TWI737305B - Combination of quetiapine and 5-fluorouracil, kits comprising the same and uses thereof - Google Patents

Abstract

This invention discloses a combination of quetiapine and 5-fluorouracil, kits comprising the same and their uses in the treatment of liver cancer.

Description

Contains the combination of quetiapine and 5-fluorouracil, kits and their uses

The present invention relates to combinations and kits containing quetiapine and 5-fluorouracil (5-fluorouracil, 5-FU), and their uses.

Liver cancer is the second most fatal cancer in the world, and about 80% of patients are hepatocellular carcinoma (HCC). Liver cancer is mainly caused by the abnormal proliferation of liver cells. Although the underlying pathogenesis is not fully understood, it is currently known that liver cancer and chronic viral hepatitis infection (for example, chronic hepatitis B and chronic viral hepatitis infection) Conditions such as chronic hepatitis C, cirrhosis and non-alcoholic steatohepatitis caused by alcohol abuse, and exposure to toxins [for example, aflatoxin] are highly correlated.

Currently, chemotherapeutic drugs used clinically to treat liver cancer mainly include cisplatin, 5-fluorouracil (5-fluorouracil, 5-FU), vincristine and doxorubicin. However, the cure rate of these chemotherapeutic drugs for liver cancer is still quite limited. The main reasons include: individual differences in patients, serious side effects of anti-cancer drugs, and drug resistance of cancer cells. ) And metastatic ability.

In VC Chen, et al. (2019), Int. J. Cancer , 144(10):2428-2439, VC Chen et al. used data from the Taiwan National Health Insurance Research Database to conduct ethnic-based cases A population-based case-control study, and the analysis results show that the use of antipsychotics is negatively correlated with the risk of hepatocellular carcinoma, and that antipsychotics can reduce liver cells in a dose-dependent manner The incidence of cancer. In addition, VC Chen et al. further discovered that quetiapine, which belongs to atypical antipsychotics, can induce apoptosis in human liver cancer cell lines Huh7 and HepG2 in vitro, and Inhibit the proliferation and invasion of these cell lines.

Despite the above, relevant researchers in the field are still committed to developing active components that can be used to treat liver cancer.

Summary of the invention

Therefore, in the first aspect, the present invention provides a combination comprising quetiapine and 5-fluorouracil.

In the second aspect, the present invention provides a use of a combination supply containing quetiapine and 5-fluorouracil for the preparation of a medicine for the treatment of liver cancer.

In a third aspect, the present invention provides a kit comprising quetiapine and 5-fluorouracil, and the kit is used for simultaneous, alternate or sequential administration of quetiapine and 5-fluorouracil.

In a fourth aspect, the present invention provides a method for treating liver cancer in an individual, which comprises administering quetiapine and 5-fluorouracil to the individual.

Detailed description of the invention

It should be understood that if any previous case publication is quoted here, the previous case publication does not constitute a recognition: in Taiwan or any other country, the previous case publication forms a common general in the art. Part of knowledge.

For the purpose of this specification, it will be clearly understood that the word "comprising" means "including but not limited to", and the word "comprises" has a corresponding meaning.

Unless otherwise defined, all technical and scientific terms used in this article have meanings commonly understood by those familiar with the art of the present invention. A person familiar with the art will recognize that many methods and materials similar or equivalent to those described herein can be used to implement the present invention. Of course, the present invention is by no means restricted by the described methods and materials.

As used herein, the term "liver cancer" is a liver malignant tumor that can be caused by the following factors: chronic viral hepatitis (for example, chronic hepatitis B and chronic hepatitis C), liver Liver toxins [for example, aflatoxin], non-alcoholic steatohepatitis, or alcoholic liver disease.

As used herein, "treating" or "treatment" means preventing, reducing, alleviating, ameliorating, relieving, or controlling ( controlling) one or more clinical signs of a disease or disorder, and lowering, stopping or reversing the severity of a condition or symptom being treated The progression of severity.

In the development of drugs that can be used to treat liver cancer, the applicant unexpectedly discovered that quetiapine and the chemotherapy drug 5-fluorouracil (5-fluorouracil, 5-FU) can mutually enhance each other's anti-liver cancer efficacy, so The combination of quetiapine and 5-fluorouracil is expected to be useful for the treatment of liver cancer.

Therefore, the present invention provides a combination comprising quetiapine and 5-fluorouracil.

As used herein, the term "combination" refers to the combined use of two or more active ingredients for the treatment of a disease for co-administering to an individual in need of the treatment. In other words, the active ingredients can be combined into a single dosage form or administered simultaneously in separate dosage forms. Alternatively, the active ingredients can also be administered in separate dosage forms alternately or sequentially, with a predetermined period of time apart from each other. In some specific examples of the present invention, quetiapine and 5-fluorouracil are in separate dosage forms.

Preferably, quetiapine can be manufactured into a dosage form suitable for oral administration by using techniques well known to those skilled in the art. This includes, but is not limited to: sterile powder, lozenge (tablet), tablet (troche), lozenge, pill, capsule, dispersible powder or granule, solution, suspension, emulsion Emulsion, syrup, elixir, slurry and the like.

Preferably, 5-fluorouracil can be manufactured into a dosage form suitable for parenteral administration, that is, injection (for example, sterile Aqueous solution (sterile aqueous solution or dispersion)]. The route of parenteral administration can be selected from the group consisting of: intraperitoneal injection, intrapleural injection, intramuscular injection, intravenous injection , Intraarterial injection, intraarticular injection, intrasynovial injection, intrathecal injection, intracranial injection, intraepidermal injection , Subcutaneous injection, intradermal injection, intralesional injection, and sublingual administration.

According to the present invention, for the manufacture of different dosage forms, those who are familiar with the technology can choose suitable pharmaceutically acceptable carriers according to their professional qualities and routine techniques, which include, but are not limited to: solvents, buffers Buffer, emulsifier, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, wetting agent, lubricant , Absorption delaying agents, liposomes and the like.

In some specific examples of the present invention, quetiapine is in a dosage form suitable for oral administration, and 5-fluorouracil is in a dosage form suitable for parenteral administration.

According to the present invention, the molar ratio of 5-fluorouracil to quetiapine can be 1:2 to 1:200. In some specific examples, the molar ratio of 5-fluorouracil to quetiapine is 1:4 to 1:100. In an exemplary embodiment, the molar ratio of 5-fluorouracil to quetiapine is 1:100.

The present invention also provides a kit comprising quetiapine and 5-fluorouracil, and the kit is used for simultaneous, alternate or sequential administration of quetiapine and 5-fluorouracil.

As used herein, the term "kit" means a packaged product containing active ingredients for treating a disease. The set may include one or more boxes or containers for containing the above-mentioned active ingredients, as well as labels and/or instructions for indicating the use of the contents contained in the set.

In some specific examples of the present invention, quetiapine and 5-fluorouracil are in separate dosage forms, that is, contained in separate boxes or containers.

In view of the synergistic effect of quetiapine and 5-fluorouracil on the effect of anti-liver cancer, the present invention also provides a combination supply as described above for the preparation of a medicine for the treatment of liver cancer.

The present invention also provides a method for treating liver cancer in an individual, which comprises administering to the individual a combination as described above.

According to the present invention, the dosage and frequency of administration of quetiapine and 5-fluorouracil will vary depending on the following factors: the severity of the disease to be treated, the route of administration, and the age, physical condition and physical condition of the individual to be treated reaction. Generally speaking, the combination can be administered as a single dose or divided into several doses.

Detailed description of the preferred embodiment

The present invention will be further described with respect to the following embodiments, but it should be understood that these embodiments are for illustrative purposes only, and should not be construed as limitations on the implementation of the present invention. Examples General experimental materials: 1. Source and culture of human liver cancer cell line HepG2:

The human liver cancer cell line HepG2 (ATCC ^® HB-8065) used in the following examples was purchased from the American Type Culture Collection (ATCC).

HepG2 cells use Dulbecco's Modified Eagle's Medium (DMEM) (purchased from gibco, catalog number 11885-084) [with 10% Fetal Bovine Serum (FBS)] , And incubate in an incubator set at 37°C and 5% CO ₂ . After that, the medium was replaced with fresh medium approximately every 2 days. When the cell density reaches about 80-90% confluence, cell subculture is performed. 2. Quetiapine (quetiapine) and anticancer drugs (anticancer drugs):

The quetiapine (trade name Silekang film-coated tablets) used in the following examples was purchased from Taiwan AstraZeneca Co., Ltd., 5-fluorouracil (5-fluorouracil, 5-FU ) Was purchased from Nam Kwong Chemical Pharmaceutical Co., Ltd. (brand name Haofu injection), Bevacizumab (Bevacizumab) was purchased from Roche Pharmaceutical Co., Ltd. (brand name Avastin), vincristine ( Vincristine) was purchased from Pfizer Pharmaceutical Co., Ltd. (trade name Vincristine), and the above-mentioned drugs were all provided by Chiayi Chang Gung Memorial Hospital. General experimental methods: 1. Statistical analysis:

In the following examples, the experiments of each group are repeated at least 3 times, and the obtained experimental data are expressed in mean. The difference between the experimental data of each group is by using Prism 5 software (GraphPad Software, San Diego, CA) to perform two-way analysis of variance, two-way ANOVA, and then to perform Taki Test (Tukey's test) to assess the differences between groups. If the result of the statistical comparison is p <0.05, it represents statistical significance. Example 1. Evaluation of the effectiveness of the combination of quetiapine and different anti-cancer drugs on anti-hepatocellular carcinoma

In order to understand the efficacy of the combination of quetiapine and three commonly used clinical anticancer drugs (including 5-fluorouracil, bevacizumab, and vincristine) on liver cancer, the applicant conducted the following experiment. A. The ineffective concentration of different anticancer drugs for HepG2 cells (ineffective concentration)

The applicant used 5-fluorouracil, bevacizumab, and vincristine to perform the following experiments respectively, in order to find out the concentration of these anticancer drugs that have no inhibitory effect on the growth of HepG2 cells, that is, the ineffective concentration.

The HepG2 cells subcultured in accordance with item 1 of the above "General Experimental Materials" were cultured at a rate of 5×10 ³ cells/well in 96-containing DMEM medium (supplemented with 10% FBS) of 0.1 mL. In the well culture plate, the culture was then carried out in an incubator (37° C., 5% CO ₂ ) for 24 hours to allow HepG2 cells to adhere to the culture plate. Then, after removing the liquid in each well, the following different concentrations of anticancer drugs (prepared in DMEM medium, including 5-fluorouracil with concentrations of 0.001, 0.005, 0.01, 0.02, and 0.05 mM, respectively) Bevacizumab at 0.125, 0.25, 0.5, and 1.0 μg/mL, and vincristine at concentrations of 0.25, 0.5, 1.0, and 2.0 nM, respectively, were added to the cells in each well, and further cultured for 48 hours. In addition, cells treated with DMEM medium without any anti-cancer drugs were used as a control group.

(dimethyl sulfoxide, DMSO)並予以混合均勻，然後於570 nm的波長下以一ELISA讀取機(ELISA reader)(型號為EnSpire™ Multimode Plate Reader，廠牌為PerkinElmer)來讀取各井的吸光值(OD ₅₇₀)。 After that, the liquid in each well was removed, and 200 μL of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide {3-[4,5-Dimethylthiazol- 2-yl]-2,5-diphenyltetrazolium bromide} (MTT, 5 mg/mL) was added to each well and incubated for 4 hours. Next, add 200 μL of dimethyl sulfoxide

(dimethyl sulfoxide, DMSO) and mix well, and then use an ELISA reader (EnSpire™ Multimode Plate Reader, brand PerkinElmer) at a wavelength of 570 nm to read the absorbance of each well (OD ₅₇₀ ).

The cell viability percentage (%) is _{calculated by substituting the measured absorbance value (OD 570} ) into the following formula (1): formula (1) : A=( B/C) ×100 where: A= Percentage of cell viability (%) B= OD _{570 absorbance measured in each well C= OD 570} absorbance measured in the control group

After that, analyze the obtained experimental data in accordance with the method described in item 1 "Statistical Analysis" of the "General Experimental Methods" above. The experimental results showed that the ineffective concentrations of 5-fluorouracil, bevacizumab, and vincristine for HepG2 cells were 0.005 mM, 1.0 μg/mL, and 2.0 nM, respectively (data not shown). B. The inhibitory effect of the combination of quetiapine and different anticancer drugs on HepG2 cells

In order to understand the difference in the anti-liver cancer efficacy of the combination of quetiapine and the above three anticancer drugs compared with quetiapine alone, the applicant obtained the ineffective concentration of the three anticancer drugs on HepG2 cells based on item A above , Choose lower concentrations of 5-fluorouracil (0.001 mM), bevacizumab (0.25 μg/mL) and vincristine (1.0 nM) to compare with different concentrations of quetiapine (0.1 and 0.2 mM) Combine, and then use the 6 combinations obtained to treat HepG2 cells respectively, and perform cell viability analysis with reference to the operating procedure described in item A above. In addition, HepG2 cells treated with quetiapine (0.1 and 0.2 mM) alone were used as a positive control group, and cells treated with DMEM medium without quetiapine and any anticancer drugs were used as a negative control group.

After that, analyze the obtained experimental data in accordance with the method described in item 1 "Statistical Analysis" of the "General Experimental Methods" above. The results obtained are shown in Figures 1 to 3, respectively.

Figure 1 shows the growth inhibitory effect of different concentrations of quetiapine alone and its combined use with 0.001 mM 5-fluorouracil on HepG2 cells. It can be seen from Figure 1 that the single use of quetiapine can effectively inhibit the growth of HepG2 cells and exhibit a dose-dependent effect. In contrast, quetiapine has a 5- The combination of fluorouracil has a significant improvement in the growth inhibitory effect of HepG2 cells, which means that the combination of quetiapine and 5-fluorouracil can exhibit a synergic effect on the ability of anti-liver cancer, which makes 5 -The dosage of fluorouracil can be greatly reduced.

Figures 2 and 3 respectively show the combined use of quetiapine and 0.25 μg/mL bevacizumab, the combined use of quetiapine and 1.0 nM vincristine and the growth inhibitory effect of quetiapine alone on HepG2 cells. It can be seen from Figure 2 and Figure 3 that although quetiapine alone can effectively inhibit the growth of HepG2 cells, the combination of quetiapine and ineffective concentrations of bevacizumab or vincristine has no effect on the growth of HepG2. Increase, but antagonistic effect (antagonistic effect) appears. In particular, the combination of bevacizumab and 0.1 mM quetiapine cannot even exhibit the effect of anti-liver cancer. C. The inhibitory effect of the combination of different concentrations of quetiapine and 5- fluorouracil on HepG2 cells

In order to further verify the synergistic effect of the combination of quetiapine and 5-fluorouracil in inhibiting the growth of HepG2 cells, the applicant chose higher concentrations of 5-fluorouracil (including 0.005, 0.01, 0.02, and 0.05 mM) to compare with Different concentrations of quetiapine (0.1 and 0.2 mM) were combined, and the 8 combinations obtained were subjected to cell viability analysis with reference to the procedure described in item B above. In addition, the positive control group and the negative control group are also as described in item B above.

After that, analyze the obtained experimental data in accordance with the method described in item 1 "Statistical Analysis" of the "General Experimental Methods" above. The results obtained are shown in Figures 4 to 7 respectively.

Figures 4 to 7 respectively show the growth inhibitory effects of different concentrations of quetiapine alone and its combined use with 0.005, 0.01, 0.02, and 0.05 mM 5-fluorouracil on HepG2 cells. It can be seen from Figure 4 to Figure 7 that compared with quetiapine alone, the anti-hepatocellular cancer activity of the combination of 0.005 to 0.05 mM 5-fluorouracil and 0.1 to 0.2 mM quetiapine is significantly improved, showing that quintiapine Tiapine and higher concentrations of 5-fluorouracil can also exhibit excellent synergistic effects in anti-liver cancer activity when used in combination.

Based on the above, the applicant believes that the combination of quetiapine and 5-fluorouracil can be used as a combination therapy for the treatment of liver cancer, and it is beneficial to reduce the clinical dose of 5-fluorouracil, and even reduce it to the conventional wisdom. The low dose with no therapeutic effect can effectively avoid the possible side effects of 5-fluorouracil.

All patents and documents cited in this specification are incorporated into this case as reference materials in their entirety. If there is a conflict, the detailed description of the case (including definitions) will prevail.

Although the present invention has been described with reference to the above specific specific examples, it is obvious that many modifications and changes can be made without departing from the scope and spirit of the present invention. Therefore, it is intended that the present invention is only limited by the scope of the patent application attached hereto.

The above and other objectives, features and advantages of the present invention will become apparent with reference to the following detailed description and preferred embodiments and the attached drawings. Among them: Figure 1 shows that HepG2 cells are used in different concentrations. Quetiapine and its combination with 0.001 mM 5-fluorouracil, the percentage of cell viability measured after treatment, where "*" means when compared with the corresponding concentration of quetiapine alone, p >0.05; Figure 2 shows the percentage of cell viability measured by HepG2 cells treated with different concentrations of quetiapine and its combination with 0.25 μg/mL bevacizumab, where "*" means when and corresponding Concentration of quetiapine alone for comparison, p >0.05; Figure 3 shows the measured cell viability of HepG2 cells treated with different concentrations of quetiapine and its combination with 1.0 nM vincristine. Percentage; Figure 4 shows the percentage of cell viability measured in HepG2 cells treated with different concentrations of quetiapine and its combination with 0.005 mM 5-fluorouracil, where "*" means when and the corresponding concentration For comparison with quetiapine alone, p >0.05; Figure 5 shows that HepG2 cells are treated with different concentrations of quetiapine and its combination with 0.01 mM 5-fluorouracil. Activity percentage, where "*" means that when compared with the corresponding concentration of quetiapine alone, p >0.05; Figure 6 shows that HepG2 cells were used with different concentrations of quetiapine and 0.02 mM 5-fluorourea. The percentage of cell viability measured after treatment with the combination of pyrimidines, where "*" means when compared with the corresponding concentration of quetiapine alone, p >0.05; and Figure 7 shows that HepG2 cells are used with different concentrations of quetiapine alone. The percentage of cell viability measured after quetiapine and its combination with 0.05 mM 5-fluorouracil are treated, where "*" means when compared with the corresponding concentration of quetiapine alone, p >0.05 .

Claims (6)

A combination of quetiapine and 5-fluorouracil is used to prepare a medicine for the treatment of liver cancer. Such as the use of claim 1, wherein quetiapine and 5-fluorouracil are in separate dosage forms. The use according to claim 2, wherein the combination is for simultaneous, alternate or sequential administration of quetiapine and 5-fluorouracil. Such as the use of claim 1, wherein quetiapine is in a dosage form for oral administration. The use of claim 1, wherein 5-fluorouracil is in a dosage form for parenteral administration. Such as the use of claim 1, wherein 5-fluorouracil and quetiapine have a molar ratio ranging from 1:2 to 1:200.

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