Application of compound or salt thereof in preparation of cancer treatment medicine

Abstract

The invention belongs to the technical field of preparation of cancer drugs, and particularly relates to application of a compound or salt thereof in preparation of cancer treatment drugs, the structure of the compound is shown in the formula I. The compound shown in the formula I can remarkably inhibit growth of tumors (such as breast cancer), and compared with a blank control group and mice treated by the compound shown in the formula I, the tumors grow more slowly and are smaller, and the cancer treatment effect is better. The material is lighter, and the life cycle is obviously prolonged; in addition, the compound shown in the formula I can be combined with Anti-PD1 to synergistically enhance the tumor inhibition effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of cancer drug preparation, and relates to the use of a specific compound or its salt in the preparation of cancer treatment drugs. Background Art

[0002] The progression of malignant tumors depends on complex biological regulatory mechanisms, including abnormal cell proliferation, angiogenesis, invasion and metastasis, and immune escape. At present, clinical treatment still mainly relies on surgery, radiotherapy, chemotherapy, and targeted drugs, but there are limitations such as drug resistance, significant toxic and side effects, and high treatment costs. In recent years, targeted treatment 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 downstream signaling pathway cascades (such as PI3K / Akt, MAPK / ERK, etc.), thereby promoting the proliferation and migration of vascular endothelial cells and accelerating tumor angiogenesis. This abnormal angiogenesis not only provides oxygen and nutrients for tumors, but also creates conditions for metastatic cells to break through the basement membrane barrier; in addition, tumor cells can also activate intracellular pro-survival pathways through autocrine signals, inhibit the expression of apoptosis-related proteins, and enhance their survival ability in a harsh microenvironment.

[0004] At the level of existing treatment technologies, RNA interference technology (RNAi) (such as siRNA / shRNA) has been tried to specifically inhibit the expression of pro-cancer genes, but it has defects such as low targeting efficiency, poor in vivo stability, and insufficient drugability; although monoclonal antibody drugs can specifically inhibit the function of specific proteins, their clinical applications are restricted due to problems such as complex production processes, large dosing amounts, and heavy economic burdens on patients. In contrast, small molecule compounds have become an important direction for targeted therapy development due to their small molecular weights, strong structural modifiability, convenient dosing methods, and controllable production costs. Summary of the Invention

[0005] The present invention has screened out a small molecule compound (hereinafter also referred to as 3628-0029, which is also the small molecule library number) from 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 object, the present invention can adopt the following technical solutions:

[0007] On the one hand, the present invention provides a use of a compound or its salt in the preparation of cancer treatment drugs, and the structural formula of the compound is shown in Formula I:

[0008]

[0009] Among them, R is selected from (CH2) n CH3, and n is an integer.

[0010] Preferably, in the compound of formula I for the above use, the value of n is 0 - 4.

[0011] More preferably, in the compound of formula I for the above use, R is CH3.

[0012] Preferably, in the above use, the salt of the compound of formula I is selected as hydrochloride.

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

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

[0015] Preferably, in the above use, the dosage forms of the cancer treatment drug include injections, liquid oral preparations, sprays, patches, tablets, powders, ointments or suppositories.

[0016] On the other hand, the present invention provides a cancer treatment drug composition, comprising Anti-PD1 and the compound of formula I as described above or its salt.

[0017] On yet another aspect, the present invention provides a cancer treatment drug, which comprises the cancer treatment drug composition of the present invention.

[0018] On yet another aspect, the present invention provides the use of the cancer treatment drug composition of the present invention in the preparation of a cancer treatment drug.

[0019] The beneficial effects of the present invention include:

[0020] (1) The compound of formula I can significantly inhibit tumor (such as breast cancer) growth. Compared with the blank control group, the tumor growth of mice treated with the compound of formula I is slower, the tumor is smaller and lighter.

[0021] (2) The compound of formula I combined with Anti-PD1 has a synergistic effect and can synergistically inhibit tumor growth. The inhibitory effect of the combination of the two on tumors is better than that of the single compound of formula I and the single Anti-PD1. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a physical picture of the tumor growth situation after the intervention of the 3628 - 0029 small molecule inhibitor;

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

[0024] Figure 3Graph showing the effects of the 3628-0029 small molecule inhibitor on the blood routine, AST, and ALT in mice. Detailed implementation mode

[0025] The examples given are for better illustration of the present invention, but the content of the present invention is not limited to the examples given. Therefore, those skilled in the art who make non-essential improvements and adjustments to the implementation mode based on the above invention content still fall within the protection scope of the present invention.

[0026] The terms used herein are only for describing specific embodiments and are not intended to limit the present disclosure. Unless having significantly different meanings in the context, the expressions in the singular form include the plural form. As used herein, it should be understood that terms such as "including", "having", "containing" are intended to indicate the existence of features, numbers, operations, components, parts, elements, materials, or combinations. The terms of the present invention are disclosed in the specification, and are not intended to exclude the possibility of the existence or addition of one or more other features, numbers, operations, components, parts, elements, materials, or their combinations. As used herein, depending on the situation, " / " can be interpreted as "and" or "or".

[0027] An embodiment of the present invention provides the use of a compound or its salt in the preparation of a cancer treatment drug, and the structural formula of the compound is shown in Formula I:

[0028]

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

[0030] It should be noted that in the present invention, through the ChemDiv / Chembridge small molecule library screening (that is, the compound when n is 0, hereinafter also referred to as 3628-0029, and the compound 3628-0029 is the number in the ChemDiv / Chembridge small molecule library, compound name: 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), a small molecule compound found to be effective in inhibiting tumor growth, can be used in the preparation of cancer therapeutic drugs; and when n in R in the above compound of formula I selects other values (i.e., R is an alkyl group), it can also effectively inhibit tumor growth; so the compound shown in formula S can be used in the preparation of cancer therapeutic drugs.

[0032] In some specific examples, in the above cancer therapeutic drug composition, the value of n in the compound shown in formula I can be 0 - 4.

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

[0034] In some specific examples, in the above use, the salt of the compound in the cancer therapeutic drug composition can be selected as hydrochloride.

[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 the salt forms well-known in the art, and generally, pharmaceutically acceptable hydrochloride can be selected.

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

[0037] It should be noted that the present invention has discovered and verified that the compound with the above structure or its salt form can effectively inhibit the growth of breast tumors, achieving the purpose of treating breast cancer, especially the breast cancer subtype triple-negative breast cancer.

[0038] In some specific examples, in the above use, the dosage forms of the cancer therapeutic drugs include injections, liquid oral preparations, sprays, patches, tablets, powders, ointments or suppositories.

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

[0040] The embodiment of the present invention also provides a cancer treatment drug composition, which includes Anti-PD1 and a compound or its salt; the structural formula of the compound is shown in Formula I:

[0041]

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

[0043] It should be noted that as described above, the compound shown in Formula I can effectively inhibit tumor growth; in addition, this compound can be used in combination with Anti-PD1 to synergistically enhance the therapeutic effect. It can be verified that the effect of the compound shown in Formula I and Anti-PD1 in the combined treatment of tumors is better than that of using the compound shown in Formula I alone and using Anti-PD1 alone. In addition, since Anti-PD1 and the compound or its salt are used separately, the cancer treatment drug composition is preferably in the form of independent packaging, that is, the cancer treatment drug composition includes Anti-PD1 and the compound or its salt in independent packaging.

[0044] In some specific examples, in the above cancer treatment drug composition, the value of n in the compound shown in Formula I 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), and the therapeutic effect is better when R is methyl than other groups.

[0046] In some specific examples, in the above cancer treatment drug composition, the salt of the compound can be selected as hydrochloride.

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

[0048] The embodiments of the present invention also provide a cancer treatment drug, which includes the cancer treatment drug composition in the present invention.

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

[0050] It should be noted that the above cancer treatment drug composition can be prepared into different dosage forms by combining with different excipients, such as injections, liquid oral preparations, sprays, patches, tablets, powders, ointments, or suppositories; the above dosage forms can be comprehensively selected according to the tumor medication location and the patient's condition; in addition, the excipients of the above different dosage forms are well-known to those skilled in the art, and the preparation methods of drugs in different dosage forms are also well-known to those skilled in the art. For example, when preparing tablets, diluents (such as starch, dextrin, sucrose, or mannitol, etc.), absorbents (calcium sulfate, calcium hydrogen phosphate, or light magnesium oxide, etc.), binders (povidone, syrup, or hydroxypropyl methylcellulose, etc.), wetting agents (water, etc.), or disintegrants (dry starch, sodium carboxymethyl starch, or cross-linked povidone, etc.) are mainly used; for example, when preparing liquid oral preparations, solubilizers, suspending agents, emulsifiers, or colorants, etc. are mainly used.

[0051] The embodiments of the present invention also provide the use of the cancer treatment drug composition in the present invention in the preparation of cancer treatment drugs.

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

[0053] It should be noted that the present invention has discovered and verified that the compound of the above structure or its salt form in combination with Anti-PD1 can effectively inhibit the growth of breast tumors and achieve the purpose of treating breast cancer, especially the triple-negative breast cancer subtype.

[0054] In some specific examples, in the above use, the dosage forms of the cancer treatment drug include injections, liquid oral preparations, sprays, patches, tablets, powders, ointments, or suppositories.

[0055] To better understand the present invention, the following specific examples are further used to clarify the content of the present invention, but the content of the present invention is not limited to the following examples.

[0056] Example 1

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

[0058] Single reagent method:

[0059] The method of preparing working reagents mainly involves diluting the reagents in the kit according to a certain ratio. For example, for reagent five, it is necessary to dilute reagent five and double distilled water at a volume ratio of 1:9 and prepare it before use. This preparation method is suitable for single-reagent detection and can ensure the stability of the reagents and the accuracy of the reaction.

[0060] Double reagent method: Use R1 and R2 separately; the specific preparation steps are as follows: Preparation of washing solution: dilute with distilled water 1:20 (for example: add 19mL of distilled water to 1mL of concentrated washing solution); the concentrated washing solution taken out of the refrigerator may have crystals, which is a normal phenomenon. You can use a 40℃ water bath to slightly heat it to completely dissolve the crystals before preparing the washing solution. Use it on the same day. Preparation of standard: Take out the standard from the kit, add 1mL of standard & sample diluent to dissolve, cover it and let it stand at room temperature for about 10 minutes. Take 7 1.5mL centrifuge tubes, mark them as S6, S5, S4, S3, S2, S1 and S0 respectively, and add 250μL of standard & sample diluent to each tube; pipette 250μL of standard from S7 into the first centrifuge tube S6, and mix them by gently pipetting; prepare biotin-labeled antibody working solution: 20 minutes before use, dilute 100× concentrated biotin-labeled antibody to 1× working concentration with biotin-labeled antibody diluent (for example: 10μL concentrated solution + 990μL diluent); prepare according to the required amount, use it on the same day, and discard the rest; in addition, prepare the enzyme conjugate working solution: 20 minutes before use, dilute 100× concentrated enzyme conjugate to 1× working concentration with enzyme conjugate diluent (for example: 10μL concentrated solution + 990μL diluent). Prepare according to the required amount, use it on the same day and discard the rest; the preparation of TMB colorimetric solution is as follows: 10 minutes before use, mix TMB colorimetric solution A and solution B in a 1:1 ratio and keep it away from light for later use.

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

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

[0063] (1) Mouse triple-negative breast cancer cells EO771 in the logarithmic growth phase were routinely digested, centrifuged, the supernatant removed, and washed twice with PBS solution (PBS solution includes 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4 and 1.8 mM KH2PO4, pH 7.2-7.4, the same below);

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

[0065] (3) Three days before inoculation, 6-8 week old female C57BC / L mice (20 g to 22 g per mouse) were ear-tagged; the prepared cell suspension was blown evenly and then inoculated subcutaneously on one side of the buttocks at 50 μl / side, i.e., 40×10 4 cells / mouse, and a total of 28 mice were inoculated.

[0066] (4) After inoculation, mice were kept in an SPF environment, and the length and width of the tumors were measured every other day starting from the third day;

[0067] (5) Starting from the 7th day after inoculation, the mice were randomly divided into 4 groups, with 7 mice in each group; 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 treatment group (3628-0029+αPD1);

[0068] Drug intervention will be carried out from the seventh day after vaccination, including:

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

[0070] The αPD1 monotherapy group was first gavaged with 100uL of blank solution, followed by intraperitoneal injection of 100uL of αPD1 solution (dissolved in PBS solution, αPD1 concentration was 2mg / mL, the same below), and the injection volume of αPD1 solution was 10mg / kg each time;

[0071] The 3628-0029 monotherapy group was first gavaged with 100uL of 3628-0029 solution (3628-0029 was dissolved in blank solution, 3628-0029 concentration was 2mg / mL, the same below); followed by intraperitoneal injection of Isotype solution;

[0072] The 3628-0029 combined with αPD1 treatment group was first gavaged with 100uL of 3628-0029 solution, followed by intraperitoneal injection of 100uL of αPD1 solution;

[0073] The drug administration frequency for the above groups was once every three days (i.e., on the 7th, 10th, 13th and 16th days after inoculation);

[0074] (6) On the 18th day after inoculation, the mice were sacrificed; the in-situ tumors were removed intact and weighed, and the growth curve graph of the mice and the tumor volume (tumor volume (mm 3 ) = long diameter × width diameter2 × 0.5) and tumor weight graph were made using GraphPad software.

[0075] (II) Results

[0076] The physical tumors of the mice in each group were as Figure 1 shown. The results showed that the tumor volume of the EO771 subcutaneous in-situ tumor formation in the 3628-0029 monotherapy group was significantly smaller than that of the blank control group and the αPD1 monotherapy group; in addition, the tumor volume of the EO771 subcutaneous in-situ tumor formation in the 3628-0029 combined with αPD1 treatment group was significantly smaller than that of the 3628-0029 monotherapy group and the αPD1 monotherapy group.

[0077] In addition, the changes in the tumor volume of the mice in each group from the 5th day to the 18th day after inoculation were as Figure 2 shown. The results showed that the tumor growth of the EO771 subcutaneous in-situ tumor formation in the 3628-0029 group was significantly slower than that of the control group; the tumor growth of the 3628-0029 combined with Anti-PD1 treatment group was slower than that of the Anti-PD1 monotherapy group.

[0078] On the 18th day after inoculation, the tumor weights of the mice in each group were as Figure 3 shown. The results showed that the tumor weight of the EO771 subcutaneous in-situ tumor formation in the 3628-0029 group was significantly lower than that of the control group, and there was a statistically significant difference in tumor weight between the 3628-0029 combined with Anti-PD1 treatment group and the Anti-PD1 monotherapy group (average 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 with the PD1 monotherapy group, the tumor inhibition rate of the 3628-0029 combined with Anti-PD1 treatment group 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 them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the purpose and scope of the technical solutions of the present invention, and they should all be covered by the scope of the claims of the present invention.

Claims

1. Use of a compound or its salt in the preparation of a cancer treatment drug, wherein the compound has a structural formula as shown in Formula I: in, R is selected from (CH2) n CH3, n is an integer.

2. The use according to claim 1, characterized in that The value of n is 0-4.

3. The use according to claim 2, characterized in that: R is CH3.

4. The use according to any one of claims 1 to 3, characterized in that The salt of the compound is preferably hydrochloride.

5. The use according to any one of claims 1 to 3, characterized in that The cancer is breast cancer.

6. The use according to claim 5, characterized in that The cancer is triple-negative breast cancer.

7. The use according to claim 1, 2, 3 or 6, characterized in that Cancer treatment drugs come in injections, oral liquids, sprays, patches, tablets, powders, creams, or suppositories.

8. A pharmaceutical composition for treating cancer, characterized in that: The method comprises Anti-PD1 and the compound or salt thereof according to any one of claims 1 to 4.

9. A cancer treatment drug, characterized in that: The cancer treatment pharmaceutical composition according to claim 8.

10. Use of the cancer therapeutic drug composition according to claim 8 in the preparation of cancer therapeutic drugs.

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