Coordination compounds, compositions and their use for treatment of cancer

Abstract

The invention provides a coordination compound as shown in a formula (I). The present invention also provides a pharmaceutical composition useful in the treatment of cancer, comprising the coordination compound of formula (I) and a pharmaceutically acceptable carrier. The invention also provides uses of the coordination compounds in inhibiting cancer cell survival or proliferation, inhibiting cancer cell migration or invasion, and treating cancerous tumors in an individual in need thereof.

Description

Technical Field

[0001] This application claims priority to U.S. Provisional Application No. 63 / 431,320, filed on December 9, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] This application contains a sequence listing, which is filed electronically in the WIPOST.26 standard format, the entire contents of which are incorporated herein by reference. The XML file was created on November 21, 2023, is named "PIEN-2PCT.xml" and is 10,000 bytes in size.

[0003] The present invention relates to a coordination compound and its use in treating proliferative diseases. Specifically, the present invention relates to a coordination compound, a pharmaceutical composition comprising the coordination compound, and a method of treating cancer using the coordination compound.

[0004] Prior Art

[0005] Cancer is a widespread and devastating disease that continues to pose a huge challenge to medicine and healthcare systems worldwide. Although there have been significant advances in our understanding of the molecular basis of cancer, there is still a need to explore more effective cancer treatments. Current cancer treatment strategies mainly include surgery, radiotherapy and chemotherapy. Surgery and radiotherapy are usually only effective for localized early-stage cancers and have limited applicability for more advanced or metastatic cancers. Conventional chemotherapy, which administers cytotoxic drugs to tumors, has long been the cornerstone of cancer treatment. Although chemotherapy has shown some success in reducing tumor size and prolonging patient survival, its effect has significant limitations. A major limitation is that cancer cells can develop resistance to chemotherapy drugs, rendering them ineffective over time.

[0006] The development of drug resistance is a common problem in cancer treatment. Cancer cells evolve mechanisms to resist the effects of chemotherapy, rendering once-effective treatments ineffective and requiring patients to undergo multiple rounds of chemotherapy with different drugs. An example is colorectal cancer (CRC), a malignancy that causes a large number of deaths worldwide and is the third leading cause of cancer-related deaths worldwide. The main treatment for advanced colorectal cancer is oxaliplatin (OXA) chemotherapy. Although numerous studies have demonstrated its potential for efficacy, drug resistance remains a major challenge, making colorectal cancer difficult to treat effectively and leading to recurrent CRC.

[0007] In addition to resistance to chemotherapy, the presence of metastatic cancer cells can also contribute to cancer recurrence, causing patients to experience tumor growth again, often in a more aggressive and drug-resistant form. Metastasis, the spread of cancer cells from the primary tumor to distant sites in the body, is a major factor in cancer-related mortality. This complex process involves cancer cells invading surrounding tissues, infiltrating the blood or lymphatic vessels, circulating through the bloodstream, extravasating at secondary sites, and subsequently growing into secondary tumors. Metastatic cancers are often highly invasive and treatment-resistant, presenting significant challenges to clinicians.

[0008] Given the above challenges, there is still an urgent unmet need for innovative solutions in cancer treatment, especially in preventing cancer progression. Summary of the Invention

[0009] The present invention relates to a coordination compound useful as an anticancer agent for inhibiting the survival, proliferation, migration, or invasion of cancer cells. The coordination compound is represented by formula (I): [XZ2(CH3CO2)6(H2O)4(OH)2]NO3. In formula (I), X and Z represent different metal ions, wherein X is a trivalent ion of chromium (Cr) or molybdenum (Mo), and Z is a trivalent ion of iron (Fe), ruthenium (Ru), or osmium (Os).

[0010] One object of the present invention is to provide a method for inhibiting the survival or proliferation of cancer cells, comprising contacting the cancer cells with an effective amount of a coordination compound represented by formula (I), wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. The coordination compound preferably comprises a trivalent chromium ion and a trivalent iron ion and is represented by formula (II).

[0011] The cytotoxicity of the coordination compound is applicable to a wide range of cancer cells. In some embodiments, the cancer is selected from colorectal cancer, lung cancer, liver cancer, pancreatic cancer, bone cancer (including osteosarcoma), brain cancer (including glioblastoma), breast cancer, ovarian cancer, cervical cancer, prostate cancer, bladder cancer (including transitional cell carcinoma), blood cancer (including leukemia and lymphoma), gastric cancer, skin cancer (including melanoma), head and neck cancer (including oral squamous cell carcinoma), or thyroid cancer. In some embodiments, the cancer cell is an invasive cancer cell. In some embodiments, the cancer cell is resistant to a chemotherapeutic drug (e.g., oxaliplatin).

[0012] In some embodiments, the coordination compound induces ER stress-mediated apoptosis in the cancer cell, thereby inhibiting the survival or proliferation of the cancer cell.

[0013] In some embodiments, the aforementioned method of inhibiting cancer cell survival or proliferation further comprises the step of contacting the cancer cell with a chemotherapeutic agent selected from oxaliplatin, irinotecan, 5-fluorouracil, gemcitabine, doxorubicin, or any combination thereof.

[0014] Another object of the present invention is to provide a method for inhibiting cancer cell migration or invasion, comprising contacting the cancer cells with an effective amount of a coordination compound of formula (I), wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. The coordination compound preferably comprises a trivalent chromium ion and a trivalent iron ion and is represented by formula (II).

[0015] The inhibitory effect of the coordination compound on cell migration or invasion is applicable to various types of cancer cells. In some embodiments, the cancer is colorectal cancer or bone cancer. In some embodiments, the cancer cell is an invasive cancer cell. In some embodiments, the cancer cell is resistant to a chemotherapy drug (e.g., oxaliplatin).

[0016] In some embodiments, the ligand compound inhibits epithelial-mesenchymal transition (EMT) of the cancer cell, thereby inhibiting the migration or invasion of the cancer cell.

[0017] Another object of the present invention is to provide a method for treating or treating cancerous tumors, comprising administering to a subject in need thereof an effective amount of a coordination compound represented by formula (I), wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. The coordination compound preferably comprises a trivalent chromium ion and a trivalent iron ion and is represented by formula (II).

[0018] In some embodiments, the cancerous tumor is selected from colorectal cancer, lung cancer, liver cancer, pancreatic cancer, bone cancer, brain cancer, breast cancer, ovarian cancer, cervical cancer, prostate cancer, bladder cancer, blood cancer, stomach cancer, skin cancer, head and neck cancer, or thyroid cancer. In some embodiments, the cancerous tumor comprises an invasive cancer cell. In some embodiments, the cancerous tumor comprises cancer cells that are resistant to a chemotherapy drug (e.g., oxaliplatin).

[0019] The present invention also relates to a pharmaceutical composition that is useful for at least treating cancer. The pharmaceutical composition comprises an effective amount of a coordination compound represented by formula (I), wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium, and a pharmaceutically acceptable carrier. The coordination compound preferably comprises a trivalent chromium ion and a trivalent iron ion and is represented by formula (II).

[0020] In some embodiments, the pharmaceutical composition further comprises an additional pharmaceutically active agent, such as a chemotherapeutic agent or an immunomodulatory agent. In some embodiments, the additional pharmaceutically active agent is a chemotherapeutic agent selected from oxaliplatin, irinotecan, 5-fluorouracil, gemcitabine, amycin, or any combination thereof.

[0021] The coordination compounds disclosed herein can target multiple factors involved in cancer progression and metastasis and are effective against chemotherapy-resistant cancer cells. Therefore, the compounds can be used in medical formulations to treat cancers, including but not limited to oxaliplatin-resistant colorectal cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Those skilled in the art will clearly understand the present invention through the following detailed description of the preferred embodiment in conjunction with the accompanying drawings, in which:

[0023] Figure 1A The coordination compound of formula (II) 1 H-NMR spectroscopy;

[0024] Figure 1B The coordination compound of formula (II) 13 C-NMR spectroscopy;

[0025] Figure 1Cis the Fourier transform infrared (FT-IR) spectrum of the coordination compound of formula (II);

[0026] Figure 2A is a schematic diagram illustrating the process of establishing oxaliplatin-resistant LoVo cells (referred to as LoVo-OXAR cells) from LoVo parental cells;

[0027] Figure 2B Light microscopy pictures showing LoVo parental cells and LoVo-OXAR cells;

[0028] Figure 2C showed the cytotoxic effect of oxaliplatin on LoVo parental cells and LoVo-OXAR cells; * and ** Respectively, p < 0.05 and p < 0.01 compared with those without oxaliplatin treatment (control group);

[0029] Figure 2D Shows the effect of the coordination compound of formula (II) on the activity of LoVo parent cells; * 、 ** and *** indicates p < 0.05, p < 0.01, and p < 0.001, respectively, compared with the group without compound treatment (control group);

[0030] Figure 2E Shows the effect of the coordination compound of formula (II) on the activity of LoVo-OXAR cells; * and ** Respectively, p < 0.05 and p < 0.01 compared with the group without compound treatment (control group);

[0031] Figure 2F The coordination compound of formula (II) was shown to have a synergistic cytotoxic effect on LoVo parental cells and LoVo-OXAR cells when used in combination with oxaliplatin (OXA), irinotecan, or 5-fluorouracil (5-FU); * and *** Respectively, p < 0.05 and p < 0.001 compared with LoVo parental cells without compound treatment (control group); # and ### Respectively, p < 0.05 and p < 0.001 compared with LoVo-OXAR cells without compound treatment (control group);

[0032] Figure 3A Shows the effect of the coordination compound of formula (II) on the activity of CL1 parental cells;

[0033] Figure 3BThe effect of the coordination compound of formula (II) on the activity of gemcitabine-resistant CL1 cells (abbreviated as CL1-GEMR cells) is shown;

[0034] Figure 3C Shows the effect of the coordination compound of formula (II) on the activity of BEAS-2B cells;

[0035] Figure 4A Shows the effect of the coordination compound of formula (II) on apoptosis of LoVo parental cells; ** indicates p < 0.01 compared with the control group without compound treatment;

[0036] Figure 4B Shows the effect of the coordination compound of formula (II) on apoptosis of LoVo-OXAR cells; ** indicates p < 0.01 compared with the group without compound treatment (control group);

[0037] Figure 4C Shown are Western blot analyses of ER stress markers in LoVo parental and LoVo-OXAR cells in the presence or absence of the coordination compound of formula (II) or oxaliplatin (OXA);

[0038] Figure 5A Shows the effect of the coordination compound of formula (II) on the migration of LoVo parental cells and LoVo-OXAR cells; * 、 ** and *** Respectively, p < 0.05, p < 0.01, and p < 0.001 compared with LoVo parental cells not treated with the compound (control group); # 、 ## and ### Respectively, p < 0.05, p < 0.01, and p < 0.001 compared with LoVo-OXAR cells without compound treatment (control group);

[0039] Figure 5B Shows the effect of the coordination compound of formula (II) on the invasiveness of LoVo parental cells and LoVo-OXAR cells; ** and *** Respectively, p < 0.01 and p < 0.001 compared with LoVo parental cells (control group) without compound treatment; # and ## Respectively, p < 0.05 and p < 0.01 compared with LoVo-OXAR cells without compound treatment (control group);

[0040] Figure 5CWestern blot analysis showing EMT markers in LoVo parental cells treated or not with the coordination compound of formula (II);

[0041] Figure 5D Western blot analysis showing EMT markers in LoVo-OXAR cells treated or not with the coordination compound of formula (II);

[0042] Figure 5E The effects of the complex of formula (II) on the expression of EMT markers in LoVo parental cells were shown, as detected by immunofluorescence staining; ** indicates p < 0.01 compared with the group without compound treatment (control group);

[0043] Figure 5F The effects of the coordination compound of formula (II) on the expression of EMT markers in LoVo-OXAR cells were shown, and these cell lines were detected by immunofluorescence staining; * and ** Respectively, p < 0.05 and p < 0.01 compared with the group without compound treatment (control group);

[0044] Figure 5G The effects of the coordination compound of formula (II) on the expression of messenger RNA (mRNA) of EMT markers in LoVo parental cells are shown; * 、 ** and *** indicates p < 0.05, p < 0.01, and p < 0.001, respectively, compared with the group without compound treatment (control group);

[0045] Figure 5H Shows the effect of the coordination compound of formula (II) on the mRNA expression of EMT markers in LoVo-OXAR cells; * 、 ** and *** indicates p < 0.05, p < 0.01, and p < 0.001, respectively, compared with the group without compound treatment (control group);

[0046] Figure 6A Representative photographs of tumor masses isolated from mice in different treatment groups on day 21 after tumor cell inoculation are shown;

[0047] Figure 6B Shows the tumor growth of mice in different treatment groups from day 1 to day 21 after tumor cell inoculation;

[0048] Figure 6C The body weights of mice in different treatment groups from day 1 to day 21 after tumor cell inoculation are shown;

[0049] Figure 6D Shows the effect of the coordination compound of formula (II) on apoptosis in tumors of mice inoculated with LoVo parental cells; ** indicates p < 0.01 compared with the group without compound treatment (control group);

[0050] Figure 6E Shows the effect of the coordination compound of formula (II) on apoptosis of tumor cells in mice inoculated with LoVo-OXAR cells; ** indicates p < 0.01 compared with the group without compound treatment (control group);

[0051] Figure 6F Representative photographs of mouse organs are shown, which demonstrate the effect of the coordination compound of formula (II) on the metastasis of LoVo parental tumors in mice of different treatment groups; arrows indicate the growth of metastatic cancer cells;

[0052] Figure 6G Representative photos of mouse organs are shown, which demonstrate the effect of the coordination compound of formula (II) on LoVo-OXAR tumor metastasis in mice of different treatment groups; arrows indicate the growth of metastatic cancer cells;

[0053] Figure 7 The effects of the coordination compound of formula (II) on the activity of HA22T parental cells and SAHA-resistant HA22T cells (referred to as HA22T-HDACiR cells) are shown; * 、 ** and *** indicates p < 0.05, p < 0.01, and p < 0.001, respectively, compared with the group without compound treatment (control group);

[0054] Figure 8 Shows the effect of the coordination compound of formula (II) on the activity of LNCaP cells;

[0055] Figure 9A Shows the effect of the coordination compound of formula (II) on the activity of 143B cells;

[0056] Figure 9B The coordination compound of formula (II) was shown to have a synergistic cytotoxic effect on 143B cells when used in combination with 5-fluorouracil (5-FU), gemcitabine, or amycin;

[0057] Figure 9C Shows the effect of the coordination compound of formula (II) on the migration of 143B cells;

[0058] Figure 9D Western blot analysis showing EMT markers in 143B cells treated or not with the ligand compound of formula (II);

[0059] Figure 10 Shows the effect of the coordination compound of formula (II) on the activity of GBM8401 cells;

[0060] Figure 11 Shows the effect of the coordination compound of formula (II) on the activity of T-47D cells and MDA-MB231 cells;

[0061] Figure 12 Shows the effect of the coordination compound of formula (II) on the activity of T24 cells;

[0062] Figure 13 Shows the effect of the coordination compound of formula (II) on the activity of SCC-25 cells;

[0063] Figure 14 Shows the effect of the coordination compound of formula (II) on the activity of CP70 cells;

[0064] Figure 15 Shows the effect of the coordination compound of formula (II) on the activity of Jurkat cells;

[0065] Figure 16 Shows the effect of the coordination compound of formula (II) on the activity of HeLa cells;

[0066] Figure 17 Shows the effect of the coordination compound of formula (II) on the activity of TT cells;

[0067] Figure 18 Shows the effect of the coordination compound of formula (II) on the activity of A431 cells;

[0068] Figure 19 Shows the effect of the coordination compound of formula (II) on the activity of MIAPaCa-2 cells and PANC-1 cells;

[0069] Figure 20A Shows the tumor growth of mice in different treatment groups after tumor cell inoculation; and

[0070] Figure 20B Shown are the body weights of mice in different treatment groups after tumor cell inoculation.

[0071] Implementation Method

[0072] The following embodiments and examples are provided to further illustrate the present invention. It should be understood that the following embodiments are not intended to limit the scope of the present invention, and that those skilled in the art may make adjustments and modifications without exceeding the scope of the appended claims.

[0073] Unless defined otherwise, all technical and scientific terms and abbreviations used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0074] definition

[0075] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "a pharmaceutically acceptable carrier" includes mixtures of pharmaceutically acceptable carriers, and "an additional pharmaceutically active agent" includes more than one pharmaceutically active agent.

[0076] Data are generally presented as mean ± standard deviation. The values provided herein are approximate, and experimental values may vary within a range of 20%, preferably within a range of 10%, and more preferably within a range of 5%. Therefore, the terms "about" and "approximately" mean within a range of 20%, preferably within a range of 10%, and more preferably within a range of 5% of a given value or range.

[0077] As used herein, the term "tumor(s)" refers to an abnormal mass of cells characterized by uncontrolled cell proliferation and growth. Tumors include solid tumors that form a noticeable mass, as well as non-solid tumors, such as leukemia, which involve abnormal cell proliferation in the blood or other body fluids. Tumors may be noncancerous (also known as benign tumors) or cancerous (also known as malignant tumors). Noncancerous tumors contain relatively slow-growing tumor cells whose proliferation is limited to their original location and therefore do not spread to other parts of a person's body. Cancerous tumors (used interchangeably with the term "cancer") contain rapidly dividing tumor cells that, when present in a person, have the potential to invade nearby tissues or spread to distant organs. Depending on the context of the present invention, the term "tumor" may specifically refer to a "cancerous tumor," and the term "tumor cell" may specifically refer to a "cancer cell."

[0078] As used herein, the term "cancer cell" may refer to a single cancer cell or a homogeneous or heterogeneous group of cancer cells. Cancer cells can be cells within an individual, cells isolated from an individual (e.g., a human), or cells derived from an isolated cancer cell. Furthermore, unless otherwise indicated, cancer cells can have a variety of origins.

[0079] As used herein, the term "coordination compound" refers to a metal complex defined by formula (I) comprising three metal ion centers, each surrounded by six ligands.

[0080] coordination compounds

[0081] The coordination compound provided herein is shown in formula (I):

[0082] [XZ2(CH3CO2)6(H2O)4(OH)2]NO3 formula (I).

[0083] X can be a trivalent ion of chromium (Cr) or molybdenum (Mo), and Z can be a trivalent ion of iron (Fe), ruthenium (Ru), or osmium (Os). Since the coordination compound can contain Cr or Mo (both Group 6 metals) and Fe, Ru, or Os (all Group 8 metals), the coordination compound can be represented by formula (II), (III), (IV), (V), (VI), or (VII) based on various combinations of central metal ions:

[0084] Formula (II): [CrFe2(CH3CO2)6(H2O)4(OH)2]NO3

[0085] Formula (III): [CrRu2(CH3CO2)6(H2O)4(OH)2]NO3

[0086] Formula (IV): [CrOs2(CH3CO2)6(H2O)4(OH)2]NO3

[0087] Formula (V): [MoFe2(CH3CO2)6(H2O)4(OH)2]NO3

[0088] Formula (VI): [MoRu2(CH3CO2)6(H2O)4(OH)2]NO3

[0089] Formula (VII): [MoOs2(CH3CO2)6(H2O)4(OH)2]NO3

[0090] In some embodiments, the coordination compound comprises a trivalent chromium ion and a trivalent iron ion and is represented by formula (II). The compound of formula (II) may have structure (a) as shown below or other structures representing different stereoisomers. In other words, the compound of formula (II) refers to any compound of formula (II) that may have different arrangements of ligands around the metal ion. Similarly, compounds of formula (III), (IV), (V), (VI), or (VII) may include various stereoisomers.

[0091]

[0092] Use of coordination compounds to inhibit the survival or proliferation of cancer cells

[0093] The present invention provides a method for inhibiting cancer cell survival or proliferation, comprising contacting the cancer cell with an effective amount of a coordination compound represented by formula (I): [XZ2(CH3CO2)6(H2O)4(OH)2]NO3, wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. As used herein, "inhibiting survival or proliferation" means that the coordination compound prevents the survival of cancer cells, or slows or stops cancer cell proliferation. Such inhibition can be assessed by methods known in the art, such as direct cell counting, monitoring cell proliferation over time using a real-time cell analysis system, determining cell viability by measuring metabolically active cells (e.g., MTT assay), assessing cell division by measuring deoxyribonucleic acid (DNA) synthesis, and measuring tumor size.

[0094] The cytotoxicity of the coordination compound has been demonstrated against a variety of cancer cells, including colorectal cancer cells, lung cancer cells, liver cancer cells, pancreatic cancer cells, bone cancer cells, brain cancer cells, breast cancer cells, ovarian cancer cells, cervical cancer cells, prostate cancer cells, bladder cancer cells, blood cancer cells, stomach cancer cells, skin cancer cells, head and neck cancer cells, and thyroid cancer cells. In some embodiments, the coordination compound exerts a cytotoxic effect against invasive cancer cells, such as invasive colorectal cancer cells. In some embodiments, the coordination compound exerts a cytotoxic effect against cancer cells that are resistant to chemotherapeutic agents, such as oxaliplatin-resistant colorectal cancer cells.

[0095] An effective amount of a coordination compound administered to cancer cells to inhibit cell survival or proliferation is an amount sufficient to cause cancer cell death or limit cancer cell proliferation. As will be appreciated by those skilled in the art, the effective amount will vary depending on a variety of factors, such as the type of cancer cell, the specific coordination compound administered, the method by which the coordination compound is delivered to the cancer cell, and the use of the coordination compound in conjunction with other anti-tumor agents.

[0096] The step of contacting the cancer cells with the coordination compound can be performed according to methods known in the art. In some embodiments, the cancer cells are contacted with the coordination compound by culturing the cancer cells in a cell culture medium supplemented with the coordination compound. In some embodiments, the cancer cells are contacted with the coordination compound by exposing the cancer cells to a composition containing the coordination compound and a pharmaceutically acceptable carrier in vitro or in vivo.

[0097] In some embodiments, the coordination compound inhibits the survival or proliferation of cancer cells by inducing apoptosis mediated by endoplasmic reticulum stress (ER stress) in cancer cells. For example, the coordination compound of formula (II) induces apoptosis in colorectal cancer cells, thereby inhibiting their survival, reducing cell number and preventing proliferation. This apoptosis-inducing effect may be due to the activated and sustained endoplasmic reticulum stress in cancer cells. This state is accompanied by an unresolved unfolded protein response (UPR) that triggers cancer cell death. The induced apoptosis inhibits the growth of existing cancer cells (including chemotherapy-resistant cancer cells) and can prevent the formation and development of chemotherapy-resistant cancer cell populations.

[0098] In some embodiments, the method of inhibiting cancer cell survival or proliferation further comprises contacting the cancer cell with a chemotherapeutic agent selected from oxaliplatin, irinotecan, 5-fluorouracil, gemcitabine, amycin, or any combination thereof. In other words, the aforementioned coordination compound can be used in combination therapy with other chemotherapeutic drugs. The coordination compound and other chemotherapeutic drugs can be co-administered to cells or individuals in need thereof simultaneously or sequentially.

[0099] Use of coordination compounds to inhibit cancer cell migration or invasion

[0100] The present invention provides a method for inhibiting cancer cell migration or invasion, comprising contacting the cancer cells with an effective amount of a coordination compound represented by formula (I): [XZ2(CH3CO2)6(H2O)4(OH)2]NO3, wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. "Inhibiting migration or invasion" herein means that, after administration of the coordination compound, the cancer cells are prevented from acquiring or reduced in their ability to migrate from their original location or to penetrate and spread into surrounding or remote environments. Such inhibition can be assessed by methods known in the art, for example, live cell imaging using a microscope, measuring cell movement across a porous membrane in a chamber system (e.g., a Transwell migration or invasion assay), assessing changes in the expression of important genes or proteins involved in cell migration and invasion pathways using molecular techniques such as quantitative polymerase chain reaction (qPCR) and western blotting, and monitoring the metastasis of cancerous tumors in an individual.

[0101] In some embodiments, the coordination compound is capable of inhibiting the migration or invasion of colorectal cancer cells or bone cancer cells. In some embodiments, the coordination compound inhibits the migration or invasion of invasive cancer cells, such as invasive colorectal cancer cells or creeping osteosarcoma cells. In some embodiments, the coordination compound is capable of inhibiting the migration or invasion of cancer cells that are resistant to chemotherapeutic agents, such as oxaliplatin-resistant colorectal cancer cells.

[0102] An effective amount of a coordination compound administered to cancer cells to inhibit cell migration or invasion is an amount sufficient to prevent the transformation of non-invasive cancer cells into migratory or invasive cells or to reduce the ability of cancer cells to migrate or invade from a primary site to a secondary site. As will be appreciated by those skilled in the art, the effective amount will vary depending on a variety of factors, such as the type of cancer cell, the specific coordination compound administered, the method by which the coordination compound is delivered to the cancer cells, and the co-use of the coordination compound with other anti-tumor agents.

[0103] In some embodiments, the coordination compound inhibits cancer cell migration or invasion by inhibiting the epithelial-mesenchymal transition (EMT) of cancer cells. For example, the coordination compound of formula (II) can induce colorectal cancer cells to express epithelial phenotype markers, such as E-cadherin and tight junction protein 1 (TJP1), and inhibit colorectal cancer cells from expressing mesenchymal phenotype markers, such as vimentin and fibronectin 1 (FN1), thereby inhibiting the migration and invasion of colorectal cancer cells. The EMT-inhibiting activity of the coordination compound may help prevent cancer metastasis and chemotherapy resistance.

[0104] Use of coordination compounds in treating cancer

[0105] The coordination compound also exhibits therapeutic effects in individuals suffering from cancer. Therefore, the present invention further provides a method for treating cancerous tumors, comprising administering to a subject in need thereof an effective amount of a coordination compound of formula (I), wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. In some preferred embodiments, the administered coordination compound is the compound of formula (II) described above.

[0106] As used herein, the term "subject" refers to a mammal. The subject can be human or non-human, including but not limited to primates, rodents, dogs, cats, cows, goats, sheep, horses, rabbits, pigs, etc. Thus, both veterinary and medical treatments and compositions are contemplated herein.

[0107] The coordination compound can be used to treat a cancerous tumor selected from colorectal cancer, lung cancer, liver cancer, pancreatic cancer, bone cancer, brain cancer, breast cancer, ovarian cancer, cervical cancer, prostate cancer, bladder cancer, blood cancer, stomach cancer, skin cancer, head and neck cancer, or thyroid cancer. In some embodiments, the cancerous tumor comprises cancer cells that are aggressive and / or resistant to chemotherapeutic agents (e.g., oxaliplatin).

[0108] An effective amount of a coordination compound administered to a subject in need thereof can be a prophylactically effective amount or a therapeutically effective amount. A "prophylactically effective amount" refers to an amount sufficient to prevent the progression of cancer, metastasis, the development of invasive or chemotherapy-resistant cancer cells, or the development of symptoms or signs associated with the foregoing in a subject. A "therapeutically effective amount" refers to an amount sufficient to stop or delay tumor growth, metastasis, the development of invasive or chemotherapy-resistant cancer cells, or the development of symptoms or signs associated with the foregoing in a subject. As will be appreciated by those skilled in the art, the effective amount will vary depending on a variety of factors, such as the type of cancer, the age, weight, physical condition and responsiveness of the subject being treated, the specific coordination compound being administered, the route of administration, the use of excipients, and the use of other pharmaceutically active agents.

[0109] The coordination compound of formula (I) can be administered to a subject by any suitable route, preferably in the form of a pharmaceutical composition suitable for that route. In some embodiments, the coordination compound is administered orally, topically, transmucosally, intravenously, or enterally. In some embodiments, the coordination compound is administered to a subject in solid or liquid form.

[0110] In some embodiments, the coordination compound of formula (I) is administered to a subject at an early stage of cancer development. In some embodiments, the coordination compound is administered to a subject at a later stage of cancer development. In some embodiments, the coordination compound is administered at least once, twice, three times, or more times per day. In some embodiments, the coordination compound is administered daily, every other day, several times per week, weekly, monthly, or less frequently to maintain an effective dose level and patient compliance. In some embodiments, administration of the coordination compound continues for several weeks, months, or longer. The frequency and duration of treatment may vary depending on a subject's response to treatment.

[0111] Pharmaceutical compositions

[0112] The present invention further provides a pharmaceutical composition for treating cancer, comprising an effective amount of a coordination compound of formula (I) and a pharmaceutically acceptable carrier, wherein X is a trivalent ion of chromium or molybdenum, and Z is a trivalent ion of iron, ruthenium, or osmium. In some preferred embodiments, the pharmaceutical composition comprises the coordination compound of formula (II) as an active ingredient.

[0113] The pharmaceutical composition can be in any suitable form, such as tablets, powders, solutions, suspensions, emulsions, liposomes, nanoparticles, or other formulations.

[0114] As used herein, the term "pharmaceutically acceptable carrier" refers to any carrier or excipient that is compatible with the coordination compound and other active ingredients (if any), and preferably stabilizes the active ingredients and is harmless to the subject to be treated. Pharmaceutically acceptable carriers can be excipients, diluents, antioxidants, and preservatives known in the art. Examples of pharmaceutically acceptable carriers include, but are not limited to, water, saline, buffers, organic solvents, hydrophilic polymers, carbohydrates, peptides, amino acids, and surfactants.

[0115] In some embodiments, the pharmaceutical composition further comprises an additional pharmaceutically active agent. The term "pharmaceutically active agent" refers to a small molecule compound or a macromolecule (such as an antibody or a fragment thereof) that has the desired pharmacological action and therapeutic effect. The pharmaceutically active agent can be a chemotherapeutic agent, an immunomodulator, or any combination thereof. Examples of chemotherapeutic agents include, but are not limited to, alkylating agents (such as cyclophosphamide, melphalan, temozolomide, carboplatin, cisplatin, and oxaliplatin), antimetabolites (such as 5-fluorouracil, 6-mercaptopurine, cytarabine, gemcitabine, and methotrexate), antitumor antibiotics (such as actinomycin-D), and oxaliplatin. ), bleomycin, daunorubicin, and amycin), topoisomerase inhibitors (such as etoposide, irinotecan, teniposide, and topotecan), mitotic inhibitors (such as docetaxel, estramustine, nocodazole, paclitaxel, and vinblastine), histone deacetylase (HDAC) inhibitors (such as vorinostat, also known as suberoylanilide hydroxamic acid (SAHA)), and steroids (such as prednisone, methylprednisolone, and dexamethasone). Examples of immunomodulators include immunostimulants (such as trastuzumab) and immunosuppressants. Example

[0116] Example 1: Preparation of coordination compound

[0117] The following process for preparing the coordination compound of formula (II) (referred to as "Compound (II)") is provided as an example to illustrate the preparation methods provided herein. Chromium (III) nitrate and iron (III) nitrate are mixed in a flask at a molar ratio of approximately 1:1 to 1:3. Alcohol containing 60-95% ethanol (the remainder being water) is added to the flask at room temperature to a weight-to-volume ratio (g / ml) of iron (III) nitrate to alcohol of approximately 1:1 to 1:3, and the mixture is stirred until all solids have dissolved. Acetic anhydride is then added to the flask to a volume ratio of acetic anhydride to alcohol of approximately 4:1 to 7:1, and the reaction mixture is stirred for approximately 2 to 6 hours. The reaction with acetic anhydride is carried out at a temperature below 70°C. The reaction mixture is then stirred for 20 to 28 hours, after which the solid precipitate of Compound (II) is collected by filtration. The solid precipitate is dried in an oven to a constant weight.

[0118] The molecular weight of compound (II) was determined to be approximately 686.11 by mass spectrometry. Elemental analysis showed that compound (II) contained approximately 7.36% chromium, approximately 19.31% iron, approximately 2.21% nitrogen, approximately 21.73% carbon, approximately 3.86% hydrogen, and approximately 44.21% oxygen (by mass). The structural characteristics of compound (II) were analyzed using 1 H nuclear magnetic resonance (NMR) spectroscopy, 13 C-NMR spectroscopy and Fourier transform infrared (FT-IR) spectroscopy were used for verification. Figure 1A Show compound (II) 1 H-NMR spectrum, where the signals at approximately 3.3±0.2 and 1.9±0.2 ppm correspond to the hydrogen of H2O and acetate groups, respectively. 13 The signals at approximately 21.2±0.5, 39.7±0.5, and 171.9±0.5 ppm in the C-NMR spectrum indicate the presence of acetate groups ( Figure 1B ). In addition, the infrared spectrum showed that the peaks were approximately 3400±10, 3200±10, 3000±10, 1683±10, 1585±10, 1428±10, 1349±10, 1292±10 and 1035

[0119] ±10cm -1 There is an absorption peak at ( Figure 1C All data support the structure of compound (II) provided herein. Compound (II) was used in Examples 2-20 for efficacy studies.

[0120] The coordination compounds of formula (III) to formula (VII) can be prepared according to the above method but using appropriate metal salts instead of chromium (III) nitrate and / or iron (III) nitrate.

[0121] Example 2: Cytotoxic effect of the coordination compound of formula (II) on colorectal cancer cells

[0122] To evaluate whether the coordination compound has the potential to inhibit the survival of cancer cells (including chemotherapy-resistant cancer cells), the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide] (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide) assay was performed to measure the activity of colorectal cancer cells treated with or without compound (II).

[0123] 2.1 Cell culture

[0124] Human colon cancer LoVo cells (hereinafter referred to as LoVo parental cells; available from the American Type Culture Collection (ATCC) under the accession number CCL-229 or from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan, China under the accession number BCRC60148) and their chemotherapy-resistant subclones were cultured in Dulbecco's modified Eagle's medium (Sigma-Aldrich, Missouri, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Utah, USA). All cell cultures were maintained at 37°C and 5% carbon dioxide. Fresh medium was replaced after 48 hours of subculture.

[0125] 2.2 Establishment of oxaliplatin-resistant colorectal cancer cells

[0126] The IC50 (half-maximal inhibitory concentration) of oxaliplatin (purchased from Sigma-Aldrich, #09512) against LoVo parental cells was determined to be approximately 15.0 μg / ml using the MTT assay. To prepare LoVo subclone cells with stable resistance to oxaliplatin, LoVo cells that survived treatment with a low concentration (e.g., 15.0 μg / ml) of oxaliplatin were exposed to a higher concentration (e.g., 25.0 μg / ml) of oxaliplatin for 24 hours, and this process was repeated until an oxaliplatin-resistant LoVo cell population (referred to as LoVo-OXAR cells) with an IC50 approximately 4 times higher than that of the LoVo parental cells (i.e., an IC50 of approximately 65 μg / ml) was obtained. Figure 2A ).like Figure 2B and Figure 2CAs shown, oxaliplatin reduced the viability of LoVo parental cells and LoVo-OXAR cells in a dose-dependent manner. LoVo-OXAR cells were significantly more resistant to oxaliplatin treatment, and at 200x magnification, LoVo-OXAR cells also showed morphological differences from LoVo parental cells.

[0127] 2.3 Effect of compound (II) on the activity of colorectal cancer cells

[0128] LoVo parental cells or LoVo-OXAR cells were cultured at a rate of 1 × 10 4 Cells were seeded at a density of 100 μg / well (triplicate) into 96-well plates and cultured for 24 hours. Cells were then treated with compound (II) at different concentrations (0, 125, 250, 500, 1000, 2000, or 4000 μg / ml) for 24 hours. After removing the culture medium, 100 μl of MTT solution (5.0 mg / ml) was added to each well and cultured at 37°C for 4 to 5 hours until a purple precipitate formed. The supernatant was removed and dimethylsulfoxide (DMSO) was added to each well to dissolve the blue formazan crystals. An ELISA reader (Molecular Devices, Palo Alto, California, USA) was used to measure the absorbance at a wavelength of 570 nm (OD570) to determine cell viability and the IC50 of compound (II). The cell viability (percentage) was calculated as follows: (OD570 of each test group / OD570 of the control group) × 100%. The mean value of cell viability was calculated from three replicates. IC50 was defined as the concentration of the test compound that caused 50% cell death in a cell population (ie, a 50% decrease in cell viability).

[0129] like Figure 2D and Figure 2E As shown, compound (II) reduced the activity of the two colorectal cancer cells in a dose-dependent manner, with IC50 values of approximately 500 μg / ml and approximately 2000 μg / ml for LoVo parental cells and LoVo-OXAR cells, respectively. This result demonstrates that the coordination compound has the ability to inhibit the survival of cancer cells, including chemotherapy-resistant cancer cells.

[0130] 2.4 Synergistic effects of compound (II) and other chemotherapeutic drugs

[0131] To further investigate the effect of compound (II) on the efficacy of chemotherapy drugs, LoVo parental cells or LoVo-OXAR cells were treated with compound (II) (500 μg / ml in LoVo parental cells; 2000 μg / ml in LoVo-OXAR cells), oxaliplatin (OXA, 20 μg / ml), irinotecan (20 μg / ml), 5-fluorouracil (5-FU, 20 μg / ml), or a combination thereof for 24 hours, and then the cell viability was measured using the MTT assay. Figure 2F As shown, the combination of commonly used chemotherapeutic drugs and compound (II) is significantly more effective in inhibiting the survival of LoVo parental cells and LoVo-OXAR cells than the use of chemotherapeutic drugs alone. This result shows that the coordination compounds provided herein can be used alone or in combination with other chemotherapeutic drugs to fight cancer.

[0132] Example 3: Cytotoxic effect of the coordination compound of formula (II) on lung cancer cells

[0133] 3.1 Cell culture

[0134] Human lung adenocarcinoma CL1 cells (Creative Biolabs, Shirley, New York, USA; hereinafter referred to as CL1 parental cells) and their chemotherapy-resistant subclones were cultured in DMEM (Gibco TM BEAS-2B cells (95102433, Sigma-Aldrich) were cultured in LHC-9 medium (Gibco TM , Thermo Fisher Scientific, Waltham, MA, USA). All cell cultures were performed at 37°C with 5% CO2 supply.

[0135] 3.2 Effect of compound (II) on lung cancer cell activity

[0136] CL1 parental cells and gemcitabine-resistant CL1 cells were treated with compound (II) at different concentrations (0, 100, 200, 400, 800 or 1600 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 3A and Figure 3B As shown, compound (II) reduced the activity of the two lung cancer cell lines in a dose-dependent manner.

[0137] 3.3 Effect of compound (II) on the activity of non-tumor lung cells

[0138] The cytotoxicity of compound (II) on non-cancerous cells was studied using non-tumor human bronchial epithelial cells BEAS-2B as a cell model. BEAS-2B cells were treated with different doses (0, 200, 400, 600, or 800 μg / ml) of compound (II) for 24 hours, and then the cell viability was measured using the MTT assay. Figure 3C As shown, compound (II) did not significantly inhibit the survival of BEAS-2B cells, indicating that the coordination compounds provided herein do not hinder the growth of non-cancerous human cells.

[0139] Example 4: The coordination compound of formula (II) induces endoplasmic reticulum stress-mediated apoptosis in colorectal cancer cells

[0140] To assess whether compound (II) induces apoptosis in cancer cells, a terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay was used to detect apoptosis. Briefly, LoVo parental cells and LoVo-OXAR cells were treated with 500 μg / ml and 2000 μg / ml of compound (II), respectively, for 24 hours. TUNEL assays were then performed (In Situ Cell Death Detection Kit; Roche Applied Science, Indianapolis, IN, USA). Nuclei were counterstained with 0.1 mg / ml 4',6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich, St. Louis, MO, USA). LoVo parental cells or LoVo-OXAR cells treated with 0 μg / ml of compound (II) served as controls. TUNEL-positive cells were examined using an Olympus CKX53 microscope (Olympus, Shinjuku-ku, Tokyo, Japan). Figure 4A and Figure 4B As shown, significantly increased apoptosis (based on the increase in green fluorescence of apoptotic cells) was observed in LoVo parental cells and LoVo-OXAR cells after administration of compound (II) compared to the control group.

[0141] Subsequently, proteins involved in compound (II)-induced apoptosis were further investigated. LoVo parental cells and LoVo-OXAR cells were treated with compound (II) (500 μg / ml for LoVo parental cells and 2000 μg / ml for LoVo-OXAR cells), oxaliplatin (25 μg / ml for LoVo parental cells and 45 μg / ml for LoVo-OXAR cells), or a combination thereof for 24 hours. The cells were then harvested and lysed for Western blotting. Untreated LoVo parental cells or LoVo-OXAR cells served as controls. Western blot analysis was performed using primary antibodies against the following proteins (San-taCruz Biotechnology Inc., Santa Cruz, CA, USA): protein kinase RNA-like endoplasmic reticulum kinase (PERK), phosphorylated PERK (p-PERK), alpha subunit of eukaryotic translation initiation factor 2 (eIF2α), phosphorylated eIF2α (p-eIF2α), activating transcription factor 4 (ATF4), proliferating cell nuclear antigen (PCNA), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and caspase 3 (Cas3). GAPDH was used as an internal control for protein expression normalization. The results showed that after treatment with compound (II), the amount of eIF2α in LoVo-OXAR cells decreased, but the phosphorylation of eIF2α increased ( Figure 4C ). It is known that phosphorylated eIF2α can inhibit the synthesis of multiple proteins involved in tumorigenesis. This result also proves that compound (II) can induce endoplasmic reticulum stress, thereby enhancing the expression of endoplasmic reticulum stress markers (including PERK, p-eIF2α and ATF4), and further activate Cas3 cleavage, ultimately leading to cell apoptosis ( Figure 4C ). Therefore, the coordination compounds provided herein can be used as inducers that aggravate the endoplasmic reticulum stress of cancer cells and induce apoptosis, thereby inhibiting cancer progression.

[0142] Example 5: Inhibitory effect of the coordination compound of formula (II) on migration and invasion of colorectal cancer cells

[0143] 5.1 Inhibition of cancer cell migration and invasion

[0144] Cancer cell migration and invasion are key activities in tumor progression and metastasis. To evaluate whether the coordination compounds have an effect on these activities, Transwell migration and invasion assays were performed on LoVo parental cells and LoVo-OXAR cells. 4 Each cell was resuspended in 200 μl of serum-free medium and added to the upper compartment of a Transwell cell culture chamber (Corning, New York, USA) (i.e., a 24-well insert with a porous membrane; pore size 8 μm). The lower chamber was filled with DMEM containing 10% FBS (as a chemotactic inducer). The cells were then cultured with compound (II) (500 μg / ml for LoVo parental cells and 2000 μg / ml for LoVo-OXAR cells), oxaliplatin (50 μg / ml), or a combination thereof. LoVo parental cells or LoVo-OXAR cells that were not treated with the compound served as controls. In the migration assay, the culture time was 48 hours. In the invasion assay, the insert was pre-coated with extracellular matrix gel (BD Biosciences, Sparks, MD, USA), and the cells were cultured for 72 hours. After each experiment, the cells on the upper surface of the porous membrane were removed, and the cells on the lower surface of the membrane were fixed and stained with Giemsa. Stained cells were counted in five randomly selected areas under a microscope (200x magnification). The invasion or migration rate was calculated as follows: Invasion (or migration) rate (%) = (average number of transmembrane cells in each experimental group / average number of transmembrane cells in the control group) × 100%. The average invasion or migration rate was calculated from three replicates.

[0145] like Figure 5A and Figure 5B As shown, either compound (II) or oxaliplatin significantly reduced the migration and invasion rates of LoVo parental cells and LoVo-OXAR cells. Furthermore, the combination of compound (II) and oxaliplatin synergistically reduced the migration and invasion rates of both cell lines to less than 50%. These results indicate that the coordination compound significantly inhibits the migration and invasion capabilities of cancer cells, including chemotherapy-resistant cancer cells.

[0146] 5.2 Inhibition of epithelial-mesenchymal transition (EMT)

[0147] Vimentin and E-cadherin are molecular markers associated with tumor progression. They play a role in EMT, a pathological process that promotes cancer cell migration and invasion, and tumor progression. Vimentin is a key component of intermediate filaments, helping to maintain cell integrity and stress resistance. E-cadherin regulates contact inhibition during cell proliferation and confluence, thereby contributing to the maintenance of the epithelial phenotype. To further investigate the effects of compound (II) on the expression of EMT markers, Western blotting, immunofluorescence analysis, and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) were performed on LoVo parental cells and LoVo-OXAR cells treated with various doses of compound (II) for 24 hours. Untreated LoVo parental cells or LoVo-OXAR cells served as controls.

[0148] 5.2.1 Western transfer

[0149] The cells treated with the indicated treatments were collected and analyzed by Western blot using primary antibodies against vimentin, E-cadherin, and GAPDH (San-taCruz Biotechnology Inc., Santa Cruz, CA, USA). GAPDH was used as an internal control. The results showed that compared with the control group, the protein expression of vimentin was significantly reduced in LoVo parental cells and LoVo-OXAR cells treated with compound (II), while the protein expression of E-cadherin (which is considered a tumor suppressor due to its role in contact inhibition) was increased ( Figure 5C and Figure 5D ).

[0150] 5.2.2 Immunofluorescence detection

[0151] LoVo parental cells or LoVo-OXAR cells were cultured at a rate of 1 × 10 4The cells were seeded into 8-well plates at a density of 100 cells / well and cultured in DMEM containing 10% FBS. Compound (II) was added or not in the culture medium, and then fixed with phosphate buffered saline (PBS) containing 4% paraformaldehyde. The cells were then permeabilized with a 0.1% sodium citrate solution containing 0.1% TritonX-100, and the holes were blocked with a blocking buffer (2% bovine serum albumin), and then immunofluorescence staining was performed. The cells were reacted with primary antibodies against vimentin and E-cadherin at 4°C for 24 hours, and then reacted with fluorescent secondary antibodies at 25°C for 1 hour, followed by DAPI staining and observation under a fluorescence microscope. Figure 5E and Figure 5F As shown, treatment with compound (II) significantly inhibited the expression of vimentin but significantly increased the expression of E-cadherin.

[0152] 5.2.3qRT-PCR

[0153] Total RNA was extracted from cells after the indicated treatments using the GeneJET RNA purification kit (ThermoFisherScientific, Lithuania), and reverse transcribed into cDNA using the GScript First-Strand Synthesis Kit, cDNA synthesis kit, and dT oligonucleotide primers according to the manufacturer's instructions. RT-PCR was performed using ORA TM The PCR products were analyzed using SEEqPCR GreenROXLMix, 2X (HighQu, USA). The primers for EMT markers (vimentin, E-cadherin, TJP1, and FN1) and GAPDH (as an internal control) were synthesized by Bio-ProTech according to the sequences in Table 1. -ΔΔCt The relative fold change of gene expression was calculated by the method. The results showed that after treatment with compound (II), the mRNA expression levels of E-cadherin and TJP1 in LoVo parental cells and LoVo-OXAR cells increased, but the mRNA expression levels of vimentin and FN1 decreased ( Figure 5G and Figure 5H TJP1 is a marker for epithelial cells and is lost during EMT and cancer progression. In contrast, FN1 is involved in the development and progression of various tumors and is a key gene in gastric cancer. These data suggest that the coordination compound may help inhibit cancer cell invasion and therefore serve as an adjunct to current chemotherapy.

[0154] Table 1

[0155]

[0156]

[0157] Example 6: Treatment of colorectal cancer in mice using the coordination compound of formula (II)

[0158] 6.1 Inhibition of tumor growth in mice

[0159] Six-week-old male NU / NU mice were purchased from LoSco Biotech Co., Ltd. (Taiwan, China) and randomly divided into four groups (Groups 1 to 4), with three mice in each group. LoVo parental cells or LoVo-OXAR cells (1×10 6 Cells (100 μl of DMEM) were subcutaneously injected into the legs of mice (day 0). Starting from day 1 after tumor cell inoculation, mice were orally administered compound (II) or PBS (Table 2). Tumor volume and mouse body weight were measured every three days. Tumor volume was measured with a caliper and calculated as [(L×W×W) / 2], where L and W represent the length and width of the tumor, respectively. All mice were sacrificed on day 21, and the tumors were excised and weighed.

[0160] Table 2

[0161]

[0162] like Figure 6A and Figure 6B As shown, approximately three weeks after administration of compound (II), tumor growth in mice injected with LoVo-OXAR cells or LoVo parental cells was significantly reduced, demonstrating that the coordination compounds provided herein can treat individual cancers, including chemotherapy-resistant cancers. In addition, except for the group inoculated with chemotherapy-resistant tumor cells, the weight of the mice remained stable; the weight loss may be due to the malignancy of the chemotherapy-resistant tumors ( Figure 6C ). However, all mice were healthy after treatment with compound (II).

[0163] In addition, LoVo parental and LoVo-OXAR tumor tissues were collected for TUNEL analysis. The results showed that administration of compound (II) to mice induced significantly higher apoptosis in both LoVo parental and LoVo-OXAR tumors ( Figure 6D and Figure 6E ).

[0164] 6.2 Inhibition of cancer metastasis in mice

[0165] Six-week-old male NU / NU mice were purchased from LoSco Biotech Co., Ltd. (Taiwan, China) and randomly divided into six groups (Groups 1 to 6), with three mice in each group. LoVo parental cells or LoVo-OXAR cells (1×10 6 100 μl of DMEM) were injected into the tail vein of mice (day 0). Starting from day 1 after tumor cell inoculation, mice were orally administered compound (II) or PBS five days a week for three weeks (Table 3). All mice were sacrificed on day 30, and metastatic cancer cells were examined in the lungs, kidneys, spleens, and hearts of the mice.

[0166] Table 3

[0167]

[0168] like Figure 6F and Figure 6G As shown, high doses of compound (II) effectively inhibited the metastasis of LoVo parental cells or LoVo-OXAR cells to the lung, kidney, spleen, and heart compared to low dose treatment. This result reveals the potential of the coordination compound to prevent tumor development and metastasis.

[0169] Example 7: Cytotoxic effect of the coordination compound of formula (II) on liver cancer cells

[0170] 7.1 Cell culture

[0171] Human hepatocellular carcinoma HA22T cells (available from BCRC, catalog number BCRC60168; hereinafter referred to as HA22T parental cells) and their chemotherapy-resistant subclones were cultured in DMEM supplemented with 10% FBS. HA22T cells resistant to SAHA (an HDAC inhibitor) were established using a screening procedure similar to that described in Example 2.2. All cell cultures were performed at 37°C and 5% CO2.

[0172] 7.2 Effect of Compound (II) on the Activity of Hepatocellular Carcinoma Cells

[0173] HA22T parental cells and SAHA-resistant HA22T cells (abbreviated as HA22T-HDACiR cells) were treated with compound (II) or SAHA (1 μM or 3 μM) at different concentrations (0, 100, 250, 500, 750, 1000 or 1250 μg / ml) for 24 hours, and the cytotoxic activity of the compounds was determined by MTT assay. Figure 7As shown, compound (II) reduced the activity of the two liver cancer cells in a dose-dependent manner, indicating that compound (II) has the potential to treat liver cancer. The IC50 value of compound (II) against HA22T parental cells is about 1000 μg / ml.

[0174] Example 8: Cytotoxic effect of the coordination compound of formula (II) on prostate cancer cells

[0175] 8.1 Cell culture

[0176] Human prostate carcinoma LNCaP cells (available from ATCC, catalog number CRL-1740) were cultured in RPMI-1640 medium (Gibco TM , ThermoFisher Scientific, Waltham, MA, USA). Cell culture was performed at 37°C with 5% carbon dioxide supply.

[0177] 8.2 Effect of Compound (II) on Prostate Cancer Cell Activity

[0178] LNCaP cells were treated with compound (II) at different concentrations (0, 100, 200, 400, 800 or 1600 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 8 As shown, compound (II) reduces the activity of prostate cancer cells in a dose-dependent manner, indicating that compound (II) has the potential to treat prostate cancer. The IC50 value of compound (II) for LNCaP cells is about 1033 μg / ml.

[0179] Example 9: Cytotoxic and migration inhibitory effects of the coordination compound of formula (II) on osteosarcoma cells

[0180] 9.1 Cell culture

[0181] Human osteosarcoma 143B cells (available from ATCC, No. CRL-8303) were cultured in DMEM supplemented with 10% FBS and 1% sodium pyruvate at 37°C and 5% carbon dioxide.

[0182] 9.2 Effect of Compound (II) on Osteosarcoma Cell Activity

[0183] 143B cells were treated with compound (II) at different concentrations (0, 62, 125, 250, 500, 1000, 2000 or 4000 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 9A As shown in the results, compound (II) reduced the activity of osteosarcoma cells in a dose-dependent manner, indicating that compound (II) has the potential to treat bone cancer. The IC50 value of compound (II) for 143B cells is about 500 μg / ml. In addition, when compound (II) (500 μg / ml) is used in combination with 5-fluorouracil (20 μg / ml), gemcitabine (20 μg / ml), or amycin (50 μg / ml), the inhibition of 143B cell survival is significantly greater than that of any of the chemotherapy drugs used alone ( Figure 9B ), indicating that the coordination compounds can be used as adjuvant anticancer drugs for existing chemotherapy drugs.

[0184] 9.3 Inhibition of osteosarcoma cell crawling and epithelial-mesenchymal transition

[0185] To investigate how compound (II) affects the migration ability of bone cancer cells, 143B cells were subjected to the Transwell migration assay described in Example 5.1. Figure 9C As shown, compound (II) inhibited the crawling of 143B cells in a dose-dependent manner. In addition, Western blot analysis showed that treatment with compound (II) significantly reduced the expression of vimentin but increased the expression of E-cadherin ( Figure 9D ), indicating that the coordination compounds have the potential to inhibit the EMT process and the development of invasive cancer cells.

[0186] Example 10: Cytotoxic effect of the coordination compound of formula (II) on glioblastoma cells

[0187] 10.1 Cell culture

[0188] Human glioblastoma multiforme GBM 8401 cells (available from BCRC, catalog number BCRC 60163) were cultured in RPMI 1640 medium supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0189] 10.2 Effect of Compound (II) on Glioblastoma Cell Activity

[0190] GBM 8401 cells were treated with compound (II) at different concentrations (0, 200, 400, 800, 1000, 1200, 1400 or 1800 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 10 As shown, compound (II) reduced the activity of glioblastoma cells in a dose-dependent manner, indicating that compound (II) has the potential to treat brain cancer.

[0191] Example 11: Cytotoxic effect of the coordination compound of formula (II) on breast cancer cells

[0192] 11.1 Cell culture

[0193] Human breast cancer cells T-47D (available from ATCC, catalog number HTB-133) and MDA-MB231 (available from ATCC, catalog number HTB-26) were cultured in RPMI 1640 medium supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0194] 11.2 Effect of Compound (II) on the Activity of Breast Cancer Cells

[0195] Compound (II) was used to treat T-47D cells and MDA-MB231 cells at different concentrations (0, 500, 700, 1000, 2000, or 3000 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. The results showed that compound (II) reduced the activity of the two breast cancer cells in a dose-dependent manner, and its cytotoxic effect was more pronounced in MDA-MB231 cells ( Figure 11 MDA-MB231 is a highly invasive triple-negative breast cancer cell line characterized by epithelial-mesenchymal transition and resistance to amycin.

[0196] Example 12: Cytotoxic effect of the coordination compound of formula (II) on bladder cancer cells

[0197] 12.1 Cell Culture

[0198] Human bladder carcinoma T24 cells (available from ATCC, catalog number HTB-4) were cultured in RPMI 1640 medium supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0199] 12.2 Effect of Compound (II) on Bladder Cancer Cell Activity

[0200] T24 cells were treated with compound (II) at different concentrations (0, 100, 200, 400, 800 or 1600 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 12 As shown, compound (II) reduced the activity of bladder cancer cells in a dose-dependent manner, indicating that compound (II) has the potential to treat bladder cancer.

[0201] Example 13: Cytotoxic effect of the coordination compound of formula (II) on oral cancer cells

[0202] 13.1 Cell culture

[0203] Human oral squamous cell carcinoma (OSCC) SCC-25 cells (available from ATCC, catalog number CRL-1628) were cultured in DMEM / F12 medium supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0204] 13.2 Effect of Compound (II) on the Activity of Oral Cancer Cells

[0205] SCC-25 cells were treated with compound (II) at different concentrations (0, 100, 200, 400, 800, 1000, 1200 or 1400 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 13 As shown, compound (II) reduced the activity of OSCC cells in a dose-dependent manner, indicating that compound (II) has the potential to treat oral cancer.

[0206] Example 14: Cytotoxic effect of the coordination compound of formula (II) on ovarian cancer cells

[0207] 14.1 Cell Culture

[0208] Human ovarian endometrioid carcinoma CP70 cells (a cisplatin-resistant cell line provided by Dr. Huang Zhiyang of Tzu Chi Hospital in Hualien, Taiwan, and derived from the ovarian cancer cell line A2780, purchased from the European Collection of Authenticated Cell Cultures (ECACC), No. 93112519) were cultured in RPMI 1640 medium supplemented with 10% FBS. Cell culture was performed at 37°C and 5% carbon dioxide.

[0209] 14.2 Effect of Compound (II) on the Activity of Ovarian Cancer Cells

[0210] CP70 cells were treated with compound (II) at different concentrations (0, 100, 200, 400, 800 or 1600 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 14 As shown, compound (II) reduced the activity of ovarian cancer cells in a dose-dependent manner, indicating that compound (II) has the potential to treat ovarian cancer.

[0211] Example 15: Cytotoxic effect of the coordination compound of formula (II) on leukemia cells

[0212] 15.1 Cell Culture

[0213] Human T-cell acute lymphoblastic leukemia (T-lymphoblastic leukemia) Jurkat cells (cisplatin-resistant, available from ATCC, catalog number TIB-512) were cultured in RPMI 1640 medium supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0214] 15.2 Effect of Compound (II) on Leukemia Cell Activity

[0215] Jurkat cells were treated with compound (II) at different concentrations (0, 100, 200, 400, 800 or 1600 μg / ml) for 24 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 15 As shown, compound (II) reduced the activity of Jurkat cells in a dose-dependent manner, indicating that compound (II) has the potential to treat leukemia.

[0216] Example 16: Cytotoxic effect of the coordination compound of formula (II) on cervical cancer cells

[0217] 16.1 Cell Culture

[0218] Human cervical carcinoma HeLa cells (available from ATCC, catalog number CCL-2) were cultured in DMEM supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0219] 16.2 Effect of Compound (II) on the Activity of Cervical Cancer Cells

[0220] HeLa cells were treated with compound (II) at different concentrations (0.5, 1, 5, 10, 50, 100, 500 or 1000 μg / ml) for 48 hours, and the cytotoxic activity of the compound was determined by MTT assay. Figure 16 As shown, compound (II) reduced the activity of metastatic HeLa cells in a dose-dependent manner, indicating that compound (II) has the potential to treat cervical cancer (including metastatic cervical cancer).

[0221] Example 17: Cytotoxic effect of the coordination compound of formula (II) on thyroid cancer cells

[0222] 17.1 Cell Culture

[0223] Human medullary thyroid carcinoma TT cells (available from ATCC, catalog number CRL-1803) were cultured in F12K medium (Gibco TM , Thermo Fisher Scientific, Waltham, MA, USA). Cell culture was performed at 37°C with 5% CO2 supply.

[0224] 17.2 Effect of Compound (II) on the Activity of Thyroid Cancer Cells

[0225] TT cells were treated with compound (II) at different concentrations (0.1, 0.5, 1, 5, 10, 50, 100, 500 or 1000 μg / ml) for 72 hours, and the cytotoxic activity of compound (II) was determined by MTT assay. Figure 17 As shown, compound (II) reduced the activity of TT cells in a dose-dependent manner, indicating that compound (II) has the potential to treat thyroid cancer.

[0226] Example 18: Cytotoxic effect of the coordination compound of formula (II) on skin cancer cells

[0227] 18.1 Cell Culture

[0228] Human epidermoid carcinoma A431 cells (available from ATCC, catalog number CRL-1555) were cultured in DMEM supplemented with 10% FBS at 37°C and 5% carbon dioxide.

[0229] 18.2 Effect of Compound (II) on the Activity of Skin Cancer Cells

[0230] Compound (II) was used at different concentrations (1, 5, 10, 50, 100, 500 or 1000 μg / ml)

[0231] A431 cells were treated for 48 hours, and the cytotoxic activity of the compounds was determined by MTT assay. Figure 18 As shown, compound (II) reduced the activity of A431 cells in a dose-dependent manner, indicating that compound (II) has the potential to treat skin cancer.

[0232] Example 19: Cytotoxicity of the coordination compound of formula (II) against pancreatic cancer cells

[0233] 19.1 Cell Culture

[0234] Human pancreatic carcinoma MIAPaCa-2 cells (available from ATCC, catalog number CRL-1420) were cultured in DMEM supplemented with 10% FBS, 1 mM sodium pyruvate, and 2.5% horse serum. Human pancreatic epithelioid carcinoma PANC-1 cells (available from ATCC, catalog number CRL-1469) were cultured in DMEM supplemented with 10% FBS and 1 mM sodium pyruvate. All cell cultures were performed at 37°C with a 5% CO2 supply.

[0235] 19.2 Effect of Compound (II) on the Activity of Pancreatic Cancer Cells

[0236] MIAPaCa-2 cells and PANC-1 cells were treated with compound (II) at different concentrations (0, 31, 62.5, 125, 250, 500, 1000 or 2000 μg / ml) for 6 days, and the cytotoxic activity of the compound was determined by MTT assay. The results showed that compound (II) reduced the activity of the two pancreatic cancer cells in a dose-dependent manner ( Figure 19 ), indicating that compound (II) has the potential to treat pancreatic cancer. The IC50 values of compound (II) for MIAPaCa-2 cells and PANC-1 cells are approximately 660 μg / ml and approximately 1700 μg / ml, respectively.

[0237] Example 20: Treatment of pancreatic cancer in mice using the coordination compound of formula (II)

[0238] Female C.B17 / SCID mice were purchased from the Laboratory Animal Center (Taiwan, China) and randomly divided into five groups (Group 1 to Group 5), with seven mice in each group. PANC-1 cells (1×10 6 The cells were mixed with equal volumes of Matrigel (BD Biosciences, Franklin Lakes, New Jersey, USA) in 50 μl PBS and injected subcutaneously into the legs of mice (day 0). 3(Day 12), mice were orally administered compound (II) or PBS, or intravenously injected with Eli Lilly's Gemzar (gemcitabine for injection) for 14 days (Table 4). Tumor volume was monitored for another 14 days before all mice were sacrificed. Tumor volume was measured with a caliper and calculated as [(L×W×W) / 2], where L and W represent the length and width of the tumor, respectively. The tumor growth inhibition rate (TGI) was calculated as follows: inhibition rate (%) = [1-(Vt / Vc)]×100, where Vt refers to the net tumor growth of mice treated with the compound, and Vc refers to the net tumor growth of mice in the control group (not treated with the compound). The net tumor growth was calculated by deducting the tumor volume measured at the time of the first treatment from the tumor volume measured subsequently.

[0239] Table 4

[0240]

[0241] like Figure 20A As shown, there was no significant change in tumor growth in each group during the 14-day treatment period. However, tumor growth inhibition was observed 14 days after the end of the treatment period. According to Table 5, treatment with compound (II) inhibited tumor growth by approximately 51% to 88%, depending on the dosing regimen. Treatment with Gemzar (a commonly used chemotherapy drug for pancreatic cancer) inhibited tumor growth by approximately 112%. These data demonstrate that compound (II) can treat pancreatic tumors. In addition, the body weights of mice in groups 3 to 5 remained stable ( Figure 20B ), indicating that the therapeutically effective dose of compound (II) is not toxic to mice.

[0242] Table 5

[0243] Group TGI (%) weight changes 1 none 107±4％ 2 112±6％ 100±4％ 3 51±8％ 107±5％ 4 51±6％ 102±3％ 5 88±6％ 103±4％

[0244] In summary, the coordination compound provided herein can effectively inhibit the survival of a variety of cancer cells (including chemotherapy-resistant cancer cells), thereby inhibiting cancer cell proliferation and tumor growth. The coordination compound can also inhibit the migration and invasion of cancer cells, helping to inhibit tumor progression and metastasis. Therefore, the coordination compound can be used to prepare drugs for treating cancer, even metastatic cancer. In addition, the synergistic effect of the coordination compound with current chemotherapy drugs (such as oxaliplatin) shows that the compound can be used to prepare drug combinations to further enhance the effect of cancer treatment.

Claims

1. A coordination compound represented by formula (I): [XZ2(CH3CO2)6(H2O)4(OH)2]NO3 formula (I), wherein X is a trivalent ion of chromium (Cr) or molybdenum (Mo), and Z is a trivalent ion of iron (Fe), ruthenium (Ru), or osmium (Os).

2. A method for inhibiting the survival or proliferation of cancer cells, comprising contacting the cancer cells with an effective amount of the coordination compound according to claim 1.

3. The method of claim 2, wherein the coordination compound is represented by formula (II): [CrFe2(CH3CO2)6(H2O)4(OH)2]NO3 formula (II).

4. The method of claim 2, wherein the cancer is selected from colorectal cancer, lung cancer, liver cancer, pancreatic cancer, bone cancer, brain cancer, breast cancer, ovarian cancer, cervical cancer, prostate cancer, bladder cancer, blood cancer, stomach cancer, skin cancer, head and neck cancer, or thyroid cancer.

5. The method of any one of claims 2 to 4, wherein the cancer cells are invasive and / or resistant to chemotherapeutic agents.

6. The method of any one of claims 2 to 5, further comprising contacting the cancer cells with a chemotherapeutic agent selected from oxaliplatin, irinotecan, 5-fluorouracil, gemcitabine, emphysema, or any combination thereof.

7. The method of any one of claims 2 to 6, wherein the coordination compound induces endoplasmic reticulum stress-mediated apoptosis of the cancer cells.

8. A method for inhibiting cancer cell migration or invasion, comprising contacting the cancer cells with an effective amount of the coordination compound of claim 1.

9. The method of claim 8, wherein the coordination compound is represented by formula (II): [CrFe2(CH3CO2)6(H2O)4(OH)2]NO3 formula (II).

10. The method of claim 8, wherein the cancer is colorectal cancer or bone cancer.

11. The method of any one of claims 8 to 10, wherein the cancer cells are invasive and / or resistant to chemotherapeutic agents.

12. The method according to any one of claims 8 to 11, wherein the coordination compound inhibits epithelial-mesenchymal transition of the cancer cell.

13. A method for treating cancerous tumors, comprising administering an effective amount of the coordination compound of claim 1 to a subject in need thereof.

14. The method of claim 13, wherein the coordination compound is represented by formula (II): [CrFe2(CH3CO2)6(H2O)4(OH)2]NO3 formula (II).

15. The method of claim 13, wherein the cancer tumor is selected from colorectal cancer, lung cancer, liver cancer, pancreatic cancer, bone cancer, brain cancer, breast cancer, ovarian cancer, cervical cancer, prostate cancer, bladder cancer, blood cancer, stomach cancer, skin cancer, head and neck cancer, or thyroid cancer.

16. The method of any one of claims 13 to 15, wherein the cancerous tumor comprises cancer cells that are invasive and / or resistant to chemotherapeutic agents.

17. A pharmaceutical composition comprising an effective amount of the coordination compound according to claim 1 and a pharmaceutically acceptable carrier.

18. The pharmaceutical composition according to claim 17, wherein the coordination compound is represented by formula (II): [CrFe2(CH3CO2)6(H2O)4(OH)2]NO3 formula (II).

19. The pharmaceutical composition of claim 17, further comprising an additional pharmaceutically active agent.

20. The pharmaceutical composition of claim 19, wherein the additional pharmaceutically active agent is a chemotherapeutic agent selected from oxaliplatin, irinotecan, 5-fluorouracil, gemcitabine, emphysema, or any combination thereof.

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