CN112972443A

CN112972443A - Anticancer medicine and its application

Info

Publication number: CN112972443A
Application number: CN202110329896.8A
Authority: CN
Inventors: 汪丽
Original assignee: Hangzhou Tianfan Biotechnology Co ltd
Current assignee: Hangzhou Tianfan Biotechnology Co ltd
Priority date: 2021-03-29
Filing date: 2021-03-29
Publication date: 2021-06-18

Abstract

The invention discloses an anticancer drug and application thereof, belonging to the technical field of medicines.A main active component of the drug is prepared from cryptobrecin S, which is a toxic fungus extract and is used for inhibiting the proliferation of colorectal cancer cells, promoting the apoptosis of the colorectal cancer cells and inducing the cell cycle arrest of the colorectal cancer cells; the medicine contains cryptomerin S with pharmaceutically effective dose and pharmaceutically acceptable carrier; the preparation forms can be selected from suspension, emulsion, tablet, capsule, granule, oral liquid or injection. The invention explores the application of the fungus extract cryptomerin S in anticancer, and provides a medicinal basis for the application of the fungus-derived anticancer drugs; the medicine provided by the invention has simple source and can better expand production.

Description

Anticancer medicine and its application

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an anticancer medicine and application thereof.

Background

Colorectal cancer is one of the common cancers in China. Among the cancer statistics published in GLOBOCAN2018, colorectal cancer is the fourth most common cancer, accounting for 6.1%. Over 70% of colorectal cancer cases are sporadic, mainly due to poor dietary hygiene habits and lifestyle. Second, the incidence increases exponentially with age. Early colorectal cancer may be free of any symptoms, and the middle and late stages may be manifested as abdominal distension, dyspepsia, followed by a change in defecation habits, abdominal pain, mucous stools or bloody stools. After tumor ulceration, blood loss and toxin absorption, symptoms such as anemia, low fever, hypodynamia, emaciation, edema of lower limbs and the like often appear. Colorectal cancer is insidious, many patients are diagnosed with advanced stage of the disease, and the 5-year survival rate of advanced metastatic colorectal cancer patients may be less than 10% with current medical approaches. The traditional treatment methods such as chemotherapy and radiotherapy have obvious side effects, and cancer cells are easy to generate resistance. Therefore, it is important to find new therapeutic approaches to the treatment of colorectal cancer.

Disclosure of Invention

The invention aims to provide an anti-cancer drug, in particular to colorectal cancer, and solves the problems in the background art that compounds are searched for from natural products of fungal metabolism to treat the colorectal cancer.

In order to achieve the purpose, the invention provides the following technical scheme: the main active ingredient of the medicine is prepared from cryptomerin S, and the structural formula of the cryptomerin S is shown as the formula (I):

preferably, the cryptomerin S is a toxic fungal extract.

Preferably, the cryptoleptin S is used for inhibiting cancer cell proliferation, promoting cancer cell apoptosis and inducing cancer cell cycle arrest.

Preferably, the medicament comprises a pharmaceutically effective amount of cryptomerin S and a pharmaceutically acceptable carrier.

Preferably, the pharmaceutical preparation is in the form of suspension, emulsion, tablet, capsule, granule, oral liquid or injection.

Preferably, the cancer is colorectal cancer.

Preferably, the use of a medicament as described in any one of the above in the manufacture of a medicament for the treatment of colorectal cancer.

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

1. the invention explores the application of fungus extract illudin S in anticancer, in particular to the application of an anti-colorectal cancer medicine from fungus, and provides a medicinal basis for the application of the anti-colorectal cancer medicine from fungus;

2. the medicine provided by the invention has simple source and can better expand production.

Drawings

FIG. 1 shows CCK8 testing the proliferation of human colorectal cancer cells SW480 and HCT116 by illedin S;

FIG. 2 shows flow cytometry to detect the effect of illedin S on apoptosis of human colorectal cancer cells SW480 and HCT 116;

FIG. 3 shows flow cytometry to examine the effect of illedin S on the cell cycle of human colorectal cancer cells SW480 and HCT 116.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

CCK8 was used in this experiment to test the effect of illedin S in inhibiting the proliferation of human colorectal cancer cells SW480 and HCT 116.

The method comprises the following steps:

collecting logarithmic phase cells, adjusting the concentration of cell suspension to 5 × 104/ml, adding into a 96-well plate, wherein each well is 100 μ l, and incubating in 5% concentration CO2 at 37 ℃; adding the illudin S to enable the final concentration to reach 0.1 mu mol/L, 0.2 mu mol/L,0.4 mu mol/L,1 mu mol/L and 2 mu mol/L, setting 4 compound holes for each group, setting a blank control group, CO-culturing for 12 hours, 24 hours and 48 hours at the temperature of 37 ℃ under the condition of 5% CO2, adding 10 mu L of CCK8 solution into each hole, observing the absorbance of each group at 450nm after continuously incubating for 4 hours in a dark place, and repeating the experiment for 3 times.

The results are shown in figure 1, 0.2 mu mol/L,0.4 mu mol/L,1 mu mol/L,2 mu mol/L of the illudin S can obviously inhibit the cell viability after 12 hours of action, 0.1 mu mol/L of the illudin S can obviously inhibit the cell viability after 48 hours of action, and the inhibitory effect of the illudin S on the viability of SW480 and HCT116 cells is increased along with the increase of time and concentration.

Example 2:

the experiment adopts a flow cytometer to detect the effect of promoting apoptosis of human colorectal cancer cells SW480 and HCT116 by the illedin S.

The method comprises the following steps:

cells in log phase were collected, seeded in 100mm dishes and after 24 hours of cell adherence, ill udin S was added to achieve final concentrations of 0.1 μmol/L, 0.6 μmol/L. Each group was plated with 4 replicate wells and a blank control group was plated and CO-incubated at 5% CO2 at 37 ℃ for 24 hours.

After treatment, cells were trypsinized, centrifuged, and collected in a flow tube; PBS washed cells 1 time; on ice, 1 × Annexin-binding buffer was prepared and used to resuspend the cells. After resuspension, 5. mu.l Annexin V and 10. mu.l PI were added to the cells, incubated at room temperature for 5 minutes in the absence of light, and apoptosis was detected on a flow cytometer.

The results are shown in FIG. 2, and the concentration-dependent promotion of apoptosis by the illudin S in SW480 and HCT116 cells.

Example 3:

the experiment adopts a flow cytometer to detect the effect of inducing cell cycle arrest of human colorectal cancer cells SW480 and HCT116 by the illedin S.

The method comprises the following steps:

collecting cells in logarithmic phase, inoculating the cells into a 100mm culture dish, and adding the illudin S after the cells adhere to the wall for 24 hours to ensure that the final concentration of the illudin S reaches 0.1 mu mol/L and 0.6 mu mol/L. Each group was plated with 4 replicate wells and a blank control group was plated and CO-incubated at 5% CO2 at 37 ℃ for 24 hours.

For the detection of the cell cycle: after treatment, cells were trypsinized, centrifuged, and collected in a flow tube; PBS washed cells 1 time; the cells were added to 1 ml of ice-cooled 70% ethanol, gently blown and mixed, and fixed at 4 ℃ for 12 hours. Centrifuging, washing cells by PBS, adding 0.5 ml of propidium iodide staining solution into each tube of cell sample after collecting the cells, slowly and fully suspending cell sediment, and carrying out dark warm bath for 30 minutes at the temperature of 37 ℃; flow detection was completed within 24 hours after staining was complete.

The results are shown in FIG. 2, where illudin S induces cell cycle arrest of SW480 and HCT116 in a concentration-dependent manner in S phase.

And (3) detection results:

the invention relates to an application of a drug in resisting colorectal cancer, in particular to an application of a toxic fungus extract, namely, illudin S in preparing drugs for inhibiting colorectal cancer cells SW480 and HCT116, wherein the illudin S is added into a suspension of human colorectal cancer cells SW480 and HCT116 in a logarithmic growth phase with the concentration of 5 x 104 cells/ml, so that the final concentration of the illudin S reaches 0.1 mu mol/L, 0.2 mu mol/L,0.4 mu mol/L,1 mu mol/L and 2 mu mol/L, the mixture is cultured for 12 hours, 24 hours and 48 hours under the condition of 5% concentration CO2 and the temperature of 37 ℃, and the inhibition effect of the illudin S on the proliferation of the human colorectal cancer cells SW480 and HCT116 is determined by adopting CCK8 according to different culture times. The results demonstrate that illedin S inhibit the proliferation of colorectal cancer cells SW480 and HCT116 both time-dependent and concentration-dependent.

The application of toxic fungus extract illudin S in preparing the medicine for inhibiting the colorectal cancer cells SW480 and HCT116 is characterized in that the illudin S is added into a suspension of the human colorectal cancer cells SW480 and HCT116 in a logarithmic growth phase with the concentration of 5 x 104 cells/ml, the final concentration of the illudin S reaches 0.1 mu mol/L and 0.6 mu mol/L, CO2 with the concentration of 5% and the temperature condition of 37 ℃ are jointly cultured for 24 hours, and the control effect of the illudin S on the apoptosis and the cell cycle of the colorectal cancer cells SW480 and HCT116 is measured by flow cytometry.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. An anticancer drug, which is characterized in that: the main active ingredient of the medicine is prepared from cryptomerin S, and the structural formula of the cryptomerin S is shown as the formula (I):

2. an anticancer agent as claimed in claim 1, wherein said cryptobrecin S is a toxic fungal extract.

3. The anticancer agent as claimed in claim 1, wherein said cryptoleptin S is used for inhibiting cancer cell proliferation, promoting cancer cell apoptosis, and inducing cancer cell cycle arrest.

4. An anticancer agent as claimed in claim 2, wherein said agent contains a pharmaceutically effective amount of cryptomerin S and a pharmaceutically acceptable carrier.

5. The anticancer agent according to claim 4, wherein the pharmaceutical formulation is suspension, emulsion, tablet, capsule, granule, oral liquid or injection.

6. An anticancer agent as claimed in any one of claims 1 to 5, wherein said cancer is colorectal cancer.

7. Use of the medicament of claim 1 for the manufacture of a medicament for the treatment of rectal cancer.