A mitochondrial-targeted hecosaponin derivative, its preparation method and application

Abstract

This invention belongs to the field of natural medicines and medicinal chemistry, and relates to a triphenylphosphine derivative of hecosinate, its preparation method, and its application. Specifically, it relates to a method for preparing a triphenylphosphine derivative by introducing it into the C3 or C12 of the hecosinate core structure and its application in the preparation of antitumor drugs. The hecosinate triphenylphosphine derivative provided by this invention has mitochondrial targeting activity and, compared with unmodified hecosinate, exhibits superior antitumor activity, and can be further used in the preparation of antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the field of natural medicines and medicinal chemistry, and relates to a mitochondrial-targeted hecosinate derivative, its preparation method, and its application. Specifically, it relates to a method for preparing a triphenylphosphine derivative by introducing it into the C3 or C12 of the hecosinate core structure and its use in the preparation of antitumor drugs. Background Technology

[0002] Cancer is a major category of diseases that seriously threatens the health of Chinese residents. On March 22, 2023, the National Cancer Center released the latest national cancer statistics in the *Chinese Journal of Oncology*. The data shows that lung cancer is the most common type of newly diagnosed cancer, accounting for more than 20% of all new cancer cases; followed by colorectal cancer, stomach cancer, liver cancer, and breast cancer. These five most common cancers account for 57.4% of all new cancer cases. Lung cancer ranks first in mortality among malignant tumors, accounting for approximately 27% of all cancer deaths; followed by liver cancer, stomach cancer, colorectal cancer, and esophageal cancer. These five types of cancer account for 69.25% of all cancer deaths.

[0003] Currently, cancer treatment strategies mainly include surgery, radiotherapy and chemotherapy, targeted therapy, and immunotherapy. Current clinical practice shows that chemotherapy remains one of the most effective methods for treating cancer, but its adverse side effects severely limit its clinical application. Therefore, developing safe and effective new drugs is particularly important for cancer treatment. Traditional Chinese medicine ingredients generally possess good biological activity and safety; therefore, developing safe and effective anti-tumor drugs from traditional Chinese medicine has become a research hotspot for many researchers.

[0004] Targeted strategies enable drug delivery to specific sites, reducing off-target effects and harmful toxicity, thereby enhancing therapeutic efficacy. A growing body of research indicates that selectively delivering drugs to specific subcellular organelles can significantly improve the efficiency of cancer treatment. Mitochondria, as one of the most important subcellular organelles in the human body, not only serve as the cell's "powerhouse," providing energy for various life activities, but also play a crucial role in regulating cellular metabolism and maintaining normal cellular function. Given the vital role of mitochondria in regulating cancer cell apoptosis, mitochondrial-targeted therapies have shown great promise in cancer treatment. Designing molecules that specifically target mitochondria is a new direction for anticancer drug development. Therefore, using natural products with good antitumor activity as lead structures, modifying and optimizing their structures to obtain derivatives with potential development value is an important approach to the discovery and development of molecularly targeted antitumor drugs.

[0005] Hecogenin (HG) is naturally found in the leaves of Agave plants, such as the traditional Chinese medicine Agave sisalana. The C12 position of the HG molecule contains a carbonyl group, a necessary condition for the synthesis of adrenocortical hormones. Therefore, it has long been used as a precursor for the synthesis of steroid hormones and steroidal anti-inflammatory drugs. In recent years, it has been found to possess pharmacological activities such as anticancer, anti-inflammatory, antifungal, antihypertensive, and anti-ulcer effects. Thus, HG has broad application value both as a drug synthesis precursor and due to its pharmacological activities. However, HG is limited by its poor water solubility and moderate antitumor activity; therefore, it is necessary to modify it to improve its water solubility and antitumor activity.

[0006] This invention uses hecosaponin as a lead compound and, by employing the principle of combination, links the mitochondrial targeting group triphenylphosphine to the C3 or C12 position of the hecosaponin molecule structure via a linking group, thus designing and synthesizing novel mitochondrial-targeting hecosaponin triphenylphosphine derivatives represented by general formula I or general formula II. Summary of the Invention

[0007] The technical problem to be solved by this invention is to prepare a series of novel triphenylphosphine derivatives of hecoside with strong antitumor activity, and further provide pharmaceutical compositions comprising said derivatives, as well as methods for preparing said derivatives. In addition, the use of said derivatives or pharmaceutical compositions in the preparation of antitumor drugs is also provided.

[0008] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0009] In a first aspect, the present invention provides a triphenylphosphine derivative of hecosaponin, represented by general formula I or general formula II, wherein the structural formula of the derivative is:

[0010]

[0011] Where n is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8.

[0012] In one specific embodiment, the hecosaponin triphenylphosphine derivative is selected from the following:

[0013]

[0014]

[0015] In a second aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned hecosaponin triphenylphosphine derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0016] In a preferred embodiment, the pharmaceutical composition is in the form of an oral dosage form or an injectable dosage form.

[0017] More preferably, the oral dosage form is selected from capsules, tablets, granules, oral liquids, sustained-release preparations, or controlled-release preparations.

[0018] In a third aspect, the present invention provides a method for preparing the above-mentioned hecosaponin triphenylphosphine derivative, the method comprising the following steps:

[0019] Hecosaponin (HG)1 reacts with bromocarboxylic acid at room temperature under DCC and DMAP conditions to generate intermediate 2a-2h, and intermediate 2a-2h reacts with triphenylphosphine under reflux to generate the target compound 3a-3h.

[0020] The reaction route is shown below:

[0021]

[0022] Alternatively, the present invention also provides a method for preparing the above-mentioned hecosaponin triphenylphosphine derivative, the preparation method comprising the following steps:

[0023] Hecosaponin (HG)1 was reacted with acetic anhydride and hydroxylamine hydrochloride to generate intermediate 4. Intermediate 4 was reacted with bromocarboxylic acid at room temperature under DCC and DMAP conditions to generate intermediate 5a-5g. Intermediate 5a-5g was reacted with triphenylphosphine under reflux to generate target compound 6a-6g.

[0024] The reaction route is shown below:

[0025]

[0026] In a fourth aspect, the present invention provides the use of the above-mentioned hecosaponin triphenylphosphine derivative or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating tumor diseases.

[0027] In a preferred embodiment, the tumor is a solid tumor or a hematologic cancer.

[0028] In a more preferred embodiment, the tumor is leukemia, lymphoma, myeloma, breast cancer, prostate cancer, melanoma, osteosarcoma, neuroblastoma, pancreatic cancer, lung cancer, Wilms' tumor, rhabdomyosarcoma, Ewing sarcoma, bladder cancer, colon cancer, liver cancer, ovarian cancer, cervical cancer, nasopharyngeal carcinoma, laryngeal cancer, stomach cancer, or thyroid cancer.

[0029] In a preferred embodiment, the tumor is gastric cancer, breast cancer, lung cancer, colon cancer, or liver cancer.

[0030] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here.

[0031] Compared with the prior art, the present invention has the following advantages:

[0032] The triphenylphosphine derivative of hecosaponin of the present invention is based on hecosaponin as a lead compound. By using the principle of combination, the mitochondrial targeting group triphenylphosphine is linked to the C3 or C12 position of the hecosaponin molecule structure through a linking group. The novel mitochondrial-targeting triphenylphosphine derivative of general formula I or general formula II is designed and synthesized.

[0033] Pharmacodynamic experiments conducted on human cancer cell lines showed that these novel coxabinogen triphenylphosphine derivatives exhibited good cytotoxic effects against a variety of human tumors. Therefore, the novel coxabinogen triphenylphosphine derivatives prepared in this invention are suitable for development into novel antitumor drugs, possessing very high potential research value and clinical application prospects. Detailed Implementation

[0034] The inventors have long been dedicated to the research and development of steroidal saponin derivatives. Through extensive screening, they unexpectedly discovered that by combining hecosinate with triphenylphosphine, a series of novel hecosinate-triphenylphosphine derivatives with good antitumor activity and low toxicity could be prepared. Based on this, the present invention was completed.

[0035] To facilitate better reading of this instruction manual, the main abbreviations used in this manual are provided below.

[0036]

[0037] Preparation Example:

[0038] Example 1:

[0039]

[0040] Intermediate 2a (37.5 mg, 0.07 mmol) and 5 mL of CH3CN were added to a 25 mL dry, round-bottomed flask. After stirring to dissolve, triphenylphosphine (128.4 mg, 0.49 mmol) was added, and the mixture was refluxed and stirred for 24 h. The reaction was monitored by thin-layer chromatography (dichloromethane:methanol volume ratio = 12:1). The reaction of the starting material was basically complete. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain the target compound 3a as a yellow powder with a yield of 67.3%.

[0041] 1 HNMR (600MHz, CDCl)3, δ): 4.75 (m, 1H, H-3), 4.34 (dd, J = 7.2, 13.4 Hz, 1H, H-16), 3.79 (s, 2H, H-2′), 3.48 (dd, J = 11.0, 6.4 Hz, 1H, H-26), 3.34 (t, J = 11.0 Hz, 1H, H-26), 2.51 (dd, J = 13.4, 7.2 Hz, 1H, H-17), 2.40 (t, J = 13.9 Hz, 1H, H-11), 2.22 (dd, J = 13.9, 5.0 Hz, 1H, H-11), 2.11 (m, 1H, H-15), 1.90 (m, 1H, H-8), 1.06 (d, J = 7.0 Hz, 3H, 21-CH3), 1.04 (s, 3H, 18-CH3), 0.93 (s, 3H, 19-CH3), 0.79 (d, J = 6.4 Hz, 3H, 27-CH3).

[0042] 13 C NMR (150 MHz, CDCl3, δ): 213.7 (C-12), 164.4 (C-1′), 135.2 (d, J C,P = 3.2 Hz, C-4″), 134.3 (d, J C,P = 10.7 Hz, C-2″, C-6″), 130.5 (d, J C,P = 13.4 Hz, C-3″, C-5″), 118.2 (d, J C,P = 88.1 Hz, C-1″), 109.4 (C-22), 79.3 (C-16), 76.6 (C-3), 67.0 (C-26), 55.8 (C-9), 55.4 (C-14), 55.2 (C-13), 53.6 (C-17), 44.6 (C-5), 42.3 (C-20), 37.9 (C-11), 36.2 (C-1), 36.1 (C-10), 34.4 (C-8), 33.2 (C-4), 32.8 (d, J C,P = 56.1 Hz, C-2′), 31.6 (C-23), 31.6 (C-7), 31.3 (C-15), 30.3 (C-25), 28.9 (C-24), 28.1 (C-6), 26.8 (C-2), 17.3 (C-27), 16.1 (C-18), 13.4 (C-21), 12.0 (C-19).

[0043] ESI-HRMS: m / z 733.4008 [M - Br] + (Calculated: C 47 H 58 O5P, 733.4016).

[0044] Example 2:

[0045]

[0046] The target compound 3b was prepared according to the synthesis method of Example 1. It was a yellow powder with a yield of 56.5%.

[0047] 1 HNMR(600MHz, CDCl3, δ):7.80-7.78(m,6H,H-2″,H-6″),7.76-7.75(m,3H,H-4″),7.71-7.68(m,6H,H-3″,H-5″),4.40(m ,1H,H-3),4.32(m,1H,H-16),3.92(m,2H,H-3′),3.48(m,1H,H-26),3.34(t,J=11.0Hz,1H,H-26),2.97(m,2H,H-2′),2. 49(dd,J=8.6,6.8Hz,1H,H-17),2.35(t,J=13.8Hz,1H,H-11),2.17(dd,J=14.3,4.9Hz,1H,H-11),2.09(m,1H,H-15),1. 86(m,1H,H-8),1.05(d,J=7.0Hz,3H,21-CH3),1.02(s,3H,18-CH3),0.84(s,3H,19-CH3),0.78(d,J=6.3Hz,3H,27-CH3).

[0048] 13 C NMR (150MHz, CDCl3, δ): 213.8 (C-12), 170.6 (C-1′), 135.2 (d, J C,P =2.8Hz,C-4″),134.1(d,J C,P =10.2Hz,C-2″,C-6″),130.6(d,J C,P =12.3Hz,C-3″,C-5″),118.2(d,J C,P=86.8Hz,C-1″),109.4(C-22),79.3(C-16),74.9(C-3),67.0(C-26),55.8(C-9 ),55.4(C-14),55.2(C-13),53.6(C-17),44.5(C-5),42.3(C-20),37.9(C-11), 36.2(C-1),36.1(C-10),34.4(C-8),33.5(C-4),31.6(C-23),31.5(C-7),31.5( C-2′),31.2(C-15),30.3(C-25),28.9(C-24),28.2(C-6),27.0(C-2),18.1(d,J C,P =54.5Hz, C-3′), 17.3(C-27), 16.1(C-18), 13.4(C-21), 12.0(C-19).

[0049] ESI-HRMS: m / z 747.4135 [M-Br] + (Calculated value: C) 48 H 60 O5P,747.4173).

[0050] Example 3:

[0051]

[0052] The target compound 3c was prepared according to the synthesis method of Example 1. It is a yellow powder with a yield of 62.4%.

[0053] 1HNMR(600MHz,CDCl3,δ):7.88-7.85(m,6H,H-2″,H-6″),7.78-7.75(m,3H,H-4″),7.69-7.66(m,6H,H-3″,H-5″),4.62(m,1H,H-3),4.31(m,1H,H-16),4.05(m,2H,H-4′),3.46(m,1H,H-26),3.33(t,J=11.1Hz,1H,H-26),2.84(m,2H,H-2′),2.49(dd,J=8.5,6.7Hz,1H,H-17),2.37(t,J=13.8Hz,1H,H-11),2.18(dd,J=14.3,5.1Hz,1H,H-11),2.08(m,1H,H-15),1.88(m,3H,H-8,H-3′),1.04(d,J=6.9Hz,3H,21-CH3),1.03(s,3H,18-CH3),0.88(s,3H,19-CH3),0.77(d,J=6.4Hz,3H,27-CH3)。

[0054] 13 C NMR(150MHz,CDCl3,δ):213.6(C-12),173.0(C-1′),135.0(d,J C,P =2.8Hz,C-4″),133.9(d,J C,P =9.7Hz,C-2″,C-6″),130.5(d,J C,P =12.3Hz,C-3″,C-5″),118.4(d,J C,P =86.4Hz,C-1″),109.4(C-22),79.3(C-16),73.7(C-3),67.0(C-26),55.9(C-9),55.5(C-14),55.2(C-13),53.6(C-17),44.6(C-5),42.3(C-20),37.9(C-11),36.3(C-1),36.2(C-10),34.4(C-8),33.8(C-4),33.6(d,J C,P =19.1Hz,C-2′),31.6(C-23),31.5(C-7),31.2(C-15),30.3(C-25),28.9(C-24),28.2(C-6),27.3(C-2),21.6(d,J C,P =51.0Hz,C-4′),18.2(d,J C,P=3.1Hz, C-3′), 17.2(C-27), 16.1(C-18), 13.4(C-21), 12.0(C-19).

[0055] ESI-HRMS: m / z 761.4291 [M-Br] + (Calculated value: C) 49 H 62 O5P,761.4329).

[0056] Example 4:

[0057]

[0058] The target compound 3d was prepared according to the synthesis method of Example 1. It was a yellow powder with a yield of 70.6%.

[0059] 1 HNMR (600MHz, CDCl3, δ): 7.77-7.75 (m, 6H, H-2″, H-6″), 7.74-7.73 (m, 3H, H-4″), 7.71-7.68 (m, 6H, H-3″, H-5″), 4.58 (m, 1H, H- 3),4.33(m,1H,H-16),3.59(m,2H,H-5′),3.47(m,1H,H-26),3.34(t,J=11.0Hz,1H,H-26),2.50(dd,J=8.8,6.9Hz,1H,H-17),2 .38(t,J=7.1Hz,2H,H-2′),2.37(t,J=13.2Hz,1H,H-11),2.20(dd,J=14.2,4.9Hz,1H,H-11),2.09(m,1H,H-15),1.97(m,2H,H- 3′),1.88(m,1H,H-8),1.05(d,J=7.1Hz,3H,21-CH3),1.03(s,3H,18-CH3),0.87(s,3H,19-CH3),0.78(d,J=6.4Hz,3H,27-CH3).

[0060] 13 C NMR (150MHz, CDCl3, δ): 213.5 (C-12), 172.7 (C-1′), 135.0 (d, J C,P =2.8Hz,C-4″),133.8(d,J C,P =10.2Hz,C-2″,C-6″),130.5(d,J C,P =12.7Hz,C-3″,C-5″),118.4(d,J C,P=85.5Hz,C-1″),109.3(C-22),79.2(C-16),73.2(C-3),67.0(C-26),55.8(C-9 ),55.4(C-14),55.2(C-13),53.6(C-17),44.5(C-5),42.2(C-20),37.8(C-11), 36.3(C-1),36.1(C-10),34.3(C-8),33.8(C-4),33.8(C-2′),31.5(C-23),31.5 (C-7),31.2(C-15),30.2(C-25),28.8(C-24),28.2(C-6),27.2(C-2),25.5(d,J C,P =17.3Hz,C-3′),22.5(d,J) C,P =51.4Hz,C-5′),22.0(d,J C,P =3.6Hz, C-4′), 17.2(C-27), 16.1(C-18), 13.3(C-21), 12.0(C-19).

[0061] ESI-HRMS: m / z 775.4469 [M-Br] + (Calculated value: C) 50 H 64 O5P,775.4486).

[0062] Example 5:

[0063]

[0064] The target compound 3e was prepared as a yellow powder with a yield of 74.6% by following the synthesis method in Example 1.

[0065] 1HNMR(600MHz,CDCl3,δ):7.79-7.76(m,6H,H-2″,H-6″),7.75-7.74(m,3H,H-4″),7.72-7.68(m,6H,H-3″,H-5″),4.65(m,1H,H-3),4.33(m,1H,H-16),3.64(m,2H,H-6′),3.48(m,1H,H-26),3.34(t,J=11.0Hz,1H,H-26),2.50(dd,J=8.7,6.7Hz,1H,H-17),2.37(t,J=13.9Hz,1H,H-11),2.27(t,J=7.6Hz,2H,H-2′),2.21(dd,J=14.2,4.9Hz,1H,H-11),2.10(m,1H,H-15),1.88(m,1H,H-8),1.87(m,2H,H-3′),1.05(d,J=7.0Hz,3H,21-CH3),1.03(s,3H,18-CH3),0.89(s,3H,19-CH3),0.78(d,J=6.3Hz,3H,27-CH3)。

[0066] 13 C NMR(150MHz,CDCl3,δ):213.8(C-12),174.1(C-1′),135.0(d,J C,P =2.7Hz,C-4″),133.8(d,J C,P =10.0Hz,C-2″,C-6″),130.6(d,J C,P =12.7Hz,C-3″,C-5″),118.5(d,J C,P =85.6Hz,C-1″),109.4(C-22),79.3(C-16),73.5(C-3),67.0(C-26),55.8(C-9),55.5(C-14),55.2(C-13),53.7(C-17),44.5(C-5),42.3(C-20),37.9(C-11),36.4(C-1),36.2(C-10),34.4(C-8),34.3(C-2′),33.9(C-4),31.6(C-23),31.6(C-7),31.3(C-15),30.3(C-25),29.8(d,J C,P =15.6Hz,C-4′),28.9(C-24),28.3(C-6),27.3(C-2),24.6(C-3′),22.5(d,J C,P =2.8Hz,C-5′),22.3(d,JC,P =53.8Hz,C-6′),17.3(C-27),16.2(C-18),13.4(C-21),12.0(C-19)

[0067] ESI-HRMS: m / z 789.4653 [M-Br] + (Calculated value: C) 51 H 66 O5P,789.4642).

[0068] Example 6:

[0069]

[0070] The target compound 3f was prepared as a yellow powder with a yield of 76.5%, following the synthesis method described in Example 1.

[0071] 1 HNMR(600MHz, CDCl3, δ):7.80-7.75(m,9H,H-2″,H-4″,H-6″),7.72-7.69(m,6H,H-3″,H-5″),4.66(m,1H,H-3),4.34(m,1H ,H-16),3.67(m,2H,H-7′),3.48(m,1H,H-26),3.34(t,J=10.8Hz,1H,H-26),2.51(dd,J=8.6,6.7Hz,1H,H-17),2.38(t,J= 13.8Hz,1H,H-11),2.24(t,J=7.4Hz,2H,H-2′),2.22(dd,J=13.8,5.3Hz,1H,H-11),2.10(m,1H,H-15),1.90(m,1H,H-8),1 .84(m,2H,H-3′),1.06(d,J=7.1Hz,3H,21-CH3),1.04(s,3H,18-CH3),0.90(s,3H,19-CH3),0.78(d,J=6.4Hz,3H,27-CH3).

[0072] 13 C NMR (150MHz, CDCl3, δ): 213.7 (C-12), 174.0 (C-1′), 134.9 (d, J C,P =2.8Hz,C-4″),133.9(d,J C,P =9.9Hz,C-2″,C-6″),130.6(d,J C,P =12.2Hz,C-3″,C-5″),118.7(d,J C,P=84.2Hz,C-1″),109.4(C-22),79.3(C-16),73.3(C-3),67.0(C-26),5 5.8(C-9),55.5(C-14),55.2(C-13),53.7(C-17),44.6(C-5),42.3(C- 20),37.9(C-11),36.4(C-1),36.2(C-10),34.6(C-2′),34.4(C-8),33 .9(C-4),31.6(C-23),31.6(C-7),31.3(C-15),30.3(C-25),30.0(d,J C,P =14.0Hz,C-5′),28.9(C-24),28.6(C-4′),28.3(C-6),27.3(C-2),24.6(C-3′),22.5(d,J C,P =4.6Hz,C-6′),22.4(d,J C,P =55.0Hz, C-7′), 17.3(C-27), 16.2(C-18), 13.4(C-21), 12.1(C-19).

[0073] ESI-HRMS: m / z 803.4799 [M-Br] + (Calculated value: C) 52 H 68 O5P, 803.4799).

[0074] Example 7:

[0075]

[0076] 3 g of the target compound was prepared according to the synthesis method of Example 1. It was a yellow powder, with a yield of 68.4%.

[0077] 1HNMR(600MHz,CDCl3,δ):7.78-7.73(m,9H,H-2″,H-4″,H-6″),7.67-7.64(m,6H,H-3″,H-5″),4.58(m,1H,H-3),4.27(m,1H,H-16),3.65(m,2H,H-8′),3.42(m,1H,H-26),3.28(t,J=11.0Hz,1H,H-26),2.44(dd,J=8.6,6.9Hz,1H,H-17),2.33(t,J=13.7Hz,1H,H-11),2.15(dd,J=14.0,5.0Hz,1H,H-11),2.13(t,J=8.1Hz,2H,H-2′),2.04(m,1H,H-15),1.82(m,1H,H-8),1.73(m,2H,H-3′),0.99(d,J=6.9Hz,3H,21-CH3),0.98(s,3H,18-CH3),0.85(s,3H,19-CH3),0.72(d,J=6.3Hz,3H,27-CH3)。

[0078] 13 C NMR(150MHz,CDCl3,δ):213.5(C-12),173.2(C-1′),135.0(d,J C,P =2.7Hz,C-4″),133.6(d,J C,P =10.0Hz,C-2″,C-6″),130.5(d,J C,P =12.3Hz,C-3″,C-5″),118.3(d,J C,P =86.6Hz,C-1″),109.2(C-22),79.1(C-16),72.9(C-3),66.8(C-26),55.7(C-9),55.3(C-14),55.1(C-13),53.5(C-17),44.4(C-5),42.1(C-20),37.7(C-11),36.2(C-1),36.0(C-10),34.5(C-2′),34.3(C-8),33.8(C-4),31.4(C-23),31.4(C-7),31.1(C-15),30.2(d,J C,P =13.8Hz,C-6′),30.1(C-25),28.8(C-24),28.7(C-4′),28.7(C-5′),28.1(C-6),27.2(C-2),24.8(C-3′),22.7(d,J C,P=50.2Hz,C-8′),22.6(d,J C,P =4.6Hz, C-7′), 17.1(C-27), 16.0(C-18), 13.2(C-21), 11.8(C-19).

[0079] ESI-HRMS: m / z 817.4904 [M-Br] + (Calculated value: C) 53 H 70 O5P, 817.4955).

[0080] Example 8:

[0081]

[0082] The target compound was prepared by the synthesis method described in Example 1 for 3 hours. It was a yellow powder with a yield of 70.3%.

[0083] 1 HNMR (600MHz, CDCl3, δ): 7.83-7.80 (m, 6H, H-2″, H-6″), 7.78-7.76 (m, 3H, H-4″), 7.69-7.66 (m, 6H, H-3″, H-5″), 4.62 (m, 1H, H- 3),4.31(m,1H,H-16),3.76(m,2H,H-9′),3.45(m,1H,H-26),3.32(t,J=11.0Hz,1H,H-26),2.48(dd,J=8.7,6.9Hz,1H,H-17),2 .36(t,J=14.1Hz,1H,H-11),2.18(dd,J=14.2,5.2Hz,1H,H-11),2.17(t,J=7.4Hz,2H,H-2′),2.08(m,1H,H-15),1.87(m,1H,H- 8),1.74(m,2H,H-3′),1.03(d,J=7.0Hz,3H,21-CH3),1.01(s,3H,18-CH3),0.89(s,3H,19-CH3),0.76(d,J=6.3Hz,3H,27-CH3).

[0084] 13 C NMR (150MHz, CDCl3, δ): 213.6 (C-12), 173.4 (C-1′), 135.1 (d, J C,P =2.9Hz,C-4″),133.8(d,J C,P =9.9Hz,C-2″,C-6″),130.5(d,J C,P=12.3Hz,C-3″,C-5″),118.5(d,J C,P =85.8Hz,C-1″),109.3(C-22),79.1(C-16),73.0(C-3),66.9(C-26),55.8(C-9),55.4(C-14),55.2(C-13),53.6(C-17),44.5(C-5),4 2.2(C-20),37.8(C-11),36.3(C-1),36.2(C-10),34.6(C-2′),34.4(C-8),33.9(C-4),31.5(C-23),31.5(C-7),31.2(C-15),30.4(d,J C,P =15.4Hz,C-7′),30.3(C-25),29.1(C-6′),29.0(C-5′),28.9(C-24),28.8(C-4′),28.2(C-6),27.3(C-2),25.0(C-3′),22.7(d,J C,P =4.8Hz,C-8′),22.6(d,J C,P =50.0Hz, C-9′), 17.2(C-27), 16.1(C-18), 13.3(C-21), 12.0(C-19).

[0085] ESI-HRMS: m / z 831.5111 [M-Br] + (Calculated value: C) 54 H 72 O5P,831.5112).

[0086] Example 9:

[0087]

[0088] The target compound 6a was prepared according to the synthesis method of Example 1. It is a yellow powder with a yield of 60.2%.

[0089] 1HNMR(600MHz,CDCl3,δ):7.86-7.83(m,6H,H-2″,H-6″),7.79-7.76(m,3H,H-4″),7.69-7.66(m,6H,H-3″,H-5″),4.64(m,1H,H-3),4.35(m,1H,H-16),4.18(m,2H,H-3′),3.47(m,1H,H-26),3.34(t,J=11.1Hz,1H,H-26),3.14(dd,J=14.6,4.3Hz,1H,H-11),3.03(td,J=6.3,13.2Hz,1H,H-2′),2.92(td,J=5.5,13.2Hz,1H,H-2′),2.40(dd,J=8.4,6.0Hz,1H,H-14),2.06(m,1H,H-15),2.01(s,3H,Ac-CH3),0.95(d,J=7.0Hz,3H,21-CH3),0.94(s,3H,18-CH3),0.86(s,3H,19-CH3),0.78(d,J=6.4Hz,3H,27-CH3)。

[0090] 13 C NMR(150MHz,CDCl3,δ):172.6(C-12),170.8(Ac-CH3),169.6(C-1′),135.2(d,J C,P =2.9Hz,C-4″),133.9(d,J C,P =9.9Hz,C-2″,C-6″),130.6(d,J C,P =12.8Hz,C-3″,C-5″),118.1(d,J C,P =86.2Hz,C-1″),109.3(C-22),79.7(C-16),73.3(C-3),67.0(C-26),56.2(C-9),55.9(C-14),53.7(C-17),47.9(C-13),44.6(C-5),42.2(C-20),36.7(C-1),36.1(C-10),34.7(C-8),33.9(C-4),31.9(C-2′),31.6(C-23),31.5(C-7),30.9(C-15),30.3(C-25),28.9(C-24),28.2(C-6),27.2(C-2),22.8(C-11),21.6(Ac-CH3),18.3(d,J C,P=55.1Hz, C-3′), 17.4(C-27), 17.3(C-18), 13.4(C-21), 12.0(C-19).

[0091] ESI-HRMS: m / z 804.4398 [M-Br] + (Calculated value: C) 50 H 63 NO6P, 804.4388). Example 10:

[0092]

[0093] The target compound 6b was prepared according to the synthesis method of Example 1. It is a yellow powder with a yield of 65.5%.

[0094] 1 HNMR (600MHz, CDCl3, δ): 7.88-7.85 (m, 6H, H-2″, H-6″), 7.77-7.75 (m, 3H, H-4″), 7.68-7.65 (m, 6H, H-3″, H-5″), 4.65 (m, 1H, H-3), 4. 37(m,1H,H-16),4.04(m,2H,H-4′),3.45(m,1H,H-26),3.33(t,J=11.0Hz,1H,H-26),3.13(dd,J=14.9,4.5Hz,1H,H-11),3.05(td,J=6 .2,13.0Hz,1H,H-2′),2.90(td,J=5.3,13.0Hz,1H,H-2′),2.53(dd,J=8.1,6.0Hz,1H,H-14),2.06(m,1H,H-15),2.00(s,3H,Ac-CH3), 1.85(t,J=6.3Hz,2H,H-3′), 1.13(d,J=7.1Hz,3H,21-CH3), 0.99(s,3H,18-CH3), 0.87(s,3H,19-CH3), 0.76(d,J=6.3Hz,3H,27-CH3).

[0095] 13 C NMR (150MHz, CDCl3, δ): 172.1 (C-12), 171.8 (C-1′), 170.7 (Ac-CH3), 135.0 (d, J C,P =2.6Hz,C-4″),133.9(d,J C,P =10.4Hz,C-2″,C-6″),130.5(d,J C,P =12.9Hz,C-3″,C-5″),118.3(d,J C,P=85.6Hz,C-1″),109.3(C-22),79.7(C-16),73.4(C-3),67.0(C-26),56.1(C-9),56.0(C-14),53.6(C -17),47.9(C-13),44.6(C-5),42.3(C-20),36.6(C-1),36.1(C-10),34.7(C-8),33.9(C-4),32.3(d,J C,P =19.6Hz,C-2′),31.7(C-23),31.6(C-7),30.9(C-15),30.3(C-25),28.9(C-24),28.2(C-6),27.2(C-2),22.8(C-11),21.7(d,J C,P =51.8Hz,C-4′),21.5(Ac-CH3),18.1(d,J C,P =2.7Hz, C-3′), 17.5(C-27), 17.2(C-18), 13.6(C-21), 11.9(C-19).

[0096] ESI-HRMS: m / z 818.4528 [M-Br] + (Calculated value: C) 51 H 65 NO6P, 818.4544). Example 11:

[0097]

[0098] The target compound 6c was prepared according to the synthesis method of Example 1. It is a yellow powder with a yield of 71.2%.

[0099] 1HNMR(600MHz,CDCl3,δ):7.87-7.84(m,6H,H-2″,H-6″),7.80-7.77(m,3H,H-4″),7.71-7.68(m,6H,H-3″,H-5″),4.66(m,1H,H-3),4.39(m,1H,H-16),3.91(m,2H,H-5′),3.48(m,1H,H-26),3.36(t,J=10.8Hz,1H,H-26),3.12(dd,J=15.2,4.9Hz,1H,H-11),2.60-2.49(m,3H,H-2′,H-14),2.08(m,1H,H-15),2.02(s,3H,Ac-CH3),1.87(t,J=6.7Hz,2H,H-3′),1.12(d,J=7.0Hz,3H,21-CH3),1.01(s,3H,18-CH3),0.89(s,3H,19-CH3),0.79(d,J=6.4Hz,3H,27-CH3)。

[0100] 13 C NMR(150MHz,CDCl3,δ):171.8(C-12),171.8(C-1′),170.8(Ac-CH3),135.0(d,J C,P =2.9Hz,c-4″),133.9(d,J C,P =10.2Hz,C-2″,C-6″),130.5(d,J C,P =12.4Hz,C-3″,C-5″),118.5(d,J C,P =85.8Hz,C-1″),109.3(C-22),79.7(C-16),73.3(C-3),67.0(C-26),56.2(C-9),55.9(C-14),53.7(C-17),48.0(C-13),44.6(C-5),42.4(C-20),36.6(C-1),36.1(C-10),34.7(C-8),33.9(C-4),31.9(C-2′),31.8(C-23),31.7(C-7),30.9(C-15),30.3(C-25),28.9(C-24),28.2(C-6),27.3(C-2),25.2(d,J C,P =16.5Hz,C-3′),22.7(C-11),22.7(d,J C,P =50.0Hz,C-5′),21.9(d,J C,P=4.3Hz, C-4′), 21.5(Ac-CH3), 17.4(C-27), 17.3(C-18), 13.4(C-21), 12.0(C-19).

[0101] ESI-HRMS: m / z 832.4698 [M-Br] + (Calculated value: C) 52 H 67 NO6P, 832.4701).

[0102] Example 12:

[0103]

[0104] The target compound 6d was prepared according to the synthesis method of Example 1. It is a yellow powder with a yield of 73.8%.

[0105] 1 HNMR (600MHz, CDCl3, δ): 7.78-7.74(m,9H,H-2″,H-4″,H-6″),7.71-7.68(m,6H,H-3″,H-5″),4.66(m,1H,H-3),4.39(m, 1H,H-16),3.59(t,J=6.8Hz,2H,H-6′),3.47(m,1H,H-26),3.34(t,J=11.1Hz,1H,H-26),3.13(dd,J=15.1,4.7Hz,1H,H-1 1),2.56(dd,J=8.4,6.0Hz,1H,H-14),2.41(t,J=6.6Hz,2H,H-2′),2.07(m,1H,H-15),2.01(s,3H,Ac-CH3),1.86(t,J=6. 6Hz, 2H, H-3′), 1.12 (d, J = 7.0Hz, 3H, 21-CH3), 1.00 (s, 3H, 18-CH3), 0.88 (s, 3H, 19-CH3), 0.77 (d, J = 6.4Hz, 3H, 27-CH3).

[0106] 13 C NMR (150MHz, CDCl3, δ): 172.4 (C-12), 171.8 (C-1′), 171.0 (Ac-CH3), 135.0 (d, J C,P =2.8Hz,C-4″),133.8(d,J C,P =10.0Hz,C-2″,C-6″),130.6(d,J C,P =12.8Hz,C-3″,C-5″),118.4(d,J C,P=86.1Hz,C-1″),109.3(C-22),79.8(C-16),73.6(C-3),67.0(C-26),56.2(C-9),55.9(C-14),53.8(C-17),48.0(C-13),44.5(C-5),4 2.4(C-20),36.5(C-1),36.1(C-10),34.8(C-8),33.9(C-4),32.6(C-2′),31.7(C-23),31.5(C-7),30.9(C-15),30.3(C-25),29.7(d,J C,P =15.9Hz,C-4′),28.9(C-24),28.2(C-6),27.3(C-2),24.3(C-3′),22.7(C-11),22.6(d,J C,P =4.4Hz,C-5′),22.3(d,J C,P =50.5Hz, C-6′), 21.6(Ac-CH3), 17.4(C-27), 17.3(C-18), 13.4(C-21), 12.0(C-19).

[0107] ESI-HRMS: m / z 846.4852 [M-Br] + (Calculated value: C) 53 H 69 NO6P, 846.4857). Example 13:

[0108]

[0109] The target compound 6e was prepared as a yellow powder with a yield of 70.4%, following the synthesis method described in Example 1.

[0110] 1HNMR(600MHz,CDCl3,δ):7.81-7.78(m,6H,H-2″,H-6″),7.75-7.72(m,3H,H-4″),7.71-7.68(m,6H,H-3″,H-5″),4.66(m,1H,H-3),4.38(m,1H,H-16),3.66(m,2H,H-7′),3.45(m,1H,H-26),3.34(t,J=11.0Hz,1H,H-26),3.12(dd,J=15.0,4.6Hz,1H,H-11),2.56(dd,J=8.2,6.1Hz,1H,H-14),2.35(t,J=7.2Hz,2H,H-2′),2.06(m,1H,H-15),2.02(s,3H,Ac-CH3),1.86(t,J=6.4Hz,2H,H-3′),1.11(d,J=7.0Hz,3H,21-CH3),1.00(s,3H,18-CH3),0.88(s,3H,19-CH3),0.77(d,J=6.3Hz,3H,27-CH3)。

[0111] 13 C NMR(150MHz,CDCl3,δ):172.3(C-12),171.7(C-1′),171.2(Ac-CH3),134.9(d,J C,P =2.6Hz,C-4″),133.8(d,J C,P =10.1Hz,C-2″,C-6″),130.6(d,J C,P =12.7Hz,C-3″,C-5″),118.5(d,J C,P =85.9Hz,C-1″),109.3(C-22),79.8(C-16),73.6(C-3),67.0(C-26),56.2(C-9),55.9(C-14),53.6(C-17),48.0(C-13),44.5(C-5),42.4(C-20),36.4(C-1),36.1(C-10),34.8(C-8),33.9(C-4),33.0(C-2′),31.7(C-23),31.5(C-7),30.9(C-15),30.3(C-25),30.0(d,J C,P =14.9Hz,C-5′),28.9(C-24),28.6(C-4′),28.2(C-6),27.3(C-2),24.2(C-3′),22.7(C-11),22.4(d,J C,P=4.0Hz,C-6′),22.3(d,J C,P =49.7Hz, C-7′), 21.6(Ac-CH3), 17.4(C-27), 17.2(C-18), 13.4(C-21), 12.0(C-19).

[0112] ESI-HRMS: m / z 860.5005 [M-Br] + (Calculated value: C) 54 H 71 N2O6P, 860.5014).

[0113] Example 14:

[0114]

[0115] The target compound 6f was prepared as a yellow powder with a yield of 72.5%, following the synthesis method described in Example 1.

[0116] 1 HNMR(600MHz,CDCl3,δ):7.86-7.83(m,6H,H-2″,H-6″),7.79-7.77(m,3H,H-4″),7.71-7.68(m,6H,H-3″,H-5″),4.65(m,1 H,H-3),4.38(m,1H,H-16),3.82(m,2H,H-8′),3.46(m,1H,H-26),3.34(t,J=11.1Hz,1H,H-26),3.13(dd,J=15.1,4.7Hz,1H ,H-11),2.57(dd,J=8.3,6.1Hz,1H,H-14),2.37(t,J=7.3Hz,2H,H-2′),2.07(m,1H,H-15),2.00(s,3H,Ac-CH3),1.86(t,J= 6.7Hz,2H,H-3′),1.12(d,J=7.1Hz,3H,21-CH3),1.01(s,3H,18-CH3),0.89(s,3H,19-CH3),0.77(d,J=6.4Hz,3H,27-CH3).

[0117] 13 C NMR (150MHz, CDCl3, δ): 172.4 (C-12), 171.7 (C-1′), 170.8 (Ac-CH3), 135.0 (d, J C,P =2.6Hz,C-4″),133.8(d,J C,P =9.8Hz,C-2″,C-6″),130.6(d,J C,P=12.7Hz,C-3″,C-5″),118.6(d,J C,P =85.6Hz,C-1″),109.3(C-22),79.8(C-16),73.3(C-3),67.0(C-26),56.3(C-9),55.9(C-14),53.7(C-17),48.0(C-13),44.6(C-5),4 2.4(C-20),36.5(C-1),36.1(C-10),34.8(C-8),33.9(C-4),33.0(C-2′),31.7(C-23),31.5(C-7),30.9(C-15),30.3(C-25),30.2(d,J C,P =15.1Hz,C-6′),29.0(C-24),28.9(C-4′),28.7(C-5′),28.2(C-6),27.3(C-2),24.6(C-3′),22.8(d,J C,P =50.0Hz,C-8′),22.7(C-11),22.7(d,J C,P =4.7Hz, C-7′), 21.6(Ac-CH3), 17.4(C-27), 17.3(C-18), 13.4(C-21), 12.0(C-19).

[0118] ESI-HRMS: m / z 874.5177 [M-Br] + (Calculated value: C) 55 H 73 N2O6P, 874.5170).

[0119] Example 15:

[0120]

[0121] 6 g of the target compound was prepared according to the synthesis method of Example 1. It was a yellow powder, with a yield of 69.6%.

[0122] 1HNMR(600MHz,CDCl3,δ):7.78-7.75(m,9H,H-2″,H-4″,H-6″),7.73-7.69(m,6H,H-3″,H-5″),4.67(m,1H,H-3),4.38(m,1H,H-16),3.55(m,2H,H-9′),3.46(m,1H,H-26),3.34(t,J=11.0Hz,1H,H-26),3.13(dd,J=14.9,4.6Hz,1H,H-11),2.58(dd,J=8.3,6.0Hz,1H,H-14),2.37(t,J=7.4Hz,2H,H-2′),2.07(m,1H,H-15),2.02(s,3H,Ac-CH3),1.86(t,J=6.4Hz,2H,H-3′),1.12(d,J=7.0Hz,3H,21-CH3),1.01(s,3H,18-CH3),0.88(s,3H,19-CH3),0.77(d,J=6.4Hz,3H,27-CH3)。

[0123] 13 C NMR(150MHz,CDCl3,δ):172.5(C-12),171.7(C-1′),171.2(Ac-CH3),135.0(d,J C,P =2.5Hz,C-4″),133.8(d,J C,P =9.8Hz,C-2″,C-6″),130.7(d,J C,P =12.7Hz,C-3″,C-5″),118.5(d,J C,P =86.0Hz,C-1″),109.3(C-22),79.8(C-16),73.6(C-3),67.0(C-26),56.3(C-9),55.9(C-14),53.6(C-17),48.0(C-13),44.5(C-5),42.4(C-20),36.5(C-1),36.1(C-10),34.8(C-8),33.9(C-4),33.1(C-2′),31.7(C-23),31.5(C-7),30.9(C-15),30.3(C-25),30.3(d,J C,P =15.5Hz,C-7′),29.0(C-24),29.0(C-6′),28.9(C-5′),28.9(C-4′),28.2(C-6),27.3(C-2),24.7(C-3′),22.6(d,J C,P=4.9Hz,C-8′),22.6(C-11),22.4(d,J C,P =51.9Hz, C-9′), 21.6(Ac-CH3), 17.4(C-27), 17.3(C-18), 13.4(C-21), 12.0(C-19).

[0124] ESI-HRMS: m / z 888.5321 [M-Br] + (Calculated value: C) 56 H 75 N2O6P, 888.5327).

[0125] Pharmacodynamic Examples:

[0126] 1. Main experimental equipment, reagents, and materials

[0127] Clean bench (1300 series A2, Thermo, Waltham, MA, USA)

[0128] Constant temperature CO2 incubator (Heracell 150i, Thermo, Waltham, MA, USA)

[0129] Microplate reader (Synergy H1, Biotek, Winooski, VT, USA)

[0130] Inverted phase contrast microscope (Primovert, Zeiss, Beijing, China)

[0131] Cell culture medium RPMI-1640, DMEM (high glucose) (HyClone, Logan, UT, USA)

[0132] Fetal bovine serum (HyClone, Logan, UT, USA)

[0133] PBS (Solarbio, Beijing, China)

[0134] CellTiter-Glo reagents (Promega, Madison, Wisconsin, USA)

[0135] DMSO (Sigma-Aldrich, Beijing, China)

[0136] Cell lines: human breast cancer cells MCF-7 (ATCC, Beijing, China), human lung cancer cells A549 (ATCC, Beijing, China), human colon cancer cells HCT116 (ATCC, Beijing, China), human gastric cancer cells MKN-45 (BNCC, Xinyang, China), and human liver cancer cells HepG2 (ATCC, Beijing, China).

[0137] 2. Experimental Methods: Cell Inhibitory Activity Assay

[0138] The above five cell types were used at 2×10 4 Cells / well were seeded into 96-well plates and allowed to adhere overnight. After adhesion, the culture medium was removed, and a series of test compounds (0.5-100 μM) prepared in DMSO were added (maintaining a final DMSO concentration of 0.1%, i.e., adding the corresponding stock solution to the cell culture medium according to the stock solution concentration, e.g., 1 μL added to 999 μL of cell culture medium). Cisplatin (0.5-100 μM) served as a positive control (dissolved in 1‰ DMSO medium served as a control). After 48 hours of treatment, the corresponding volume of CellTiter-Glo reagent solution was added to each 96-well plate and lysed at room temperature for 2 minutes, followed by incubation at room temperature for 10 minutes. The plates were then transferred to new white, non-transparent 96-well plates, and luminescence was measured using a microplate reader in cold light mode. The relative inhibition rate of each well was calculated based on the readings, and the IC50 was calculated based on the inhibition rate. 50 Value. Inhibition rate calculation method:

[0139]

[0140] Relative OD value of drug sensitivity wells = Absolute OD value of drug sensitivity wells - Absolute OD value of blank control wells 3. Experimental Results

[0141] As shown in Table 1, the pharmacological experimental results show that the triphenylphosphine derivative of the hecosaponin of the present invention has cytotoxic activity and tumor cell specificity against a variety of tumor cell lines, and can be further used to prepare anti-tumor drugs.

[0142] Table 1: IC50 values ​​of cytotoxic activity against five human cancer cell lines in Examples 1-15 50 Value (μM)

[0143]

[0144]

[0145] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A mitochondrial-targeted hecoside triphenylphosphine derivative, characterized in that, Its structural formula is shown in General Formula II: Where n is an integer selected from 5, 6, 7, and 8.

2. The hecosaponin triphenylphosphine derivative according to claim 1, characterized in that, The hecosaponin triphenylphosphine derivative is selected from the following structures: 。 3. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the derivative of claim 1 or claim 2, and a pharmaceutically acceptable carrier.

4. The pharmaceutical composition according to claim 3, characterized in that: The dosage form of the pharmaceutical composition is an oral dosage form or an injectable dosage form.

5. The pharmaceutical composition according to claim 4, characterized in that: The oral dosage form is selected from capsules, tablets, granules or oral liquids.

6. The method for preparing the hecosaponin triphenylphosphine derivative according to claim 1 or claim 2, characterized in that: The preparation method includes the following steps: Hecosaponin (HG)1 was reacted with acetic anhydride and hydroxylamine hydrochloride to generate intermediate 4. Intermediate 4 was reacted with bromocarboxylic acid at room temperature under DCC and DMAP conditions to generate intermediate 5d-5g. Intermediate 5d-5g was reacted with triphenylphosphine under reflux to generate the target compound 6d-6g. The reaction route is shown below: 。 7. The use of the hecosaponin triphenylphosphine derivative of claim 1 or claim 2, or the pharmaceutical composition of any one of claims 3 to 5, in the preparation of an antitumor drug, characterized in that: The tumor is selected from stomach cancer, breast cancer, lung cancer, colon cancer, or liver cancer.

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