Pharmaceutical composition containing photosensitizer and autophagy inhibitor and application

Abstract

The invention discloses a pharmaceutical composition containing a photosensitizer and an autophagy inhibitor, each unit dose of the pharmaceutical composition contains 1-100mg of a photosensitive drug DTP and 6-600mg of an autophagy inhibitor chloroquine phosphate, and the unit dose is the total amount of a pharmaceutical preparation used every day. The pharmaceutical composition containing the photosensitizer and the autophagy inhibitor is prepared by mixing 1-100mg of a photosensitive drug DTP and 6-600mg of an autophagy inhibitor chloroquine phosphate to prepare a liquid preparation for injection administration, such as freeze-dried powder injection, injection, large and small infusion or intravenous drip. The invention also provides an application of a pharmaceutical composition taking the photosensitive drug DTP and the autophagy inhibitor chloroquine phosphate as active ingredients in preparation of antitumor drugs. The pharmaceutical composition taking the photosensitive drug DTP and the autophagy inhibitor chloroquine phosphate as the active ingredients has a remarkable treatment effect in the anti-tumor aspect, not only has a good treatment effect on local tumors of prostatic cancer, but also shows synergistic anti-tumor activity. Combined application of the photodynamic therapy and the autophagy inhibitor is a research hot spot in the field of tumor treatment, and the research trend focuses on improvement of the treatment effect and deep understanding of an interaction mechanism between the photodynamic therapy and cell autophagy, so that the photodynamic therapy and the autophagy inhibitor can be more effectively applied to clinical treatment.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a pharmaceutical composition containing a photosensitizer and an autophagy inhibitor and its use, especially to the preparation of a pharmaceutical composition from a photosensitizing drug DTP and chloroquine, and the application of this pharmaceutical composition in anti-tumor drugs. Background Art

[0002] Prostate cancer is one of the most common malignancies in men originating from prostatic epithelial cells and is also the second most common cause of cancer-related death. [1] With the improvement of people's understanding of prostate cancer, the popularization of prostate-specific antigen (PSA) screening, the development of magnetic resonance imaging (MRI) and positron emission tomography / computed tomography (PET / CT) technologies, the diagnostic rate of prostate cancer is gradually increasing. [2]

[0003] Currently, the clinical treatment methods for prostate cancer include: drug therapy, radiotherapy, hormone therapy, surgical treatment, etc. [3-5] However, there are risks such as tumor drug resistance, toxic and side effects of drugs, urinary incontinence, pelvic pain, erectile dysfunction, etc. [6] When the tumor is small, doctors usually recommend active surveillance for patients. However, active surveillance may also cause anxiety and uneasiness in patients, and the probability of cancer development is also higher. [7] The functional impairments caused by radical surgical treatment of prostate cancer, such as urinary and sexual dysfunction, have prompted researchers to focus on minimally invasive treatment. Photodynamic therapy (PDT), as an emerging focal treatment method, has advantages such as minimally invasive, low toxic and side effects, broad-spectrum anti-cancer, and repeatable treatment. [8] It can avoid debilitating damage to the urethra and neurovascular bundles and is expected to improve the treatment effect of prostate cancer. [9]

[0004] Autophagy refers to the process by which cells degrade organelles and proteins through lysosomes under stress conditions to achieve the recycling of these components, which is crucial for maintaining the basic metabolism of cells, providing energy, and macromolecular precursors.

[10] In the field of cancer, the role of autophagy is dual, which may either promote or inhibit cancer development.

[11] Therefore, intervention measures to regulate autophagy are regarded as a potential cancer treatment strategy.

[12] Shao

[13] ​​Studies by [authors] et al. have shown that curcumin-PDT can inhibit the epithelial-mesenchymal transition (EMT) of lung cancer cells. However, autophagy induced by curcumin-PDT has a promoting effect on the EMT of lung cancer cells. Therefore, when curcumin-PDT is combined with an autophagy inhibitor, it can significantly inhibit the EMT, invasion, and metastasis of lung cancer cells. Dai

[14] Studies by [authors] et al. have confirmed that the combination of photodynamic therapy and an autophagy inhibitor can effectively inhibit the protective autophagy of oral cancer cells, thereby exacerbating cell death caused by ROS. Zhu et al

[15] The combined use of the photosensitizing drug sodium huaporfin and an autophagy inhibitor in the treatment of colorectal tumors achieved better anti-tumor effects than the photosensitizing drug porfimer sodium. The mechanism is related to the inhibition of autophagy promoting apoptosis of tumor cells. And Chen

[16] [authors] et al. combined the photosensitizing drug Chlorin e6 with the autophagy promoter celastrol in the treatment of colon cancer and also achieved good treatment effects. The mechanism of action is that the photosensitizing drug generates ROS in tumor cells to enhance oxidative stress and promote autophagy. Overactivated autophagy induces autophagic cell death, showing enhanced phototoxicity.

[0005] The photosensitizer DTP is a highly efficient and low-toxic photodynamic anti-tumor candidate drug screened and obtained by our research group in the early stage. In this invention, the photosensitizer DTP and the autophagy inhibitor chloroquine phosphate are made into a pharmaceutical composition, which has achieved unexpected effects in the treatment of prostate cancer. It not only has a good treatment effect on local tumors of prostate cancer, but also shows synergistic anti-tumor activity. The combined application of PDT and autophagy inhibitors is a research hotspot in the field of tumor treatment. This research trend focuses on improving the treatment effect and deeply understanding the interaction mechanism between photodynamic therapy and autophagy in order to be more effectively applied to clinical treatment. With the continuous in-depth research, it is expected to see more innovative treatment strategies and drug combinations in the future to improve the cure rate of tumors and the survival rate of patients, and to achieve personalized and precision medicine.

[0006] References

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[0011] [5] Sekhoacha M, Riet K, Motloung P, et al. Prostate cancer review: genetics, diagnosis, treatment options, and alternative approaches[J]. Molecules,2022,27(17):5730.

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[0013] [7] Osuchowski M, Bartusik-Aebisher D, Osuchowski F, et al. Photodynamic therapy for prostate cancer - A narrative review[J]. Photodiagnosis PhotodynTher,2021,33:102158.

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[0016]

[10] Jiang B,Feng L,Yang T,et al.Combination of chloroquine diphosphate and salidroside induces human liver cell apoptosis via regulation of mitochondrial dysfunction and autophagy[J].Mol Med Rep,2023,27(2):37.

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[0019]

[13] Shao L, Zhu Y, Liao B, et al. Effects of curcumin-mediated photodynamic therapy on autophagy and epithelial-mesenchymal transition of lung cancer cells[J]. Photodiagnosis Photodyn Ther, 2022, 38: 102849.

[0020]

[14] Dai H, Yan H, Dong F, et al. Tumor-targeted biomimetic nanoplatform precisely integrates photodynamic therapy and autophagy inhibition for collaborative treatment of oral cancer[J]. Biomater Sci, 2022, 10(6): 1456 - 1469.

[0021]

[15] Zhu B, Li S, Yu L, et al. Inhibition of autophagy with chloroquine enhanced sinoporphyrin sodium mediated photodynamic therapy-induced apoptosis in human colorectal cancer cells. Int J Biol Sci, 2019, 15(1): 12 - 23.

[0022]

[16] Chen S Y, Zhao L P, Chen Z X, et al. Self-delivery biomedicine for enhanced photodynamic therapy by feedback promotion of tumor autophagy[J]. Acta Biomater, 2023, 158: 599 - 610. Summary of the Invention

[0023] The object of the present invention is to overcome the deficiencies of the prior art and provide a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients.

[0024] The second object of the present invention is to provide the use of a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients in the preparation of an anti-tumor drug.

[0025] The object of the present invention is mainly achieved by the following technical solutions:

[0026] A pharmaceutical composition of the present invention containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients can directly mix 1 - 100 mg of the photosensitizing drug DTP and 6 - 600 mg of the autophagy inhibitor chloroquine phosphate to prepare a liquid preparation for injection, such as: freeze-dried powder for injection, injection solution, large and small infusion solutions or intravenous drip solutions. The structure of the photosensitizing drug DTP is The structure of the autophagy inhibitor chloroquine phosphate is

[0027] Each unit dose of the pharmaceutical composition of the present invention contains 1 - 100 mg of the photosensitizing drug DTP and 6 - 600 mg of the autophagy inhibitor chloroquine phosphate. The dosage can be adjusted according to many factors such as the patient's age, weight, disease type and severity, and can be appropriately changed according to the manifestations of clinical cases. The daily dose is generally 1 - 10 mg / kg of the photosensitizing drug DTP and 6 - 60 mg / kg of the autophagy inhibitor chloroquine phosphate.

[0028] The application of a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients of the present invention in the preparation of an anti-tumor drug.

[0029] The pharmaceutical composition of the present invention containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients has a significant therapeutic effect in anti-tumor. It not only has a good therapeutic effect on local tumors of prostate cancer, but also shows a synergistic anti-tumor activity. The combined use of the photosensitizing drug and the autophagy inhibitor can inhibit the protective autophagy of tumor cells, promote the apoptosis of tumor cells, and improve the overall treatment efficiency. Description of the Drawings

[0030] Figure 1 It is the anti-tumor effect of the pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients of Example 10 of the present invention on human prostate cancer cell line DU145.

[0031] Figure 2 It is the anti-tumor effect of the pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate as active ingredients of Example 11 of the present invention on human prostate cancer cell line LNCaP.

[0032] Figure 3Antitumor effect of the pharmaceutical composition with photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients in Example 12 of the present invention on human prostate cancer cell line PC-3.

[0033] Figure 4 Antitumor effect of the pharmaceutical composition with photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients in Example 13 of the present invention on mouse fibroblast cell line L-929.

[0034] Figure 5 Change in body weight of mice xenografted with human tumor cell lines treated with the pharmaceutical composition with photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients in Example 14 of the present invention.

[0035] Figure 6 Tumor growth curve of mice xenografted with human tumor cell lines treated with the pharmaceutical composition with photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients in Example 14 of the present invention.

[0036] Figure 7 Tumor mass of mice xenografted with human tumor cell lines treated with the pharmaceutical composition with photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients in Example 14 of the present invention.

[0037] Figure 8 Organ index of mice xenografted with human tumor cell lines treated with the pharmaceutical composition with photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients in Example 14 of the present invention.

[0038] The present invention will be further illustrated by the following examples, the purpose of which is only to better understand the content of the present invention rather than to limit the protection scope of the present invention:

[0039] Example 1 Infusion

[0040] Take 1 mg of photosensitizing drug DTP and 6 mg of autophagy inhibitor chloroquine phosphate, add 5 mg of polyethylene glycol, 5 mg of lecithin, 8 mg of sodium acetate, 8 mg of sodium sulfite and 9 mg of sodium chloride, dissolve in water for injection, filter, seal, sterilize and package to obtain.

[0041] Example 2 Infusion

[0042] Take 50 mg of photosensitizing drug DTP and 300 mg of autophagy inhibitor chloroquine phosphate, add 250 mg of polyethylene glycol, 250 mg of lecithin, 500 mg of sodium sulfite and 750 mg of glucose, dissolve in water for injection, filter, seal, sterilize and package to obtain. Each preparation unit contains 50 mg of photosensitizing drug DTP and 300 mg of autophagy inhibitor chloroquine phosphate, and can be used for intramuscular injection or intravenous injection, with a dosage of once every 3 days and 1 preparation unit each time.

[0043] Example 3 Infusion

[0044] Take 100 mg of photosensitive drug DTP and 600 mg of autophagy inhibitor chloroquine phosphate, add 500 mg of polyethylene glycol, 500 mg of lecithin, 800 mg of sodium acetate, 800 mg of sodium sulfite and 900 mg of sodium chloride, dissolve in water for injection, filter, seal, sterilize and package to obtain the product.

[0045] Example 4 Freeze-dried Powder for Injection

[0046] Take 10 mg of photosensitive drug DTP and 10 mg of autophagy inhibitor chloroquine phosphate, add 1.5 g of mannitol and 2.5 mg of sodium bisulfite, dissolve in 10 ml of water for injection, then add 250 mg of medicinal activated carbon, stir at room temperature for 20 min, filter to remove the activated carbon, and then filter and sterilize with a 0.22 μm filter membrane, dispense into 5-ml vials, 1 ml per vial, and freeze-dry.

[0047] Example 5 Freeze-dried Powder for Injection

[0048] Take 1 mg of photosensitive drug DTP and 6 mg of autophagy inhibitor chloroquine phosphate, add 200 mg of glucose, 4 mg of sodium bicarbonate and 3 mg of sodium sulfite, dissolve in 1 ml of water for injection, then add 100 mg of medicinal activated carbon, stir at room temperature for 20 min, filter to remove the activated carbon, and then filter and sterilize with a 0.22 μm filter membrane, dispense into 5-ml vials, 1 ml per vial, and freeze-dry.

[0049] Example 6 Freeze-dried Powder for Injection

[0050] Take 100 mg of photosensitive drug DTP and 600 mg of autophagy inhibitor chloroquine phosphate, add 4 g of glucose, 0.1 ml of glycerol and 5 mg of sodium sulfite, dissolve in 100 ml of water for injection, then add 500 mg of medicinal activated carbon, stir at room temperature for 20 min, filter to remove the activated carbon, and then filter and sterilize with a 0.22 μm filter membrane, dispense into 5-ml vials, 1 ml per vial, and freeze-dry.

[0051] Example 7 Injection

[0052] Take 1 mg of photosensitive drug DTP and 6 mg of autophagy inhibitor chloroquine phosphate, add 9 mg of sodium chloride and 1 ml of water for injection to prepare the solution, stir well, filter, seal, and sterilize by autoclaving at 115 °C for 30 min to obtain the product.

[0053] Example 8 Injection

[0054] Take 50 mg of the photosensitive drug DTP and 300 mg of the autophagy inhibitor chloroquine phosphate, add 500 mg of glucose and 50 ml of water for injection to prepare a solution, dropwise add a hydrochloric acid aqueous solution with a volume ratio of 1:1, adjust the pH value to 4.0, add 50 mg of activated carbon, boil for 30 min under stirring, filter to remove the activated carbon, and then filter and sterilize with a 0.22 μm filter membrane to obtain a clear and sterile solution. Seal it and sterilize it by autoclaving at 115 °C for 30 min to prepare it.

[0055] Example 9 Injection

[0056] Take 100 mg of the photosensitive drug DTP and 600 mg of the autophagy inhibitor chloroquine phosphate, add 900 mg of sodium chloride and 100 ml of water for injection to prepare a solution, stir well, filter, seal it, and sterilize it by autoclaving at 115 °C for 30 min to prepare it.

[0057] In vitro anti-tumor effect (DU145) of the pharmaceutical composition with the photosensitive drug DTP and the autophagy inhibitor chloroquine phosphate as active ingredients

[0058] In vitro anti-tumor evaluation of the pharmaceutical composition with the photosensitive drug DTP and the autophagy inhibitor chloroquine phosphate as active ingredients on human prostate cancer cells DU145 includes the following steps:

[0059] Take human prostate cancer cells DU145 in the logarithmic growth phase, digest them with trypsin, resuspend them in RPMI 1640 medium containing 10% fetal bovine serum, and inoculate them in a 96-well plate at a density of 1×10 4 cells / well, and incubate them in a cell culture incubator for 24 h. Discard the medium, add 100 μl of drug solutions with different concentrations to each well. The single-use concentrations of the photosensitive drug DTP are 0.64 μM, 0.32 μM, 0.16 μM, 0.08 μM, 0.04 μM, and 0.02 μM in sequence; the single-use concentrations of the autophagy inhibitor chloroquine phosphate are 40 μM, 20 μM, 10 μM, 5 μM, 2.50 μM, and 1.25 μM in sequence; the combined-use group drug concentrations are 0.64 μM photosensitive drug DTP + 40 μM autophagy inhibitor chloroquine phosphate, 0.32 μM photosensitive drug DTP + 20 μM autophagy inhibitor chloroquine phosphate, 0.16 μM photosensitive drug DTP + 10 μM autophagy inhibitor chloroquine phosphate, 0.08 μM photosensitive drug DTP + 5 μM autophagy inhibitor chloroquine phosphate, 0.04 μM photosensitive drug DTP + 2.50 μM autophagy inhibitor chloroquine phosphate, and 0.02 μM photosensitive drug DTP + 1.25 μM autophagy inhibitor chloroquine phosphate in sequence. The control group uses blank culture medium instead of the drug solution, with 4 sub-wells for each concentration, and incubate them in a cell culture incubator for 24 h. Irradiate with a laser with a wavelength of 650 nm, and the energy density is 6 J / cm 2, the light exposure time was 20 min. Meanwhile, a control experiment with only the drug solution added and no light exposure was set up. Incubate in a cell incubator for 24 h, aspirate the liquid in the wells, add 10 μL of MTT (5 mg / mL) and 100 μL of blank culture medium to each well, and continue culturing for 4 h. Discard the culture medium, and add 150 μL of DMSO to each well to dissolve the color crystals. Use an enzyme-linked immunosorbent assay (ELISA) reader to detect the light absorption value of each well at a wavelength of 490 nm, calculate the cell survival rate, and the results are shown in Figure 1 .

[0060] As can be seen from the figure, the photosensitizing drug DTP alone and the combination of the photosensitizing drug DTP and the autophagy inhibitor chloroquine phosphate both have good photodynamic antitumor activity, and there is an obvious dose-dependent relationship. Under light exposure conditions, the tumor inhibition rate of 0.64 μM photosensitizing drug DTP is 84.10%, the tumor inhibition rate of 40 μM autophagy inhibitor chloroquine phosphate is 20.39%, while the tumor inhibition rate of the combination of 0.64 μM photosensitizing drug DTP + 40 μM autophagy inhibitor chloroquine phosphate is 91.74%, showing an obvious synergistic antitumor effect, and the synergy index is 0.91.

[0061] Example 11 In vitro antitumor effect of a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate (LNCaP)

[0062] In vitro antitumor evaluation of a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate on human prostate cancer cells LNCaP (see Figure 2 ), the experimental subject was human prostate cancer cells LNCaP, and the experimental procedure was the same as that in Example 10.

[0063] As can be seen from the figure, the photosensitizing drug DTP alone and the combination of the photosensitizing drug DTP and the autophagy inhibitor chloroquine phosphate both have good photodynamic antitumor activity, and there is an obvious dose-dependent relationship. Under light exposure conditions, the tumor inhibition rate of 0.64 μM photosensitizing drug DTP is 81.23%, the tumor inhibition rate of 40 μM autophagy inhibitor chloroquine phosphate is 21.14%, while the tumor inhibition rate of the combination of 0.64 μM photosensitizing drug DTP + 40 μM autophagy inhibitor chloroquine phosphate is 88.28%, showing an obvious synergistic antitumor effect, and the synergy index is 0.93.

[0064] Example 12 In vitro antitumor effect of a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate (PC-3)

[0065] In vitro antitumor evaluation of a pharmaceutical composition containing a photosensitizing drug DTP and an autophagy inhibitor chloroquine phosphate on human prostate cancer cells PC-3 (see Figure 3 ), the experimental subject was human prostate cancer cells PC-3, and the experimental procedure was the same as that in Example 10.

[0066] As can be seen from the figure, the photosensitizing drug DTP alone and in combination with the autophagy inhibitor chloroquine phosphate both have good photodynamic antitumor activity, and there is an obvious dose-dependent relationship. Under light irradiation, the tumor inhibition rate of 0.64 μM photosensitizing drug DTP is 91.50%, the tumor inhibition rate of 40 μM autophagy inhibitor chloroquine phosphate is 24.95%, while the tumor inhibition rate of the combination of 0.64 μM photosensitizing drug DTP + 40 μM autophagy inhibitor chloroquine phosphate is 95.82%, showing an obvious synergistic antitumor effect, and the synergy index is 0.88.

[0067] Example 13 In vitro cell evaluation (L-929) of a pharmaceutical composition containing the photosensitizing drug DTP and the autophagy inhibitor chloroquine phosphate

[0068] In vitro cell evaluation of a pharmaceutical composition containing the photosensitizing drug DTP and the autophagy inhibitor chloroquine phosphate on mouse fibroblast L-929 (see Figure 4 ), the experimental subject was mouse fibroblast L-929, and the experimental procedure was the same as that in Example 10.

[0069] As can be seen from the figure, the phototoxicity of the photosensitizing drug DTP alone and in combination with the photosensitizing drug DTP and the autophagy inhibitor chloroquine phosphate to mouse fibroblasts is less than that to prostate cancer cells. Under light irradiation, the tumor inhibition rate of 0.64 μM photosensitizing drug DTP is 44.43%, the tumor inhibition rate of 40 μM autophagy inhibitor chloroquine phosphate is 11.75%, and the tumor inhibition rate of the combination of 0.64 μM photosensitizing drug DTP + 40 μM autophagy inhibitor chloroquine phosphate is 61.95%.

[0070] Example 14 Therapeutic effect of a pharmaceutical composition containing the photosensitizing drug DTP and the autophagy inhibitor chloroquine phosphate on xenograft mice with human tumor cell lines

[0071] Take the human prostate cancer cell line DU145 in the logarithmic growth phase and prepare a cell suspension of 5×10 7 cells / ml under sterile conditions. Select 24 Balb / c-nu nude mice at 4-6 weeks of age and weighing 16-18 g, and inoculate the cell suspension into the right posterior dorsal part of the nude mice at 0.2 ml per mouse. Wait for the tumor to grow to 100 mm 3After that, they were randomly divided into 4 groups according to body weight, with 6 animals in each group. Control group: 200 μl of normal saline was injected into the tail vein once every 2 days for 3 consecutive times; ALA group: 200 μl of photosensitizing drug ALA was intraperitoneally injected at a dosage of 100 mg / kg once every 2 days for 3 consecutive times; DTP group: 200 μl of photosensitizing drug DTP was injected into the tail vein at a dosage of 10 mg / kg once every 2 days for 3 consecutive times; DTP+CQ group: The pharmaceutical composition prepared in Example 7 containing photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients was injected into the tail vein once every 2 days for 3 consecutive times. At 3 h after administration in the ALA group, and at 24 h and 48 h after administration in the DTP group and DTP+CQ group, the experimental animals were irradiated with light using an Intense 650 nm PDT system, only irradiating the local tumor, and other parts were shielded from light. The light intensity was 0.15 W, the irradiation time was 10 min, and the light energy density was 100 J / cm 2 . Every 2 days, the body weight changes of the mice were recorded (see Figure 5 ), and the maximum diameter and transverse diameter of the tumor were measured with a vernier caliper, and the tumor volume V = 1 / 2 (maximum diameter × transverse diameter 2 ), and the tumor growth curve was plotted (see Figure 6 ). On the 21st day after the first administration, all experimental animals were sacrificed, and the tumors (see Figure 7 ), heart, liver, spleen, lung, and kidney were dissected and weighed separately. The tumor growth inhibition rate was calculated: IR(%) = (1 - average tumor weight of the treatment group / average tumor weight of the Control group) × 100% and the organ index (see Figure 8 ): OI (mg / g) = organ weight (mg) / animal body weight (g).

[0072] It can be seen from Figure 5 , Figure 8 that there were no statistically significant differences in body weight changes and organ indices among the experimental animals in each group (p>0.05), indicating that the experimental animals had good tolerance to the pharmaceutical composition containing photosensitizing drug DTP and autophagy inhibitor chloroquine phosphate as active ingredients. It can be seen from Figures 6-7 that the tumor growth of the experimental animals in the ALA group (tumor inhibition rate 43.74%), DTP group (tumor inhibition rate 76.35%) and DTP+CQ group (tumor inhibition rate 96.23%) was inhibited to varying degrees. The tumor inhibition rate of the DTP+CQ group was significantly better than that of the DTP group, showing a synergistic and enhanced anti-tumor therapeutic effect.

Claims

1. A pharmaceutical composition with photosensitizer DTP and autophagy inhibitor chloroquine phosphate as active ingredients, characterized in that The photosensitizer DTP 1-100 mg and the autophagy inhibitor chloroquine phosphate 6-600 mg are mixed to prepare a liquid preparation for injection, such as freeze-dried powder injection, injection, large and small infusion or intravenous drip.

2. The pharmaceutical composition according to claim 1, characterized in that The chemical structure of the photosensitizer DTP is 3. The pharmaceutical composition according to claim 1, characterized in that The chemical structure of the autophagy inhibitor chloroquine phosphate is 4. The pharmaceutical composition according to claim 1, characterized in that Each unit dose contains 1-100 mg of photosensitizer DTP and 6-600 mg of autophagy inhibitor chloroquine phosphate. The dosage can be adjusted according to many factors such as the patient's age, weight, disease type and severity, and can be appropriately changed according to the manifestations of clinical cases. The daily dose is generally 1-10 mg / kg of photosensitizer DTP and 6-60 mg / kg of autophagy inhibitor chloroquine phosphate.

5. Use of a pharmaceutical composition comprising a photosensitizer DTP and an autophagy inhibitor chloroquine phosphate as active ingredients according to claim 1, 2, 3 or 4 in the preparation of an anti-tumor drug.

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