CN112603989A

CN112603989A - Application of vinblastine derivative in preparation of drugs for inhibiting tumor metastasis

Info

Publication number: CN112603989A
Application number: CN202110051439.7A
Authority: CN
Inventors: 张冬梅; 陈河如; 叶文才
Original assignee: Jinan University
Current assignee: Jinan University
Priority date: 2017-07-18
Filing date: 2017-07-18
Publication date: 2021-04-06
Also published as: CN107349415A

Abstract

The invention discloses an application of vinblastine derivatives in preparation of drugs for inhibiting tumor metastasis. The vinblastine derivative comprises vinblastine dipeptide and physiologically acceptable salt thereof; the vinblastine dipeptide is a compound obtained by condensing a vinblastine hydrazinolysis compound and N-benzyloxycarbonyl dipeptide; the catharanthine hydrazinolysis compound is a compound obtained by reacting a catharanthine compound with hydrazine hydrate; the vinblastine compound is vinblastine, vinorelbine, vinflunine or vincristine. The vinblastine dipeptide and physiologically acceptable salts thereof can remarkably inhibit invasion, migration and horizontal movement capability of tumor cells in vitro, remarkably inhibit far-end metastasis of tumors in vivo, and can be applied to treatment of patients with middle and late malignant tumors accompanied with far-end metastasis.

Description

Application of vinblastine derivative in preparation of drugs for inhibiting tumor metastasis

The application is a divisional application with application date of 2017, month 07 and 18, application number of 201710587131.8, and invention name of application of vinblastine derivatives in preparation of drugs for inhibiting tumor metastasis.

Technical Field

The invention belongs to the field of medicines, and particularly relates to an application of vinblastine derivatives in preparation of a medicine for inhibiting tumor metastasis.

Background

Tumor metastasis is an important feature of malignancy and is also the leading cause of death in most malignant patients. Therefore, inhibiting tumor metastasis has a crucial clinical significance for saving the life of malignant tumor patients, and has become one of the strategies for developing antitumor drugs.

The metastasis of tumor cells from the primary lesion to the distant tissue involves several ways: (1) directly spread to adjacent parts to erode primary organs; (2) cancer cells are transferred to all levels of lymph nodes from near to far along with lymph node drainage, and distant metastasis occurs; (3) cancer cells directly enter blood vessels and are transferred to remote tissues such as lung, bone, brain and the like along with blood flow to form secondary tumor lesions. On the one hand, epithelial-mesenchymal transition (EMT) of tumor cells plays an important role in tumor metastasis, enhancing the motility, migration and invasion ability of tumor cells, thereby promoting tumor metastasis (Prieto-garcia. e, et al. med oncol.2017,34(7): 122.). On the other hand, tumor blood vessels also provide the necessary transport channels for the metastasis of tumor cells. Therefore, targeting tumor cells to inhibit cancer cell EMT and targeting tumor vessels are two important therapeutic strategies for inhibiting tumor metastasis.

Currently, inhibitors targeting tumor cell EMT are primarily focused on the TGF- β pathway, such as inhibitors SD-093 and LY-580276 (subaramanian.g., et al. cancer res.2004,64(15):5200-11.) that competitively bind to the ATP site of TGF- β RI kinase; TGF-. beta./ALK 5 specific inhibitors EW-7203(Park CY, et al. cancer Sci.2011,102(10):1889-96.), EW-719(Park CY, et al. Eur J cancer.2011,47(17):2642-53.), and EW-7197(Jin CH, et al. J Med Chem.2014,57(10):4213-38.), etc. However, these inhibitors have been only in clinical trials, and there is no clinically effective EMT inhibitor drug on the market.

Therapeutic agents that target tumor vessels mainly include two major classes (McKeage MJ, et al. cancer 2010,116(8): 1859-71.). One class is Angiogenesis Inhibitors (AIs), mainly the VEGF monoclonal antibodies bevacizumab (bevacizumab) and VEGFR2 tyrosine kinase inhibitors Apatinib, Vandetanib, etc. AIs mainly inhibit the neogenesis of tumor blood vessels, and although they can promote the normalization of tumor blood vessels and reduce tumor metastasis, they have no significant effect on blood vessels formed by tumors, so they can only slow down the progression of tumors, cannot effectively kill formed tumors, and have poor curative effect on middle and late stage tumors. Therefore, the clinical application of AIs is significantly limited and the patient benefits to a lesser extent. The other is tumor Vascular Disruptors (VDAs), which can selectively damage the formed blood vessels in the central region of the tumor and induce large-area necrosis of the tumor center, thereby exerting good anti-tumor effect. However, the degree of vascular maturation in the tumor margin region is high (relative high pericyte coverage) and this part of the vessels is insensitive to VDAs, and therefore, significant survival circles remain in the tumor margin region after VDAs treatment (Tozer GM, et al nat Rev cancer.2005,5(6): 423-35.; Tozer GM, et al cancer res.2008,68(7): 2301-11.; Nguyen L, et al bmc cancer 2012,12: 522.). On one hand, the residual tumor survival circle leads to the enhancement of the EMT of tumor cells due to oxygen deficiency factors (Fifis T, et al. cancer Med.2013,2(5): 595-; on the other hand, the survival circle is also a major cause of rapid relapse after discontinuation of VDAs therapy. In conclusion, the search for the medicine which can inhibit the tumor cell EMT, destroy the tumor blood vessel and overcome the tumor survival circle has very important therapeutic significance.

Vinblastine compounds such as vinblastine, vincristine and its derivatives vinorelbine and vinflunine, which are both bisindole alkaloids, are separated from Catharanthus roseus of Apocynaceae. Pharmacological action research shows that the vinblastine and the derivatives thereof belong to cytotoxic drugs, mainly inhibit polymerization of tubulin, hinder formation of spindle microtubules and arrest cell nucleus division in the metaphase. Vinblastine and its derivatives have broad-spectrum antitumor activity, and can be used for treating Hodgkin's disease, chorioepithelioma, acute leukemia, breast cancer, ovarian cancer, testis cancer, head and neck cancer, oropharyngeal cancer, and monocytic leukemia (Wilson L, et al, Ann N Y Acad Sci.1975,253: 213-31). Recently, Angelo Vacca et al have discovered that vinblastine significantly inhibits angiogenesis at the cellular level in non-toxic amounts (Vacca A, et al, blood 1999,94(12): 4143-55.); giannoula Klement et al showed that the continuous administration of low doses of vinblastine inhibited the neogenesis of tumor vessels (Klement G, et al. J Clin invest.2000,105(8): R15-24.); james Moore et al demonstrated that vincristine was able to inhibit the growth of tumor neovasculature (Moore J, et al. J Peditar Surg.2001,36(8): 1273-6.); on the other hand, Anna Kruczynskia et al found that vinflunine could inhibit tumor angiogenesis, destroy the generated tumor vessels, and significantly inhibit the metastasis of experimental malignant tumors (Kruczynski A, et al Eur J cancer.2006,42(16): 2821-32.). Like many chemotherapy drugs used clinically, vinblastine drugs also have many serious side effects in the course of disease treatment, such as myelosuppression, myalgia, and adverse reactions such as malignant vomiting (Bonnetre J, et al. Ann Oncol.2001,12(12): 1683-91.18.; Spiller M, et al. Pediatr Blood cancer.2005,45(3):344-6.), which greatly limits their clinical application. One of the effective ways to reduce the toxic and side effects of the drugs is to modify the structure of the drugs to make the drugs become prodrugs, and selectively act on genes, enzymes, signal transduction factors and the like of focus target cells by utilizing the difference in molecular biology between focus tissues and normal tissues, thereby achieving the purpose of targeted therapy. In recent years, a great deal of research shows that Fibroblast activation protease (FAP alpha) is specifically expressed on the surfaces of injured tissues and more than 90% of tumor tissues activated fibroblasts and pericytes (Ramirez-Montagut T, et al. oncogene 2004,23(32): 5435-46.).

Research shows that after the vinblastine compound is subjected to hydrazinolysis and dipeptide derivatization to form an FAP alpha enzyme activated prodrug, the vinblastine compound has good antitumor, angiogenesis inhibiting and vascular damaging activities in vivo and in vitro, such as the application of Chinese invention patents (CN 201310138241.8; CN201310738860.0) and PCT patents (PCT/CN 2014/000192). However, the application and research of the vinblastine derivative in the aspect of inhibiting tumor metastasis are not reported yet.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provides the application of the vinblastine derivative in preparing the drugs for inhibiting tumor metastasis.

The purpose of the invention is realized by the following technical scheme: an application of vinblastine derivatives in preparing medicine for inhibiting tumor metastasis is disclosed, wherein the vinblastine derivatives comprise vinblastine dipeptide and physiologically acceptable salt thereof.

The vinblastine dipeptide is a compound obtained by condensing a vinblastine hydrazinolysis compound and N-benzyloxycarbonyl dipeptide (namely, the vinblastine dipeptide is a compound obtained by condensing the vinblastine hydrazinolysis compound formed by the reaction of the vinblastine compound and hydrazine hydrate and then condensing the vinblastine hydrazinolysis compound and the N-benzyloxycarbonyl dipeptide); preferably one or more of vinblastine plus dipeptide (BX-CCJ), vinorelbine plus dipeptide (BX-CCRB), vinflunine plus dipeptide (BX-CCFN) and vincristine plus dipeptide (BX-CCXJ), and the structural formula is shown in formulas II-V:

wherein Z-GP represents benzyloxycarbonyl glycyl prolyl.

The synthetic route of the vinblastine derivative is as follows:

the catharanthine hydrazinolysis compound is a compound obtained by reacting a catharanthine compound with hydrazine hydrate; preferably catharanthine hydrazinolysis (JJ-CCJ), vinorelbine hydrazinolysis (JJ-CCRB), vinflunine hydrazinolysis (JJ-CCFN) or vincristine hydrazinolysis (JJ-CCXJ).

The vinblastine compound is preferably vinblastine (CCJ), vinorelbine (CCRB), vinflunine (CCFN) or vincristine (CCXJ).

The N-benzyloxycarbonyl dipeptide is preferably N-benzyloxycarbonyl glycyl proline (Z-GP-OH); the structural formula of the N-benzyloxycarbonyl glycyl proline is shown as a formula I:

the tumors include but are not limited to malignant tumors such as breast cancer, lung cancer, liver cancer, stomach cancer, cervical cancer, ovarian cancer, esophageal cancer, colorectal cancer, liver cancer, nasopharyngeal cancer, brain cancer, bone cancer and the like; preferably breast cancer.

The tumor metastasis includes all stages of tumor metastasis, including early stage, middle stage and late stage of tumor metastasis.

Compared with the prior art, the invention has the following advantages and effects: the catharanthine compound, the catharanthine dipeptide and the physiologically acceptable salt thereof can obviously inhibit the invasion, migration and horizontal movement capability of tumor cells in vitro and the distal metastasis of tumors in vivo, have better drug effect than the catharanthine compound, and can be applied to the treatment of patients with middle and late malignant tumors accompanied with the distal metastasis.

Drawings

FIG. 1 is a graph comparing the inhibitory effect of vinblastine derivatives on the invasion capacity of TGF-beta 1-induced human breast cancer cells MDA-MB-231: (^***P<A 0.001vs blank control group,^###P<0.001vs TGF-. beta.1 group,^#P<0.01vs TGF-. beta.1 group).

FIG. 2 is a graph comparing the inhibitory effect of vinblastine derivatives on TGF- β 1-induced migration ability of human breast cancer cells MDA-MB-231: (^***P<A 0.001vs blank control group,^###P<0.001vs TGF-. beta.1 group,^#P<0.01vs TGF-. beta.1 group).

FIG. 3 is a graph comparing the inhibitory effect of vinblastine derivatives on TGF- β 1-induced horizontal motility of human breast cancer cells MDA-MB-231: (^***P<A 0.001vs blank control group,^###P<0.001vs TGF-. beta.1 group,^#P<0.01vs TGF-. beta.1 group).

FIG. 4 is a graph showing the effect of vinblastine derivatives on TGF- β 1-induced expression of human breast cancer cell MDA-MB-231 epithelial mesenchymal transition-related proteins (II)^***P<A 0.001vs blank control group,^###P<0.001vs TGF-. beta.1 group,^#P<0.01vs TGF-. beta.1 group); wherein, the graph A is epithelial cell cadherin, the graph B is N-cadherin, and the graph C is vimentin.

Fig. 5 is a comparison graph of the inhibitory effect of vinblastine derivatives on MDA-MB-231 breast cancer nude mouse orthotopic transplantation tumor lung metastasis (n ═ 8, mean ± SEM,^###P<0.001vs blank;^#P<0.01vs blank); wherein, the diagram A is H&E staining to detect the number of lung tissue metastases of nude mice, and B is H&E, staining to detect the area of a lung tissue metastasis of a nude mouse, a graph C is a detection result of epithelial markers (epithelial cell cadherins) and interstitial markers (N-cadherins and fibronectin) of a lung tissue of the mouse before and after administration by using an immunohistochemical method, and a graph D is a graph D for detecting changes of the epithelial markers (epithelial cell cadherins) and the interstitial markers (N-cadherins and vimentins) in a tumor tissue of the mouse before and after the action of the vinblastine derivative by using an immunoblotting method.

Fig. 6 is a graph comparing the inhibitory effect of vinblastine derivatives on MDA-MB-231 lung metastasis of breast cancer in nude mice (n-8, mean ± SEM,^###P<0.001vs blank;^#P<0.01vs blank); wherein, the diagram A is H&E, staining and detecting the number of lung tissue metastasis foci of the nude mice; FIG. B is H&E, staining and detecting the area of the lung tissue metastasis of the nude mice; and the graph C is the detection result of epithelial markers (epithelial cell cadherins) and mesenchymal markers (N-cadherins and fibronectin) of the lung tissues of the mice before and after administration by using an immunohistochemical method, and the graph D is the effect of the vinblastine derivative on the epithelial markers (epithelial cell cadherins) and the mesenchymal markers (N-cadherins and vimentin) in the lung tissues of the mice detected by using an immunoblotting method.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.

The vinblastine compounds related in the embodiment of the invention are vinblastine (CCJ), vinorelbine (CCRB), vinflunine (CCFN) and vincristine (CCXJ) in turn, and the vinblastine compounds are hydrazinolyzed to obtain the vinblastine compounds; the catharanthine hydrazinolysis compound is as follows: catharanthine hydrazinolysis (JJ-CCJ), vinorelbine hydrazinolysis (JJ-CCRB), vinflunine hydrazinolysis (JJ-CCFN) and vincristine hydrazinolysis (JJ-CCXJ);

the vinblastine dipeptide related in the embodiment of the invention is a compound which is obtained by reacting a vinblastine compound with hydrazine hydrate to form a hydrazinolysis vinblastine compound and then condensing the hydrazinolysis vinblastine compound with N-benzyloxycarbonyl dipeptide (N-benzyloxycarbonyl glycyl proline (Z-GP-OH)); the vinblastine dipeptide is vinblastine dipeptide (BX-CCJ), vinorelbine dipeptide (BX-CCRB), vinflunine dipeptide (BX-CCFN) and vincristine dipeptide (BX-CCXJ);

the structure and nomenclature of the compounds are as follows:

example 1 inhibitory Effect of vinblastine derivatives on TGF-. beta.1-induced invasive Capacity of human Breast cancer cells MDA-MB-231

The experimental method comprises the following steps: human breast cancer cells MDA-MB-231 cells (purchased from American type culture Collection, ATCC) in logarithmic growth phase were digested, centrifuged, counted, and counted at 2X 10⁴Each of the cells was inoculated in a Transwell chamber plated with Matrigel matrix gel (BD Co.), and the cells were resuspended in a serum-free medium. Each well is 100 μ L, a blank control group, a TGF-beta 1 group (TGF-beta 12 ng/mL) and a drug adding group are arranged, the drug adding groups are BX-CCJ, BX-CCRB, BX-CCFN, BX-CCXJ, CCJ, CCRB, CCFN and CCXJ (6.25nM) and TGF-beta 1(2ng/mL) are treated together, and 700 μ L of 1640 culture solution containing 10% newborn bovine serum is added into the lower chamber. After 48 hours of incubation, the culture was aspirated, washed once with PBS buffer, fixed with 4% paraformaldehyde at room temperature for 30 minutes, stained with Giemsa dye for 30 minutes, wiped off the cells on the upper membrane of the chamber with a cotton swab, washed once with PBS, observed microscopically and counted for the number of cells.

The experimental results are as follows: compared with a blank control group, the TGF-beta 1 group obviously increases the invasion effect of human breast cancer cells MDA-MB-231; compared with the factor group, the vinblastine derivative obviously inhibits the invasion of human breast cancer cells MDA-MB-231, and the drug effect of the vinblastine derivative is superior to that of the vinblastine compound (shown in figure 1).

Example 2 inhibitory Effect of vinblastine derivatives on TGF-. beta.1-induced migration Capacity of human Breast cancer cells MDA-MB-231

The experimental method comprises the following steps: MDA-MB-231 cells in logarithmic growth phase are taken, washed and centrifuged by PBS, and the cells are resuspended in serum-free medium. 2 x 10 to⁴Each/mL cell was inoculated into a Transwell chamber at 100. mu.L per well, and a blank control group, a TGF-beta 1 (TGF-. beta.12 ng/mL) group, and an addition group were set, wherein the addition group was BX-CCJ, BX-CCRB, BX-CCFN, BX-CCXJ, CCJ, CCRB, CCFN, CCXJ (6.25nM) and TGF-. beta.1 (2ng/mL) were treated together, and 700. mu.L of 1640 culture medium containing 10% newborn bovine serum was added to the lower chamber. Standing at 37 deg.C and containing 5% CO₂After culturing for 48h, the culture solution was discarded, washed once with PBS, fixed with 4% paraformaldehyde at room temperature for 30 minutes, stained with Giemsa dye for 30 minutes, wiped off the cells on the upper membrane of the chamber with a cotton swab, washed once with PBS, observed under a microscope and counted the number of cells.

The experimental results are as follows: compared with a blank control group, the TGF-beta 1 group obviously increases the migration effect of human breast cancer cells MDA-MB-231; compared with the factor group, the vinblastine derivative obviously inhibits the migration of human breast cancer cell MDA-MB-231, and the drug effect is superior to that of the vinblastine compound (shown in figure 2).

Example 3 inhibitory Effect of vinblastine derivatives on TGF-. beta.1-induced horizontal motility of human Breast cancer cells MDA-MB-231

The experimental method comprises the following steps: MDA-MB-231 cells in logarithmic growth phase were taken, washed with PBS and digested. Will be 5X 10⁵And (3) inoculating each/mL of cells into a six-well plate, starving for 8h by using a serum-free culture medium when the cell density reaches 90%, drawing a cross trace by using a 10-microliter pipettor, washing by using PBS, and taking pictures, wherein a blank control group, a TGF-beta 1 (TGF-beta 12 ng/mL) group and a medicine adding group are arranged, the medicine adding group is BX-CCJ, BX-CCRB, BX-CCFN, BX-CCXJ, CCJ, CCRB, CCFN, CCXJ (6.25nM) and TGF-beta 1(2ng/mL) are jointly treated, and the pictures are taken in the same visual field after 8 h. Results were counted using Image Plus 5 software, averaged, and the number of migrated cells was counted.

The experimental results are as follows: compared with a blank control group, the TGF-beta 1 group remarkably increases the horizontal movement capacity of human breast cancer cells MDA-MB-231; compared with the TGF-beta 1 group, the vinblastine derivative obviously inhibits the horizontal movement capability of human breast cancer cell MDA-MB-231, and the drug effect is superior to that of the vinblastine compound (see figure 3).

Example 4 Effect of vinblastine derivatives on TGF- β 1-induced expression of proteins associated with epithelial-mesenchymal transition in human Breast cancer cells MDA-MB-231

The experimental method comprises the following steps: MDA-MB-231 cells in logarithmic growth phase are taken, a blank control group, a TGF-beta 1 (TGF-beta 12 ng/mL) group and a dosing group are arranged, after the dosing group is treated for 48 hours by BX-CCJ, BX-CCRB, BX-CCFN, BX-CCXJ, CCJ, CCRB, CCFN and CCXJ (6.25nM), TGF-beta 1(2ng/mL) is added into the TGF-beta 1 group and the dosing group at the same time, and the effect is carried out for 1 hour. Lysing cellular proteins with RIPA lysate; carrying out BCA protein quantification; taking 40-50 mu g of protein samples to carry out SDS-PAGE gel electrophoresis, and separating the proteins according to the difference of molecular weight; transferring the protein band on the gel to a PVDF membrane; blocking the protein band in 5% Bovine Serum Albumin (BSA); primary antibody (purchased from Cell Signaling Technology, CST): diluting epithelial cell cadherin, vimentin and N-cadherin according to the volume ratio of 1:1000, and incubating overnight at 4 ℃; incubation secondary antibody (purchased from Cell Signaling Technology, CST); protein bands are shown by ECL color developing liquids A and B, and gray scale analysis is carried out by using Quantity One software.

The experimental results are as follows: the TGF-beta 1 group has decreased epithelial cell cadherin expression compared to a blank control group; expression of N-cadherin and vimentin is increased. The expression of cadherin of epithelial cells is increased in the administration group compared with the TGF-beta 1 group; decreased expression of N-cadherin and vimentin; the vinca alkaloid derivative can obviously inhibit the reduction of the expression of the epithelial marker epithelial cell cadherin induced by TGF-beta 1 and inhibit the increase of the expression of the mesenchymal marker N-cadherin and vimentin after the vinca alkaloid derivative acts, and the function of the vinca alkaloid derivative in inhibiting the epithelial mesenchymal transition related protein is superior to that of the vinca alkaloid compound (see figure 4).

Example 5 Effect of vinblastine derivatives on MDA-MB-231 Breast cancer nude mouse orthotopic transplantation tumor Lung metastasis

The experimental method comprises the following steps: MDA-MB-231 cells in logarithmic growth phase were digested, resuspended in Matrigel (diluted 1:1 by volume) diluted in pre-cooled PBS, and cell density was adjusted to 1X 10⁷After one/mL, 0.15 mL/female BALB/C nu/nu nude mice (purchased from Guangdong provincial medical laboratory animal center) at the age of 6 weeks were inoculated into the left mammary fat pad. The tumor mass is 100mm long³～200mm³Size, nude mice were randomly divided into 9 groups of 10 mice each, which were separately administered by tail vein injection with 2. mu. mol/kg of physiological saline, BX-CCJ, BX-CCRB, BX-CCFN, BX-CCXJ, CCJ, CCRB, CCFN, CCXJ groups. The administration is carried out 1 time every other day, the experiment is finished after 10 times of continuous administration, and tumor tissues and lung tissues are stripped after blood sampling is carried out on nude mice by heart. The lung tissue of the nude mice was stained with hematoxylin-eosin (Hay) and analyzed for the size and number of metastases. Lung tissue was immunohistochemically stained by ABC method. The expression of the epithelial cadherins as epithelial cell marker proteins and the N-cadherins and fibronectin as mesenchymal cell markers in the lung tissues of mice in a blank control group and a drug administration group was analyzed. Taking a half of tumor tissues of a nude mouse, homogenizing, cracking tissue proteins, and respectively detecting the expression conditions of epithelial cell cadherin, N-cadherin and vimentin in the tumor tissues of the nude mouse by using a western blotting method.

The experimental results are as follows: the vinblastine derivative remarkably inhibits epithelial-mesenchymal transition of the breast cancer of a nude mouse, and further inhibits lung metastasis of the nude mouse in-situ transplantation tumor of the MDA-MB-231 breast cancer of the nude mouse; the drug effect is better than that of the vinblastine compound (see figure 5).

Example 6 inhibitory Effect of vinblastine derivatives on nude mouse MDA-MB-231 Breast cancer metastasis

The experimental method comprises the following steps: MDA-MB-231 cells in logarithmic growth phase are digested and resuspended in serum-free DMEM blank medium, and cell density is adjusted to 1 × 10⁷After one/mL, 0.1 mL/tail vein was injected into 6-week-old female BALB/C nu/nu nude mice. Two weeks later, the nude mice were randomly divided into 9 groups of 10 mice each, physiological saline, BX-CCJ, BX-CCRB, BX-CCFN, BX-CCXJ, CCJ, CCRB, CCFN, CCXJ groups, respectively, and administered at a dose of 2. mu. mol/kg. Septum caudal veinThe administration is carried out for 1 time, the experiment is finished after 10 times of continuous administration, the lung tissue is stripped after blood sampling is carried out on a nude mouse through the heart, the lung tissue of the nude mouse is stained by hematoxylin-eosin, and the size and the number of the metastasis foci are analyzed. Immunohistochemical staining was performed by the ABC method. The expression of the epithelial cadherins as epithelial cell marker proteins and the N-cadherins and fibronectin as mesenchymal cell markers in the lung tissues of mice in a blank control group and a drug administration group was analyzed. Half of the lung tissues of the nude mice are taken to be homogenized and the tissue protein is cracked, and the expression conditions of epithelial cell cadherin, N-cadherin and vimentin in the lung tissues of the nude mice are detected by a western blotting method (see figure 6).

The experimental results are as follows: the vinblastine derivative obviously inhibits epithelial-mesenchymal transition of the breast cancer of a nude mouse, further inhibits lung metastasis of the breast cancer MDA-MB-231 of the nude mouse, and has better drug effect than the vinblastine compound.

The experimental results show that the vinblastine derivative can obviously inhibit the breast cancer metastasis in vitro and in vivo, and can be applied to the treatment of early and late breast cancer metastasis.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. An application of vinblastine derivatives in preparing drugs for inhibiting tumor metastasis is characterized in that: the vinblastine derivative is vinblastine dipeptide and physiologically acceptable salt thereof;

the vinblastine dipeptide is one or more of vinblastine plus dipeptide, vinorelbine plus dipeptide, vinflunine plus dipeptide and vincristine plus dipeptide, and the structural formula is shown as formulas II-V:

wherein Z-GP represents benzyloxycarbonyl glycyl prolyl;

the vinblastine dipeptide is a compound shown in formula I obtained by condensing a vinblastine hydrazinolysis compound and N-benzyloxycarbonyl dipeptide:

2. the use of the vinca alkaloid derivative according to claim 1 in the preparation of a medicament for inhibiting tumor metastasis, wherein: the tumor is breast cancer.