Pharmaceutical composition for preventing and / or treating tumor malignant polymenal effusion and application thereof
Through the combined use of LIN28B inhibitor and PARP inhibitor, the treatment problem of malignant serous effusion is solved, effective effusion inhibition and survival prolongation is achieved, and a safe and efficient treatment plan is provided.
Patent Information
- Application Number
- CN202510447248.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-10
- Publication Date
- 2025-07-04
AI Technical Summary
The prior art lacks effective treatment methods to deal with malignant serous effusion, resulting in short survival and poor prognosis of patients, and lack of specificity of existing treatment methods, resulting in low response rates and high side effects.
The combination of LIN28B inhibitor and PARP inhibitor is used, specifically through the combination of siLIN28b and BMN673, using viral or nonviral vectors, combined with liposome vectors for targeted delivery, increasing the concentration of the drug at the lesion site and reducing side effects on normal tissues.
It significantly inhibits the formation of multi-membrane effusion, prolongs the patient's survival, improves the treatment effect while reducing side effects, and has good application prospects.
Smart Images
Figure CN120242031A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a pharmaceutical composition for preventing and / or treating malignant multi-pleural cavity effusion of tumors and its application. Background Art
[0002] Serous multi-pleural cavity effusion refers to the pathological accumulation of body fluids, most commonly in the pleural cavity, peritoneal cavity, and pericardial cavity. Accurate diagnosis is crucial for determining the root cause of the disease, appropriate treatment, and predicting the prognosis of patients. Malignant serous effusions (MSE) are characterized by excessive fluid accumulation in the pleural, peritoneal, or pericardial cavities, usually caused by primary tumors invading the body cavity and serosa, leading to local metastasis or systemic spread. Studies have shown that malignant pleural effusion (MPE) is most commonly found in lung cancer (37.5%), breast cancer (16.8%), and malignant lymphoma (11.5%), while malignant ascites (MA) is common in ovarian cancer (37%), hepatobiliary pancreatic tumors (21%), and gastric cancer (18%). MSE often endangers life and is associated with poor prognosis and high recurrence rates.
[0003] However, the current understanding of the molecular mechanism of MSE is limited, and there is a lack of standardized treatment methods, which poses a major clinical challenge. Current methods mainly focus on reducing the tumor burden and actively draining the effusion to prevent disease progression. Intracavitary chemotherapy is commonly used, but they lack specificity for cancer cells, resulting in a low response rate and a high incidence of side effects.
[0004] Therefore, there is an urgent need to develop more effective and safer therapeutic drugs and delivery strategies to improve the prognosis of patients with advanced cancer and MSE. Summary of the Invention
[0005] To solve the problem of the lack of effective drugs for treating malignant multi-pleural cavity effusion in the prior art, the present invention discloses a pharmaceutical composition for preventing and / or treating malignant multi-pleural cavity effusion of tumors and its application. The pharmaceutical composition includes: a LIN28B inhibitor and a PARP inhibitor. And it has been proven by in vivo experiments that the pharmaceutical composition provided by the present invention can effectively inhibit the formation of multi-pleural cavity effusion, achieving the effect of preventing and / or treating malignant multi-pleural cavity effusion of tumors, which is beneficial to improving the survival period of patients.
[0006] The present invention first provides a pharmaceutical composition for preventing and / or treating malignant multi-pleural cavity effusion of tumors, including: a LIN28B inhibitor and a PARP inhibitor.
[0007] Preferably, the inhibitor includes: siRNA or a gene knockout vector targeting the coding gene, or a protein antibody targeting the protein, or any one or a combination of multiple other inhibitors capable of inhibiting protein expression.
[0008] Preferably, the LIN28B inhibitor is siLIN28b, the nucleotide sequence of the sense strand is shown in SEQ ID NO: 1, and the nucleotide sequence of the antisense strand is shown in SEQ ID NO: 2.
[0009] Preferably, the PARP inhibitor is BMN673.
[0010] Preferably, the mass ratio of the siLIN28b to BMN673 is 1:500 - 1:1000.
[0011] Preferably, the siLIN28b is delivered by a viral vector, and the viral vector includes any one or more of an adeno-associated virus vector, an adenovirus vector, or a retrovirus vector.
[0012] Preferably, the siLIN28b is delivered by a non-viral vector, and the non-viral vector includes any one or more of a plasmid vector, a liposome vector, or a cationic polymer vector.
[0013] Preferably, the siLIN28b is delivered by a liposome vector, and the liposome vector is modified with folic acid polyethylene glycol phospholipid.
[0014] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
[0015] On the other hand, the present invention also provides the use of the aforementioned pharmaceutical composition in the preparation of a drug for preventing and / or treating malignant multi-pleural effusion of tumors.
[0016] Compared with the prior art, the beneficial effects of the present invention at least include:
[0017] The present invention provides a pharmaceutical composition for preventing and / or treating malignant multi-pleural effusion of tumors, comprising: a LIN28B inhibitor and a PARP inhibitor. And through in vivo experiments, it is proved that after co-administering the LIN28B inhibitor (such as siLIN28b) and the PARP inhibitor (such as BMN673), it can synergistically inhibit the formation of multi-pleural effusion, achieve the purpose of preventing and / or treating malignant multi-pleural effusion of tumors, and improve the survival period of patients. It can be used in the preparation and application of drugs for preventing and treating malignant multi-pleural effusion of tumors, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1 It shows the synthesis flow chart of DSSP in Example 1 of the present invention. In the figure, Formula I is 2,2'-thiobis(ethylamine), Formula II is acryloyl chloride, Formula III represents Compound 1, Formula IV represents spermine, and Formula V represents DSSP.
[0019] Figure 2A1H NMR spectrum of Compound 1 prepared in Example 1.
[0020] Figure 2B 13C NMR spectrum of Compound 1 prepared in Example 1.
[0021] Figure 2C 1H NMR spectrum of DSSP prepared in Example 1.
[0022] Figure 2D 13C NMR spectrum of DSSP prepared in Example 1.
[0023] Figure 2E 1H NMR spectrum of spermine used in Example 1.
[0024] Figure 2F 13C NMR spectrum of spermine used in Example 1.
[0025] Figure 3 Particle size distribution and transmission electron microscopy (TEM) images of siLIN28b / DSSP nanoparticles and siLIN28b / DSSP@lip-PEG-FA materials; where:
[0026] a shows the particle size distribution and transmission electron microscopy (TEM) images of siLIN28b / DSSP nanoparticles;
[0027] b shows the particle size distribution and transmission electron microscopy (TEM) images of siLIN28b / DSSP@lip-PEG-FA materials.
[0028] Figure 4 Shows the release of siLIN28b from siLIN28b / DSSP nanoparticles under different conditions.
[0029] Figure 5 Shows the ascites height (a) and quantitative analysis (b) of each group of mice detected by ultrasound.
[0030] Figure 6 Shows the change in ascites production (a) and statistical chart (b) of each group of mice.
[0031] Figure 7 Shows the comparison of ascites formation time and survival curves of each group of mice, where:
[0032] a is the comparison of ascites formation time;
[0033] b is the comparison of survival curves.
[0034] Figure 8 Shows the safety evaluation of the combined administration of siLIN28b / DSSP@lip-PEG-FA and BMN673, where:
[0035] a is the result of HE staining;
[0036] b is the test results of multiple biochemical serum indicators. Detailed implementation manners
[0037] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments.
[0038] As mentioned above, patients with malignant serous cavity effusion (i.e., tumor malignant polyserositis effusion) have a short median survival period, poor prognosis, and high fatality rate. Currently, there is still a lack of effective therapeutic drugs. Therefore, there is an urgent need in the art for new and effective drugs for treating and alleviating malignant serous cavity effusion to improve the survival period and quality of life of patients with this complication.
[0039] Based on the above, through a large number of studies and experiments, the present invention finally provides a pharmaceutical composition for preventing and / or treating tumor malignant polyserositis effusion, including: a LIN28B inhibitor and a PARP inhibitor. LIN28B (Lin-28 Homolog B) is a highly conserved RNA-binding protein and belongs to the LIN28 family (including LIN28A and LIN28B); the PARP (Poly(ADP-ribose)polymerase) gene family encodes poly(ADP-ribose) polymerases, which are a class of enzymes involved in various cellular processes such as DNA damage repair, gene expression regulation, and apoptosis. Among them, PARP1 and PARP2 are the most well-known members.
[0040] In some embodiments, the LIN28B inhibitor includes: siRNA of the coding gene of the LIN28B or a gene knockout vector, or a protein antibody of the LIN28B, or any one or a combination of multiple other inhibitors capable of inhibiting the expression of the LIN28B. As a specific example, the LIN28B inhibitor is siLIN28b, and the nucleotide sequence of its sense strand is as shown in SEQ ID NO: 1, and the nucleotide sequence of its antisense strand is as shown in SEQ ID NO: 2.
[0041] The PARP inhibitor includes: siRNA of the coding gene of the PARP or a gene knockout vector, or a protein antibody of the PARP, or any one or a combination of multiple other inhibitors capable of inhibiting the expression of the PARP. As a specific example, the PARP inhibitor is BMN673. BMN673 (Talazoparib) is a highly potent orally active PARP1 / 2 inhibitor.
[0042] In some embodiments, the mass ratio of the siLIN28b to BMN673 is 1:500 - 1:1000, and further, the mass ratio is 1:660.
[0043] In some embodiments, the siLIN28b is delivered by a viral vector, and the viral vector includes any one or more of an adeno-associated virus vector, an adenovirus vector, or a retrovirus vector.
[0044] In other embodiments, the siLIN28b is delivered by a non-viral vector, and the non-viral vector includes any one or more of a plasmid vector, a liposome vector, or a cationic polymer vector. As a specific example, the siLIN28b is delivered by a liposome vector, and the liposome vector is modified with folic acid polyethylene glycol phospholipid (DSPE-PEG-FA). Among them, folic acid (FA) as a targeting ligand can enable the liposome to specifically bind to the receptor on the surface of the target cell, achieve targeted delivery of siRNA, increase the concentration of the drug at the lesion site, and reduce the side effects on normal tissues; the polyethylene glycol (PEG) modification can increase the water solubility and steric hindrance of the liposome, reduce its recognition and clearance by the reticuloendothelial system, extend the half-life in blood circulation, and enable more liposomes to reach the target tissue or organ; the thiol group enables these components to bind, thereby generating a functional drug carrier with targeting ability.
[0045] In some embodiments, the pharmaceutical composition further includes a pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" refers to a carrier for the administration of therapeutic agents, including various excipients and diluents. This term refers to such pharmaceutical carriers: they are not necessary active ingredients themselves and have no excessive toxicity after administration. Suitable pharmaceutically acceptable carriers are well-known to those of ordinary skill in the art. A full description of pharmaceutically acceptable carriers can be found in Remington’s Pharmaceutical Sciences. Pharmaceutically acceptable carriers in the composition may contain liquids, such as water, phosphate buffer solution, ringer solution, physiological saline, balanced salt solution, glycerol, or sorbitol, etc. Additionally, auxiliary substances may also be present in these carriers, such as lubricants, glidants, wetting agents or emulsifiers, pH buffering substances, and stabilizers, such as albumin, etc.
[0046] On the other hand, the present invention also provides the use of the aforementioned pharmaceutical composition in the preparation of a drug for preventing and / or treating malignant multi-pleural effusion of tumors.
[0047] The experimental process and results of the present invention will be described in detail below. Unless otherwise specified, the reagents or materials used in the embodiments of the present invention are purchased from commercial products. The main reagents used include: 2,2'-thiobis(ethylamine) (purchased from Jiangsu Aikon Biopharmaceutical R & D Co., Ltd., product number 871-76-1), acryloyl chloride (purchased from Tianjin Xiensi Biochemical Technology Co., Ltd., product number 814-68-6), spermine (purchased from Tianjin Xiensi Biochemical Technology Co., Ltd., product number 71-44-3), N,N'-methylenebisacrylamide (purchased from Tianjin Xiensi Biochemical Technology Co., Ltd., product number: 110-26-9), N,N-dimethylethylenediamine (purchased from Tianjin Xiensi Biochemical Technology Co., Ltd., product number 108-00-9), folic acid-conjugated polyethylene glycol phospholipid (DSPE-PEG-FA) (purchased from Aladdin, product number B2224303), dioleoylphosphatidylethanolamine (DOPE) (purchased from Ruixi Biology, product number 4004-05-1), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) (purchased from Merck & Co., product number 816-94-4), cholesterol (purchased from Tianjin Xiensi Biochemical Technology Co., Ltd., product number 57-88-5), BMN673 (purchased from Selleck Biotechnology Co., Ltd.).
[0048] Experimental animals: C57BL / 6, 6-8 weeks old, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. All animals were housed in a pathogen-free animal facility. Food and water were supplied at a controlled temperature (22 ± 2 °C), and the light-dark cycle was 12 hours. All animal experiments were conducted in accordance with the ethical approval number A2023097-003 of Shanghai Jiao Tong University.
[0049] Example 1 Preparation and related characterization of siLIN28b / DSSP@lip-PEG-FA material
[0050] It should be noted that the siLIN28b / DSSP@lip-PEG-FA material provided in this example, whether in terms of material selection, structural design or specific preparation method, is only a specific example. In fact, according to different application requirements, different structural designs and preparation methods can be adopted to prepare the delivery material, as long as the prepared delivery material can complete the in vivo delivery of the LIN28B inhibitor (siLIN28b in this example).
[0051] Next, the preparation process of the LIN28B inhibitor delivery material provided in this example will be described in detail.
[0052] 1. Synthesis of redox-sensitive polyamine (DSSP)
[0053] As Figure 1As shown, in this embodiment, spermine is synthesized into poly-spermine by two-step reactions using a monothiol bond to prepare DSSP. It should be noted that DSSP will be mixed with siLIN28b and then encapsulated in liposomes later, because DSSP can form a stable complex with siLIN28b, thereby improving the in vivo stability of siLIN28b. It can be understood that in other embodiments, other substances with similar functions can also be mixed with siLIN28b for delivery, or siLIN28b can be directly delivered alone without mixing with substances such as DSSP. The present invention does not limit this. The specific synthesis process of DSSP in this embodiment is as follows:
[0054] Dissolve 0.025 mol of 2,2'-thiobis(ethylamine) in 50 mL of dichloromethane (DCM) in a three-necked flask, and then cool the solution to 0 - 5 °C in an ice-water bath. Under the protection of N2 atmosphere, simultaneously slowly drop 5 mL of 0.1 mol acryloyl chloride dichloromethane solution and 10 mL of 0.2 mol NaOH aqueous solution into the reaction flask. After stirring for 1 h, heat to room temperature and react for 12 h. After the reaction is completed, evaporate dichloromethane under reduced pressure, wash with water to remove the generated salts, extract the reaction product with ethyl acetate, and remove the reaction solvent under reduced pressure to obtain a crude product of thiocystamine bisacrylamide (TCBA). Purify the aforementioned crude product by silica gel column chromatography. The eluent is: n-hexane:ethyl acetate = 40:1. Collect the first spot and spin-dry to obtain a yellow liquid (7.3 g), which is compound 1 with a yield of 85%.
[0055] Dissolve 2 mmol of spermine (SP) in 10 mL of 10% v / v methanol aqueous solution, and then add it to a three-necked flask equipped with a condenser under the protection of N2 atmosphere at 50 °C. Subsequently, add an equimolar methanol solution of compound 1 (i.e., 2 mmol of compound 1 dissolved in 10 mL of 10% v / v methanol aqueous solution), and react in the dark. The reactant is dialyzed for 2 days to remove unreacted spermine, and then freeze-dried to obtain a light yellow solid product DSSP.
[0056] As Figures 2A - 2F shown, proton nuclear magnetic resonance ( 1 1H NMR) and carbon nuclear magnetic resonance ( 13 13C NMR) are used to identify the structures of the prepared compound 1 and DSSP. The results show that the method of this embodiment successfully synthesizes compound 1 and DSSP. The spectral analysis is as follows:
[0057] Confirmation of the structure of compound 1: According to Figure 2AAs shown, it can be seen that the methylene groups at δ2.718 (d, J = 6.0 Hz, 2H) and δ3.536 (d, J = 6.0 Hz, 2H) are coupled with each other, and it is inferred to be the -S-CH2-CH2-NH- structure. The unsaturated double bonds at δ6.273 (d, J = 16.8 Hz, 1H), δ6.177 (dd, J = 16.8 Hz and 10.2, 1H) and δ5.640 (d, J = 10.2 Hz, 1H) are coupled with each other, and it is inferred to be the -CH=CH2 structure; combined with Figure 2B the two unsaturated carbons (δ130.82 and 126.91), two saturated carbons (δ38.91 and 32.01), and one amide carbon (δ166.08) in
[0058] further verify the above structural fragments, and the structure of the obtained compound is confirmed to be Compound 1. Figure 2A The structural fragment of -S-CH2-CH2-NH- (δ3.536 and δ2.718) in Figure 2C is basically consistent with the methylene groups δ3.387 and δ2.704 in the 1H-NMR of 1 ; the two saturated carbons (δ38.91 and 32.01) and one amide carbon (δ166.08) in Figure 2B are basically consistent with (δ37.454, δ31.148) and (δ162.257) in Figure 2D . Therefore, it is determined that the structural fragments of -S-CH2-CH2-NH- and amide also exist in DSSP. According to the spermine structural fragment (δ2.619, δ2.577, δ2.547, δ1.595 and δ1.472) in Figure 2E is basically consistent with the methylene groups δ2.990, δ2.911, δ2.849, δ1.797 and δ1.639 in Figure 2C ; δ48.932, δ46.657, δ32.026 and δ26.863 in Figure 2F are basically consistent with δ48.412, δ46.256, δ27.972 and δ25.534 in Figure 2D . Therefore, it is determined that the spermine structural fragment of -NH-CH2-CH2-CH2-NH-CH2-CH2- exists in DSSP. In addition, the unsaturated carbon-carbon double bond in Compound 1 disappears to generate two methylene groups, and Figure 2F δ39.146 in Figure 2D becomes δ45.786 in
[0059] 2. Preparation of liposome carriers
[0060] Cholesterol and phospholipids (phospholipids include DOPE and DSPC) were dissolved in ethanol at a molar ratio of 1:1 (or a mass ratio of 2:1), dried under nitrogen, and then 10% m / v glucose solution was added for incubation and esterification to obtain a blank liposome carrier.
[0061] 3. Design of the siLIN28b sequence
[0062] In this example, a siRNA interfering with Lin28B, designated as siLin28b, was designed. It contains a sense strand and an antisense strand, and the specific sequences are as follows:
[0063] Nucleotide sequence of the sense strand: 5’-GGAUGUAUUUGUACACCAAtt-3’ (SEQ ID NO: 1);
[0064] Nucleotide sequence of the antisense strand: 5’-UUGGUGUACAAAUACAUCCtt-3’ (SEQ ID NO: 2).
[0065] The above designed sequences were submitted to a biological company for synthesis. Among them, tt at the 3’ end represents the overhang.
[0066] 4. Preparation of the siLIN28b / DSSP@lip-PEG-FA material
[0067] The previously obtained siLin28b was mixed with DSSP at a mass ratio of 1:4 (using PBS buffer solution), and siLin28b / DSSP nanoparticles were obtained by vortex oscillation. Then, the prepared blank liposomes were mixed with the siLin28b / DSSP nanoparticles, and they were extruded 20 times through a liposome extruder with 0.45 μm and 0.22 μm microporous membranes, and then DSPE-PEG-FA was added and incubated at room temperature for 10 minutes to obtain siLin28b / DSSP@lip-PEG-FA.
[0068] 5. Material characterization
[0069] (1) Particle size analysis and TEM characterization
[0070] The above-prepared siLin28b / DSSP nanoparticles and siLin28b / DSSP@lip-PEG-FA materials were subjected to particle size analysis and TEM characterization. The results are shown in Figure 3 a and b. Compared with the siLin28b / DSSP nanoparticles (designated as siRNA / DSSP NPs in the figure), the particle size distribution of the siLin28b / DSSP@lip-PEG-FA material (designated as siRNA / DSSP@lip-PEG-FA in the figure) showed a slight increase.
[0071] Transmission electron microscopy (TEM) images showed that siLIN28b / DSSP@lip-PEG-FA had an additional membrane layer on the outside.
[0072] (2) Evaluation of the ability of siLIN28b / DSSP nanoparticles to release siRNA in an intelligent response in vitro
[0073] By labeling siRNA (i.e., siLin28b) with FAM fluorescent dye, at 37 °C, siLIN28b / DSSP nanoparticles were added to PBS buffer, PBS buffer containing 10 mM GSH, or PBS buffer containing 1 mM H2O2, and kept under gentle stirring. Then, aliquots (500 μL) were collected at predetermined time intervals (0, 2, 4, 8, 12, 24 hours). The sample collection operation was as follows: ultracentrifugation at 40000×g for 30 minutes, and after taking 500 μL of the supernatant, the supernatant was immediately replaced with an equal volume of fresh buffer to maintain the sink conditions. After sample collection, the cumulative release of FAM-siRNA was determined by measuring the fluorescence intensity (λex / λem = 488 / 520 nm) using a fluorescence microplate reader (BioTek Synergy H1).
[0074] The results were as Figure 4 shown, siLIN28b / DSSP nanoparticles could rapidly release siRNA (i.e., siLIN28b) under redox conditions; while under PBS conditions, less than 20% of the siRNA was released within 24 hours. It was shown that the siLIN28b / DSSP nanoparticles prepared in the present invention could react with the abundant hydrogen peroxide in the tumor microenvironment to achieve responsive degradation.
[0075] Example 2 Therapeutic effect of the combination of siLIN28b and BMN673 on malignant pleural effusion of tumors
[0076] The siLin28b / DSSP@lip-PEG-FA material prepared in Example 1 and the PARP inhibitor (BMN673 in this example) were used to evaluate the in vivo anti-tumor effect and the therapeutic effect on malignant pleural effusion of tumors by single administration or combined administration. Specifically, it included:
[0077] 1×10 7Eight ID8 cells (ovarian cancer cell line) were intraperitoneally injected into C57BL / 6 mice (each mouse weighing approximately 20 g - 21 g) to establish a mouse model of malignant pleural effusion. On the 10th day after injection, the baseline abdominal fluorescence was evaluated using the IVIS Lumina II optical imaging system. Subsequently, the mice were randomly divided into four groups and treated accordingly: the solvent control group (PBS group), the siLin28b group, the BMN673 group, and the combined administration group of siLin28b + BMN673. Among them, the dosage of siLin28b (i.e., siLin28b / DSSP@lip-PEG-FA material) was 10 nmol / mouse (10 nmol refers to the dosage of the active ingredient siLin28b of the material), and the dosage of BMN673 was 0.33 mg / kg. Subsequently, tumor progression was monitored through the IVIS Lumina II system, while the formation of ascites and the abdominal height of the mice were monitored through ultrasound imaging, and survival analysis was determined by measuring the abdominal circumference. The endpoint was defined as twice the normal abdominal circumference of healthy mice.
[0078] Figure 5 For the ultrasonic detection of the ascites height and quantitative analysis of the mice on the 45th day after administration, the results showed that by the 45th day, the abdominal circumference of the mice in the PBS group was twice that of the other groups, and obvious fluid accumulation was observed in the peritoneal cavity, resulting in organ compression; after the administration of siLin28b or BMN673 alone, the amount of ascites decreased to a certain extent; but after the combined administration, the amount of fluid accumulation in the peritoneal cavity of the mice was the least, and the abdominal height was the lowest, with the abdominal height not exceeding 10 mm.
[0079] Figure 6 For the photographed images of the ascites of the mice collected on the 45th day after administration and the quantitative analysis data, the results further demonstrated the therapeutic effect of the combined administration of siLin28b and BMN673 on malignant pleural effusion. Specifically, it was found that by the 45th day, the amount of ascites in the mice in the siLin28b + BMN674 combined administration group was the smallest, while the ascites volume of the mice in the other groups exceeded 1 ml, especially the ascites of the mice in the solvent control group was the most, up to 10 ml.
[0080] Figure 7 For the comparison of the survival analysis curves of the mice in each group. Survival analysis was performed by recording the time when the mice developed ascites and the time when the abdominal circumference of the mice was twice that of normal mice (the endpoint of mouse death). The mice in the PBS group developed ascites on the 13th day, while the mice in the siLin28b group and the BMN673 group developed ascites on the 24th day and the 22nd day respectively, and the mice in the siLin28b + BMN673 combined administration group developed ascites on the 28th day ( Figure 7a). In addition, except for the group receiving combined administration of siLin28b + BMN673, mice in other groups showed mortality on the 45th day. The PBS group showed complete mortality, while mice treated with siLin28b + BMN673 did not die at 45 days. Figure 7 b), further demonstrating that the combined administration of siLin28b and BMN673 can synergistically treat malignant pleural effusion of tumors and prolong the survival period of mice.
[0081] In addition, it should be specifically pointed out that in the anti-tumor activity detection experiment of the combined administration of siLin28b and BMN673 in the present invention, although the combined administration of siLin28b and BMN673 can inhibit the growth of tumors to a certain extent, the inhibitory effect is very weak, extremely unobvious, far less prominent than the effect exerted by these two drugs in the treatment of malignant pleural effusion of tumors.
[0082] In addition, after the experiment of the present invention was completed, the main organs (heart, liver, spleen, lung and kidney) and tumors were also collected for biocompatibility evaluation. The results are as Figure 8 shown. The results of the tissue organ safety and serum biochemical indexes of the mice in the PBS group and the combined administration group indicate that the treatment with siLin28b combined with BMN673 is safe and non-toxic.
[0083] In summary, the present invention provides a pharmaceutical composition for preventing and / or treating malignant pleural effusion of tumors, including: a LIN28B inhibitor and a PARP inhibitor. It is demonstrated by in vivo experiments that after the combined administration of a LIN28B inhibitor (such as siLin28b) and a PARP inhibitor (such as BMN673), it can synergistically treat malignant pleural effusion of tumors, improve the survival period of patients, and can be used in the preparation and application of drugs for the prevention and treatment of malignant pleural effusion of tumors, having good application prospects.
[0084] Although the content of the present invention has been introduced in detail through the above preferred embodiments, it should be recognized that the above description should not be considered as a limitation of the present invention. After those skilled in the art have read the above content, various modifications and substitutions of the present invention will be obvious. Therefore, the protection scope of the present invention should be defined by the appended claims.
Claims
1. A pharmaceutical composition for preventing and / or treating malignant multi - pleural effusion of tumors, characterized in that, Comprising: LIN28B inhibitor and PARP inhibitor.
2. The pharmaceutical composition for preventing and / or treating malignant multi - pleural cavity effusion of tumors according to claim 1, characterized in that, The inhibitor includes: any one or a combination of multiple kinds of siRNA or gene knockout vectors targeting the coding gene, or protein antibodies targeting the protein or other inhibitors capable of inhibiting protein expression.
3. The pharmaceutical composition for preventing and / or treating malignant multi - pleural cavity effusion of tumors according to claim 1, wherein, The LIN28B inhibitor is siLIN28b, the nucleotide sequence of its sense strand is as shown in SEQ ID NO: 1, and the nucleotide sequence of its antisense strand is as shown in SEQ ID NO:
2.
4. The pharmaceutical composition for preventing and / or treating malignant multi - pleural cavity effusion of tumors according to claim 3, characterized in that, The PARP inhibitor is BMN673.
5. The pharmaceutical composition for preventing and / or treating malignant multi-pleural cavity effusion of tumors according to claim 4, characterized in that, The mass ratio of the siLIN28b and BMN673 is 1:500 - 1:1000.
6. The pharmaceutical composition for preventing and / or treating malignant multi - pleural cavity effusion of tumors according to claim 3, wherein, The siLIN28b is delivered by a viral vector, and the viral vector includes any one or more of an adeno-associated virus vector, an adenovirus vector, or a retrovirus vector.
7. The pharmaceutical composition for preventing and / or treating malignant multi - pleural cavity effusion of tumors according to claim 3, wherein The siLIN28b is delivered by a non-viral vector, and the non-viral vector includes any one or more of a plasmid vector, a liposome vector, or a cationic polymer vector.
8. The pharmaceutical composition for preventing and / or treating malignant multi-pleural cavity effusion of tumors according to claim 7, characterized in that, The siLIN28b is delivered by a liposome vector, and the liposome vector is modified with folic acid polyethylene glycol phospholipid.
9. The pharmaceutical composition for preventing and / or treating malignant multi-pleural effusion of tumors according to claim 3, wherein The pharmaceutical composition further includes a pharmaceutically acceptable carrier.
10. Use of the pharmaceutical composition according to any one of claims 1 - 9 in the preparation of a drug for preventing and / or treating malignant multi-pleural effusion of tumors.