Use of compounds or their salts in the preparation of cancer therapeutic drugs

Abstract

This invention belongs to the field of cancer drug preparation technology, specifically relating to the use of compounds or their salts in the preparation of cancer therapeutic drugs. The structure of the compound is shown in Formula I. The compound shown in Formula I can significantly inhibit tumor (e.g., breast cancer) growth. Compared with the blank control group, the tumors of mice treated with the compound shown in Formula I grow more slowly, are smaller and lighter, and have a significantly longer survival period. In addition, the compound shown in Formula I can be used in combination with Anti-PD1 to synergistically enhance the tumor inhibition effect.

Description

Technical Field

[0001] This invention belongs to the field of cancer drug preparation technology, specifically the use of compounds or their salts in the preparation of cancer therapeutic drugs. Background Technology

[0002] The progression of malignant tumors depends on complex biological regulatory mechanisms, including abnormal cell proliferation, angiogenesis, invasion and metastasis, and immune evasion. Currently, clinical treatment still primarily relies on surgery, radiotherapy, chemotherapy, and targeted drugs, but these methods have limitations such as drug resistance, significant toxic side effects, and high treatment costs. In recent years, targeted therapy strategies targeting key regulatory factors in the tumor microenvironment have gradually become a research hotspot.

[0003] Studies have shown that tumor cells can activate G protein-coupled receptors in endothelial cells by secreting specific signaling molecules, triggering a cascade of downstream signaling pathways (such as PI3K / Akt, MAPK / ERK, etc.), thereby promoting vascular endothelial cell proliferation and migration, and accelerating tumor angiogenesis. This abnormal angiogenesis not only provides oxygen and nutrients to tumors but also creates conditions for metastatic cells to breach the basement membrane barrier. In addition, tumor cells can also activate intracellular pro-survival pathways through autocrine signaling, inhibiting the expression of apoptosis-related proteins and enhancing their survival ability in harsh microenvironments.

[0004] At the level of existing therapeutic technologies, RNA interference (RNAi) technology (such as siRNA / shRNA) has been attempted to specifically inhibit the expression of oncogenes, but it suffers from drawbacks such as low targeting efficiency, poor in vivo stability, and insufficient drug-likeness. While monoclonal antibody drugs can specifically inhibit the function of specific proteins, their clinical application is limited by issues such as complex manufacturing processes, large dosages, and heavy economic burden on patients. In contrast, small molecule compounds, due to their advantages of small molecular weight, high structural modifiability, convenient administration methods, and controllable production costs, have become an important direction for targeted therapy development. Summary of the Invention

[0005] This invention screened a small molecule compound (hereinafter also referred to as 3628-0029, which is also the small molecule library number) through the ChemDiv / Chembridge small molecule library, which can effectively inhibit the growth of tumors (such as breast cancer) and can be used in the preparation of cancer treatment drugs.

[0006] To achieve the above objectives, the present invention can adopt the following technical solutions:

[0007] This invention provides the use of a compound or its salt in the preparation of a cancer therapeutic drug, the compound having the structural formula shown in Formula I:

[0008]

[0009] Wherein, R is selected from (CH2). n CH3, where n is an integer.

[0010] Preferably, in the compound of Formula I used for the above purposes, the value of n is 0-4.

[0011] More preferably, in the compound shown in Formula I above, R is CH3.

[0012] Preferably, in the above-described uses, the salt of the compound represented by Formula I is selected as a hydrochloride salt.

[0013] Preferably, in the above-mentioned use, the cancer is breast cancer.

[0014] More preferably, in the above-mentioned use, the cancer is triple-negative breast cancer.

[0015] Preferably, in the above-mentioned uses, the dosage form of the cancer treatment drug includes injections, liquid oral preparations, sprays, patches, tablets, powders, ointments, or suppositories.

[0016] Another aspect of the present invention provides a cancer treatment pharmaceutical composition comprising Anti-PD1 and the compound of Formula I above or a salt thereof.

[0017] In another aspect, the present invention provides a cancer treatment medicament comprising the cancer treatment medicament composition of the present invention.

[0018] In another aspect, the present invention provides the use of the cancer therapeutic pharmaceutical composition of the present invention in the preparation of a cancer therapeutic pharmaceutical.

[0019] The beneficial effects of this invention include:

[0020] (1) The compound shown in Formula I can significantly inhibit tumor (e.g. breast cancer) growth. Compared with the blank control group, the tumors of mice treated with the compound shown in Formula I grew more slowly, and the tumors were smaller and lighter.

[0021] (2) The compound shown in Formula I, when combined with Anti-PD1, has a synergistic effect and can synergistically inhibit tumor growth. The combined effect of the two in inhibiting tumor growth is better than the inhibitory effect of either the compound shown in Formula I alone or Anti-PD1 alone. Attached Figure Description

[0022] Figure 1 This is a photograph of tumor growth after intervention with the 3628-0029 small molecule inhibitor.

[0023] Figure 2 This is a time-volume curve of tumor growth after intervention with the small molecule inhibitor 3628-0029;

[0024] Figure 3The figure shows the effects of the 3628-0029 small molecule inhibitor on blood routine tests and AST and ALT levels in mice. Detailed Implementation

[0025] The illustrated embodiments are provided to better illustrate the present invention, but are not intended to limit the scope of the invention to the illustrated embodiments. Therefore, non-essential improvements and adjustments made to the embodiments by those skilled in the art based on the above description of the invention still fall within the protection scope of the present invention.

[0026] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit this disclosure. Singular expressions include plural expressions unless they have a distinct meaning in the context. As used herein, it should be understood that terms such as “comprising,” “having,” “including,” are intended to indicate the presence of features, numbers, operations, components, parts, elements, materials, or combinations thereof. The terminology of the invention is disclosed in the specification and is not intended to exclude the possibility that one or more other features, numbers, operations, components, parts, elements, materials, or combinations thereof may be present or added. As used herein, “ / ” may be interpreted as “and” or “or,” depending on the context.

[0027] This invention provides the use of a compound or its salt in the preparation of a cancer therapeutic drug, the compound having the structural formula shown in Formula I:

[0028]

[0029] Wherein, R is selected from (CH2). n CH3, where n is an integer.

[0030] It should be noted that in this invention, small molecule molecules were screened using the ChemDiv / Chembridge small molecule library. (That is, the compound when n is 0, hereinafter also referred to as 3628-0029. Compound 3628-0029 is the number in the ChemDiv / Chembridge small molecule library, and the compound name is: ethyl)

[0031] 2-{2-[(4,6-dioxo-5-phenyl-1,4,5,6-tetrahydropyrimidin-2-yl)sulfanyl]acetamido}-1,3-benzothiazole-6-carboxylate (molecular formula: C22 H18 N4 O5 S2, InChI Key: GIERRJVQQQJDIC-UHFFFAOYSA-N) has been found to be a small molecule compound that can effectively inhibit tumor growth, and can be used in the preparation of cancer treatment drugs. Furthermore, when the n in R of the compound shown in Formula I is chosen to be other values ​​(i.e., R is an alkyl group), it can also effectively inhibit tumor growth. Therefore, the compound shown in Formula S can be used in the preparation of cancer treatment drugs.

[0032] In some specific examples, the value of n in the compound represented by Formula I in the above-mentioned cancer treatment drug composition can be 0-4.

[0033] It should be noted that n in the compound shown in Formula I above can be 0-4, that is, R can be methyl, ethyl, propyl, butyl or pentyl.

[0034] In some specific examples, the salt of the compound in the above-mentioned cancer treatment drug composition may be a hydrochloride salt.

[0035] It should be noted that preparing the compound with the above structure into a salt form can increase its solubility, stability and bioavailability. The salt form can be a salt form known in the art, and generally a pharmaceutically acceptable hydrochloride salt can be selected.

[0036] In some specific examples, the cancer mentioned above can be breast cancer, especially triple-negative breast cancer.

[0037] It should be noted that this invention has discovered and verified that compounds with the above-described structure or their salts can effectively inhibit the growth of breast tumors, thereby achieving the goal of treating breast cancer, especially the triple-negative breast cancer subtype.

[0038] In some specific examples, the dosage forms of cancer treatment drugs mentioned above include injections, liquid oral solutions, sprays, patches, tablets, powders, ointments, or suppositories.

[0039] It should be noted that the compounds or their salts with the above-mentioned structures can be combined with different excipients to prepare different dosage forms, such as injections, liquid oral preparations, sprays, patches, tablets, powders, ointments, or suppositories. The above dosage forms can be selected comprehensively according to the tumor site and the patient's condition. In addition, the excipients for the above-mentioned different dosage forms are all known to those skilled in the art, and the methods for preparing different dosage forms are also known to those skilled in the art. For example, the preparation of tablets mainly uses diluents (such as starch, dextrin, sucrose, or sucrose), absorbents (calcium sulfate, calcium hydrogen phosphate, or light magnesium oxide), binders (povidone, syrup, or hydroxypropyl methylcellulose), wetting agents (water, etc.), or disintegrants (dry starch, sodium hydroxymethyl starch, or crospovidone, etc.); the preparation of liquid oral preparations mainly uses solubilizers, suspending agents, emulsifiers, or colorants.

[0040] This invention also provides a cancer treatment pharmaceutical composition comprising Anti-PD1 and a compound or a salt thereof; the compound has the structural formula shown in Formula I:

[0041]

[0042] Wherein, R is selected from (CH2). n CH3, where n is an integer.

[0043] It should be noted that, as described above, the compound shown in Formula I can effectively inhibit tumor growth; furthermore, this compound can be used in combination with Anti-PD1 to synergistically enhance the therapeutic effect. Verification has shown that the combined therapeutic effect of the compound shown in Formula I and Anti-PD1 on tumors is superior to the therapeutic effect of using the compound shown in Formula I alone or using Anti-PD1 alone. Furthermore, since Anti-PD1 and the compound or its salts are used separately, the cancer treatment drug composition is preferably in individually packaged form, i.e., the cancer treatment drug composition includes individually packaged Anti-PD1 and the compound or its salts.

[0044] In some specific examples, the value of n in the compound represented by Formula I in the above-mentioned cancer treatment drug composition can be 0-4.

[0045] It should be noted that R in the compound shown in Formula I can be methyl, ethyl, propyl, butyl, or pentyl, etc., preferably methyl (CH3). When R is methyl, the therapeutic effect is better than other groups.

[0046] In some specific examples, the salt of the compound in the above-mentioned cancer treatment drug composition may be a hydrochloride salt.

[0047] It should be noted that preparing the compound with the above structure into a salt form can increase its solubility, stability and bioavailability. The salt form can be a salt form known in the art, and generally a pharmaceutically acceptable hydrochloride salt can be selected.

[0048] This invention also provides a cancer treatment drug comprising the cancer treatment drug composition of this invention.

[0049] In some specific examples, the dosage forms of the aforementioned cancer treatment drugs may include injections, liquid oral solutions, sprays, patches, tablets, powders, ointments, or suppositories.

[0050] It should be noted that the above-mentioned cancer treatment drug compositions can be prepared into different dosage forms by combining different excipients, such as injections, liquid oral preparations, sprays, patches, tablets, powders, ointments, or suppositories. The above dosage forms can be selected comprehensively according to the tumor site and the patient's condition. In addition, the excipients of the above-mentioned different dosage forms are all known to those skilled in the art, and the methods for preparing different dosage forms are also known to those skilled in the art. For example, the preparation of tablets mainly uses diluents (such as starch, dextrin, sucrose, or sucrose), absorbents (calcium sulfate, calcium hydrogen phosphate, or light magnesium oxide), binders (povidone, syrup, or hydroxypropyl methylcellulose), wetting agents (water, etc.), or disintegrants (dry starch, sodium hydroxymethyl starch, or crospovidone, etc.); for example, the preparation of liquid oral preparations mainly uses composting agents, suspending agents, emulsifiers, or coloring agents.

[0051] This invention also provides the use of the cancer therapeutic pharmaceutical composition of this invention in the preparation of cancer therapeutic pharmaceuticals.

[0052] In some specific examples, the cancer mentioned above can be breast cancer, especially triple-negative breast cancer.

[0053] It should be noted that this invention has discovered and verified that compounds with the above-described structure or their salts, in combination with Anti-PD1, can effectively inhibit the growth of breast tumors and achieve the goal of treating breast cancer, especially the triple-negative breast cancer subtype.

[0054] In some specific examples, the dosage forms of cancer treatment drugs mentioned above include injections, liquid oral solutions, sprays, patches, tablets, powders, ointments, or suppositories.

[0055] To better understand the present invention, specific examples are provided below to further illustrate the content of the present invention, but the content of the present invention is not limited to the examples below.

[0056] Example 1

[0057] In the following examples, the single-reagent method and the two-reagent method are as follows:

[0058] Single-reagent method:

[0059] The preparation method for working reagents mainly involves diluting the reagents in the kit according to a certain ratio. For example, for Reagent 5, it needs to be diluted with double-distilled water at a volume ratio of 1:9, and prepared fresh each time it is used. This preparation method is suitable for single-reagent detection and can ensure the stability of the reagents and the accuracy of the reaction.

[0060] Two-reagent method: Apply R1 and R2 separately; the specific preparation steps are as follows: Washing buffer preparation: Dilute with distilled water 1:20 (e.g., 1 mL concentrated washing buffer added to 19 mL distilled water); concentrated washing buffer taken from the refrigerator may have crystals, which is normal. You can use a 40℃ water bath to slightly heat and completely dissolve the crystals before preparing the washing buffer. Use on the same day. Standard preparation: Take the standard from the kit, add 1 mL of standard & sample diluent to dissolve, cover and let stand at room temperature for about 10 minutes. Take seven 1.5 mL centrifuge tubes and label them S6, S5, S4, S3, S2, S1, and S0 respectively. Add 250 μL of standard and sample diluent to each tube. Transfer 250 μL of standard from tube S7 to the first centrifuge tube S6 and gently mix. Prepare the biotin-labeled antibody working solution: 20 minutes before use, dilute 100× concentrated biotin-labeled antibody to 1× working concentration with biotin-labeled antibody diluent (e.g., 10 μL concentrate + 990 μL diluent). Prepare according to the required amount and use on the same day; discard any remaining solution. Prepare the enzyme conjugate working solution as follows: 20 minutes before use, dilute 100× concentrated enzyme conjugate to 1× working concentration with enzyme conjugate diluent (e.g., 10 μL concentrate + 990 μL diluent). Prepare according to the required amount, use on the same day, and discard any remaining solution; the TMB colorimetric solution is prepared as follows: 10 minutes before use, mix TMB colorimetric solution A and solution B in a 1:1 ratio, and store in the dark for later use.

[0061] In the following example, small molecule compound 3628-0029 was purchased from the Chem-Div compound library.

[0062] (I) Construction of a mouse breast cancer xenograft model

[0063] (1) Take EO771 triple-negative breast cancer cells from mice in the logarithmic growth phase, digest them routinely, centrifuge them, remove the supernatant, and wash them twice with PBS solution (PBS solution includes 137mM NaCl, 2.7mM KCl, 10mM Na2HPO4 and 1.8mM KH2PO4, pH 7.2–7.4, the same below);

[0064] (2) Resuspend cells in PBS solution, count cells, and adjust the cell density to 1×10⁻⁶. 7 / ml;

[0065] (3) Three days before inoculation, female C57BC / L mice aged 6-8 weeks (20g-22g per mouse) were ear-tagged; after the prepared cell suspension was well mixed, cells were subcutaneously inoculated into one side of the buttock at 50μl / side, i.e., 40×10⁶ cells / side. 4 Cells / animal, a total of 28 animals were inoculated.

[0066] (4) Mice were housed in an SPF-grade environment after inoculation, and the length and width of the tumor were measured every other day starting from day 3.

[0067] (5) Starting from day 7 after inoculation, mice were randomly divided into 4 groups of 7 mice each. The groups were: blank control group (Veh.+Isotype), αPD1 (Bioxcell,#BE0146) monotherapy group (Veh.+αPD1), 3628-0029 monotherapy group (3628-0029+Isotype) and 3628-0029 combined with αPD1 therapy group (3628-0029+αPD1);

[0068] Drug intervention began on the seventh day after vaccination, including:

[0069] The blank control group was first gavaged with 100uL of blank solution (the blank solution was 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (all volume fractions, the same below), and then intraperitoneally injected with an equal volume of Isotype solution (isotype antibody, no specific effect, dissolved in PBS solution, Isotype concentration of 2mg / mL) from the αPD1 group.

[0070] The αPD1 monotherapy group was first administered 100uL of blank solution by gavage, followed by an intraperitoneal injection of 100uL of αPD1 solution (dissolved in PBS solution, αPD1 concentration of 2mg / mL, the same below). The injection dose of αPD1 solution was 10mg / kg each time.

[0071] The 3628-0029 monotherapy group was first administered 100 μL of 3628-0029 solution by gavage (3628-0029 was dissolved in a blank solution, and the concentration of 3628-0029 was 2 mg / mL, the same below); then Isotype solution was injected intraperitoneally.

[0072] In the 3628-0029 combined with αPD1 treatment group, 100uL of 3628-0029 solution was first administered by gavage, followed by 100uL of αPD1 solution injected intraperitoneally.

[0073] All groups were given medication every three days (i.e., on days 7, 10, 13 and 16 after vaccination, respectively).

[0074] (6) On day 18 post-inoculation, the mice were euthanized; the in situ tumors were completely removed and weighed, and the mouse growth curve and tumor volume (tumor volume (mm)) were plotted using GraphPad software. 3 ( = length diameter × width diameter 2 × 0.5) and tumor weight diagram.

[0075] (II) Results

[0076] Tumor samples from each group of mice, as shown below Figure 1 As shown, the results indicated that the tumor volume of EO771 after subcutaneous in situ tumor formation in the 3628-0029 monotherapy group was significantly smaller than that in the blank control group and the αPD1 monotherapy group; in addition, the tumor volume of EO771 after subcutaneous in situ tumor formation in the 3628-0029 combined with αPD1 therapy group was significantly smaller than that in the 3628-0029 monotherapy group and the αPD1 monotherapy group.

[0077] In addition, the changes in tumor volume in each group of mice from day 5 to day 18 post-inoculation are as follows: Figure 2 As shown, the results indicated that tumor growth was significantly slower in the 3628-0029 group after subcutaneous in situ tumor formation with EO771 compared to the control group; and tumor growth was even slower in the 3628-0029 group treated with Anti-PD1 combined with Anti-PD1 compared to the Anti-PD1 monotherapy group.

[0078] On day 18 post-inoculation, the tumor weights of mice in each group were as follows: Figure 3 As shown, the results indicated that the tumor weight after subcutaneous in situ tumor formation with EO771 in the 3628-0029 group was significantly lower than that in the control group. Furthermore, the tumor weight difference between the 3628-0029 combined with Anti-PD1 treatment group and the Anti-PD1 monotherapy group was statistically significant (mean tumor weight: blank control group: 1.44 mg, PD1 monotherapy group: 0.54 g, 3628-0029 monotherapy group: 0.94 g, 3628-0029 combined with Anti-PD1 treatment group: 0.1 mg, p<0.05). Compared to the PD1 monotherapy group, the tumor inhibition rate of the 3628-0029 combined with Anti-PD1 treatment group was increased by 30.5%.

[0079] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. Use of a compound or a salt thereof in the manufacture of a therapeutic drug for triple negative breast cancer, wherein the compound has a structure as shown in Formula I:

2. Use according to claim 1, characterized in that, The salt of the compound is selected from hydrochloride.

3. Use according to claim 1 or 2, characterized in that, The dosage form of the therapeutic drug for cancer includes injection, liquid oral agent, spray, patch, tablet, powder, paste or suppository.

4. A pharmaceutical composition for the treatment of cancer, characterized by, The therapeutic drug for cancer comprises Anti-PD1 and the compound or a salt thereof, wherein the compound has a structure as shown in Formula I:

5. A cancer therapeutic drug characterized by, The therapeutic drug composition for cancer of claim 4.

6. Use of the therapeutic drug composition for cancer of claim 5 in the manufacture of a therapeutic drug for triple negative breast cancer.

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