 3H, NCH ₂ CH ₃ ), 0.88 (t, J = 7.2, 2.1 Hz, 3H, H-18′′′). ¹³ C NMR data are shown in Figure 11. HRMS calculated for C ₅₀ H ₇₅ N ₂ O ₉ 847.5473 , found 847.5605[M+H] ⁺ .

Example 14:

In vitro antitumor activity test:

1. Experimental samples and experimental methods

Activity experiments were carried out using the compounds prepared in Examples 1-33.

Using MTS method, with doxorubicin and etoposide as positive controls, the _IC50 values of compounds 1-33 against doxorubicin-resistant human breast cancer cells (ADR-MCF-7) were determined.

The principle of MTS assay for detecting cell activity: MTS is a new MTT analog, the full name is 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxy-methoxyphenyl)-2-(4-sulfopheny)- 2H-tetrazolium, is a yellow dye. Succinate dehydrogenase in the mitochondria of living cells can metabolize and reduce MTS to generate soluble formazan (Formazan) compounds. The content of formazan can be measured at 490nm with a microplate reader. Under normal circumstances, the amount of formazan produced is proportional to the number of viable cells, so the number of viable cells can be inferred from the optical density OD value.

experimental method:

1) Sample configuration: Take 1 mg of each compound prepared in Examples 1-33, dissolve it in 1 mL of a weakly acidic buffer (pH 5.5-7) with a certain pH value, and store it as a stock solution under refrigeration at the corresponding pH. The blank buffer was used as diluent and blank control.

2). Seeding cells: prepare a single cell suspension with a culture medium containing 10% fetal bovine serum (DMEM or RMPI1640), and seed 3000~15000 cells per well into a 96-well plate, with a volume of 100ul per well, adherent cells Inoculate and culture 12-24 hours in advance.

3). Add the solution of the compound to be tested, the final volume of each well is 200ul, and each treatment has 3 duplicate wells.

4). Color development: after culturing at 37 degrees Celsius for 48 hours, the adherent cells discard the medium in the wells, add 20ul MTS solution and 100ul medium to each well; discard 100ul culture supernatant for suspended cells, and add 20ul MTS solution to each well; 3 blank duplicate wells (a mixture of 20 ul MTS solution and 100 ul culture medium) were incubated for 2 to 4 hours to allow the reaction to proceed fully and then measure the light absorption value.

5). Colorimetry: select the wavelength of 492nm, read the light absorption value of each well with a multi-function microplate reader (MULTISKAN FC), record the results, and draw the cell growth curve with the concentration as the abscissa and the cell viability as the ordinate after data processing. The _IC50 values of the compounds were calculated using the two-point method (Reed and Muench method). 6). Positive control compounds: Doxorubicin and etoposide were set as positive compounds in each experiment, and the cell growth curve was drawn with the concentration as the abscissa and the cell viability as the ordinate, and the two-point method (Reed and Muench method) was used. Calculate the _IC50 value of the compound.

2. Experimental results (see Figure 6)

3. Conclusion

Compounds 7-12, 14, 16-22, 26, 28-29, 31-32 have obvious effects of reversing multidrug resistance of tumor cells, and have anti-canine breast cancer cell effects, and can be used as anti-tumor drugs in humans. A study of drug reversal agents 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 male, all Kunming mice were kept under strict conditions, in which the temperature was 22±1°C, and the humidity was 55±5%, which satisfies the mice's free access to food and water.

1.2 Test operation

110 healthy mice (half male and female) were randomly divided into 11 groups, each with 10 mice in the experimental group and 10 mice in the blank group. 200, 210, 220 mg/kg) and aconitine (dose of 0.8, 0.4, 0.2, 0.1, 0.05 mg/kg, respectively) were intraperitoneally administered, and the mice were fasted for 8-10 hours before the oral administration. , but not water. The blank group was injected with the same dose of solvent. Before the test, the drug was formulated to a certain concentration, so that the dosage was 0.02 mL/g body weight. After administration, the general health status, poisoning performance and death process of the mice were observed and recorded, and the observation time was 14 days. At 14 days, the mice were sacrificed and necropsy for data collection.

1.3 Experimental results

After calculation, the LD ₅₀ of compound 7 administered intraperitoneally to mice was 192.944 mg/kg, and its 95% confidence limit was 179.692-201.337 mg/kg; the LD ₅₀ of aconitine administered intraperitoneally to mice was 0.2 mg/kg kg acute toxicity test. The internal organs of the undead mice were observed after dissection, and it was found that there was no obvious abnormality in the general observation of the organs of aconitine linoleate (7) (see Figure 13). The aconitine (0.2 mg/kg) group and aconitine linoleate (7) (192.944 mg/kg) were taken for slice observation, and it was found that except for the liver and lungs with slight changes, other organs such as heart, spleen, Compared with the blank group, the kidney and thymus had no significant changes in the two drug groups. The H&E staining results are shown in the appendix.

The results of liver pathological sections are shown in Figure 14. Compared with the blank group, the liver tissue of the aconitine medium-dose group was significantly damaged, with moderate diffuse vacuolar degeneration and punctate necrosis. Punctate necrosis was occasionally seen in the high-dose ester (7) group.

From Figure 15, it can be seen from the pathological section results of lung tissue that compared with the blank group, the alveolar septum in the aconitine medium-dose group was significantly thickened ^[66] , and the number of alveoli decreased; aconitine linoleate ( 7) The alveolar septum was slightly thickened in the high dose group, but it was significantly improved compared with the middle dose group of aconitine.

1.4 Conclusion

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

Claims (6)

1. A compound of formula I or a salt thereof

The method is characterized in that: r₀Hydrogen, methyl and ethyl; r₁，R₃，R₄，R₈Hydrogen, hydroxy, C1-6 alkoxy such as methoxy or ethoxy, C1-6 alkanoyloxy such as acetyl; r₂Is a hydroxyl group; r₆，R₉，R₁₀Is hydrogen, hydroxy or C1-6 alkanoyloxy such as acetyl, R₅Is a saturated or unsaturated or halogen-containing long-chain fatty acyl oxygen containing 8-24 carbon atomsExamples of the group include octanoyl group, oleoyloxy group, linolenoyloxy group, oleoyloxy group, palmitoyloxy group, stearoyloxy group, eicosapentaenoic acid acyloxy group, 10-fluoroctearoyloxy group, 9, 13-difluorooctadecanoyloxy group, 10-bromooctadecanoyloxy group, 9, 13-dibromooctadecanoyloxy group, R₇Is hydrogen or hydroxy or benzoyloxy or p-halobenzoyloxy; x is a pharmaceutically commonly used acid radical, such as chlorine, bromine, trifluoroformyloxy and the like.

2. Use according to claim 1, characterized in that: the C1-6 alkoxy is selected from methoxy or ethoxy; the C1-6 alkanoyloxy is selected from acetyl; the long-chain fatty acyloxy is selected from octanoyl, linoleoxy, oleoyloxy, palmitoyloxy, hard acyloxy, 9, 13-difluoro octadecanoyloxy, 10-bromo octadecanoyloxy and 9, 13-dibromo octadecanoyloxy.

3. Use according to claim 1, characterized in that: the R is₀Preferably hydrogen and ethyl; r₁，R₃，R₄，R₈Preferably methoxy, R₂Preferably hydroxy, R₆，R₉，R₁₀Preferably hydrogen or hydroxy or acetyl, R₅Preferably a saturated or unsaturated or halogen-containing long-chain fatty acyloxy group having 8 to 18 carbon atoms such as octanoyl, oleoyloxy, palmitoyloxy, stearoyloxy, R₇Preferably, it is p-methoxybenzoyloxy, and X is preferably chlorine.

4. Use according to claim 1, characterized in that it is selected from the following compounds:

5. the compound of claim 1, wherein the compound is prepared by using natural diterpene alkaloids as raw materials, chemically changing substituents in the structure, and introducing 8-bit long-chain fatty acid ester, and the preparation method comprises the following steps:

。

6. the compound of claim 1, wherein the compound is used for preparing an anti-tumor multidrug resistance reversal agent and an anti-canine breast cancer preparation.

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