CN112263591A

CN112263591A - Composition for treating cancer and application thereof

Info

Publication number: CN112263591A
Application number: CN202011244260.5A
Authority: CN
Inventors: 周文虎; 王丽彤; 余梦真; 彭颖
Original assignee: Central South University
Current assignee: Central South University
Priority date: 2020-11-10
Filing date: 2020-11-10
Publication date: 2021-01-26

Abstract

The invention discloses a composition for treating cancer and application thereof. The invention combines the A medicine and the B medicine for preparing the medicine for treating cancer. The dithiocanadine has longer half-life, and the efficiency of forming CuET in a tumor part after the dithiocanadine is combined with oral copper gluconate is higher, and the anti-tumor activity is stronger. After the dithiocarbazinate and the copper sulfate are combined at a tumor part to form CuET, the tumor cells can be effectively killed, and the growth of solid tumors is inhibited.

Description

Composition for treating cancer and application thereof

Technical Field

The invention relates to the technical field of biological medicines, in particular to a composition for treating cancer and application thereof.

Background

Malignant tumor is a serious disease seriously threatening the life safety of human beings, and the morbidity and the mortality are high. At present, chemotherapy is one of the main means for treating tumors clinically, and common chemotherapeutic drugs include adriamycin, paclitaxel, cisplatin and the like. However, non-selective distribution of chemotherapeutic drugs in the body can cause major toxic and side effects, and long-term administration can cause drug resistance. Therefore, the high-efficiency and low-toxicity treatment method has important practical significance in treating the tumor. In recent years, research and development of molecular targeted drugs have become a hot field for research and development of antitumor drugs. The molecular targeted medicine mainly aims at key targets of pathophysiology generation and development of malignant tumor to perform therapeutic intervention without affecting the activity of normal cells. At present, some molecular targeted drugs have already shown better curative effect in the corresponding tumor treatment. Disulfiram (DSF) is a commonly used clinical drug for abstinence, the metabolite sodium dithionate and copper ions form a complex (CuET for short), can specifically recognize nuclear localization protein 4(NPL4) and induce aggregation of the nuclear localization protein, interfere with the important pathway p97-NPL4-UFD1 pathway of tumor cell growth, further cause damaged protein degradation and ubiquitinated protein aggregation, finally induce apoptosis, and the anti-tumor mechanism of the disulfiram forms molecular targeting characteristics. However, CuET itself has very poor solubility and cannot be taken orally directly. Must be loaded in nano material or other materials to be utilized by human body, resulting in high medication cost.

By analyzing the pharmacodynamic mechanism of DSF, Cu can be found²⁺Is the key point of exerting the drug effect. At the same time, DSF itself is inactive and needs to be metabolized into DTC in vivo to interact with Cu²⁺Binding, rapid plasma clearance of DSF with short half-life, with Cu²⁺The CuET forming efficiency at the tumor part is low, and the CuET has certain toxic and side effects.

Disclosure of Invention

In order to solve the above problems, it is an object of the present invention to provide a composition for treating cancer and use thereof.

In order to achieve the above object, the present invention provides a composition for treating cancer, comprising an agent a and an agent B, wherein the agent a is dithiocarbazinate and the agent B is copper gluconate.

The composition for treating cancer as described above, further, the dosage of said drug A is 0.1 mg/kg-50 mg/kg, and the dosage of said drug B is 0.1 mg/kg-5 mg/kg.

Based on one general technical concept, the present invention provides a use of the composition for treating cancer in the preparation of a medicament for treating cancer.

In addition, the medicine for treating cancer is one or more of medicines for treating solid tumors such as plasmacytoma, pancreatic cancer, multiple myeloma, acute lymphatic leukemia, ovarian cancer, non-small cell lung cancer, colorectal cancer, breast cancer, malignant glioma and the like.

The above application, further, the method of the application is: the sodium dithiocarbazate is orally taken firstly, and the copper sulfate is orally taken after 5-10 hours.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a composition for treating cancer, namely a composition of sodium dithionite and copper sulfate. Compared with DSF, DTC has longer half-life, and has higher efficiency of forming CuET in a tumor part after being combined with oral copper gluconate, and stronger antitumor activity. After the DTC and the copper sulfate are combined at a tumor part to form CuET, the CuET can effectively kill tumor cells and inhibit the growth of solid tumors.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 is a pharmacokinetic profile of Disulfiram (DSF) and sodium Dithionate (DTC) in an example of the invention;

FIG. 2 shows the content of CuET produced at the tumor site after oral administration of DSF, DTC, DSF + copper gluconate (CuGlu) and DTC + CuGlu, respectively, in the present invention.

FIG. 3 is a graph showing the tumor growth curves of mice after oral administration of DSF, DTC, DSF + CuGlu, and DTC + CuGlu, respectively, in the present example.

FIG. 4 is a graph showing tumor growth curves of mice after oral administration of DTC and CuGlu at different ratios, respectively, in the examples of the present invention.

FIG. 5 is a graph of tumor growth in mice following oral administration of varying doses of DTC and CuGlu in an example of the invention.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.

Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art; all reagents used in the examples are commercially available unless otherwise specified.

The percentage "%" referred to in the present invention means mass% unless otherwise specified; but the percentage of the solution, unless otherwise specified, refers to the grams of solute contained in 100ml of the solution.

The weight parts in the invention can be the weight units known in the art such as mu g, mg, g, kg, and the like, and can also be multiples thereof, such as 1/10, 1/100, 10, 100, and the like.

In the following examples, details of the equipment and manufacturer used are given in Table 1:

TABLE 1 name of main instrument and manufacturer

Name of instrument	Manufacturer of the product
		CP225D model electronic balance	Siderelius de Sartorius (Sartorius) Co
BP224S type electronic balance	Siderelius de Sartorius (Sartorius) Co

In the following examples, the names of the main reagents and the manufacturers used are shown in Table 2:

TABLE 2 Main reagent names and manufacturers

Name of reagent	Manufacturer of the product
		Sodium dithiocarbinate	Sigma Co USA
Disulfiram	Sigma Co USA
		Gluconic acid copper salt	Aladdin reagents Ltd

Example 1

A composition for treating cancer comprises sodium dithionite and copper gluconate.

The use of the composition for treating cancer of example 1 in the preparation of a medicament for treating cancer, comprising: the sodium dithiocarbazate is orally taken, and the copper dose of copper gluconate is orally taken after 5-10 hours.

Experimental example 2:

the plasma half-life of DSF and DTC was examined by the following specific steps:

the SD rats were randomly divided into two groups, a DSF administration group and a DTC administration group, each of 6 rats. The DSF administration group was gavage at a dose of 50 mg/kg. The DTC administration group was administered by intragastric administration at a dose of 50 mg/kg. Collecting blood in orbit at 0, 1, 2, 4, 8, 12, 24, and 48h, adding 100 μ L heparin sodium into the collected blood, and storing in refrigerator at-80 deg.C. After the sample collection is completed, 200 mu L of each blood sample is taken out and added into a 1.5ml EP tube, 10 mu L of 100mM GSH solution is added for incubation for 10min, then the mixture is centrifuged at 3000rpm for 10min to obtain supernatant plasma, 100 mu L of methanol is continuously added into the plasma for vortex 120s to extract the drug, the mixture is centrifuged at 10000rpm for 5min, and an organic phase nitrogen blower is taken for drying completely and then is added into a quantitative volume dissolving HPLC-MS/MS for sample injection analysis.

Fig. 1 is a pharmacokinetic profile of DSF and DTC, showing that DTC has a longer plasma half-life.

Experimental example 3:

sodium dithionate and copper gluconate were examined for the formation of CuET in mouse tumors.

The investigation steps are as follows:

(1) establishing a tumor-bearing nude mouse model: collection of logarithmic growth of M231 cells dispersed in PBS at a cell density of 1X 10⁷A volume of 100. mu.L of the mixture was mixed with matrigel and injected into the axillary region of BALB/c nude mice (female, 6 weeks). Female BALB/c nude mice, 6 weeks old, were purchased from Calvens laboratory animals, Inc., Changzhou.

(2) The tumor-bearing nude mice are randomly divided into four groups, namely a DSF group, a DTC group, a DSF + CuGlu group and a DTC + CuGlu group, wherein each group comprises 6 mice.

DsF or DTC group were administered separately by gavage at a dose of 50 mg/kg.

DSF + CuGlu and DTC + CuGlu groups DSF or DTC was administered by gavage at 9:00 in the morning and CuGlu was administered at 19:00 in the evening (dose: 1 mg/kg).

And taking out tumors in the mice of each group after 24h, and detecting the content of CuET in each group by HPLC-MS/MS sample injection analysis.

FIG. 2 shows the amount of CuET present in tumors in mice of each group after administration of DSF, DTC, DSF + CuGlu, and DTC + CuGlu. As can be seen from fig. 2, the CuET content of the DTC + CuGlu group was highest, indicating that the combination of DTC + CuGlu produced CuET more efficiently.

Example 4:

the tumor inhibition effect of sodium dithionate and copper gluconate is examined. The method comprises the following specific steps:

putting lotus in the airCulturing nude tumor mouse until tumor grows to 200mm³On the left and right, the tumor-bearing nude mice were randomly divided into four groups, which were DSF, DTC, DSF + CuGlu, DTC + CuGlu, and 6 mice per group. PBS, DSF, DTC, DSF + CuGlu, DTC + CuGlu (dose: DSF or DTC 50 mg/kg; CuGlu: 1mg/kg) were orally administered for 0, 3, 6, 9 days, respectively. Mice were weighed every two days and tumor volumes were measured with a vernier caliper to day 14, and the antitumor efficiency of each group was compared by the relative volume of the tumors of each group. Tumor volume calculation formula: length x width²/2。

FIG. 3 is a graph of tumor volume change after oral administration of PBS, DSF, DTC, DSF + CuGlu, DTC + CuGlu on

days

0, 3, 6, and 9, respectively. As can be seen from FIG. 3, DSF, DTC, DSF + CuGlu, and DTC + CuGlu all had a certain tumor-inhibiting effect. The two groups added with CuGlu have obviously enhanced tumor inhibition effect, and the curative effect of the DTC + CuGlu group is stronger than that of the DSF + CuGlu group. The results indicate that DTC is the optimal combination with CuGlu.

Example 5:

investigating the influence of the mass ratio of DTC to CuGlu on the anti-tumor efficacy, comprising the following steps:

the tumor-bearing nude mice are randomly divided into four groups, 6 mice are respectively provided with DTC and CuGlu with different proportions (the mass ratio of DTC to CuGlu is respectively 50: 0.5, 50: 1, 50: 2 and 50: 5). The administration time interval, the number of administrations and the evaluation index were referred to example 3.

FIG. 4 is a graph of tumor volume change after oral administration of PBS and DTC + CuGlu at different mass ratios on

days

0, 3, 6, and 9, respectively. As can be seen from FIG. 4, the DTC + CuGlu groups of each ratio have significant tumor-inhibiting effects, with the therapeutic effects of 50: 1, 50: 2 and 50: 5 being slightly different. The results show that DTC and CuGlu are combined at a mass ratio of 50: 1 to exert obvious anti-tumor efficacy, and the curative effect cannot be improved by continuously increasing the dosage of CuGlu.

Example 6:

examining the change curve of the tumor volume after DTC + CuGlu with different doses, which comprises the following steps:

tumor-bearing nude mice were randomly divided into four groups of 6 mice each. The fixed DTC and CuGlu are administered in a mass ratio of 50: 1. DTC and CuGlu were given at different doses (DTC and CuGlu at doses of 5mg/kg and 0.1mg/kg, 10mg/kg and 0.2mg/kg, 20mg/kg and 0.4mg/kg, 50mg/kg and 1mg/kg, respectively). The administration time interval, the number of administrations and the evaluation index were referred to example 3.

FIG. 5 is a graph of tumor volume change after oral administration of PBS and different doses of DTC + CuGlu on

days

0, 3, 6, and 9, respectively. As can be seen from FIG. 5, the antitumor effect of each group was increased with the increase of the dose, and the antitumor effect was strongest at the doses of DTC and CuGlu of 50mg/kg and 1mg/kg, respectively.

It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.

Claims

1. A composition for treating cancer comprises a drug A and a drug B, wherein the drug A is dithiocanadine, and the drug B is copper gluconate.

2. The composition for treating cancer according to claim 1, wherein the dosage of the drug a is 0..1mg/kg to 50mg/kg, and the dosage of the drug B is 0..1mg/kg to 5 mg/kg.

3. Use of a composition according to claim 1 or 2 for the treatment of cancer in the manufacture of a medicament for the treatment of cancer.

4. The use of claim 3, wherein the cancer treatment is one or more of plasmacytoma, pancreatic cancer, multiple myeloma, acute lymphoblastic leukemia, ovarian cancer, non-small cell lung cancer, colorectal cancer, breast cancer, and glioblastoma.

5. The use according to claim 3, characterized in that the method of application is: : the sodium dithiocarbinate is firstly orally taken, and the copper sulfate is then orally taken after 5 to 10 hours of interval.