A pharmaceutical composition for improving the treatment effect of pancreatic cancer
By inducing ICD in pancreatic cancer cells through ES-CU combined with L-OHP and activating the immune activity of DCs, the problem of poor efficacy of existing pancreatic cancer immunotherapy has been solved, achieving significant anti-tumor effects and a low-cost treatment plan.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- ZHEJIANG CANCER HOSPITAL
- Filing Date
- 2026-01-26
- Publication Date
- 2026-05-15
AI Technical Summary
Existing technologies are insufficient to effectively activate the immune system of pancreatic cancer patients, resulting in high recurrence rates after chemotherapy and surgery. Traditional immune-enhancing therapies have short-lived effects, and in vitro engineered DCs therapy is costly and difficult to promote.
Elisclomol (ES) and copper chloride (CuCl2) combined with oxaliplatin (L-OHP) were used to induce immunogenic cell death (ICD) in tumor cells, activate the immune activity of dendritic cells (DCs), and form thermal tumors to enhance the anti-tumor immune response.
It significantly inhibits pancreatic cancer tumor progression, activates DCs and CD8+ T cells, increases the apoptosis rate of pancreatic cancer cells, enhances anti-tumor immune response, and reduces the risk of tumor recurrence.
Smart Images
Figure CN121550254B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of cancer drugs, specifically relating to a pharmaceutical composition that enhances the therapeutic effect of pancreatic cancer. Background Technology
[0002] Pancreatic cancer is a malignant tumor with an extremely poor prognosis. Pancreatic ductal adenocarcinoma (PDAC) is the main type of pancreatic cancer. In recent years, the incidence and mortality rates of pancreatic cancer have been rising globally, posing a serious threat to public health. Chemotherapy and surgery remain the main treatment options for pancreatic cancer. However, approximately 80-85% of pancreatic cancer patients have distant metastases or other reasons that make the primary lesion unresectable. Even among the small percentage of pancreatic cancer patients with R0 surgical resection opportunities, nearly three-quarters will relapse within two years after surgery, indicating that even patients undergoing surgical resection may still have micrometastases. Therefore, the prognosis for pancreatic cancer patients with surgical options remains very poor, with a 5-year survival rate of only 20%.
[0003] To actively improve the prognosis of pancreatic cancer patients, medical researchers and frontline clinicians have been actively exploring various systemic therapies, including improved surgery, combined radiotherapy and chemotherapy, the development of targeted therapy, and immunotherapy regimens. However, these efforts have only provided limited improvements in the survival rate of pancreatic cancer patients. The main reason for this is that pancreatic cancer develops an immunosuppressive tumor microenvironment (TME), making it resistant to traditional targeted and cytotoxic therapies (such as radiotherapy). Even with immunomodulatory therapies targeting T cells (such as PD-1 antibodies), the response to pancreatic cancer is always short-lived.
[0004] Immunotherapy using immune checkpoint antibodies to directly enhance T cells appears insufficient for long-term improvement of immunosuppressive tumor cell endothelial growth factor (TME). A 2024 study by Krishnan K et al. in *Science* showed that activation of conventional dendritic cells (DCs) can promote immunotherapy for pancreatic cancer, and tumor antigen-stimulated in vitro engineered DCs can serve as an immune vaccine to enhance the sensitivity of pancreatic cancer to immunotherapy. However, the acquisition of tumor antigen-stimulated in vitro engineered DCs is limited by patient condition and sample size; only a portion of patients can yield sufficient quantities of qualified DCs for in vitro immune activation. Furthermore, in vitro engineered DC therapy is expensive, hindering widespread adoption. Recent studies have shown that antigens released during tumor cell immunogenic cell death (ICD) can promote DC maturation and induce CD8+. + T-cell immune responses promote tumor immunotherapy and inhibit tumor progression and metastasis. Summary of the Invention
[0005] This invention induces the intracellular diffusion cell (ICD) of tumor cells, activates the immune activity of dendritic cells (DCs), transforms cold pancreatic cancer tumors into hot pancreatic cancer tumors, and triggers a strong anti-tumor immune response to achieve the desired immunotherapy effect for pancreatic cancer. Finding a suitable tumor ICD inducer to activate DCs is crucial. This invention uses a mixture of elixclomol (ES) and copper chloride (Elesclomol-CuCl2, ES-CU) combined with oxaliplatin (L-OHP) to induce ICD in pancreatic cancer cells to achieve the goal of pancreatic cancer immunotherapy.
[0006] The purpose of this invention is to provide a pharmaceutical composition that can induce ICD in tumor cells, activate the immune activity of DCs, and transform cold pancreatic cancer tumors into hot pancreatic cancer tumors, thereby improving the therapeutic effect of pancreatic cancer.
[0007] The technical solution adopted by the present invention to achieve the above objectives is as follows:
[0008] A pharmaceutical composition comprising L-OHP, ES, and CU, wherein the mass ratio of L-OHP to ES is 1:0.0005-5, and the mass ratio of ES to CU is 1:0.05-0.5. This invention provides a simpler and more effective way to activate autologous tumor immunity. Given the limited efficacy of current immune checkpoint inhibitors targeting T cells alone and the difficulty in widely promoting in vitro engineered DC therapy, this invention's pharmaceutical composition can bridge the gap between innate and adaptive immunity through chemotherapy, significantly activating tumor immunity and achieving better therapeutic effects. The pharmaceutical compositions of this invention are all based on combinations of existing chemotherapeutic drugs, compounds, and chemical substances, resulting in low cost, high safety, and ease of promotion.
[0009] Preferably, the pharmaceutical composition further contains a medical solvent.
[0010] More preferably, the medical solvent includes at least one of ethanol, propylene glycol, glycerin, 1,3-butanediol, polyethylene glycol, and physiological saline.
[0011] More preferably, the concentration of L-OHP is 4-200 μg / mL.
[0012] Preferably, the pharmaceutical composition further contains an aminosalicylic acid derivative, which is prepared by reacting 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole. In addition to using L-OHP, ES, and CU, this invention can further incorporate an aminosalicylic acid derivative having the structures of acetamidosalicylic acid and aminoethylindole. The combined use of L-OHP, ES, CU, and the aminosalicylic acid derivative can increase the apoptosis rate of pancreatic cancer cells and simultaneously enhance hsp90 expression, exhibiting a good inhibitory effect on pancreatic cancer.
[0013] More preferably, in the preparation of the aminosalicylic acid derivative, the amount of 3-(2-aminoethyl)indole used is 80-120 wt% of 4-acetamidosalicylic acid.
[0014] More preferably, the mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.005-0.03.
[0015] More preferably, the aminosalicylic acid derivative is prepared using the condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; or, the aminosalicylic acid derivative is prepared using the catalyst 1-hydroxybenzotriazole.
[0016] Preferably, in the preparation of the aminosalicylic acid derivative, 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole are added to dichloromethane, followed by the addition of condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and catalyst 1-hydroxybenzotriazole, and then triethylamine. The reaction is carried out at a temperature of 20-40°C for 4-16 hours. After adding water, the layers are separated. The organic phase is washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution, then dried with anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel chromatography to obtain the aminosalicylic acid derivative.
[0017] More preferably, in the preparation of aminosalicylic acid derivatives, the amount of 4-acetamidosalicylic acid used is 1-5 wt% of dichloromethane.
[0018] More preferably, in the preparation of the aminosalicylic acid derivative, the amount of 3-(2-aminoethyl)indole used is 80-120 wt% of 4-acetamidosalicylic acid.
[0019] More preferably, in the preparation of the aminosalicylic acid derivative, the amount of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride used is 100-150 wt% of 4-acetaminosalicylic acid.
[0020] More preferably, in the preparation of the aminosalicylic acid derivative, the amount of 1-hydroxybenzotriazole used is 80-100 wt% of 4-acetamidosalicylic acid.
[0021] More preferably, in the preparation of the aminosalicylic acid derivative, the amount of triethylamine used is 100-150 wt% of 4-acetamidosalicylic acid.
[0022] More preferably, in the preparation of the aminosalicylic acid derivative, the purification reagent in the silica gel chromatography is a mixture of dichloromethane and methanol, wherein the dichloromethane and methanol are mixed in a volume ratio of 1:0.01-0.05.
[0023] Preferably, the pharmaceutical composition comprises L-OHP, ES, and CU. The mass ratio of L-OHP to ES is 1:0.0005-5, and the mass ratio of ES to CU is 1:0.05-0.5.
[0024] More preferably, the pharmaceutical composition further includes an aminosalicylic acid derivative, wherein the mass ratio of L-OHP to the aminosalicylic acid derivative is 1:0.005-0.03.
[0025] More preferably, the pharmaceutical composition further includes 6-hydroxynicotinic acid, with the mass ratio of L-OHP to 6-hydroxynicotinic acid being 1:0.001-0.006. When L-OHP, ES, and CU are used as pharmaceutical compositions, an aminosalicylic acid derivative can be added; furthermore, 6-hydroxynicotinic acid can be added. The combined use of 6-hydroxynicotinic acid and the above pharmaceutical composition can further increase the cell apoptosis rate of pancreatic cancer and further increase the expression of hsp90, thus exhibiting a good inhibitory effect on pancreatic cancer.
[0026] Preferably, the mass ratio of L-OHP to ES is 1:0.079, and the mass ratio of ES to CU is 1:0.34.
[0027] The present invention discloses an injection solution comprising the above-described pharmaceutical composition.
[0028] This invention discloses the use of the above-mentioned pharmaceutical composition in the preparation of a medicament that enhances the therapeutic effect of pancreatic cancer.
[0029] The beneficial effects of this invention include:
[0030] Significantly inhibits the progression of subcutaneous pancreatic cancer xenografts: In a mouse model of subcutaneous pancreatic cancer xenografts, ES-CU combined with L-OHP significantly inhibited the progression of pancreatic cancer tumors and promoted cancer cell death compared with L-OHP alone.
[0031] Activating anti-tumor immunity in subcutaneous pancreatic cancer xenografts: In a mouse model of subcutaneous pancreatic cancer xenografts, ES-CU combined with L-OHP induced activation of dendritic cells (DCs), immune infiltration, and CD8 activation. + T-cell infiltration.
[0032] In vitro, it has a significant killing effect on pancreatic cancer cell lines: compared with L-OHP alone, ES-CU combined with L-OHP can increase the killing effect on cells, specifically manifested as inhibited proliferation, increased apoptosis, and decreased migration ability of pancreatic cancer cell lines (mouse cell line Panc02 and human cell line ASPC-1).
[0033] Significantly induces ICD in pancreatic cancer: Compared with L-OHP monotherapy and ES-CU, ES-CU combined with L-OHP can significantly induce ICD in pancreatic cancer cell lines (mouse cell line Panc02 and human cell line ASPC-1), manifested by increased expression of ICD-related proteins and increased release of DAMPs after pancreatic cancer cell death.
[0034] In vitro, it can significantly activate DCs and CD8. + T cells: Compared with L-OHP monotherapy and ES-CU, ES-CU combined with L-OHP induces the release of multiple inducing factors from ICDs, enhancing the immune activity of DCs, and presenting antigens to T cells, promoting CD8. + T cell activation enhances anti-tumor immunotherapy.
[0035] This invention is a pharmaceutical composition that can induce ICD in tumor cells, activate the immune activity of DCs, and transform cold pancreatic cancer tumors into hot pancreatic cancer tumors, thereby enhancing the therapeutic effect of pancreatic cancer. Attached Figure Description
[0036] Figure 1 This is a data graph showing the treatment of subcutaneous pancreatic cancer xenografts in mice using ES-CU combined with L-OHP.
[0037] Figure 2 This image shows the effect of ES-CU combined with L-OHP in treating subcutaneous pancreatic cancer xenografts in mice.
[0038] Figure 3 The image shows the effect of ES-CU combined with L-OHP on activating anti-tumor immunity in mouse subcutaneous pancreatic cancer xenografts.
[0039] Figure 4 This image shows the effect of ES-CU combined with L-OHP on inhibiting the proliferation and migration of pancreatic cancer cell lines.
[0040] Figure 5 This figure shows the effect of ES-CU combined with L-OHP in inhibiting the proliferation and migration of pancreatic cancer cell lines and promoting apoptosis.
[0041] Figure 6 Figure 1 shows an experiment on the inhibition of pancreatic cancer cell line migration by ES-CU combined with L-OHP.
[0042] Figure 7This is a protein expression diagram of ICD induced in pancreatic cancer cell lines by ES-CU combined with L-OHP.
[0043] Figure 8 This image shows the effect of ES-CU combined with L-OHP in inducing ICD in pancreatic cancer cell lines.
[0044] Figure 9 This is a flow cytometry result of pancreatic cancer cell lines induced by ES-CU combined with L-OHP.
[0045] Figure 10 An activation index map for pancreatic cancer ICD induced by ES-CU combined with L-OHP to promote DC activation.
[0046] Figure 11 A diagram of inflammatory cytokines associated with ES-CU combined with L-OHP-induced pancreatic cancer ICD to promote DC activation.
[0047] Figure 12 A signaling pathway diagram for ES-CU combined with L-OHP to induce pancreatic cancer ICD to promote DC activation.
[0048] Figure 13 This image shows the effect of ES-CU combined with L-OHP on enhancing CD8+ T cell activity after promoting DC activation.
[0049] Figure 14 This is a graph showing the cell apoptosis rate.
[0050] Figure 15 This is a graph representing HSP90. Detailed Implementation
[0051] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0052] The concepts involved in this application will first be described with reference to the accompanying drawings. It should be noted that the following descriptions of various concepts are only for the purpose of making the content of this application easier to understand and do not constitute a limitation on the scope of protection of this application; furthermore, the embodiments and features in the embodiments of this application can be combined with each other unless otherwise specified. This application will now be described in detail with reference to the accompanying drawings and embodiments.
[0053] The basic research content of this invention is as follows:
[0054] Study 1. Efficacy of ES-CU combined with L-OHP in treating subcutaneous pancreatic cancer xenografts in mice.
[0055] Six C57BL / 6J mice in each group were subcutaneously inoculated with 1×10⁶ murine pancreatic cancer cell line Panc02 cells. 6 Tumors formed approximately 4 weeks after the tumor developed, with LOHP 4 mg / kg / animal and ES (10 mg / kg / animal) and CU (1 mg / kg / animal) administered intraperitoneally every 4 days. Tumor size was measured and recorded every 4 days. Figure 1 As shown in Figure A, the mice in the ES-CU combined with L-OHP group experienced a slightly more significant decrease in body weight, but all were within 10%. Figure 1 As shown in Figure B, in terms of tumor growth rate, the combination therapy group significantly inhibited tumor progression, such as... Figure 1 C and Figure 2 As shown in Figure A; HE pathological staining in the combined drug group also indicated that the tumor tissue damage was more severe in the combined drug group, such as... Figure 2 As shown in Figure B; and the proportion of cancer cells undergoing apoptosis is higher, such as Figure 1 As shown in Figure C; the above results indicate that ES-CU combined with L-OHP is significantly effective in treating subcutaneous pancreatic cancer xenografts in mice.
[0056] Study 2. ES-CU combined with L-OHP activates anti-tumor immunity in mouse subcutaneous pancreatic cancer xenografts.
[0057] Based on the multicolor immunofluorescence staining results of mouse tumor tissues from Study 1, we can see that the key ICD protein CALR was significantly upregulated in the ES-CU combined with L-OHP group. Figure 3 As shown in Figure A; simultaneously, DC-related activators (CD11c, CD80, CD86) were significantly upregulated, and CD8... + The proportion of T cell infiltration also increased, such as Figure 3 As shown in Figure B, this suggests that ES-CU combined with L-OHP can significantly activate the anti-tumor immunity of subcutaneous pancreatic cancer xenografts in mice.
[0058] Study 3. ES-CU combined with L-OHP inhibits the proliferation and migration of pancreatic cancer cell lines and promotes apoptosis.
[0059] The pancreatic cancer cell lines were Panc02 and ASPC-1. The Panc02 and ASPC-1 pancreatic cancer cell lines were divided into two groups of 1×10⁻⁶ cells / year. 5Cells / well were seeded in 6-well plates. The next day, the cells were treated with a combination of ES-CU and L-OHP. The in vitro drug intervention dosage for Panc02 was: L-OHP: 100 μg / ml, ES: 78.9 ng / ml, CU: 26.89 ng / ml. L-OHP, ES, and CU were added sequentially without pre-mixing. The intervention time was 48 hours. The in vitro drug intervention dosage for ASPC-1 was: L-OHP: 20 μg / ml, ES: 15.78 ng / ml, CU: 5.38 ng / ml. L-OHP, ES, and CU were added sequentially without pre-mixing. The intervention time was 48 hours. Results showed that the combined administration of ES-CU and L-OHP significantly inhibited the proliferation of pancreatic cancer cell lines. Figure 4 As shown in Figure A; the mortality rate of each cell line also increased significantly, such as... Figure 4 China BC and Figure 5 As shown in Figure A; the migration ability of the cell line was also significantly inhibited, such as... Figure 5 China BC and Figure 6 As shown in Figures AB, the above results indicate that ES-CU combined with L-OHP can significantly inhibit the proliferation and migration of pancreatic cancer cell lines and promote apoptosis.
[0060] Study 4. ES-CU combined with L-OHP induces ICD in pancreatic cancer cell lines.
[0061] Following the cell line culture and intervention procedures outlined in Study 3, and through relevant experimental techniques, we found that the ICD-related proteins were significantly upregulated in the ES-CU combined with L-OHP group. Figure 7 As shown in Figure A; compared with ES-CU monotherapy and L-OHP monotherapy, the combination therapy group showed a significant increase in the key ICD protein CALR, as shown in Figure A. Figure 7 As shown in BC; the levels of ATP and HMGB1, two key factors released after ICD, were also significantly increased in the cell supernatant, such as Figure 8 As shown in Figure A; the combination therapy group also showed upregulation of ICD-related genes compared to other groups, such as... Figure 8 As shown in Figure B; when ROS in each group was detected by immunofluorescence and flow cytometry, the accumulation of ROS required to induce ICD was also observed. The immunofluorescence results are shown in Figure B. Figure 8 As shown in Figure C, the flow cytometry results are as follows: Figure 9 As shown, this suggests that ES-CU combined with L-OHP induces ICD in pancreatic cancer cell lines.
[0062] Study 5. ES-CU combined with L-OHP induces pancreatic cancer ICD to promote DC activation.
[0063] After 48 hours of intervention, the supernatant of Panc02 cell culture following intervention induced by the DC culture was mixed with DC culture medium at a 1:1 ratio. This study found that DCs showed a significant upregulation of activation markers (CD80 and CD86), such as... Figure 10 As shown in AC, the levels of related inflammatory cytokines IL-1β, IL-18, and TNF-α were significantly increased in the cell supernatant, such as... Figure 11 As shown; the signaling pathways that activate DCs are mainly NF-κB and MAPK, such as NF-κB Figure 12 As shown in A, MAPK is as follows Figure 12 As shown in Figure B. The above results indicate that S-CU combined with L-OHP induces pancreatic cancer ICDs to promote DC activation.
[0064] Study 6. ES-CU combined with L-OHP promotes enhanced CD8 activation after DC activation. + T cell activity
[0065] In our study of co-culturing activated DCs with T cells, we found that, compared with ES-CU monotherapy and L-OHP monotherapy, the activated DCs significantly increased CD8 levels. + T cell ratio, such as Figure 6 As shown in A; CD8 + The proportion of activated T cells (mainly CD25 and CD69) also increased significantly, such as Figure 6 As shown in BC; this suggests that ES-CU combined with L-OHP promotes the activation of DCs and enhances CD8. + T cell activity.
[0066] Example 1: A pharmaceutical composition
[0067] Pharmaceutical composition: L-OHP, ES, and CU. The mass ratio of L-OHP to ES is 1:0.079, and the mass ratio of ES to CU is 1:0.34.
[0068] Example 2: A pharmaceutical composition
[0069] Preparation of aminosalicylic acid derivatives: 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole were added to dichloromethane, followed by the addition of condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and catalyst 1-hydroxybenzotriazole, and then triethylamine. The mixture was reacted at 30°C for 8 hours. After adding water, the mixture was separated into layers. The organic phase was washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution, dried with anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel chromatography to obtain aminosalicylic acid derivatives. The amounts of 4-acetaminosalicylic acid used were 3 wt% of dichloromethane, 100 wt% of 3-(2-aminoethyl)indole, 120 wt% of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 90 wt% of 1-hydroxybenzotriazole, and 130 wt% of triethylamine. In silica gel chromatography purification, the purification reagent was a mixture of dichloromethane and methanol, used at a volume ratio of 1:0.03.
[0070] Pharmaceutical composition: L-OHP, ES, CU and aminosalicylic acid derivative. The mass ratio of L-OHP to ES is 1:0.079, the mass ratio of ES to CU is 1:0.34, and the mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.02.
[0071] Example 3: A pharmaceutical composition
[0072] Preparation of aminosalicylic acid derivatives: 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole were added to dichloromethane, followed by the addition of condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and catalyst 1-hydroxybenzotriazole, and then triethylamine. The mixture was reacted at 30°C for 8 hours. After adding water, the mixture was separated into layers. The organic phase was washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution, dried with anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel chromatography to obtain aminosalicylic acid derivatives. The amounts of 4-acetaminosalicylic acid used were 3 wt% of dichloromethane, 100 wt% of 3-(2-aminoethyl)indole, 120 wt% of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 90 wt% of 1-hydroxybenzotriazole, and 130 wt% of triethylamine. In silica gel chromatography purification, the purification reagent was a mixture of dichloromethane and methanol, used at a volume ratio of 1:0.03.
[0073] Pharmaceutical composition: L-OHP, ES, CU and aminosalicylic acid derivative. The mass ratio of L-OHP to ES is 1:0.079, the mass ratio of ES to CU is 1:0.34, and the mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.006.
[0074] Example 4: A pharmaceutical composition
[0075] Preparation of aminosalicylic acid derivatives: 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole were added to dichloromethane, followed by the addition of condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and catalyst 1-hydroxybenzotriazole, and then triethylamine. The mixture was reacted at 30°C for 8 hours. After adding water, the mixture was separated into layers. The organic phase was washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution, dried with anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel chromatography to obtain aminosalicylic acid derivatives. The amounts of 4-acetaminosalicylic acid used were 3 wt% of dichloromethane, 100 wt% of 3-(2-aminoethyl)indole, 120 wt% of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 90 wt% of 1-hydroxybenzotriazole, and 130 wt% of triethylamine. In silica gel chromatography purification, the purification reagent was a mixture of dichloromethane and methanol, used at a volume ratio of 1:0.03.
[0076] Pharmaceutical composition: L-OHP, ES, CU, aminosalicylic acid derivative and 6-hydroxynicotinic acid. The mass ratio of L-OHP to ES is 1:0.079, the mass ratio of ES to CU is 1:0.34, the mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.02, and the mass ratio of L-OHP to 6-hydroxynicotinic acid is 1:0.005.
[0077] Example 5: A pharmaceutical composition
[0078] Preparation of aminosalicylic acid derivatives: 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole were added to dichloromethane, followed by the addition of condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and catalyst 1-hydroxybenzotriazole, and then triethylamine. The mixture was reacted at 30°C for 8 hours. After adding water, the mixture was separated into layers. The organic phase was washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution, dried with anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel chromatography to obtain aminosalicylic acid derivatives. The amounts of 4-acetaminosalicylic acid used were 3 wt% of dichloromethane, 100 wt% of 3-(2-aminoethyl)indole, 120 wt% of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 90 wt% of 1-hydroxybenzotriazole, and 130 wt% of triethylamine. In silica gel chromatography purification, the purification reagent was a mixture of dichloromethane and methanol, used at a volume ratio of 1:0.03.
[0079] Pharmaceutical composition: L-OHP, ES, CU, aminosalicylic acid derivative and 6-hydroxynicotinic acid. The mass ratio of L-OHP to ES is 1:0.079, the mass ratio of ES to CU is 1:0.34, the mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.02, and the mass ratio of L-OHP to 6-hydroxynicotinic acid is 1:0.002.
[0080] Comparative Example 1: A pharmaceutical composition
[0081] The difference between this comparative example and Example 2 lies in the pharmaceutical composition.
[0082] Pharmaceutical composition: L-OHP, ES, CU and aminosalicylic acid derivative. The mass ratio of L-OHP to ES is 1:0.079, the mass ratio of ES to CU is 1:0.34, and the mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.0005.
[0083] Comparative Example 2: A pharmaceutical composition
[0084] The difference between this comparative example and Example 2 lies in the pharmaceutical composition.
[0085] Pharmaceutical composition: aminosalicylic acid derivative.
[0086] Experimental example:
[0087] This invention involves incorporating the drug compositions of Examples 1-5 and Comparative Example 1 into pancreatic cancer cells for intervention, testing the inhibitory effects of the drug compositions of Examples 1-5 and Comparative Example 1 on pancreatic cancer cells. The Panc02 cell line was used for testing; the Panc02 pancreatic cancer cell line was seeded in 6-well plates (1×10⁻⁶ cells / well). 5 (cells / well), and on the second day, intervention was performed using the drug compositions of Examples 1-5 and Comparative Example 1, wherein: the in vitro drug intervention doses for Panc02 were: L-OHP: 100 μg / ml, ES: 200 nM; CU: 200 nM; L-OHP, ES, and CU were added one at a time, without pre-mixing, and the intervention time was 48 h. A control group was set up, in which only the solvent was added.
[0088] The apoptosis rates of the pharmaceutical compositions of Examples 1-5 and Comparative Example 1 after PancO2 intervention in this invention are as follows: Figure 7 As shown, S1 is Example 1, S2 is Example 2, S3 is Example 3, S4 is Example 4, S5 is Example 5, D1 is Comparative Example 1, D2 is Comparative Example 2, and B is the control group. This invention first studied L-OHP and ES-CU. ES-CU alone could not significantly increase the apoptosis rate of Panco2 cells, indicating that ES-CU had virtually no inhibitory effect on pancreatic cancer cells. However, when L-OHP and ES-CU were used in combination, it was found that they could increase the apoptosis rate of Panco2 cells, and the effect of the combined use of L-OHP and ES-CU significantly exceeded that of L-OHP, indicating that ES-CU can increase the apoptosis rate of L-OHP cells. In addition to the effect of P, this invention also uses an aminosalicylic acid derivative prepared by the reaction of 4-acetaminosalicylic acid and 3-(2-aminoethyl)indole. By combining the aminosalicylic acid derivative with L-OHP and ES-CU, the apoptosis rate of Panco2 cells can be improved. The amount of aminosalicylic acid derivative used needs to meet a certain range. If the amount used is too small, the apoptosis rate of Panco2 cells cannot be improved. Furthermore, using only the aminosalicylic acid derivative will also fail to improve the apoptosis rate of Panco2 cells. In addition, 6-hydroxynicotinic acid can be added to the pharmaceutical composition. The use of 6-hydroxynicotinic acid further improves the apoptosis rate of Panco2 cells.
[0089] The drug compositions of Examples 1-5 and Comparative Example 1 in this invention affect Hsp90 expression after Panco2 intervention as follows: Figure 8As shown, S1 is Example 1, S2 is Example 2, S3 is Example 3, S4 is Example 4, S5 is Example 5, D1 is Comparative Example 1, D2 is Comparative Example 2, and B is the control group. This invention first studied L-OHP and ES-CU. ES-CU alone could not significantly increase Hsp90 expression in Panco2 cells, indicating that ES-CU had little effect on increasing Hsp90 expression in pancreatic cancer cells. However, when L-OHP and ES-CU were used in combination, it was found that it could increase Hsp90 expression in Panco2 cells, and the effect of the combination of L-OHP and ES-CU significantly exceeded that of L-OHP, indicating that ES-CU has the effect of increasing L-OHP expression. In addition to the effects of OHP, this invention also utilizes an aminosalicylic acid derivative prepared by the reaction of 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole. By combining the aminosalicylic acid derivative with L-OHP and ES-CU, the expression of Hsp90 in Panc02 can be increased. The amount of aminosalicylic acid derivative used needs to meet a certain range. If the amount used is too small, the expression of Hsp90 in Panc02 cannot be increased. Furthermore, using only the aminosalicylic acid derivative also cannot increase the expression of Hsp90 in Panc02. In addition, 6-hydroxynicotinic acid can be added to the pharmaceutical composition. The use of 6-hydroxynicotinic acid further increases the expression of Hsp90 in Panc02.
[0090] The embodiments and / or implementation methods described above are merely preferred embodiments and / or implementation methods for implementing the technology of the present invention, and are not intended to limit the implementation methods of the technology of the present invention in any way. Any person skilled in the art can make some modifications or alterations to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as the technology or embodiments that are substantially the same as the present invention.
[0091] This document uses specific examples to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the methods and core ideas of this application. The above descriptions are only preferred embodiments of this application. It should be noted that due to the limitations of written expression, while there are objectively infinite specific structures, those skilled in the art can make several improvements, modifications, or changes without departing from the principles of this application, and can also combine the above technical features in an appropriate manner. These improvements, modifications, changes, or combinations, or the direct application of the inventive concept and technical solution to other situations without modification, should all be considered within the scope of protection of this application.
Claims
1. A pharmaceutical composition comprising: L-OHP, ES, and CU, wherein the mass ratio of L-OHP to ES is 1:0.0005-5, the mass ratio of ES to CU is 1:0.05-0.5, and L-OHP is oxaliplatin, ES is ilisimo, and CU is copper chloride; The pharmaceutical composition includes an aminosalicylic acid derivative, which is prepared by reacting 4-acetamidosalicylic acid with 3-(2-aminoethyl)indole. In the preparation of the aminosalicylic acid derivative, 4-acetamidosalicylic acid and 3-(2-aminoethyl)indole are added to dichloromethane, followed by the addition of condensing agent 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and catalyst 1-hydroxybenzotriazole, and then triethylamine. The reaction is carried out at 20-40°C for 4-16 hours. After adding water, the layers are separated. The organic phase is washed successively with saturated sodium bicarbonate solution and saturated sodium chloride solution, then dried with anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel chromatography to obtain the aminosalicylic acid derivative. In the preparation of the aminosalicylic acid derivative, the amount of 3-(2-aminoethyl)indole used is 80-120 wt% of 4-acetamidosalicylic acid; The pharmaceutical composition comprises 6-hydroxynicotinic acid, and the mass ratio of L-OHP to 6-hydroxynicotinic acid is 1:0.001-0.
006.
2. The pharmaceutical composition according to claim 1, characterized in that: The pharmaceutical composition also contains a medical solvent.
3. The pharmaceutical composition according to claim 2, characterized in that: The medical solvent includes at least one of ethanol, propylene glycol, glycerin, 1,3-butanediol, polyethylene glycol, and physiological saline.
4. The pharmaceutical composition according to claim 2, characterized in that: The concentration of L-OHP is 4-200 μg / mL.
5. The pharmaceutical composition according to claim 1, characterized in that: The mass ratio of L-OHP to aminosalicylic acid derivative is 1:0.005-0.
03.
6. The pharmaceutical composition according to claim 1, characterized in that: The mass ratio of L-OHP to ES is 1:0.079, and the mass ratio of ES to CU is 1:0.
34.
7. An injection solution, comprising: The pharmaceutical composition according to any one of claims 1-6.