Application of cobalt complex in preparation of medicine for treating pancreatic cancer

Abstract

The invention discloses application of a cobalt complex in preparation of a medicine for treating pancreatic cancer. According to the invention, a series of small molecule compounds are artificially synthesized through a machine learning algorithm assisted drug design, and the pancreatic cancer resisting effect of the small molecule compounds is evaluated in an animal model. The cobalt complex Co68 can effectively induce macrophages in a tumor microenvironment to secrete type I interferon, so that the tumor microenvironment of pancreatic cancer is remodeled. In addition, the Co68 remarkably increases immune infiltration of natural killer cells, killer CD8 + T cells and the like in the pancreatic cancer tumor microenvironment, and further enhances the anti-tumor effect. Therefore, the cobalt complex Co68 has a wide application prospect in the field of treatment of cold tumors such as pancreatic cancer.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and particularly relates to the application of a cobalt complex in the preparation of a medicine for treating pancreatic cancer. Background Art

[0002] Pancreatic cancer is known as the "king of cancer" because of its difficulty in early diagnosis, high drug resistance and low survival rate after surgery. According to statistics, about half a million people are diagnosed with pancreatic cancer each year worldwide, and the five-year survival rate of pancreatic cancer is usually only about 10%. The disease has no obvious symptoms in the early stages and is usually advanced when diagnosed, which makes patients miss the best time for surgery and leads to extremely limited intervention measures. Its unique biological characteristics give pancreatic cancer a strong invasive and metastatic ability. Forecasts show that in the next decade, pancreatic cancer will become the second most lethal tumor in cancer-related deaths.

[0003] Currently, the treatment of pancreatic cancer mainly includes surgical resection, chemotherapy, radiotherapy, and targeted therapy for EGFR, angiogenesis, tumor microenvironment and hypoxia. However, these treatment strategies have not significantly improved the long-term prognosis of patients, and the treatment of pancreatic cancer still faces great challenges. First, surgical resection is the only potential cure, but due to the dual biological characteristics of pancreatic cancer: on the one hand, the rapid growth and spread of tumor cells, and on the other hand, the complexity of the surrounding pancreatic structure, many patients have lost the opportunity for surgery when they are diagnosed. In addition, although chemotherapy and radiotherapy can prolong survival, multidrug resistance, toxic side effects and effects on the tumor microenvironment limit the efficacy of these treatment options. Based on these challenges, the focus of research has gradually shifted to immunotherapy. Although immunotherapy has made significant progress in other types of cancer, pancreatic cancer faces considerable obstacles due to its complex and highly immunosuppressive tumor microenvironment. The interaction between tumor-associated fibroblasts, immune cells and tumor cells in the tumor microenvironment leads to the escape of tumor cells from the immune system, which has prevented immunotherapy from producing breakthrough effects so far.

[0004] Therefore, the research and treatment of pancreatic cancer still face many challenges, and more basic and clinical research is needed to promote progress in the fight against this malignancy. Summary of the invention

[0005] In order to solve at least some of the technical problems in the above-mentioned prior art, the present invention uses a new convolutional neural network model, VDLIN, to screen small molecule compounds that can inhibit the positive feedback signal axis of IL-1β and PGE2 in the pancreatic cancer microenvironment. The small molecule compounds of the present invention can activate macrophages in the tumor microenvironment to secrete IFN-I, converting cold tumors into hot tumors, thereby providing new ideas and targeted drugs for the treatment of pancreatic cancer. Specifically, the present invention includes the following contents.

[0006] In a first aspect of the present invention, a cobalt complex Co68 is provided for use in the preparation of a drug for treating or improving pancreatic cancer, wherein the structural formula of the cobalt complex Co68 is shown in Formula I:

[0007]

[0008] In certain embodiments, the use of the cobalt complex Co68 according to the present invention in the preparation of a drug for treating or improving pancreatic cancer, wherein the tBu The PNP ligand and the cobalt salt react to obtain the cobalt complex Co68.

[0009] In certain embodiments, the use of the cobalt complex Co68 according to the present invention in the preparation of a drug for treating or improving pancreatic cancer, wherein the treatment or improvement includes at least one of the following:

[0010] (1) Inhibit the proliferation or growth of tumor cells;

[0011] (2) Kill tumor cells or induce apoptosis of tumor cells;

[0012] (3) reduce or decrease tumor cell volume and / or weight;

[0013] (4) Increase or enhance the number of immune cells and induce immune cells to enter the tumor microenvironment;

[0014] (5) Promote or enhance the tumor-killing function of immune cells;

[0015] (6) Modulating the immunosuppressive microenvironment;

[0016] (7) Inhibit the metastasis of pancreatic cancer;

[0017] (8) Prolong survival time.

[0018] In certain embodiments, according to the use of the cobalt complex Co68 described in the present invention in the preparation of a drug for treating or improving pancreatic cancer, the immune cells include one or more of natural killer cells, cytotoxic T lymphocytes, tumor infiltrating lymphocytes, macrophages, cytokine-induced killer cells, dendritic cells combined with cytokine-induced killer cells, chimeric antigen receptor T cells and T cell receptor chimeric T cells.

[0019] In a second aspect of the present invention, a pharmaceutical composition is provided, wherein the pharmaceutical composition comprises a cobalt complex Co68 and a pharmaceutically acceptable carrier, and the structural formula of the cobalt complex Co68 is shown in Formula I:

[0020]

[0021] In certain embodiments, according to the pharmaceutical composition of the present invention, the pharmaceutically acceptable carrier comprises at least one of a diluent, a filler, an absorbent, a wetting agent, a binder, a disintegrant, a lubricant, a sweetener, a preservative and an antioxidant.

[0022] The third aspect of the present invention provides the use of the cobalt complex Co68 in the preparation of a drug for activating or promoting type I interferon, wherein the structural formula of the cobalt complex Co68 is as shown in Formula I:

[0023]

[0024] In certain embodiments, the cobalt complex Co68 according to the present invention is used in the preparation of a drug for activating or promoting type I interferon, wherein the activation or promotion of type I interferon includes promoting or increasing the expression of interferon-related genes.

[0025] In a fourth aspect of the present invention, a cobalt complex Co68 is provided for use in the preparation of a drug for treating pancreatic cancer in combination with other drugs, wherein the structural formula of the cobalt complex Co68 is as shown in Formula I:

[0026]

[0027] In certain embodiments, the cobalt complex Co68 according to the present invention is used in the preparation of a drug for treating pancreatic cancer in combination with other drugs, wherein the other drugs include chemotherapeutic agents and / or immune checkpoint inhibitors.

[0028] The present invention discloses the use of a cobalt complex in the preparation of a drug for treating pancreatic cancer. The present invention artificially synthesizes a series of small molecule compounds through machine learning algorithm-assisted drug design, and evaluates their anti-pancreatic cancer effects in animal models. The cobalt complex Co68 of the present invention can effectively induce macrophages in the tumor microenvironment to secrete type I interferon, thereby reshaping the tumor microenvironment of pancreatic cancer. In addition, Co68 significantly increases the number of natural killer cells (such as but not limited to NK cells) and cytotoxic CD8 T cells in the pancreatic cancer tumor microenvironment. + The immune infiltration of T cells further enhances the anti-tumor effect. Therefore, the cobalt complex Co68 has broad application prospects in the treatment of cold tumors such as pancreatic cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The therapeutic effect of Co68 on pancreatic cancer in tumor-bearing mice and gene knockout mice is shown.

[0030] Figure 2 The therapeutic effect of Co68 on pancreatic cancer in knockout mice is shown.

[0031] Figure 3The results show that Co68 and other therapeutic agents act separately or together to treat pancreatic cancer in tumor-bearing mice.

[0032] Figure 4 Shown are the results of single-cell transcriptome analysis of PBS- and Co68-treated Pan02 tumor models.

[0033] Figure 5 The results of the study on the mechanism of action of Co68 and STING agonist DMXAA on pancreatic cancer are shown. DETAILED DESCRIPTION

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that the upper and lower limits of the scope and each intermediate value therebetween are specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as commonly understood by those of ordinary skill in the art to which the invention belongs. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0037] application

[0038] In one aspect of the present invention, a cobalt complex Co68 is provided for use in the preparation of a drug for treating or improving pancreatic cancer, wherein the structural formula of the cobalt complex Co68 is as shown in Formula I:

[0039]

[0040] The cobalt complex Co68 used in the present invention can be prepared by the method described in Chinese patent application CN107964028A, the content of which is incorporated herein by reference in its entirety. As an example, the cobalt complex in the present invention is prepared by the following method:

[0041] will dissolvetBu A solution of PNP ligand (0.361 g, 1.0 mmol) in tetrahydrofuran (THF) (5 ml) was added to a solution of cobalt dichloride (0.115 g, 0.90 mmol) in THF (5 ml). After the addition of the ligand, the reaction was stirred at room temperature (25°C) for 10 hours, and then the solution was concentrated to 2 ml, 15 ml of degassed n-hexane was added, and after stirring for 1 hour, argon was filtered, and the filter cake was dried to obtain a blue solid. The cobalt complex Co68 can be analyzed and verified by nuclear magnetic resonance and infrared determination. The cobalt complex Co68 used in the present invention tBu The PNP ligand is bis[2-(di-tert-butylphosphino)ethyl]amine, and the reaction equation is as follows:

[0042]

[0043] In the present invention, the treatment or improvement includes at least one of the following:

[0044] (1) Inhibit the proliferation or growth of tumor cells;

[0045] (2) Kill tumor cells or induce apoptosis of tumor cells;

[0046] (3) reduce or decrease tumor cell volume and / or weight;

[0047] (4) Increase or enhance the number of immune cells and induce immune cells to enter the tumor microenvironment;

[0048] (5) Promote or enhance the tumor-killing function of immune cells;

[0049] (6) Modulating the immunosuppressive microenvironment;

[0050] (7) Inhibit the metastasis of pancreatic cancer;

[0051] (8) Prolong survival time.

[0052] In the present invention, the term "treat or improve" refers to therapeutic treatment and preventive or prophylactic measures, the purpose of which is to prevent or slow down (reduce) undesirable physiological changes or disorders, such as the progression of tumors. Beneficial or desired clinical results include, but are not limited to, the following, whether detectable or undetectable, including relief of symptoms, reduction in disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, and relief (whether partial or complete). Those in need of treatment include those who already have cancer or tumor-related diseases or those who need to prevent or improve cancer or tumor-related diseases.

[0053] In the present invention, the immune cells include natural killer cells (such as but not limited to NK cells), cytotoxic T lymphocytes (such as but not limited to CD8+ T cells), tumor infiltrating lymphocytes, macrophages, cytokine-induced killer cells, dendritic cells combined with cytokine-induced killer cells, chimeric antigen receptor T cells and T cell receptor chimeric T cells.

[0054] A therapeutically effective amount of the cobalt complex Co68 of the present invention can be administered to a subject in need thereof, such as a mammal (eg, a human).

[0055] The administration mode of the drug of the present invention is not particularly limited, and representative administration modes include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical administration. Accordingly, the drug of the present invention can be prepared into various clinically acceptable dosage forms, including oral dosage forms, injection dosage forms, topical dosage forms or external dosage forms, etc.

[0056] The therapeutically effective amount of the present invention refers to the effective dosage considered in medicine, that is, the amount of the active compound (i.e., the cobalt complex Co68) is sufficient to significantly improve the condition without causing serious side effects. The daily dosage of the cobalt complex Co68 is usually 0.01-200 mg / Kg, preferably 0.5-200 mg / Kg or 0.5-180 mg / Kg, or 0.5-150 mg / Kg, or 0.5-120 mg / Kg, or 0.5-100 mg / Kg, or 0.5-50 mg / Kg, or 0.5-40 mg / Kg, or 0.5-30 mg / Kg, most preferably 0.5-25 mg / Kg. Exemplary effective dosages are, for example, 0.5 mg / Kg, 0.75 mg / Kg, 0.95 mg / Kg, 1 mg / Kg, 1.25 mg / Kg, 1.5 mg / Kg, 1.75 mg / Kg, 2 mg / Kg, 2.5 mg / Kg, 2.75 mg / Kg, 3 mg / Kg, 3.25 mg / Kg, 3.5 mg / Kg, 3.75 mg / Kg, 4 mg / Kg, 4.25 mg / Kg, 4.5 mg / Kg, 4.75 mg / Kg, 5 mg / Kg, 5.25 mg / Kg, 5.5 mg / Kg, 5.75 mg / Kg, 6 mg / Kg, 6.25 mg / Kg, 6.5 mg / Kg, 6.75 15mg / Kg, 16mg / Kg, 17mg / Kg, 18mg / Kg, 19mg / Kg, 20mg / Kg, 21mg / Kg, 22mg / Kg, 23mg / Kg, 24mg / Kg, 25mg / Kg. It can be administered as a single dose once a day, can be administered multiple times a day, or can be used at intervals.

[0057] Pharmaceutical composition

[0058] One aspect of the present invention provides a pharmaceutical composition comprising a cobalt complex Co68 and a pharmaceutically acceptable carrier.

[0059] In the present invention, "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or medium, which includes at least one of a diluent, a filler, an absorbent, a wetting agent, a binder, a disintegrant, a lubricant, a sweetener, a preservative and an antioxidant. Examples of diluents include, but are not limited to, physiological saline, aqueous buffer solutions, solvents, dispersion vehicles, etc.; fillers include, but are not limited to starch, lactose, mannitol, microcrystalline cellulose, etc.; absorbents include, but are not limited to, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents include, but are not limited to, water, ethanol, etc.; binders include, but are not limited to, hydroxypropyl methylcellulose, povidone, microcrystalline cellulose, etc.; disintegrants include, but are not limited to, cross-linked sodium carboxymethyl cellulose, cross-linked povidone, surfactants, low-substituted hydroxypropyl cellulose, etc.; lubricants include, but are not limited to, magnesium stearate, talc, polyethylene glycol, sodium lauryl sulfate, micropowdered silica gel, talc, etc.; sweeteners include, but are not limited to, sucralose, acesulfame, saccharin, sucrose, xylitol, mannitol, sorbitol, glucose, fructose, aspartame, etc.; preservatives include, but are not limited to, parabens, chlorobutanol, phenol, sorbic acid, etc.; antioxidants include, but are not limited to, ascorbic acid, methionine, etc.

[0060] In the present invention, pharmaceutically acceptable carriers participate in carrying or delivering a medicament from one organ or part of the body to another organ or part of the body. Each carrier is "acceptable" in the sense that it is compatible with the other ingredients of the formulation (e.g., Co68) and does not harm the patient.

[0061] The present invention further provides use of the cobalt complex Co68 in preparing a drug for activating or promoting type I interferon.

[0062] In a preferred embodiment, the activating or promoting type I interferon includes promoting or increasing the expression of interferon or its related genes.

[0063] In a preferred embodiment, the interferon or its related gene includes at least one of IFNA4, IFNB1 and EGR1.

[0064] Joint Application

[0065] One aspect of the present invention provides the use of the cobalt complex Co68 in the preparation of a drug for treating pancreatic cancer in combination with other drugs.

[0066] In the present invention, the other drugs include at least one of chemotherapeutic agents, immune checkpoint inhibitors, anti-tumor drugs and immunomodulators. Among them, chemotherapeutic agents include but are not limited to cisplatin, etoposide, floxuridine, capecitabine, mitomycin, methotrexate, vinblastine, cyclophosphamide, dacarbazine, ifosfamide, topotecan, docetaxel, temozolomide, paclitaxel, doxorubicin, camptothecin, etc.; immune checkpoint inhibitors include but are not limited to BK1 inhibitors, PDK1 inhibitors, CTLA-4 inhibitors, anti-PD-1 antibodies, etc.; anti-tumor drugs include but are not limited to gemcitabine, irinotecan, fluorouracil, oxaliplatin, carboplatin, etc.; immunomodulators include but are not limited to epidermal growth factor inhibitors, statins, metformin, angiotensin receptor blockers, thalidomide, lenalidomide, pomalidomide, prednisone, etc.

[0067] Example 1

[0068] This example shows the evaluation of the therapeutic effect of Co68 on pancreatic cancer in tumor-bearing mice and gene knockout mice.

[0069] 1. The therapeutic effect of Co68 on pancreatic cancer in C57BL / 6 mice

[0070] 1.1 Experimental methods

[0071] 5 × 10 5 PBS suspension of mouse pancreatic cancer cells (Pan02 cells) was added. When the tumor grew to a palpable size, tumors of equal size were selected and divided into two groups. The experimental group began to be intraperitoneally injected with Co68 in saline solution at a dose of 25 mg / kg, and the control group was intraperitoneally injected with an equal volume of solvent once every two days, and the size of the tumor was measured with a vernier caliper. After 15 days, the mice were killed with CO2, and the mice were dissected to obtain their complete tumors and photographed and recorded. Tumor survival curves were drawn according to the measured size, and the tumors were weighed, the volume was measured, and statistical analysis was performed. Experimental group n = 10, control group n = 10.

[0072] 1.2 Experimental Results

[0073] The tumor sizes in the experimental and control groups were Figure 1 A is shown in Figure 1. The tumor survival curve is drawn according to the measured tumor size, and the results are shown in Figure 1. Figure 1 The tumors were weighed, their volumes were measured and statistically analyzed. Figure 1 As shown in C and D.

[0074] The results showed that Co68 could significantly reduce the size, volume and weight of tumors in pancreatic cancer mice.

[0075] 2. The therapeutic effect of Co68 on pancreatic cancer in IFNAR KO C57BL / 6 mice

[0076] 2.1 Experimental methods

[0077] IFNAR KO C57BL / 6 mice aged 6-8 weeks were injected subcutaneously in the groin with 5×10 5 Pan02 cells were suspended in PBS. When the tumor grew to a palpable size, tumors of the same size were selected and divided into two groups. The experimental group began to be intraperitoneally injected with Co68 in saline solution at a dose of 25 mg / kg, and the control group was intraperitoneally injected with an equal volume of solvent once every two days, and the size of the tumor was measured with a vernier caliper. After 15 days, the mice were killed with CO2, and the mice were dissected to obtain their complete tumors and photographed and recorded. Tumor survival curves were drawn according to the measured size, and the tumors were weighed, measured in volume, and statistically analyzed. Experimental group n = 10, control group n = 10.

[0078] 2.2 Experimental Results

[0079] The tumor sizes in the experimental and control groups were Figure 1 The tumor survival curve was drawn according to the measured tumor size, and the results were shown in Figure 1 The tumors were weighed, their volumes were measured and statistically analyzed. Figure 1 G and H are shown.

[0080] The experimental results showed that Co68 had no significant therapeutic effect on pancreatic cancer tumors in IFNAR KO mice, indicating that Co68 plays a role in treating pancreatic cancer through the type I interferon pathway.

[0081] 3. The therapeutic effect of Co68 on pancreatic cancer in TLR4 KO C57BL / 6 mice

[0082] 3.1 Experimental methods

[0083] 5 × 10 TLR4 KO C57BL / 6 mice were injected subcutaneously in the groin of 6-8 week old mice. 5 Pan02 cells were suspended in PBS. When the tumor grew to a palpable size, tumors of equal size were selected and divided into two groups. The experimental group was injected with Co68 in saline solution at a dose of 25 mg / kg, and the control group was injected with an equal volume of solvent once every two days. After 15 days, the mice were killed with CO2, and the mice were dissected to obtain their complete tumors and photographed and recorded. The tumors were weighed, the volume was measured, and statistical analysis was performed. The experimental group n = 10, and the control group n = 10.

[0084] 3.2 Experimental Results

[0085] The tumor sizes in the experimental and control groups were Figure 2The tumors were weighed, their volumes were measured and statistically analyzed. Figure 2 As shown in B and C.

[0086] The experimental results showed that Co68 had no significant therapeutic effect on pancreatic cancer tumors in TLR4 KO mice, indicating that Co68 plays a therapeutic role in pancreatic cancer through the downstream signaling pathway mediated by the TLR4 receptor.

[0087] 4. The therapeutic effect of Co68 on pancreatic cancer in IRF3 KO C57BL / 6 mice

[0088] 4.1 Experimental methods

[0089] 5 × 10 5 Pan02 cells were suspended in PBS. When the tumor grew to a palpable size, tumors of equal size were selected and divided into two groups. The experimental group was injected with Co68 in saline solution at a dose of 25 mg / kg, and the control group was injected with an equal volume of solvent once every two days. After 15 days, the mice were killed with CO2, and the mice were dissected to obtain their complete tumors and photographed and recorded. The tumors were weighed, the volume was measured, and statistical analysis was performed. The experimental group n = 10, and the control group n = 10.

[0090] 4.2 Experimental Results

[0091] The tumor sizes in the experimental and control groups were Figure 2 The tumors were weighed, their volumes were measured and statistically analyzed. Figure 2 As shown in E and F.

[0092] The results showed that Co68 had no significant therapeutic effect on pancreatic cancer tumors in IRF3 KO mice, indicating that Co68 plays a therapeutic role in pancreatic cancer through the IRF3-mediated downstream interferon signaling pathway.

[0093] Example 2

[0094] This example shows the comparative effect of Co68 and other anti-tumor drugs in treating pancreatic cancer.

[0095] 1. Experimental Methods

[0096] 5 × 10 5PBS suspension of Pan02 cells was added. When the tumor grew to a palpable size, tumors of equal size were selected and divided into two groups. The experimental group began to be intraperitoneally injected with DMXAA in saline solution at a dose of 25 mg / kg, and the control group was intraperitoneally injected with an equal volume of solvent once every two days, and the size of the tumor was measured with a vernier caliper. After 15 days, the mice were killed with CO2, and the mice were dissected to obtain their complete tumors and photographed and recorded. The tumors were weighed, the volume was measured, and statistical analysis was performed. The tumor survival curve was drawn according to the measured size. Experimental group n = 10, control group n = 10.

[0097] 2. Experimental results

[0098] The tumor sizes in the experimental and control groups were Figure 3 The tumors were weighed, their volumes were measured and statistically analyzed. Figure 3 The tumor survival curve was drawn according to the measured tumor size, and the results were shown in Figure 3 As shown in C.

[0099] The experimental results showed that although DMXAA could significantly reduce the size, volume and weight of pancreatic cancer tumors in mice, Co68 had a significantly better therapeutic effect on pancreatic cancer than DMXAA.

[0100] Example 3

[0101] This example shows the effect of Co68 combined with other anti-tumor drugs in the treatment of pancreatic cancer.

[0102] 1. Experimental Methods

[0103] 5 × 10 5 PBS suspension of Pan02 cells was added. When the tumor grew to a palpable size, tumors of the same size were selected and divided into four groups: PBS group, Co68 treatment group, anti-PD-1 treatment group, and combined treatment (Combo, Co68 + anti-PD-1) group. The Co68 treatment group and the Combo group were intraperitoneally injected with Co68 saline solution at a dose of 25 mg / kg once every two days, and the PBS group was intraperitoneally injected with an equal volume of solvent at the same time; the anti-PD-1 group and the Combo group were intraperitoneally injected with 200 μg of anti-PD-1 on the 3rd, 7th, and 11th days, and the PBS group was intraperitoneally injected with 200 μg of IgG2a isotype antibody at the same time. The size of the tumor was measured with a vernier caliper every two days. After 15 days, the mice were killed with CO2, and the mice were dissected to obtain their complete tumors and photographed and recorded. The tumors were weighed, the volume was measured, and statistical analysis was performed. The tumor survival curve was drawn according to the measured size. N = 8 mice in each group.

[0104] 2. Experimental results

[0105] The tumor sizes in the experimental and control groups were Figure 3 The tumors were weighed, their volumes were measured and statistically analyzed. Figure 3 The tumor survival curve was drawn according to the measured tumor size, and the results were shown in Figure 3 As shown in F.

[0106] The results showed that the therapeutic effect of Co68 on pancreatic cancer was better than the anti-PD-1 antibody widely used in clinical practice, and the combined effect of Co68 and anti-PD-1 antibody in the treatment of pancreatic cancer was better than that of a single component.

[0107] Example 4

[0108] This example shows the mechanism of action of Co68 in treating pancreatic cancer.

[0109] 1. Single-cell transcriptome analysis

[0110] 1.1 Experimental methods

[0111] Single cell transcriptome analysis was performed on Pan02 tumor models treated with PBS and Co68. The specific steps include: A) using tSNE dimensionality reduction algorithm to reduce the dimension and cluster into 24 subgroups ( Figure 4 A) ; B) query the marker genes of different cell subsets for annotation ( Figure 4 CAFs represent tumor-associated fibroblasts, TAMs represent tumor-associated macrophages, and TANs represent tumor-associated neutrophils ( Figure 4 C) ; C) Macroscopic comparison of cell subpopulations in PBS group and Co68 group; D) Counting of the number of cells in each subpopulation; Flow cytometry was used to verify the results of single cell analysis ( Figure 4 F).

[0112] 1.2 Experimental Results

[0113] The results showed that TANs subsets, CAFs subsets and T cell subsets had significant differences ( Figure 4 D). The tumor subpopulation Cancer Cell 1 decreased in the Co68-treated group, while CD8 T cells and NK cells increased significantly in the Co68-treated group ( Figure 4 E). Compared with the PBS group, the CD8 T and NK cells in the Co68-treated group increased significantly, but the CD4 T cells decreased significantly. Among them, the CD8 T cells in the Co68 group upregulated more genes related to T cell activation, such as upregulating the expression of type II interferon ( Figure 4In addition, in addition to the significant upregulation of TAM in the Co68 group, the proportion of cells in the TANs1 group was also significantly upregulated after Co68 treatment ( Figure 4 H).

[0114] In this example, macrophage subpopulations in the pancreatic cancer microenvironment were sub-clustered and divided into IL1B high expression subpopulation, S100A10 high expression subpopulation and HSP high expression subpopulation ( Figure 4 By enrichment analysis of the IL1B subgroup with the largest number, it was found that the upregulated genes in the Co68 group were mainly in the signaling pathways related to type I interferon activation, and the downregulated genes were mainly genes related to cholesterol metabolism and synthesis ( Figure 4 In addition, through the analysis of cell communication in the subpopulation with high expression of IL1B, it was found that the Co68 group did significantly upregulate IFN-I related signals and reduce IL1B signals ( Figure 4 K), among which, the upregulated IFN-I signal in the Co68 group was mainly between the subpopulation with high expression of IL1B and CD4T, CD8T and endothelial cells ( Figure 4 of L).

[0115] 2. Comparative analysis of the effects of Co68 and STING agonist DMXAA on pancreatic cancer

[0116] Single-cell transcriptome analysis of Pan02 tumor models treated with DMXAA and Co68 showed that Co68 treatment could significantly affect the proportion of tumor-associated neutrophils (TANs). Figure 5 A). Co68 significantly upregulated the proportion and number of TANs1 subpopulation ( Figure 5 B). Further differential analysis revealed that Co68 significantly increased the expression of ISGs in the neutrophil subpopulation TANs1 ( Figure 5 C). Enrichment analysis found that Co68-induced ISGs high-expression TANs1 was significantly enriched in immune-related signaling pathways such as innate immune response activation, pattern receptor recognition, and IFNA response ( Figure 5 D). By detecting the transcriptome of single cells treated with Co68 and DMXAA, the dimensionality reduction clustering results of cell subpopulations were obtained, with a total of 21 subpopulations ( Figure 5 E). The cell subsets are annotated into major categories, including tumor cells, neutrophils, macrophages, and fibroblasts ( Figure 5 F). Sub-clustering of macrophages based on marker genes ( Figure 5 G). The macrophages in the tumor microenvironment were divided into four subgroups, such as the subgroup with high expression of IL1B, the proliferative subgroup with high expression of MKI67, etc. ( Figure 5By differential analysis of the subgroups with high expression of IL1B, it was found that Co68 treatment upregulated more genes related to type I interferon compared with the DMXAA group, but downregulated genes related to inflammation ( Figure 5 L) Through enrichment analysis of genes upregulated by Co68 compared with DMXAA treatment, it was found that the main signaling pathways were related to innate immune regulation, TNF production, and immune activation ( Figure 5 of L).

[0117] It can be seen that Co68 exerts its significant anti-tumor effect by upregulating the IFN-I signal of TAMs in the tumor microenvironment and reducing the IL1B signal produced by TAMs, thereby cutting off the positive feedback signal axis between IL1B of TAMs and PGE2 of pancreatic cancer tumor cells.

[0118] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of the cobalt complex Co68 in the preparation of a drug for treating or improving pancreatic cancer, characterized in that: The structural formula of the cobalt complex Co68 is shown in Formula I:

2. The use of the cobalt complex Co68 according to claim 1 in the preparation of a drug for treating or improving pancreatic cancer, characterized in that: By making tBu The PNP ligand and the cobalt salt react to obtain the cobalt complex Co68.

3. The use of the cobalt complex Co68 according to claim 1 in the preparation of a drug for treating or improving pancreatic cancer, characterized in that: The treatment or improvement includes at least one of the following: (1) Inhibit the proliferation or growth of tumor cells; (2) Kill tumor cells or induce apoptosis of tumor cells; (3) reduce or decrease tumor cell volume and / or weight; (4) Increase or enhance the number of immune cells and induce immune cells to enter the tumor microenvironment; (5) Promote or enhance the tumor-killing function of immune cells; (6) Modulating the immunosuppressive microenvironment; (7) Inhibit the metastasis of pancreatic cancer; (8) Prolong survival time.

4. The use of the cobalt complex Co68 according to claim 3 in the preparation of a drug for treating or improving pancreatic cancer, characterized in that: The immune cells include one or more of natural killer cells, cytotoxic T lymphocytes, tumor infiltrating lymphocytes, macrophages, cytokine-induced killer cells, dendritic cells combined with cytokine-induced killer cells, chimeric antigen receptor T cells and T cell receptor chimeric T cells.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises a cobalt complex Co68 and a pharmaceutically acceptable carrier, and the structural formula of the cobalt complex Co68 is shown in Formula I:

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutically acceptable carrier includes at least one of a diluent, a filler, an absorbent, a wetting agent, a binder, a disintegrant, a lubricant, a sweetener, a preservative and an antioxidant.

7. Use of the cobalt complex Co68 in the preparation of a drug for activating or promoting type I interferon, characterized in that: The structural formula of the cobalt complex Co68 is shown in Formula I:

8. The use according to claim 7, characterized in that The activating or promoting type I interferon includes promoting or increasing the expression of interferon.

9. Use of the cobalt complex Co68 in the preparation of a drug for treating pancreatic cancer in combination with other drugs, characterized in that: The structural formula of the cobalt complex Co68 is shown in Formula I:

10. Use of the cobalt complex Co68 according to claim 9 in the preparation of a drug for treating pancreatic cancer in combination with other drugs, characterized in that: The other drugs include chemotherapeutic agents and / or immune checkpoint inhibitors.

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