Pharmaceutical composition and application thereof

Abstract

The invention discloses a pharmaceutical composition, which contains carboxamide triazole and bisphosphate which have the same or different specifications and are administered simultaneously or respectively, and the bisphosphate is at least one of clodronate sodium, alendronate sodium, risedronate sodium and zoledronic acid. Researches show that the anti-tumor and anti-inflammatory effects of the carboxamidotriazole combined with the diphosphate are better than those of single drugs DES and ZOA; meanwhile, compared with the single administration of the zoledronic acid with the concentration of 0.1 mg / kg or the single administration of the CAI with the concentration of 40 mg / kg and 80 mg / kg, the combined administration of the zoledronic acid with the concentration of 0.1 mg / kg and the combined administration of the CAI with the concentration of 40 mg / kg and 80 mg / kg can significantly inhibit the growth and proliferation of human lung cancer A549-luc xenotransplantation tumors in bone metastasis of B-NDG mice, and has an unexpected combined synergistic effect. And the zoledronic acid and CAI combined administration group can significantly prolong the survival time of the human lung cancer A549-luc cells in a non-small cell lung cancer bone metastasis model animal established in a B-NDG mouse. The traditional Chinese medicine composition has the effects of remarkable combined synergism and less toxic and side effects.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a pharmaceutical composition and use thereof. Background Art

[0002] Carboxyamide triazole is the world's first small molecule immunotherapy drug that regulates the inflammatory microenvironment of tumors. It has regulatory effects on multiple cytokines (TNF-α, IL-6, etc.) and multiple signaling pathways (NF-κB and MAPKs) in the inflammatory microenvironment. On the one hand, carboxyamide triazole regulates the tumor microenvironment to exert anti-tumor effects by downregulating the production of cytokines such as TNF-α and IL-6 in tumor-associated macrophages (TAMs). The applicant's previous in vivo and in vitro tests have shown that carboxyamide triazole has a significant inhibitory effect on dozens of solid tumors such as lung cancer, breast cancer, liver cancer, kidney cancer, and ovarian cancer; and the data from the Phase III clinical trial of 495 first-line advanced non-small cell lung cancer patients showed that the objective efficacy of the carboxyamide triazole test group increased by 38.4%; and it can reduce the risk of disease progression by 31%. The clinical trial successfully achieved the preset primary endpoint, with significant efficacy, few adverse reactions, and high safety. On the other hand, carboxyamide triazole plays an anti-autoimmune role by inhibiting the activity of NF-κB and MAPKs and reducing the release of inflammatory cytokines such as TNF-α and IL-6 in inflammatory tissues. It can be used to treat autoimmune diseases such as rheumatoid arthritis, psoriasis, and inflammatory bowel disease. Therefore, carboxyamide triazole has a broad-spectrum application prospect in anti-tumor and anti-autoimmune diseases.

[0003] Bisphosphonates (BPs) are a new class of drugs for treating metabolic bone diseases developed in the past 20 years. They have a significant inhibitory effect on bone resorption and are mainly used to prevent and treat bone metabolic diseases such as osteoporosis, osteitis deformans, and cancerous bone pain and hypercalcemia caused by bone metastasis of malignant tumors. Bisphosphonates can be divided into three generations based on the time of their launch and the strength of their effects. The first generation comprises nitrogen-free bisphosphonates synthesized in the 1970s and 1980s, such as etidronate and clodronate. However, they suffer from significant toxic side effects, and long-term use can lead to abnormal bone mineralization, some of which can cause rickets or induce pathological fractures. The second generation comprises nitrogen-containing bisphosphonates, such as pamidronate and tiludronate, which contain amino groups in their molecular structures, significantly enhancing their anti-bone resorption effects. However, their toxic side effects include local tissue necrosis and thrombophlebitis at the injection site. The third generation comprises nitrogen-containing imidazole derivatives with heterocyclic structures, which were launched in the early 2000s and include clodronate, alendronate, risedronate, and zoledronic acid. They have the strongest anti-bone resorption effects with fewer toxic side effects.

[0004] Zoledronic acid, a third-generation bisphosphonate, is clinically used primarily to treat cancerous bone metastases and metabolic bone diseases. Studies have shown that zoledronic acid primarily acts on osteoclasts. Zoledronic acid not only directly adsorbs onto the surface of trabecular bone, preventing osteoclast damage and bone resorption, but also directly interferes with osteoclast growth, inducing apoptosis and ultimately lowering blood calcium levels. Zoledronic acid also inhibits the synthesis and release of pain-inducing substances such as prostaglandins and inflammatory mediators in osteoclasts, thereby alleviating bone pain.

[0005] Currently, zoledronic acid has limited clinical efficacy as a single agent for the treatment of cancer bone metastasis. It is highly toxic when used in combination with chemotherapy drugs, and its combination with targeted drugs is mainly targeted at patients with specific target mutations, with a small audience. Moreover, zoledronic acid alone cannot directly inhibit the occurrence, development, and metastasis of tumor cells. Therefore, it is necessary to develop other combination drugs to obtain a combination drug composition with better efficacy and higher safety. Summary of the Invention

[0006] The present invention aims to overcome at least one of the above-mentioned defects of the prior art and provide a pharmaceutical composition to achieve the purpose of combined synergy with fewer toxic and side effects.

[0007] A pharmaceutical composition contains carboxyamidotriazole and bisphosphonate of the same or different strengths, which are administered simultaneously or separately. The bisphosphonate is at least one of clodronate sodium, alendronate sodium, risedronate sodium and zoledronic acid.

[0008] Preferably, the bisphosphonate is zoledronic acid.

[0009] Furthermore, the dosage of carboxyamide triazole for adults is 1-10 mg / kg; the dosage of zoledronic acid for adults is 2-4 mg / time.

[0010] Furthermore, the mass ratio of the carboxyamide triazole to zoledronic acid is 15 to 150:1.

[0011] Furthermore, the pharmaceutical composition of the present invention contains carboxyamidotriazole and zoledronic acid in unit preparations of the same or different specifications for simultaneous or separate administration, and a pharmaceutically acceptable carrier.

[0012] Preferably, the pharmaceutical composition is an oral or external preparation.

[0013] Furthermore, the oral preparation is one of tablets, capsules, and granule solid dispersions.

[0014] Furthermore, the external preparation is one of a cream, an ointment, a gel, and a patch.

[0015] The present invention also protects the use of the pharmaceutical composition as described above for treating solid tumors, wherein the solid tumor is at least one of lung cancer, liver cancer, breast cancer, colon cancer, prostate cancer, and kidney cancer.

[0016] The present invention also protects the use of the pharmaceutical composition as described above for treating bone metastasis of solid tumors, wherein the solid tumor is at least one of lung cancer, liver cancer, breast cancer, colon cancer, prostate cancer, and kidney cancer.

[0017] The present invention also protects the use of the pharmaceutical composition as described above for anti-inflammatory purposes. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is the TRAP staining image of Raw264.7 cells in the vehicle control group in Example 1.

[0019] Figure 2 This is the TRAP staining image of Raw264.7 cells in the LPS-induced model group in Example 1.

[0020] Figure 3 This is a TRAP staining image of Raw264.7 cells in the denosumab positive control group in Example 1.

[0021] Figure 4 This is a TRAP staining image of Raw264.7 cells in the zoledronic acid positive control group in Example 1.

[0022] Figure 5 This is the TRAP staining image of Raw264.7 cells in the CAI 10 μM group in Example 1.

[0023] Figure 6 This is the TRAP staining image of Raw264.7 cells in the CAI 40 μM group in Example 1.

[0024] Figure 7 This is the TRAP staining image of Raw264.7 cells in the CAI 80 μM group in Example 1.

[0025] Figure 8 This is the TRAP staining image of Raw264.7 cells in the CAI10μM+Dishu group in Example 1

[0026] Figure 9 This is the TRAP staining image of Raw264.7 cells in the CAI 40 μM + Dishu group in Example 1.

[0027] Figure 10 This is the TRAP staining image of Raw264.7 cells in the CAI 80 μM + Dishu group in Example 1.

[0028] Figure 11This is the TRAP staining image of Raw264.7 cells in the CAI 10 μM + zoledronic acid group in Example 1.

[0029] Figure 12 This is the TRAP staining image of Raw264.7 cells in the CAI 40 μM + zoledronic acid group in Example 1.

[0030] Figure 13 This is the TRAP staining image of Raw264.7 cells in the CAI 80μM + zoledronic acid group in Example 1

[0031] Figure 14 This is a statistical chart of TRAP cell counts in Raw264.7 cells after combined use of CAI and denosumab in Example 1.

[0032] Figure 15 This is a statistical chart of TRAP cell counts in Raw264.7 cells after combined use of CAI and zoledronic acid in Example 1.

[0033] Figure 16 TNF-a content in Raw264.7 cells after combined use of CAI and denosumab in Example 1.

[0034] Figure 17 TNF-a content in Raw264.7 cells after combined use of CAI and zoledronic acid in Example 1.

[0035] Figure 18 This is a line graph of the total bioluminescent flux of the test substance in Example 2 in the A549-luc xenograft tumor model.

[0036] Figure 19 This is a line graph of the relative total bioluminescence flux of the test substance in Example 2 in the A549-luc xenograft tumor model.

[0037] Figure 20 This is a bioluminescent total flux image of the test substance in Example 2 in the A549-luc xenograft tumor model.

[0038] Figure 21 This is a line graph of the body weight of the test substance in Example 2 in the A549-luc xenograft tumor model.

[0039] Figure 22 This is a line graph of the body weight change rate of the test substance in Example 2 in the A549-luc xenograft tumor model.

[0040] Figure 23 This is the Kaplan-Meiyer curve obtained from the statistics of Example 2. DETAILED DESCRIPTION

[0041] The drawings in the embodiments provide a more detailed description of the technical solutions in the embodiments of the present invention. In the drawings, the same or similar reference numerals throughout represent the same or similar elements or elements with the same or similar functions. The described embodiments are part of the embodiments of the present invention, not all of them. The embodiments described below with reference to the drawings are exemplary and are intended to be used to explain the present invention, and should not be understood as limiting the present invention. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. The embodiments of the present invention are described in detail below with reference to the drawings.

[0042] It should be noted that if the embodiments of the present application involve directional indications (such as up, down, left, right, front, back, etc.), the directional indications are only used to explain the relative position relationship, movement status, etc. between the various components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indications will also change accordingly.

[0043] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present application, the descriptions of "first", "second", etc. are only for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of such features. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the fact that they can be implemented by ordinary technicians in this field. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0044] Examples 1 and 2 analyzed the effects of carboxyamidotriazole (CAI) and denosumab (DES) or zoledronic acid (ZOA) alone or in combination on lipopolysaccharide (LPS)-induced osteoclast differentiation of RAW264.7 cells and tumor necrosis factor-a (TNF-a).

[0045] Mouse monocyte-macrophage RAW264.7 cells were purchased from Gibco Biotechnology. The cell culture medium used was Gibco, supplemented with 10% fetal bovine serum. The ELISA kit was purchased from Ecosai Biotechnology Co., Ltd., lot number 022C-0808A. The TRAP kit was commercially available, lot number CR2305034, manufactured by Servicebio.

[0046] Test Substance Information Sheet

[0047] CAI was homemade, and zoledronic acid, denosumab, vinorelbine, and cisplatin were purchased commercially. The test substance information is as follows:

[0048] Detection indicators:

[0049] Osteoclast formation: tartrate acid phosphatase staining (TRAP staining) was used to count the number of osteoclasts; tumor necrosis factor-α in the cell supernatant was detected by ELISA. Example 1

[0050] This example mainly examines the effects of three doses of CAI (10 μM), CAI (40 μM), and CAI (80 μM) on tumor necrosis factor α (TNF-α) and osteoclast formation in RAW264.7 cells induced by LPS to differentiate into osteoclasts.

[0051] RAW264.7 cells were induced to differentiate into osteoclasts using LPS, and the induction results were identified by tartrate-resistant acid phosphatase (TRAP) staining. RAW264.7 cells were cultured in groups; the blank control group (CTRL) received no intervention, the model group was treated with LPS (150 ng / mL), and the other eight groups were treated with 10, 40, and 80 μM CAI; 0.1 mg / mL DES; 1 μM ZOA; CAI (10 μM) + DES; CAI (40 μM) + DES; CAI (80 μM) + DES; CAI (10 μM) + ZOA; CAI (40 μM) + ZOA; CAI (80 μM) + ZOA. On day 5 of culture, tumor necrosis factor-α (TNF-α) levels in the supernatant were measured by ELISA, and osteoclast formation was observed in the remaining cell plates using TRAP staining.

[0052] The experimental data were statistically analyzed using SPSS 13.0, and the images were analyzed using GraphPad Prism 6 software. The statistical method of factor variance analysis was used, and the experimental results were expressed as mean ± standard deviation. =Indicates the difference between groups. The t-test was used for comparison. *P < 0.05 indicates a significant difference, **P < 0.01 indicates a relatively significant difference. For tartrate-resistant acid phosphatase staining, five randomly selected fields of view in each group were photographed under a microscope, and positive multinucleated cells (≥ 3 nuclei) were counted.

[0053] Test results

[0054] TRAP staining test results (x100) Figures 1 to 13 The cell counting results are shown in Table 1 and Figures 14-15 The statistical results of TNF-α are shown in Table 2 and Figures 16-17 shown. Table 1 TRAP cell counts in Raw264.7 cells after CAI and denosumab / zoledronic acid were used alone or in combination Note: Experimental data are expressed as mean ± standard deviation (x ± SD). * There was a statistically significant difference compared with the LPS group Table 2 TNF-a levels in Raw264.7 cells after single or combined use of CAI and denosumab / zoledronic acid Note: Experimental data are expressed as mean ± standard deviation (x ± SD). * There was a statistically significant difference compared with the LPS group

[0055] TRAP staining results showed that CAI-10μM, 40μM, and 80μM significantly reduced LPS (150ng / ml)-induced osteoclast counts. The inhibitory effect of CAI-10μM on osteoclasts was comparable to that of DES and ZOA alone, with CAI-40μM exhibiting statistically significant superiority over ZOA. CAI-80μM significantly reduced osteoclast counts compared to DES and ZOA, with statistically significant differences. The combination of CAI-10μM with DES or ZOA showed no significant advantage. The combination of CAI-40μM and 80μM with DES or ZOA was more effective than DES or ZOA alone. ELISA results for inflammatory factors showed that CAI-10μM, 40μM, and 80μM significantly reduced TNF-α levels. Compared with DES and ZOA, CAI-10μM and 40μM showed no significant advantage. CAI-80μM exhibited lower TNF-α levels, but this was not statistically significant. There was no obvious advantage after the combination of CAI-10μM, and the TNF-α content was also lower after the combination of CAI-40μM and 80μM, and there was a statistical difference.

[0056] The experimental results showed that single-agent CAI-10μM, 40μM, and 80μM could inhibit LPS-induced osteoclast formation, and the single-agent effects or combined use of CAI-40μM and 80μM were superior to the pharmacodynamic effects of single-agent DES and ZOA; the anti-inflammatory effects of CAI-80μM and its combination were significantly better than those of single-agent DES and ZOA. Example 2

[0057] This example uses a human lung cancer A549-luc cell B-NDG mouse bone metastasis model to evaluate the antitumor activity of carboxyamidotriazole (CAI) combined with zoledronic acid.

[0058] A total of 100 animals were inoculated in this experiment, with 64 animals divided into groups. Immediately after grouping, the test substance, zoledronic acid, was administered via tail vein injection three times per week for nine consecutive doses (IV, TIW × 3 weeks); the test substance, denosumab, was administered via tail vein injection twice per week for six consecutive doses (IV, BIW × 3 weeks); the test substance, vinorelbine, was administered via intraperitoneal injection once per week for three consecutive doses (IP, QW × 3 weeks); the test substance, cisplatin, was administered via tail vein injection once per week for three consecutive doses (IV, QW × 3 weeks); and the test substance, CAI, was administered orally once daily for 21 consecutive doses (PO, QD × 21). Animals were euthanized by CO2 if they died, met animal welfare criteria, or reached the end of the experiment (Day 60), and tumor tissues were collected.

[0059] Evaluation indicators:

[0060] Bioluminescence (Radiance) is the number of photons emitted per pixel per A549-luc cell, expressed in photons / second / cm² / steradian. Specific values ​​were quantified using Living Image Software.

[0061] Total bioluminescence flux (Total Flux) is the sum of the bioluminescence values ​​of all pixels within the selected region (ROI), expressed in photons / second. The total bioluminescence flux at the tumor inoculation site is used in the following calculations.

[0062] The tumor growth inhibition rate (%TGITF) was calculated as follows: %TGITF = (TFC-TFT) / TFC × 100%, where TFC is the average total bioluminescence flux of the negative control group and TFT is the average total bioluminescence flux of the treatment group;

[0063] The relative total flux (RTF) was calculated as follows: RTF = TFt / TF0, where TF0 is the total bioluminescence flux at the time of grouping and TFt is the total bioluminescence flux at each measurement.

[0064] The relative tumor proliferation rate (%T / CRTF) was calculated as follows: %T / CRTF = TRTF / CRTF × 100%, where TRTF is the average RTF of the treatment group and CRTF is the average RTF of the negative control group;

[0065] The median survival time (MST) is the survival time corresponding to when the cumulative survival rate is 50% (the day of group administration is recorded as Day 0). The specific value is calculated by GraphPad Prism 6 software (San Diego, CA, USA);

[0066] The relative life extension rate (%T / CMST) was calculated as follows: TMST / CMST × 100%, where TMST is the MST of the treatment group and CMST is the MST of the negative control group;

[0067] The calculation formula of animal body weight change rate (%BWC) is: (BWt-BW0) / BW0×100%, where BWt is the animal weight at each measurement and BW0 is the animal weight at grouping.

[0068] Statistical analysis

[0069] In this study, experimental data were expressed as Mean ± SEM.

[0070] Tumor growth curves were plotted with time as the X-axis and the common logarithm of total bioluminescence flux (p / s) (lgTF) as the Y-axis. Animal weight change curves were plotted with time as the X-axis and animal weight (g) as the Y-axis. Comparisons between groups were performed using a two-tailed t-test, with P < 0.05 considered significant and P < 0.01 considered extremely significant (GraphPad Prism 6).

[0071] The Kaplan-Meier method was used to draw survival curves, and the log-rank test was used for comparison between groups. P < 0.05 was considered a significant difference (GraphPad Prism 6).

[0072] Evaluation of antitumor activity of test substances

[0073] In this experiment, the effects of each test substance alone or in combination on the total tumor bioluminescence flux of human lung cancer A549-luc cells in the non-small cell lung cancer bone metastasis model established in B-NDG mice are shown in Table 5. Figures 18-20 shown. Table 5. Antitumor activity of the test substances in the A549-luc xenograft tumor model

[0074] *: P < 0.05 compared with the model group, ***: P < 0.001 compared with the model group, &: P < 0.05 compared with the zoledronic acid 0.1 mg / kg group, &&: P < 0.01 compared with the zoledronic acid 0.1 mg / kg group, ###: P < 0.001 compared with the CAI HD group.

[0075] After the administration on Day 21, the average total bioluminescence flux of the zoledronic acid 0.1 mg / kg single-dose group was 7.75×10 9 ±2.07×10 9p / s, tumor growth inhibition rate %TGI TF The total bioluminescence flux of the vehicle control group (2.03×10 10 ±1.93×10 9 There was a significant difference between the two groups (P<0.001); the relative tumor proliferation rate %T / C RTF It was 37.08% (P<0.001).

[0076] The average total bioluminescence flux of the denosumab 10 mg / kg single-dose group was 1.35×10 10 ±1.52×10 9 p / s, %TGI TF The total bioluminescence flux of the 2477 cells was 33.23%, which was significantly different from that of the vehicle control group (P<0.05); %T / C RTF It was 67.70% (P<0.05).

[0077] The average total bioluminescence flux of the 4 mg / kg vinorelbine and 2 mg / kg cisplatin combined group was 1.54×10 10 ±2.62×10 9 p / s, %TGI TF The total bioluminescence flux of the 24.09% group was not significantly different from that of the vehicle control group (P>0.05); %T / C RTF It was 74.58% (P>0.05).

[0078] The average total bioluminescence flux of the CAI 40 mg / kg single-dose group was 1.40×10 10 ±1.61×10 9 p / s, %TGI TF The total bioluminescence flux of the 2477 cells was 31.01%, which was significantly different from that of the vehicle control group (P<0.05); %T / C RTF It was 69.80% (P<0.05).

[0079] The average total bioluminescence flux of the zoledronic acid 0.1 mg / kg and CAI 40 mg / kg combined administration group was 4.75×10 9 ±5.13×10 8 p / s, %TGI TF The total bioluminescence flux of the 2477 cells was 76.56%, which was significantly different from that of the vehicle control group (P<0.001); %T / C RTF It was 25.25% (P<0.001).

[0080] The average total bioluminescence flux of the denosumab 10 mg / kg and CAI 40 mg / kg combined group was 1.36×1010 ±1.85×10 9 p / s, %TGI TF The total bioluminescence flux of the 247 cells was 32.87%, which was significantly different from that of the vehicle control group (P<0.05); %T / C RTF It was 69.47% (P<0.05).

[0081] The average total bioluminescence flux of the 4 mg / kg vinorelbine, 2 mg / kg cisplatin, and 40 mg / kg combined administration group was 1.40 × 10 10 ±3.17×10 9 p / s, %TGI TF The total bioluminescence flux of the 2477 cells was 31.16%, which was not significantly different from that of the vehicle control group (P>0.05); %T / C RTF It was 65.08% (P<0.05).

[0082] After Day 28, animal deaths occurred and some animals were removed from the experimental group for animal welfare reasons. Therefore, the data from Day 28 were selected for statistical analysis.

[0083] On Day 28, the average total bioluminescence flux of the zoledronic acid 0.1 mg / kg single-dose group was 1.16×10 10 ±3.11×10 9 p / s, tumor growth inhibition rate %TGI TF The total bioluminescence flux was 40.21% compared with the average total bioluminescence flux of the vehicle control group (1.93×10 10 ±2.83×10 9 There was no significant difference between the two groups (P>0.05); the relative tumor proliferation rate %T / C RTF It was 56.70% (P<0.05).

[0084] The average total bioluminescence flux of the denosumab 10 mg / kg single-dose group was 1.71×10 10 ±2.21×10 9 p / s, %TGI TF The total bioluminescence flux of the 10-well plate was 11.43%, which was not significantly different from that of the vehicle control group (P>0.05); %T / C RTF It was 92.09% (P>0.05).

[0085] The average total bioluminescence flux of the 4 mg / kg vinorelbine and 2 mg / kg cisplatin combined group was 1.29×10 10 ±2.50×10 9 p / s, %TGI TFThe total bioluminescence flux of the 2477 cells was 33.50%, which had no significant difference compared with the average total bioluminescence flux of the vehicle control group (P>0.05); %T / C RTF 67.46% (P>0.05)

[0086] The average total bioluminescence flux of the CAI 40 mg / kg single-dose group was 1.43×10 10 ±1.55×10 9 p / s, %TGI TF The total bioluminescence flux of the 2477 cells was 26.14%, which was not significantly different from that of the vehicle control group (P>0.05); %T / C RTF It was 75.89% (P>0.05).

[0087] The average total bioluminescence flux of the zoledronic acid 0.1 mg / kg and CAI 40 mg / kg combined administration group was 3.52×10 9 ±3.57×10 8 p / s, %TGI TF The total bioluminescence flux of the 2-well plate was 81.80%, which was significantly different from that of the vehicle control group (P<0.001); %T / C RTF It was 20.64% (P<0.001).

[0088] The average total bioluminescence flux of the denosumab 10 mg / kg and CAI 40 mg / kg combined group was 1.43×10 10 ±2.09×10 9 p / s, %TGI TF The total bioluminescence flux of the 2477 cells was 26.32%, which was not significantly different from that of the vehicle control group (P>0.05); %T / C RTF It was 79.03% (P>0.05).

[0089] The average total bioluminescence flux of the 4 mg / kg vinorelbine, 2 mg / kg cisplatin, and 40 mg / kg combined administration group was 1.11×10 10 ±1.69×10 9 p / s, %TGI TF The relative tumor proliferation rate (%T / C) was 42.66%, which was significantly different from the average total bioluminescence flux of the vehicle control group (P<0.05). RTF It was 57.44% (P<0.05).

[0090] In summary, the combined administration of zoledronic acid 0.1 mg / kg and CAI 40 mg / kg can significantly inhibit the growth and proliferation of human lung cancer A549-luc xenograft tumors in bone metastases of B-NDG mice compared with the administration of zoledronic acid 0.1 mg / kg alone or CAI 40 mg / kg alone (P<0.01, P<0.001), showing a combined synergistic effect.

[0091] Effects of the test substances on the body weight of tumor-bearing animals

[0092] In this experiment, the effects of the test substances alone or in combination on the body weight of the non-small cell lung cancer bone metastasis model established in B-NDG mice using human lung cancer A549-luc cells are shown in Table 6. Figures 21-22 After Day 28, animal deaths occurred and some animals were removed from the experimental group for animal welfare reasons. Therefore, the data from Day 28 were selected for statistical analysis.

[0093] The experimental results showed that on Day 28, the average animal weight of the vehicle control group, the zoledronic acid alone group, the denosumab alone group, the vinorelbine and cisplatin combination group, the CAI alone group, the zoledronic acid and CAI combination group, the denosumab and CAI combination group, and the vinorelbine, cisplatin, and CAI combination group decreased by 2.45%-10.03% (0.56g-2.24g) compared with the start of the experiment on Day 0. No drug-related animal deaths were observed during the experiment. Table 6 Effects of the test substances on animal body weight in the A549-luc xenograft tumor model

[0094] Evaluation of life-extending activity after administration of the test substance

[0095] In this experiment, the survival analysis results of each experimental group are shown in Table 7 and Figure 23 .

[0096] On Day 35, animals began to die or reached the animal welfare endpoint. At the end of the experiment (Day 60), the MST of the vehicle control group was 47.5 days, with a survival rate of 1 / 8.

[0097] The MST of the zoledronic acid alone group was 53 days, the survival rate was 1 / 8, and the life extension rate was 112%, which was not significantly different from that of the vehicle control group (P>0.05).

[0098] The MST of the denosumab monotherapy group was 51 days, the survival rate was 0 / 8, and the life extension rate was 107%, which was not significantly different from that of the vehicle control group (P>0.05).

[0099] The MST of the vinorelbine and cisplatin combination group was 49.5 days, the survival rate was 2 / 8, and the life extension rate was 104%, which was not significantly different from that of the vehicle control group (P>0.05).

[0100] The MST of the CAI alone administration group was 54 days, the survival rate was 1 / 8, and the life extension rate was 114%, which had no significant difference compared with the solvent control group (P>0.05).

[0101] In the zoledronic acid and CAI combined administration group, the MST exceeded 60 days and the survival rate was 5 / 8, which was significantly different from that in the vehicle control group (P<0.05).

[0102] The MST of the denosumab and CAI combined administration group was 56 days, the survival rate was 1 / 8, and the life extension rate was 118%, which was not significantly different from the solvent control group (P>0.05).

[0103] The MST of the vinorelbine, cisplatin and CAI combined administration group was 56.5 days, the survival rate was 3 / 8, and the life extension rate was 119%, which had no significant difference compared with the solvent control group (P>0.05).

[0104] The experimental results showed that the combined administration of zoledronic acid and CAI could significantly prolong the survival time of animals in the non-small cell lung cancer bone metastasis model established in B-NDG mice with human lung cancer A549-luc cells. Table 7. Effects of the test substances on the life-extending activity of the A549-luc xenograft tumor model

[0105] *:P<0.05: compared with the model group

[0106] The results above demonstrate that the combined anti-tumor and anti-inflammatory effects of carboxyamidotriazole and zoledronic acid are superior to those of single-agent DES and ZOA. Furthermore, the combined administration of zoledronic acid 0.1 mg / kg and CAI 40 mg / kg significantly inhibited the growth and proliferation of bone metastases of human lung cancer A549-luc xenografts in B-NDG mice compared with either zoledronic acid 0.1 mg / kg or CAI 40 mg / kg alone (P<0.01, P<0.001), demonstrating an unexpected synergistic effect. Furthermore, the combined administration of zoledronic acid and CAI significantly prolonged the survival of animals in the B-NDG mouse model of non-small cell lung cancer bone metastasis established with human lung cancer A549-luc cells.

[0107] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the above preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent replacements of the technical solutions of the present invention may be made without departing from the spirit and scope of the technical solutions of the present invention. Those skilled in the art may also make other changes within the spirit of the present invention and apply them to the design of the present invention, as long as they do not deviate from the technical effects of the present invention. These changes made in accordance with the spirit of the present invention should all be included in the scope of protection claimed by the present invention.

Claims

1. A pharmaceutical composition, characterized in that The invention contains carboxyamide triazole and bisphosphonate of the same or different specifications for simultaneous or separate administration, wherein the bisphosphonate is at least one of clodronate sodium, alendronate sodium, risedronate sodium and zoledronic acid.

2. The pharmaceutical composition according to claim 1, characterized in that The bisphosphonate is zoledronic acid.

3. The pharmaceutical composition according to claim 2, characterized in that The dosage of the carboxyamide triazole for adults is 1-10 mg / kg; the dosage of zoledronic acid for adults is 2-4 mg / time.

4. The pharmaceutical composition according to claim 1, characterized in that The mass ratio of the carboxyamide triazole to zoledronic acid is 15 to 150:

1.

5. The pharmaceutical composition according to claim 1, characterized in that The invention contains carboxyamidotriazole and zoledronic acid in unit preparations of the same or different specifications for simultaneous or separate administration, and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical composition is an oral or external preparation.

7. The pharmaceutical composition according to claim 5, characterized in that The oral preparation is one of tablets, capsules, and granule solid dispersions; the external preparation is one of creams, ointments, gels, and patches.

8. Use of the pharmaceutical composition according to any one of claims 1 to 7 for treating solid tumors, characterized in that: The solid tumor is at least one of lung cancer, liver cancer, breast cancer, colon cancer, prostate cancer, and kidney cancer.

9. Use of the pharmaceutical composition according to claim 1 for treating bone metastasis of solid tumors, characterized in that: The solid tumor is at least one of lung cancer, liver cancer, breast cancer, colon cancer, prostate cancer, and kidney cancer.

10. Use of the pharmaceutical composition according to any one of claims 1 to 7 for anti-inflammatory purposes.

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