STAT3 small-molecule inhibitor as well as screening method and application thereof

Abstract

The invention discloses a screening method of a small molecule STAT3 inhibitor and application of the small molecule STAT3 inhibitor. The screening method comprises the following steps: obtaining an STAT3 protein-small molecule compound crystal structure from a Protein Data Bank database, and carrying out pretreatment on the STAT3 protein-small molecule compound crystal structure; a SiteMap module is adopted to search a protein surface binding pocket, and a pocket at an SH2 structural domain is selected to screen an STAT3 inhibitor; selecting the center of a butt joint box and setting the size of the butt joint box; selecting and downloading a to-be-screened compound library; compound library preprocessing, multi-mode molecular docking and MM / GBSA free energy calculation are performed in sequence in a virtual screening workflow mode, and sorting is performed according to combination free energy values from small to large; and a molecular dynamics simulation method of Binding Pose Metadynamics is adopted for further screening, so that a compound with potential activity is obtained.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a virtual screening method and application for STAT3 small molecule inhibitors. Background Technology

[0002] Signal transducers and activators of transcription (STATs) are an important class of transcription factors involved in multiple biological processes, including cell cycle, differentiation, proliferation, and apoptosis, and possess dual functions of signal transduction and transcriptional regulation. The STAT family consists of seven members—STAT1, STAT2, STAT3, STAT4, STAT5A, STAT5B, and STAT6—which are highly similar in structure and function. These STAT protein isoforms share several common domains: the N-terminal domain (NTD), the coil-coil domain (CCD), the DNA-binding domain (DBD), the linker domain (LD), the Src homology 2 domain (SH2), and the transcription activation domain (TAD). Each domain plays a different role in signal transduction and gene transcription activation. (J. Dong, et al., J. Med. Chem., 2021, 64, 8884-8915; G. Miklossy, et al., Nat. Rev. Drug Discovery, 2013, 12, 611-629.) Despite their high structural homology, STAT3 typically acts on different cellular processes. Studies have shown that STAT3 plays a crucial role in the early, advanced stages of cancer and in the tumor microenvironment. Abnormal activation of STAT3 is closely related to tumor cell proliferation, invasion, and metastasis, and has therefore attracted considerable attention (Johnson, DE; O'Keefe, RA; Grandis, JR, Nat. Rev. Clin. Oncol. 2018, 15, 234-248.). Hyperphosphorylation of STAT3 has been detected in various tumor cell lines and clinical samples, including breast cancer, pancreatic cancer, and gastric cancer. Therefore, inhibiting STAT3 activation or expression is an effective approach for treating various cancers.

[0003] Continuously activated STAT3 is crucial for the development of various cancers, including breast cancer, colorectal cancer, pancreatic cancer, and gastric cancer, making it an ideal drug target. Although no STAT3 inhibitor has yet been approved for the market, several candidates (such as Napabucasin, STA-21, CDDO-Me, TTI-1-1, Nicolasamide, Pyrimethamine, and WP1066) have successfully entered early-stage clinical trials, which has inspired our interest in exploring more effective STAT3 inhibitors (J. Wang, et al., RSC Med. Chem., 2025, 16, 2848.).

[0004] High-throughput virtual screening (HTVS) is a cutting-edge biopharmaceutical technology that uses computational algorithms to identify bioactive molecules from a vast library of compounds (NMTripathi, et al., Eur. J. Med. Chem., 2022, 243, 114766.). This method is widely used in the screening of lead compounds and plays a crucial role in the early stages of drug development. With the resolution of the three-dimensional structures of a large number of biomolecules (including proteins), computer-aided drug design based on receptor structures is becoming increasingly feasible (T. Zhu, et al., J. Med. Chem., 2013, 56, 6560-6572.). Compared with traditional drug screening methods, virtual screening not only significantly shortens the drug development cycle but also reduces related costs (D. Steadman, et al., J. Med. Chem., 2022, 65, 562-578.). Therefore, establishing an efficient and rapid screening method is of great significance for discovering anti-tumor drugs targeting STAT3. Disrupting the interactions within the SH2 domain has been shown to effectively inhibit STAT3 activity; however, this approach still faces challenges in drug discovery, limiting its clinical application (Y. Zhong, et al., Acta Pharmacol. Sin., 2022, 43, 1013-1023; H. Chen, et al., Eur. J. Med. Chem., 2014, 82, 195-203; J. Wang, et al., Adv. Sci., 2022, 9, e2200169.). Summary of the Invention

[0005] To address the aforementioned technical problems and shortcomings in the field, this invention provides a virtual screening method and application for STAT3 inhibitors with anti-tumor efficacy.

[0006] This invention provides a rapid virtual screening method for anti-tumor drugs targeting STAT3. This screening process yielded novel STAT3 inhibitors with significant inhibitory effects on tumor cell proliferation, STAT3 transcription, and cell metastasis. This invention can provide a reference for screening STAT3 inhibitors with anti-tumor effects.

[0007] This invention provides a rapid screening scheme for anti-tumor drugs targeting STAT3. The screened compounds have significant anti-tumor effects and STAT3 inhibitory effects, and can be used to develop drugs for treating anti-tumor or STAT3-inhibition-related diseases.

[0008] The specific technical solution is as follows:

[0009] In a first aspect, the present invention provides a method for screening small molecule STAT3 inhibitors, comprising the following steps:

[0010] (1) Selection and pretreatment of STAT3 protein crystal structure: The crystal structure of STAT3 protein-small molecule complex was obtained from the Protein Data Bank database and the crystal structure of STAT3 protein-small molecule complex was pretreated.

[0011] (2) Searching for protein surface binding pockets: The SiteMap module was used to search for protein surface binding pockets, and pockets at the SH2 domain were selected for screening STAT3 inhibitors;

[0012] (3) Generate docking box: Select the center of the docking box and set the size of the docking box;

[0013] (4) Select and download the compound library to be screened;

[0014] (5) Virtual screening workflow for compounds: The compounds are preprocessed in the virtual screening workflow mode, followed by multi-mode molecular docking and MM / GBSA free energy calculation. The compounds are sorted from smallest to largest according to their binding free energy values.

[0015] (6) Further screening of active molecules using molecular dynamics simulation: Compounds with potential activity were further screened using the molecular dynamics simulation method of Binding Pose Metadynamics.

[0016] This invention uses a SiteMap module to automatically search for protein surface binding pockets and selects pockets at the SH2 domain for screening STAT3 inhibitors. Compared to other binding pockets, compounds obtained through SH2 domain screening exhibit higher hit rates for STAT3 inhibition and cell proliferation inhibition activities.

[0017] In step (1), the crystal structure of the STAT3 protein-small molecule complex can be the crystal structure of the complex of human STAT3 protein and small molecule SD-36.

[0018] In step (1), the Protein Preparation Workflow module can be used to preprocess the crystal structure of the STAT3 protein-small molecule complex.

[0019] In step (1), the pretreatment may include one or more of the following: deleting solvent molecules, hydrogenation, repairing missing residues and side chains, protonating protein residues, optimizing hydrogen bond networks, minimizing energy, and repairing incorrect atom types.

[0020] Preferably, in step (3), the Receptor Grid Generation module is used to generate the docking box. The docking box is set with the SH2 domain found in the SiteMap as the center and the pocket as the center. The size of the box is set to

[0021] In step (4), you can select the ChemDiv compound library and download all the molecular 3D structures therein.

[0022] Preferably, in step (5), the virtual screening workflow for compounds is performed in the Virtual Screening Workflow module: the compound library is first filtered by the QikProp module to ensure that the screened compounds have good pharmacokinetic characteristics and physicochemical properties. The filtered molecules are protonated and stereochemical information is obtained through the LigPrep module, and then they enter the molecular docking stage. The screened compounds are obtained by molecular docking screening through three stages: high throughput virtual screening (HTVS), standard precision screening (SP), and extra precision screening (XP). The binding free energy of these screened compounds is calculated by the MM / GBSA method, and the binding free energy is calculated according to the MM / GBSA ΔG. bind The score (binding free energy) values ​​are sorted from smallest to largest.

[0023] Preferably, in step (6), the molecular dynamics simulation uses the Binding Pose Metadynamics module. Each protein ligand docking complex undergoes a 1×10ns molecular dynamics simulation. Compounds with CompScore values ​​<0 are screened and then subjected to a 10×10ns molecular dynamics simulation. The molecules are sorted from smallest to largest according to their CompScore values, and the top-ranked molecules are selected as compounds with potential activity.

[0024] Secondly, the present invention provides the use of compounds D1, D3, D4, D5, D8, D11, D12, D13, D14, D16, D17, D18, D19, or pharmaceutically acceptable salts thereof having the following structures in the preparation of STAT3 inhibitors:

[0025]

[0026] Thirdly, the present invention provides the use of compounds D1, D3, D4, D5, D8, D11, D12, D13, D14, D16, D17, D18, D19, or pharmaceutically acceptable salts thereof having the following structures in the preparation of medicaments for treating and / or preventing diseases associated with abnormal STAT3 activity:

[0027]

[0028] In the third aspect of the application, the diseases include breast cancer, stomach cancer, prostate cancer, lung cancer, pancreatic cancer, etc.

[0029] Compared with the prior art, the beneficial effects of this invention are as follows:

[0030] The screening scheme for antitumor drugs targeting STAT3 described in this invention is simple, rapid, and efficient, with a high probability of identifying positive compounds. The screened compounds exhibit significant antitumor and STAT3-inhibiting effects. Attached Figure Description

[0031] Figure 1 This is a schematic diagram of the STAT3 protein surface binding pockets and their main parameter scores obtained by using the SiteMap module in Example 1.

[0032] Figure 2 The graph shows the results of the tumor cell proliferation inhibition activity verification of the compound in Example 2 and the positive control drugs Napa (Napabucasion) and BP (BP-1-102).

[0033] Figure 3 This is a graph showing the results of STAT3 transcriptional repression activity verification of the compound in Example 3 and the positive control drug Napa.

[0034] Figure 4 The graph shows the results of the analysis of the anti-metastatic effect of the compound and the positive control drug Napa on tumor cells in Example 4. Detailed Implementation

[0035] The present invention will be further described below with reference to the accompanying drawings and specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Operating methods not specifically specified in the following embodiments are generally performed under conventional conditions or as recommended by the manufacturer.

[0036] Example 1:

[0037] Virtual screening process:

[0038] A method for screening small molecule STAT3 inhibitors, comprising:

[0039] Step (1), Selection and preprocessing of STAT3 protein crystal structure: The crystal structure of the complex of human STAT3 protein and small molecule was obtained from the Protein Data Bank database. This structure is the crystal structure of the complex of STAT3 protein and small molecule SD-36 (PDB: 6NJS). The protein ligand complex structure was preprocessed using the Protein Preparation Workflow module, specifically including solvent molecule removal, hydrogenation, repair of missing residues and side chains, protonation of protein residues, optimization of hydrogen bond network, and energy minimization.

[0040] Step (2), searching for protein binding pockets: The SiteMap module was used with default parameters to search for binding pockets of the protein processed in step (1), and a total of 5 binding pockets (Site1-Site5) were obtained. Figure 1 ) Site4, located in the SH2 domain, was selected for screening STAT3 inhibitors.

[0041] Step (3), generate docking boxes: Use the Receptor Grid Generation module to generate docking boxes, centering the docking boxes around Site4 and the pocket, and setting the box size to [size missing]. All other parameters are set to default.

[0042] Step (4), select and download the compound library to be screened: select the ChemDiv compound library and download the 3D structures of all small molecules in the compound library from its official website (https: / / www.chemdiv.com / ), totaling 1,666,973 molecules (as of March 2024).

[0043] Step (5), Virtual Screening Workflow for Compounds: The virtual screening workflow for compounds is carried out in the Virtual ScreeningWorkflow module: The ChemDiv 3D database is first filtered by the QikProp module to ensure that the compounds obtained by screening have good pharmacokinetic characteristics and physicochemical properties; the filtered molecules are protonated by the LigPrep module to obtain stereochemical information (obtained from the 3D configuration of each small molecule); then, the molecular docking stage is entered, and 1666 compounds are obtained by molecular docking screening in three stages: high throughput virtual screening (HTVS) to retain the top 10% of conformational molecules, standard precision screening (SP) to retain the top 10% of conformational molecules, and extra precision screening (XP) to retain the top 10% of conformational molecules; the bounding free energy of the screened molecules is calculated by the MM / GBSA method, and the bounding free energy is calculated according to the MM / GBSA ΔG. bind The compounds are sorted by score from smallest to largest, and the top 160 compounds are selected for further analysis. This virtual screening workflow sequentially performs compound library preprocessing, multi-mode molecular docking, MM / GBSA calculation and re-scoring, and sorts the compounds by binding free energy from smallest to largest, ultimately obtaining compounds that can be used for further screening.

[0044] Step (6) further screens active molecules using molecular dynamics simulations: Binding Pose Metadynamics (BPMD) is an enhanced sampling method that can effectively assess the binding stability of ligands to proteins in solution. Simulations are performed in the Binding Pose Metadynamics module, with each protein-ligand complex undergoing a 1×10 ns molecular dynamics simulation. Compounds with CompScore values ​​<0 are then subjected to another 10×10 ns molecular dynamics simulation. On a single GPU (NVIDIA RTX 4090), the simulation time for each complex is approximately 5 hours; simulation times vary depending on the GPU model. Based on the CompScore values ​​obtained from the simulation (CompScore = PoseScore - 5×PersScore), molecules are sorted from smallest to largest, and the top-ranked molecules are purchased and their activity verified experimentally. Ultimately, 13 compounds with the following structures (D1, D3, D4, D5, D8, D11, D12, D13, D14, D16, D17, D18, D19) were successfully purchased for activity verification.

[0045]

[0046] Figure 1 The table shows the STAT3 protein surface binding pockets Site1-Site5 found using the SiteMap module. Ligand is the protein protoligand binding site. The table lists the scores of the main parameters for each pocket, including SiteScore, Dscore, Volume, and Balance.

[0047] Example 2:

[0048] Cytotoxicity evaluation:

[0049] Cell viability was evaluated using the CCK8 assay to detect the inhibitory effect of the compound on the proliferation of human breast cancer cells MDA-MB-231 and MDA-MB-468. The specific procedure is as follows:

[0050] Tumor cells in logarithmic growth phase were collected and prepared into a cell suspension. 100 μL / well (approximately 3000 cells) was seeded into 96-well plates, with three replicates. The plates were incubated at 37°C with 5% CO2 for 24 h. The test compound was dissolved in DMSO to prepare a 10 mM stock solution, which was then diluted with cell culture medium. After removing the plate, different concentrations of the test sample were added to each well, with three replicates. The plates were then incubated in a CO2 incubator for another 72 h. The plate was then removed, and 10 μL of CCK8 solution was added to each well. The plates were incubated at 37°C with 5% CO2 for 1–2 h. The absorbance (OD value) at 450 nm was measured using a microplate reader.

[0051] The cell proliferation inhibition rate of the drug is calculated based on the OD value, using the following formula:

[0052] Cell viability (%) = (OD) sample -OD blank ) / (OD control -OD blank )×100%;

[0053] Cell inhibition rate (%) = (OD) control -OD sample ) / (OD control -OD blank )×100%;

[0054] OD sample : OD value of the experimental well;

[0055] OD control : OD value of the reference hole;

[0056] OD blank : OD value of the blank hole.

[0057] The half-maximum inhibitory concentration (IC50) was obtained by fitting the inhibition rate at each concentration. 50 value.

[0058] Figure 2 The activity verification results of the compounds screened in Example 1 and the positive control drugs Napa (Napabucasion) and BP (BP-1-102) are shown in the figure. Figure 2 Results showed that at a test concentration of 50 μM, 7 out of 13 compounds (D4, D8, D11, D14, D17, D18, and D19) exhibited strong inhibitory activity against the proliferation of MDA-MB-231 tumor cells (inhibition rate >50%); and 4 compounds (D4, D8, D17, and D18) exhibited strong inhibitory activity against the proliferation of MDA-MB-468 tumor cells (inhibition rate >50%). Compounds with an initial inhibition rate >50% were subjected to IC50 assay. 50 Tests showed that D4, D8, D14, and D18 had an effect on the IC50 of MDA-MB-231 cells. 50 The values ​​were 8.66, 3.81, 10.51, and 2.83 μM, respectively. D8 and D18 showed inhibitory effects comparable to and superior to the positive control drug Napa. The IC50 values ​​of D4, D8, and D18 on MDA-MB-468 cells were also observed. 50 The values ​​were 5.83, 2.75, and 5.51 μM, respectively. D8 showed comparable inhibitory activity to the positive control drug Napa and was superior to the positive control BP. Figure 2 B).

[0059] Example 3:

[0060] Luciferase reporter gene assay:

[0061] STAT3(Luc)HEK293 cells are a stable polyclonal HEK293 cell line containing multiple STAT3 binding sites in its genome. The transcriptional activation level of STAT3 can be detected with high sensitivity by measuring the activity of firefly luciferase (Luc). Logarithmically growing STAT3(Luc)HEK293 cells were prepared into a cell suspension and seeded at 100 μL per well (approximately 20,000 cells) into black 96-well plates, with three replicates. The plates were incubated at 37°C with 5% CO2 for 24 h. The test compound was dissolved in DMSO to prepare a 10 mM stock solution, which was then diluted with cell culture medium to the required concentration. After removing the 96-well plate, 20 ng / mL IL-6 was added to each well to stimulate cells for 30 min to activate STAT3 transcription. Different concentrations of the test compound were then added. Cell-free wells were included as blank controls. The plates were incubated in a CO2 incubator for 24 h. After removing the 96-well plate and equilibrating it to room temperature for 5-10 minutes, add 100 μL of firefly luciferase assay reagent to each well under dark conditions. Incubate at room temperature for 5 minutes, then perform chemiluminescence detection using a multi-functional microplate reader with chemiluminescence detection capabilities. Obtain the half-maximal inhibitory concentration (IC50) based on the inhibition rates of the tested compounds at various concentrations. 50 value.

[0062] Figure 3 The diagram shows the verification results of the STAT3 transcriptional repression activity of the compounds screened in Example 1 and the positive control drug Napa. Figure 3 Results showed that at a test concentration of 50 μM, 8 out of 13 compounds (D1, D3, D4, D11, D12, D13, D17, and D18) exhibited STAT3 transcriptional repression activity >50%. Compounds with an initial inhibition rate >50% were subjected to IC50 assay. 50 Test results show that ICs D1, D3, D4, D11, D17, and D18... 50 All values ​​were below 20 μM, with D18 exhibiting the strongest STAT3 transcriptional repression activity at 1.62 μM, superior to the positive control Napa. Figure 3 B).

[0063] Example 4:

[0064] Analysis of cell anti-metastasis effect:

[0065] MDA-MB-231 cells were used at 4 × 10 5Cells were seeded at a density of 5 × 10⁶ cells / well in 6-well plates and pretreated with different concentrations of the compound. The matrix gel was thawed overnight at 4°C, and pipette tips and centrifuge tubes were pre-chilled. The matrix gel concentration was diluted to 200–300 μg / mL with basal medium. 100 μL of matrix gel was added to the upper chamber of each Transwell compartment, and the compartment was incubated for 1–2 hours to solidify. The supernatant was discarded. Basal medium was added to the compartment, and the compartment was incubated at 37°C for hydration. The liquid in the compartment was removed, and the compartment was checked for any liquid passing through it. If no liquid was found, cells could be seeded. Complete medium was added to the lower chamber, and the compartment was carefully placed on top using forceps. Pretreated MDA-MB-231 cells were seeded at a density of 5 × 10⁶ cells / well. 4 Cells were seeded at a density in the upper chamber and treated with different concentrations of compounds, then cultured in serum-free medium for 48 hours. After removing unmigrated cells, the cells on the bottom surface of the filter membrane were fixed with 4% paraformaldehyde and stained with crystal violet. After rinsing, the cells were photographed under a microscope, with multiple fields of view taken randomly for each group.

[0066] Figure 4 The results show the validation of the tumor cell metastasis inhibitory ability of the highly active compounds screened in Examples 2 and 3 and the positive control drug Napa. The results indicate that compounds D4, D8, and D18 at a concentration of 1.25 μM can significantly inhibit the invasive ability of MDA-MB-231 breast cancer cells, with effects comparable to Napa.

[0067] In summary, STAT3 plays a crucial role in the early, advanced stages of cancer and in the tumor microenvironment. Abnormal STAT3 activation is closely related to tumor cell proliferation, invasion, and metastasis. Hyperphosphorylation of STAT3 has been detected in various tumor cell lines and clinical samples, including breast cancer, pancreatic cancer, and gastric cancer. Therefore, inhibiting STAT3 activation or expression is an effective approach for treating various cancers. This invention provides a rapid screening scheme for anti-tumor drugs targeting STAT3. The screened compounds exhibit significant anti-tumor activity and STAT3 inhibitory activity. The compounds obtained in this invention can be used as lead compounds for further optimization and development into drugs for treating malignant tumors or diseases related to abnormal STAT3 expression.

[0068] Furthermore, it should be understood that after reading the above description of the present invention, those skilled in the art can make various alterations or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A method for screening small molecule STAT3 inhibitors, characterized in that, Including the following steps: (1) Selection and pretreatment of STAT3 protein crystal structure: The crystal structure of STAT3 protein-small molecule complex was obtained from the Protein Data Bank database and the crystal structure of STAT3 protein-small molecule complex was pretreated. (2) Searching for protein surface binding pockets: The SiteMap module was used to search for protein surface binding pockets, and pockets at the SH2 domain were selected for screening STAT3 inhibitors; (3) Generate docking box: Select the center of the docking box and set the size of the docking box; (4) Select and download the compound library to be screened; (5) Virtual screening workflow for compounds: The compounds are preprocessed in the virtual screening workflow mode, followed by multi-mode molecular docking and MM / GBSA free energy calculation. The compounds are sorted from smallest to largest according to their binding free energy values. (6) Further screening of active molecules using molecular dynamics simulation: Compounds with potential activity were further screened using the molecular dynamics simulation method of Binding Pose Metadynamics.

2. The screening method for small molecule STAT3 inhibitors according to claim 1, characterized in that, In step (1): The crystal structure of the STAT3 protein-small molecule complex is the crystal structure of the complex of human STAT3 protein and small molecule SD-36; The crystal structure of the STAT3 protein-small molecule complex was pretreated using the Protein Preparation Workflow module; The pretreatment includes one or more of the following: solvent molecule removal, hydrogenation, repair of missing residues and side chains, protonation of protein residues, optimization of hydrogen bond network, energy minimization, and repair of incorrect atom types.

3. The screening method for small molecule STAT3 inhibitors according to claim 1, characterized in that, In step (3), the Receptor Grid Generation module is used to generate docking boxes. The docking boxes are set with the SH2 domain found in the SiteMap as the center and the pocket as the center. The size of the box is set to 4. The screening method for small molecule STAT3 inhibitors according to claim 1, characterized in that, In step (4), select the ChemDiv compound library and download all the molecular 3D structures in it.

5. The screening method for small molecule STAT3 inhibitors according to claim 1 or 4, characterized in that, In step (5), the virtual screening workflow for compounds is carried out in the Virtual Screening Workflow module: the compound library is first filtered by the QikProp module to ensure that the screened compounds have good pharmacokinetic characteristics and physicochemical properties. The filtered molecules are protonated and stereochemical information is obtained by the LigPrep module, and then they enter the molecular docking stage. The molecular docking screening is carried out in three stages: high-throughput screening, standard precision screening, and high-precision screening to obtain the screened compounds. The binding free energy of these screened compounds is calculated by the MM / GBSA method, and the binding free energy is calculated according to the MM / GBSA ΔG. bind The score values ​​are sorted from smallest to largest.

6. The screening method for small molecule STAT3 inhibitors according to claim 1, characterized in that, In step (6), the molecular dynamics simulation uses the Binding Pose Metadynamics module. Each protein ligand docking complex undergoes a 1×10ns molecular dynamics simulation. Compounds with CompScore values ​​<0 are screened and then subjected to a 10×10ns molecular dynamics simulation. The molecules are sorted from smallest to largest according to their CompScore values, and the top-ranked molecules are selected as compounds with potential activity.

7. The use of compounds D1, D3, D4, D5, D8, D11, D12, D13, D14, D16, D17, D18, D19, or pharmaceutically acceptable salts thereof having the structures shown below in the preparation of STAT3 inhibitors:

8. The use of compounds D1, D3, D4, D5, D8, D11, D12, D13, D14, D16, D17, D18, D19, or pharmaceutically acceptable salts thereof having the structures shown below in the preparation of medicaments for the treatment and / or prevention of diseases associated with abnormal STAT3 activity:

9. The application according to claim 8, characterized in that, The diseases mentioned include breast cancer, stomach cancer, prostate cancer, lung cancer, and pancreatic cancer.

Images