CN110179806B - Application of sulfa drugs in the preparation of antitumor drugs - Google Patents

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an application of sulfonamide medicines in preparation of antitumor medicines. The application is the application of the sulfonamide drug as a tumor multidrug resistance reversal agent or an anti-tumor drug sensitizer in combination with a chemotherapeutic drug in the following aspects: 1) The application in preparing eukaryotic tumor cell proliferation inhibitor; 2) The application in preparing the medicine for preventing and/or treating tumor. The invention has the advantages that: the sulfonamides are used clinically for many years, have high safety, low price and convenient use, can be used as a synergist of tumor chemotherapeutic drugs, can improve the sensitivity of drug-resistant tumors to the chemotherapeutic drugs, reduce the toxicity and improve the chemotherapeutic effect, and have wide application prospect.

Description

Application of sulfa drugs in the preparation of antitumor drugs

technical field

The invention belongs to the technical field of medicine, and in particular relates to the application of sulfa drugs in the preparation of antitumor drugs.

Background technique

Tumors, especially malignant tumors, are major diseases that seriously threaten human life and health, and the mortality rate is extremely high. According to the latest data released by the World Health Organization, 8.8 million people die from cancer every year in the world, accounting for nearly one-sixth of the total annual deaths in the world, and most of the dead are in low- and middle-income countries. With the deteriorating environment of human beings, the incidence of malignant tumors is increasing year by year.

At present, chemotherapy is still one of the main means of treating malignant tumors. Although the research on tumor treatment has made significant progress in recent years, only less than 50% of tumors are sensitive to chemotherapy, and more than 50% of tumors are resistant to chemotherapy drugs. Chemotherapy resistance of tumors, especially multidrug resistance (MDR) has become the most common and difficult problem of tumor treatment failure. Therefore, for more than half a century, combination chemotherapy has been the mainstream of cancer chemotherapy, and single drug is only used occasionally in a few cases. Using one or several non-toxic drugs combined with chemotherapy drugs to increase the sensitivity of tumor cells to chemotherapy drugs and reduce the side effects of chemotherapy drugs is of great significance to improve the survival rate of patients.

Sulfonamides are the first class of chemically synthesized antibacterial drugs, which once played an important role in the treatment of infectious diseases. These drugs compete with p-aminobenzoic acid (PABA) in the biosynthesis of bacterial dihydrofolate and bind to enzymes, affecting the synthesis of nucleic acid precursors, thereby inhibiting bacterial growth and reproduction. Since mammalian cells can directly utilize exogenous folic acid, sulfa drugs have no inhibitory effect on mammalian cells. The classic sulfanilamide antibacterial drugs with p-aminobenzenesulfonamide structure are the first synthetic antibacterial drugs. They have been used in clinical practice for more than 80 years. They have the advantages of wide antibacterial spectrum, stable properties, easy use, and no food consumption during production. .

After a long period of high selection, some highly toxic sulfonamides were gradually eliminated, such as sulfathiazole (ST), sulfamethazine (SM), and sulfamethoxazine (SMP). Only 3 to 4 kinds are actually used clinically, such as sulfadiazine (SD), sulfamethoxazole (SIZ), sulfamethoxazole (SMZ), and sulfadoxine (SDM). Although many bacteria are resistant to sulfa drugs, sulfa drugs are still the main choice for treatment of sulfa-sensitive bacteria because sulfa drugs are cheap, easy to use, and will not cause intestinal flora disorders like broad-spectrum antibiotics. , and its potency is equivalent to or higher than that of antibiotics. In recent years, the development of other medical applications of sulfa compounds has become an extremely active research field.

No report on the anti-tumor synergistic effect of classic sulfa compounds was found after searching the literature and domestic and foreign patents.

Contents of the invention

One of the purposes of the present invention is to provide a new drug application of sulfa drugs.

The new drug application of sulfa drugs provided by the present invention is: the application of sulfa drugs in the following aspects:

1) the application in the preparation of eukaryotic tumor cell proliferation inhibitors; 2) the application in the preparation of drugs for preventing and/or treating tumors.

Specifically, the application is the application of sulfonamide drugs as tumor multidrug resistance reversing agent or antitumor drug sensitizer, or in the preparation of tumor multidrug resistance reversing agent or antitumor drug sensitizer.

More specifically, the application is the application of sulfonamides as tumor multidrug resistance reversal agents or antitumor drug sensitizers combined with chemotherapy drugs in the following aspects: 1) Application in the preparation of eukaryotic tumor cell proliferation inhibitors ; 2) application in the preparation of drugs for the prevention and/or treatment of tumors.

In the application, the sulfa drugs include but are not limited to: sulfadiazine (SD), sulfisoxazole (SIZ), sulfamethoxazole (SMZ), sulfadoxine (SDM), etc.;

Sulfadiazine (SD), the molecular formula is C ₁₀ H ₁₀ N ₄ O ₂ S, and its chemical structure is as follows:

Sulfaisoxazole (SIZ), the molecular formula is C ₁₁ H ₁₃ N ₃ O ₃ S, and its chemical structure is as follows:

Sulfamethoxazole (SMZ), the molecular formula is C ₁₀ H ₁₁ N ₃ O ₃ S, and its chemical structure is as follows:

Sulfadoxine (SDM), molecular formula C ₁₂ H ₁₄ N ₄ O ₄ S, its chemical structure is as follows:

The eukaryote is a mammal; the tumor includes brain tumors, breast cancer, gastrointestinal tumors, lung cancer, liver cancer, pancreatic cancer, cervical cancer, ovarian cancer, kidney cancer, gallbladder cancer, melanoma, hematological system tumors .

Specifically, the tumor is a human glioma, and the tumor cells are U251 or U251/TMZ human glioma cells;

The chemotherapeutic drugs can be: temozolomide, doxorubicin, gemcitabine, 5-fluorouracil, cytarabine, cyclophosphamide, paclitaxel, platinum, daunorubicin, epirubicin, vinorelbine, vincristine , at least one of etoposide and mitomycin.

Another object of the present invention is to provide a drug combination preparation for treating tumors.

The drug combination preparation for treating tumor provided by the present invention includes the sulfonamide drug and the chemotherapeutic drug.

The sulfa drugs can be administered by any known administration methods.

The chemotherapeutic drug can be administered by any known method of administration.

The present invention finds that the sulfonamide drugs themselves have little effect on tumor growth, but can significantly enhance the anti-tumor effect of chemotherapy drugs and reduce the toxic and side effects of chemotherapy drugs when used in combination with commonly used anti-tumor chemotherapeutic drugs. In addition, it can improve the therapeutic effect of chemotherapy drugs on multidrug-resistant tumors.

There is an essential difference between the present invention and the existing patents or documents. It starts from the search for high-efficiency and low-toxicity chemotherapeutic synergists. Through high-throughput drug screening technology, it is found that sulfonamide compounds can enhance the anti-tumor effect of commonly used chemotherapeutic drugs, which will fight against tumors. The development of synergists and the combination therapy of clinical tumors are of great significance.

The advantages of the present invention are: sulfonamides have been used clinically for many years, have high safety, low price, and are convenient to use. Chemotherapy effect, broad application prospects.

Description of drawings

Figure 1 shows the effects of sulfadiazine (SD), sulfisoxazole (SIZ), sulfamethoxazole (SMZ), and sulfadoxine (SDM) on the growth of U251 and U251/TMZ human glioma cells at different concentrations .

Figure 2 shows the effects of 100 μM sulfadiazine (SD), sulfisoxazole (SIZ), sulfamethoxazole (SMZ), and sulfadoxine (SDM) on the inhibition of U251 cell growth by different concentrations of temozolomide and cisplatin, respectively.

Detailed ways

The present invention will be described below through specific examples, but the present invention is not limited thereto.

The experimental methods used in the following examples are conventional methods unless otherwise specified; the reagents and biological materials used in the following examples can be obtained from commercial sources unless otherwise specified.

Embodiment 1, MTT method detects the influence of sulfonamides on the anti-human glioma cell growth of chemotherapeutics

U251 and TMZU251 cells (U251/TMZ) in the logarithmic growth phase were inoculated in a 96-well culture plate, 100 μL per well (containing 3000 cells), and different concentrations (0, 50, 100, 150, 200, 250 μM) of sulfadiazine (SD), sulfadiazine, and Dimethoxazole (SIZ), sulfamethoxazole (SMZ), sulfadoxine (SDM) acted for 48 hours. Add 10 μL of MTT solution (5 mg/mL) to each well, continue to incubate for 4 hours, discard the culture medium, add DMSO to each well, vibrate for 10 minutes with a micro-mixing oscillator, and use the fully automatic enzyme-linked immunosorbent assay instrument on Measure the absorbance value at 570nm, and calculate the inhibition rate (%). The results are shown in Figure 1, indicating that within the concentration range tested, none of the four drugs showed significant antitumor activity on U251 cells, while for U251/TMZ cells, the four drugs only showed certain growth inhibition at a concentration of 200 μM role (Figure 1).

Next, we studied the effects of four sulfonamides on the antitumor effects of two common chemotherapeutic drugs (temozolomide (TMZ), cisplatin). U251 cells were routinely cultured, and cells in the proliferating phase were collected and planted in 96-well plates. 100 μM SD, SIZ, SMZ or SDM were added to each well, and different concentrations of chemotherapy drugs (TMZ: 62.5, 125, 250, 500, 1000 μM; cisplatin: 12.5, 25 , 50, 100, 200 μM), and combined (SD, SIZ, SMZ or SDM + different concentrations of chemotherapy drugs) for 48 hours. The above-mentioned MTT method was used to detect its effect on cell proliferation, and the inhibition rate (%) was calculated respectively. The results are shown in Figure 2. It can be seen that SD, SIZ, SMZ, and SDM can all significantly enhance the anti-glioma effect of TMZ and cisplatin.

Example 2. Sulfonamides reverse the drug-resistant effect of human glioma drug-resistant cell line U251/TMZ on TMZ

Take the human glioma drug-resistant cell line U251/TMZ in the logarithmic growth phase, inoculate 3000 cells/well in a 96-well cell culture plate, cultivate overnight in a constant temperature incubator, and then add 100 μM SD, SIZ, SMZ or SDM, different concentrations of TMZ (62.5, 125, 250, 500, 1000μM), and the combination (SD, SIZ, SMZ or SDM+ different concentrations of TMZ) acted for 48h. The above-mentioned MTT method was used to detect its effect on cell proliferation, the inhibition rate and IC ₅₀ were calculated, and the reversal index=IC ₅₀ (TMZ)/IC ₅₀ (TMZ+SD) was calculated respectively. The reversal indices of SIZ, SMZ and SDM on the growth of TMZ-resistant U251/TMZ cells were calculated by the same method.

The results are shown in Table 1. It can be seen that SD, SIZ, SMZ, and SDM can reverse the TMZ-resistant effect of the human glioma drug-resistant cell line U251/TMZ to varying degrees.

Table 1. Sulfonamides reverse the drug resistance of human glioma drug-resistant cell line U251/TMZ to TMZ

Example 3. Sulfonamides reversed the resistance of human glioma drug-resistant cell line U251/TMZ to cisplatin

Take the human glioma drug-resistant cell line U251/TMZ in the logarithmic growth phase, inoculate 4000 cells/well in a 96-well cell culture plate, culture overnight, and then add SD, SIZ, SMZ or SDM (100 μM ), different concentrations of cisplatin (12.5, 25, 50, 100, 200 μM) and combination (SD, SIZ, SMZ or SDM + different concentrations of cisplatin) for 48 hours. The above-mentioned MTT method was used to detect its effect on cell proliferation, the inhibition rate and IC ₅₀ were calculated, and the reversal index was calculated = IC ₅₀ (cisplatin)/IC ₅₀ (cisplatin+SD). The reversal indices of SIZ, SMZ and SDM on the growth of cisplatin-resistant U251/TMZ were calculated by the same method.

The results are shown in Table 2. It can be seen that SD, SIZ, SMZ, and SDM can reverse the cisplatin-resistant effect of the human glioma drug-resistant cell line U251/TMZ to varying degrees.

Table 2. Sulfonamides reversed the resistance of human glioma drug-resistant cell line U251/TMZ to cisplatin

Example 4, Detection of the influence of sulfa drugs combined with TMZ on the growth of human glioma xenografts in nude mice

Establishment of tumor-bearing nude mouse model: U251 and U251/TMZ cells in the logarithmic growth phase were collected under sterile conditions, and diluted with normal saline to a cell suspension with a content of 1×10 ⁷ /ml. The right axilla of nude mice was subcutaneously inoculated with 0.2ml cell suspension to prepare the glioma xenograft model.

After the tumor body grew to ^100mm3 , they were randomly divided into groups, 10 in each group, and were given positive control TMZ (50mg/kg, intraperitoneal injection), SD group (100mg/kg, intragastric administration), SIZ group (100mg/kg, intragastric administration) ), SMZ group (100mg/kg, gavage), SDM group (100mg/kg, gavage), TMZ (50mg/kg, intraperitoneal injection)+SD group (100mg/kg, gavage), TMZ (50mg/kg , intraperitoneal injection)+SIZ group (100mg/kg, intragastric injection), TMZ (50mg/kg, intraperitoneal injection)+SMZ group (100mg/kg, intragastric injection), TMZ (50mg/kg, intraperitoneal injection)+SDM group ( 100mg/kg, administered orally), administered three times a week, after 2 weeks of continuous administration, the tumor was stripped and weighed, and the tumor inhibition rate and body weight change were calculated. The test results are shown in Tables 3 and 4, wherein the tumor inhibition rates of groups 2-10 are the ratios of the tumor weights of each group to the tumor weight of group 1.

Table 3. Effects of sulfa drugs combined with TMZ on the growth of human glioma U251 transplanted tumors in nude mice

^* P<0.05, compared with the model group. ^# P<0.05, compared with TMZ alone group. ^▲ P<0.05, compared with the sulfa monotherapy group.

Table 4. Effects of sulfa drugs combined with TMZ on the growth of human glioma U251/TMZ transplanted tumors in nude mice

^* P<0.05, compared with the model group. ^# P<0.05, compared with TMZ alone group. ^▲ P<0.05, compared with the sulfa monotherapy group.

The results in Tables 3 and 4 above all showed that the mice in the model group were gradually sluggish in action, their hair was disheveled, and their diet and water intake decreased. The mice in the TMZ group showed symptoms of toxic reactions such as anorexia, abdominal distension, listlessness, and weight loss; while the sulfa monotherapy group and the combination group had mild symptoms, and the anorexia, abdominal distension, and weight loss were not obvious. There was a statistically significant difference in tumor weight between the TMZ group, the combined group and the model group (P<0.05). SD, SIZ, SMZ, SDM combined with TMZ group can produce significantly higher tumor inhibitory effect than TMZ group and sulfonamide alone group. The tumor inhibition rates of SD, SIZ, SMZ, and SDM combined with TMZ on U251 xenografts were 58.12%, 59.40%, 50.85%, and 61.54%, respectively; the tumor inhibition rates on U251/TMZ xenografts were 60.31%, 59.14%, and 57.20%, respectively. % and 61.87%. This shows that sulfonamide combined with TMZ can significantly increase the anti-glioma effect of TMZ. From the results of body weight changes in nude mice, it can be seen that the combination group significantly inhibited tumor growth, and had little effect on body weight, and its effect on mouse body weight was significantly lower than that of TMZ group, indicating that it has low toxicity and better tolerance.

It shows that the combined use of sulfonamide drugs and TMZ can increase the tumor-inhibiting effect of TMZ and reduce its side effect of reducing body weight.

Claims (2)

1. The application of sulfamethoxazole SMZ in preparing medicine for reversing the drug-resistant effect of human glioma drug-resistant cell strain U251 on TMZ is provided.

2. Application of Sulfamethoxazole SIZ in preparation of medicine for reversing the drug-resistant effect of human glioma drug-resistant cell strain U251 on cisplatin.

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