CN112920121B - Application of small molecular compound 1-benzyl imidazole in treatment of breast cancer - Google Patents
Application of small molecular compound 1-benzyl imidazole in treatment of breast cancer Download PDFInfo
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- CN112920121B CN112920121B CN202110126433.1A CN202110126433A CN112920121B CN 112920121 B CN112920121 B CN 112920121B CN 202110126433 A CN202110126433 A CN 202110126433A CN 112920121 B CN112920121 B CN 112920121B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
- C07D233/58—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring nitrogen atoms
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
- A61K31/4174—Arylalkylimidazoles, e.g. oxymetazolin, naphazoline, miconazole
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention discloses application of a small molecular compound 1-benzyl imidazole in treating breast cancer. The biological experiment and the pharmacological research show that: the 1-benzyl imidazole has no killing effect on breast Cancer cells, but can inhibit the expression of tumor stem cell (CSC) related proteins SOX2, Nanog and ALDH1, and in vivo and in vitro experiments show that the 1-benzyl imidazole inhibits dryness, invasion, migration and metastasis of the breast Cancer cells. In addition, 1-benzylimidazole enhances the chemotherapeutic sensitivity of the first-line clinical drug doxorubicin. The invention is helpful for the mechanism-to-clinical research of the 1-benzyl imidazole, and provides more choices for treating breast cancer patients.
Description
Technical Field
The invention relates to development of a small molecule compound for resisting breast cancer, in particular to application of a small molecule compound 1-benzyl imidazole in treating breast cancer.
Background of the study
Currently, breast cancer is still one of the most common malignant tumors in women, and the incidence rate and the fatality rate of breast cancer both increase year by year and present a low-age state. The international scheme for treating breast cancer is still combination of surgery and postoperative chemotherapy, radiotherapy, endocrine therapy or targeted therapy, but metastasis and recurrence often cause treatment failure and are the main causes of death of breast cancer patients. The tumor stem cell (CSC) was first proposed in Reya, 2001, and it was believed that a small proportion of cells with stem cell characteristics, called CSC, were present in tumor tissues [3] . CSC, like normal stem cells, have uncontrollable reproductive capacity, continuous self-renewal capacity and multi-directional differentiation potential. CSC has recently been considered as the primary cell for tumorigenesis, recurrence and metastasis, and many CSC or CSC-like cells have been identified and isolated in breast cancer [4] . Breast Cancer Stem Cell (BCSC) can be passed through differentThe approach causes the recurrence and the metastasis of the breast cancer, and in vivo and in vitro researches find that the inhibition of the expression of ABC transporter in BCSC can enhance the sensitivity of BCSC to chemotherapeutic drugs [5] (ii) a In addition, BCSC is in a stationary phase for a long time and has strong repair capability, so that BCSC can avoid the striking effect of chemotherapy and radiotherapy and cause the recurrence of breast cancer [6] (ii) a In addition, the tumor microenvironment places the BCSC in a special hypoxic environment, and although hypoxia can induce Epithelial-Mesenchymal Transition (EMT) of any tumor cell, only the BCSC obtains migration potential, which is more favorable for invasion and metastasis [7-9] . The above facts indicate that BCSC can cause breast cancer recurrence and metastasis. Therefore, elimination of BCSC inevitably provides a new idea for solving the problems of recurrence and metastasis of breast cancer and achieving the purpose of curing the breast cancer.
Disclosure of Invention
In order to solve the problem of metastasis and relapse of a breast cancer patient in clinic, the invention aims to provide the application of a small molecular compound 1-benzyl imidazole in preparing a medicine for treating breast cancer, and the application of anti-breast cancer and clinical drug resistance treatment is achieved by targeting CSCs.
The specific technical scheme of the invention is as follows:
1-benzyl imidazole as a small molecule inhibitor for resisting breast cancer, which has the following chemical structural formula:
the application of 1-benzyl imidazole or pharmaceutically acceptable salts thereof in preparing medicines for treating and preventing breast cancer.
The anti-breast cancer small molecule inhibitor 1-benzyl imidazole achieves the effect of resisting breast cancer by weakening sternness related proteins SOX2, ALDH1, Naong and the like of breast cancer stem cells.
The anti-breast cancer small molecule inhibitor 1-benzyl imidazole achieves the effect of resisting breast cancer by weakening the proportion of the stem cell population of breast cancer stem cells, namely CD44-/CD24 +.
The anti-breast cancer small molecule inhibitor 1-benzyl imidazole achieves the effect of resisting breast cancer by inhibiting lung metastasis of breast cancer.
The use of 1-benzylimidazole or a pharmaceutically acceptable salt thereof in combination with a second therapeutic agent in the manufacture of a medicament for the treatment or prevention of breast cancer.
Further, the second therapeutic agent is doxorubicin.
The anti-breast cancer small molecule inhibitor 1-benzyl imidazole achieves the effect of resisting breast cancer by increasing the chemotherapy sensitivity of clinical first-line drug adriamycin.
Further, the therapeutically effective amount of 1-benzylimidazole, or a pharmaceutically acceptable salt thereof, and the therapeutically effective amount of the second therapeutic agent are administered simultaneously or separately, not sequentially.
Further, the breast cancer is a breast cancer cell line, preferably MCF-7, MDA-MB-231, MCF-7 ADR (Adriamycin-resistant cell line).
Further, the pharmaceutically acceptable salt thereof is a salt of 1-benzylimidazole with any of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, trifluoroacetic acid, propionic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, and the like.
The 1-benzylimidazole or a pharmaceutically acceptable salt thereof may be administered one or more times daily.
The method of administering 1-benzylimidazole can be determined comprehensively according to the activity and toxicity of the drug, the tolerance of the patient and the like.
The active ingredient of the anti-breast cancer medicine is the 1-benzyl imidazole or the pharmaceutically acceptable salt thereof, and the anti-breast cancer medicine further comprises a pharmaceutically acceptable carrier or auxiliary material. Compared with the prior technical conditions, the invention aims to:
the 1-benzyl imidazole is a small molecule inhibitor with anti-breast cancer activity screened based on the principle of computer-aided drug design and the principle of interaction between drugs and targets.
The invention discovers that the 1-benzyl imidazole can enhance the sensitivity of clinical first-line chemotherapy drug adriamycin and reduce the IC50 value of the adriamycin. Through further research, 1-benzyl imidazole inhibits invasion and metastasis of breast cancer cells by weakening CSCs dryness-related proteins SOX2, ALDH1 and Nanog thereof; and the 1-benzyl imidazole can reduce the proportion of stem cells and further kill breast cancer stem cells, thereby achieving the aim of curing the recurrence and metastasis of breast cancer.
The invention is beneficial to the clinical transformation of 1-benzyl imidazole into breast cancer stem cell weakening sternness protein action mechanism in a concentration-dependent manner. Provides a new strategy for the drug resistance and the relapse and metastasis of clinical treatment. Has great significance for treating breast cancer patients. Unless otherwise indicated, the following terms used in the specification and claims shall have the following meanings for the purposes of this application.
By "pharmaceutically acceptable" is meant that it is used to prepare pharmaceutical compositions that are generally safe, non-toxic, and neither biologically nor otherwise undesirable, and include that they are acceptable for human pharmaceutical use.
Drawings
FIG. 1; MTT (methyl thiazolyl tetrazolium) experiment for detecting 1-benzyl imidazole pairs MCF-7, MDA-MB-231 and MCF ADR Breast cancer cell lines have no cytotoxic effects.
FIG. 2; MTT (methyl thiazolyl tetrazolium) experiment for detecting 1-benzyl imidazole in MCF-7, MDA-MB-231 and MCF ADR The breast cancer cell line can enhance the chemotherapy sensitivity of the adriamycin and can reduce the IC50 value of the adriamycin.
FIG. 3; western-blot detection shows that 1-benzyl imidazole down-regulates expression of sternness related proteins ALDH1, SOX2 and Nanog in MCF-7 and MDA-MB-231 breast cancer cell lines.
FIG. 4; flow cytometry detection of 1-benzylimidazole in MCF-7, MDA-MB-231 breast cancer cell lines down-regulates the dry cell population CD44-/CD24 +.
FIG. 5; MDA-MB-231 cells are injected into a nude mouse body through tail veins, 1-benzyl imidazole is administrated in the tail veins after 8 days, and lung breast cancer metastasis of the mouse is obviously reduced by Micro-CT detection.
Detailed Description
The embodiments described below in further detail with reference to the accompanying drawings and the detailed description are provided as illustrations of the method of the invention and are not intended to limit the remainder of the disclosure in any way.
Embodiment 1: determination of cell viability
Respectively treating MCF-7, MDA-MB-231 and MCF with 1-benzyl imidazole with a certain concentration gradient ADR And (3) detecting the activity of the cells by adopting an MTT (methyl thiazolyl tetrazolium) experimental technology.
One day before adding 1-benzyl imidazole, taking 3-5X 10 cells in logarithmic growth phase 3 Plated in 96-well plates.
And preparing a series of medicines with concentration gradient by using the culture medium after the cells are attached to the wall and the density is 30% -70%.
Setting blank group, negative control group and administration group. Setting 3-5 concentration gradients, each concentration having 3 parallel multiple holes, changing the solution for cells before adding medicine, adding 200ul of medicine-containing culture medium, and incubating in an incubator for 24h, 48h and 72 h.
And after the specified time is reached, adding 20ul of MTT working solution into each hole, continuously incubating for 4h, sucking out the culture solution, adding 150ul of DMSO into each hole, placing in a micro-oscillator, oscillating at a low speed for 5min to fully dissolve crystals, and selecting 570nm on a multifunctional enzyme-labeling instrument to measure the OD value.
IC50 calculations were performed using graphpad 5.0.
The MTT method detects the influence of 1-benzyl imidazole on three breast cancer cell lines, as shown in figure 1, has no influence on normal breast cancer cells, but can specifically aim at breast cancer stem cells, enhance the sensitivity of chemotherapeutic drugs and reduce the IC50, as shown in figure 2.
Embodiment 2: western blotting method
(1) Extraction of Total protein from 1-Benzylimidazole-treated MCF-7, MDA-MB-231 cells
Firstly, the cells are paved in a 6-well plate one day before administration, so that the cell density is 60%;
② a control group and an administration group are arranged. The next day, after the cells adhere to the wall, configuring the drug concentration by using a culture medium, and adding the drug concentration into a six-hole plate to enable the final concentration to be 1 mu M, 2 mu M and 5 mu M; standing and culturing for 48 h;
③ sucking off the culture medium in the six-hole plate, washing with PBS for three times, collecting cells, and adding RIPA and PMSF according to the cell amount (RIPA: PMSF is 100: 1);
and fourthly, shaking once every five minutes for 40 min.
12000rpm, centrifuging for 15min, taking the supernatant, and sucking 5ul for BCA protein quantification.
(2) Protein concentration determination-BCA quantification method
Preparing BSA standard substances of 2000ug/ml, 1500ug/ml, 1000ug/ml, 750ug/ml, 500ug/ml, 250ug/ml, 125ug/ml and 25 ug/ml;
adding 25ul of BSA standard substance and a sample to be detected into a 96-well plate;
adding 200ul of BCA working solution into each hole, and fully and uniformly mixing the solution in a micro oscillator;
fourthly, incubating for 30min at 37 ℃, cooling to room temperature and standing for 2 h;
measuring the absorbance at 595nm by using a multifunctional enzyme-labeling instrument;
drawing a linear function according to the absorbance and the concentration of the standard substance, and calculating the protein concentration of the sample by the linear function;
seventhly, adding 5X Loading buffer according to the total volume of the protein, boiling in a boiling water bath for five minutes, and preserving at-80 ℃.
(3) SDS-PAGE electrophoresis
Installing a glue making frame, preparing lower layer separation glue, and mixing a lower layer solution and a lower layer buffer solution according to the ratio of 1: 1. The sizing solution and sizing buffer were mixed as 1: 1, preparing upper layer glue;
sample application is carried out, the total protein amount is 30ug, the concentrated gel is used for 60v and 45min, the separation gel is used for 90v and 2.5h, after electrophoresis is finished, the glass plate is pried open, and the gel is taken out.
(4) Rotary film
Cutting gel according to the molecular weight of a pre-dyed marker, measuring the length and the width of the gel, cutting a PVDF membrane with the same length and width of the gel, putting the PVDF membrane into methanol for soaking, and then transferring the PVDF membrane into 1 Xelectrotransfer buffer solution for soaking for a plurality of minutes. Placing the prepared gel, the PVDF membrane, the filter paper and the spongy cushion in sequence, wherein the prepared gel, the PVDF membrane, the filter paper and the spongy cushion are respectively a spongy cushion, two layers of filter paper, gel, the PVDF membrane, the filter paper and the spongy cushion, and bubbles are prevented from being generated between the PVDF membrane and the gel as much as possible in the placing process;
secondly, after the electrophoresis tank is assembled, the electrophoresis tank is connected with a power supply of an DYY-7C electrophoresis apparatus, and the power supply is 200mA for 1 hour.
(5) Immunoblotting reactions
Firstly, taking out the PVDF membrane, putting the PVDF membrane into a rapid sealing liquid, and sealing the PVDF membrane on a horizontal shaking table for 30 min;
preparing a primary antibody by using a primary antibody diluent, diluting the primary antibody according to the antibody specification in a dilution ratio, putting the PVDF membrane into an antibody incubation box after sealing, and incubating overnight at 4 ℃;
thirdly, washing the PVDF membrane by TBST for three times, and ten minutes each time;
fourthly, after the first membrane washing is finished, preparing a secondary antibody by using a secondary antibody diluent, wherein the dilution ratio of the secondary antibody is 1: 5000; putting the PVDF membrane into an antibody incubation box for secondary antibody incubation, and incubating for 45min at normal temperature;
washing the PVDF membrane by TBST for three times, wherein each time is ten minutes;
and sixthly, after the second film washing is finished, preparing an ECL developing solution, and carrying out exposure development according to the program set by Tanon.
As shown in FIG. 3, the Western blotting results showed that the xerosis-associated proteins ALDH1, SOX2 and Nanog were down-regulated in a concentration-dependent manner after administration in MCF-7 and MDA-MB-231 cells. It is shown that 1-benzyl imidazole inhibits breast cancer cells by inhibiting dryprotein.
Embodiment 3: flow cytometry detection of CD44-/CD24+ Dry cell population
One day ahead, cells in logarithmic growth phase were taken, trypsinized, centrifuged, resuspended, plated in six-well plates, control and dosing groups were set, at least three parallel replicate wells per group. DMSO and 1-benzylimidazole concentration gradients were configured with the media. Administration at 5% CO 2 And (5) standing and culturing for 48h in an incubator.
After 48h, the cells were washed three times with PBS, trypsinized, centrifuged at 500g for 5min and finally resuspended in 200ul of PBS.
Set up ISO group, double dye group. 5ul of CD44-ISO and CD24+ ISO are respectively added into the ISO component, 5ul of CD44-/CD24+ is respectively added into the double-dyeing component, the mixture is fully mixed after the addition is finished, and the mixture is incubated for 30min on ice in a dark place.
The proportional population of CD44+/CD 24-stem cells for each set was detected using a FACS Calibur flow cytometer (BD Biosciences, USA).
As shown in fig. 4, flow cytometry results showed that 1-benzylimidazole down-regulated the CD44+/CD 24-proportion population of stem cells in a concentration-dependent manner, and further showed that 1-benzylimidazole could inhibit the development of breast cancer by inhibiting CSCs.
Embodiment 4: Micro-CT detection of lung breast cancer metastasis of mice
200L of the solution is mixed into 10 multiplied by 10 6 The individual cell suspensions were injected via tail vein into 6-week-old BALB/c, nude mice. The experimental group was injected with 1-benzylimidazole and the control group with DMSO.
After 8 weeks of breast cancer cell injection, the heart and lung of the nude mouse are scanned by a micro-CT imaging system, and the metastasis of the breast cancer cells in the lung of the mouse is observed by CTAn software.
As shown in fig. 5, 1-benzylimidazole significantly reduced lung metastatic nodules of breast cancer compared to the control group, thereby inhibiting metastasis and recurrence of breast cancer.
The above description is only an embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that are not thought of through the inventive work should be included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope defined by the claims.
Claims (1)
1.1-benzylimidazole or a pharmaceutically acceptable salt thereof in combination with a second therapeutic agent, which is doxorubicin, for use in the preparation of a medicament for the treatment and prevention of breast cancer.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4755526A (en) * | 1984-06-18 | 1988-07-05 | Eli Lilly And Company | Method of inhibiting aromatase |
| CN104650155A (en) * | 2015-02-10 | 2015-05-27 | 江苏省原子医学研究所 | Ruthenium complex as well as preparation method and use thereof |
| CN106478734A (en) * | 2016-10-18 | 2017-03-08 | 浙江大学 | A kind of have N-heterocyclic carbine ruthenium compound of pyrazoles sense dough of active anticancer and its preparation method and application |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4755526A (en) * | 1984-06-18 | 1988-07-05 | Eli Lilly And Company | Method of inhibiting aromatase |
| CN104650155A (en) * | 2015-02-10 | 2015-05-27 | 江苏省原子医学研究所 | Ruthenium complex as well as preparation method and use thereof |
| CN106478734A (en) * | 2016-10-18 | 2017-03-08 | 浙江大学 | A kind of have N-heterocyclic carbine ruthenium compound of pyrazoles sense dough of active anticancer and its preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| Efficient substrate screening and inhibitor testing of human CYP4Z1 using permeabilized recombinant fission yeast;Qi Yan等;《Biochemical Pharmacology》;20170923;第146卷;第174-187页 * |
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