CN114748478A - Composition of wogonin and irinotecan and application of composition in preparation of anti-colorectal cancer drugs - Google Patents

Abstract

The invention discloses a composition of wogonin and irinotecan and application thereof in preparing medicaments for resisting colorectal cancer. The invention provides a composition of wogonin and irinotecan, wherein the two chemical components in the composition have obvious synergistic effect on the aspect of anti-colorectal cancer and produce unexpected technical effect of 1+1 & gt 2, so the composition of wogonin and irinotecan has the prospect of developing into anti-colorectal cancer medicaments.

Description

Composition of wogonin and irinotecan and application of composition in preparation of anti-colorectal cancer drugs

Technical Field

The invention belongs to the field of medicines, and particularly relates to a composition of wogonin and irinotecan and application of the composition in preparation of medicaments for resisting colorectal cancer.

Background

Colorectal cancer (CRC) is one of the most prevalent cancers worldwide, with the fourth largest cancer type worldwide. According to epidemiological data statistics, in 2020, 193 million CRC (accounting for about 10 percent and the third place), 94 ten thousand death (accounting for about 9.4 percent and the second place) are newly diagnosed worldwide, and the trend of rapid growth and youthfulness is shown, thereby seriously threatening the health of human beings. Surgical treatment remains the most effective treatment at present, but the risk of recurrence and metastasis remains. Radiotherapy and chemotherapy as adjuvant therapy of surgery can reduce local recurrence rate, but can cause adverse reactions such as emesis, enteritis, alopecia, anorexia, leukopenia, and immunity decline. Although the clinical treatment scheme of the colon cancer is continuously improved, the 5-year survival rate of the chemotherapy, the radiotherapy and other comprehensive treatments mainly based on the operation still does not exceed 40 percent.

Wogonin is one of the main active ingredients of traditional Chinese herbal medicine scutellaria baicalensis, is a flavonoid compound derived from natural plants and separated from scutellaria baicalensis roots, and has various pharmacological effects of resisting inflammation, allergy, virus, convulsion, blood vessel protection, thrombosis and tumor and the like. Irinotecan (Irinotecan, CPT-11) is a semisynthetic water-soluble camptothecin derivative, CPT-11 and a metabolite SN38 thereof are DNA topoisomerase I inhibitors, and a compound formed by the CPT-11, the topoisomerase I and DNA can cause DNA single-strand breakage, prevent DNA replication and inhibit RNA synthesis, is a first-line medicament for late colorectal cancer, and can also be used for postoperative adjuvant chemotherapy.

At present, no research at home and abroad shows that the wogonin and the irinotecan have synergistic colon cancer activity, and no report that the wogonin and the irinotecan are combined to prepare the anti-colorectal cancer medicament exists.

Disclosure of Invention

The invention aims to provide a composition of wogonin and irinotecan and application thereof in preparing medicaments for resisting colorectal cancer.

The above purpose of the invention is realized by the following technical scheme:

a composition comprises wogonin and irinotecan.

Preferably, the ratio of the amount of wogonin to irinotecan is 0.125:1 to 1.5: 1.

The application of the composition in preparing anti-colorectal cancer drugs.

Preferably, the medicine takes the composition as an active ingredient for resisting colorectal cancer, and also contains pharmaceutically acceptable carriers or auxiliary materials, so as to prepare pharmaceutically acceptable dosage forms.

More preferably, the carrier or adjuvant is a solid, liquid or semi-solid.

More preferably, the dosage forms include tablets, capsules, pills, transdermal microneedle preparations and injections.

Has the advantages that:

the invention provides a composition of wogonin and irinotecan, wherein the two chemical components in the composition have obvious synergistic effect on the aspect of anti-colorectal cancer and produce unexpected technical effect of 1+1 & gt 2, so the composition of wogonin and irinotecan has the prospect of developing into anti-colorectal cancer medicaments.

Detailed Description

The following examples are given to illustrate the essence of the present invention, but not to limit the scope of the present invention.

First, experimental material

Reagent: DMSO (Sigma-Aldrich, USA); penicillin-streptomycin (Hyclone, usa); 0.5% pancreatin-EDTA solution (Boshide, Wuhan doctor de bioengineering, Inc.); PBS solution (Hyclone, usa); FBS (Gibco, usa); DMEM medium (Gibco, USA), MTT (Kyoki Biotechnology Co., Ltd. of Jiangsu Kayki).

Consumable material: 100mm cell culture dishes, cell cryopreservation tubes, 96-well plates (Corning, USA), 15ml/50ml centrifuge tubes (cantonhou jiert biofiltration gmbh).

Medicine preparation: wogonin (CAS: 632-85-9) was purchased from Chengdu-Ruifensi Biotech limited with a purity of 99%; irinotecan (CAS: 97682-44-5) was purchased from Shanghai Allantin Biotechnology Ltd, purity: 99 percent.

Cell: human colon cancer cell HCT116 (purchased from Nanjing Kebai Biotech Co., Ltd.).

Second, Experimental methods

1. HCT116 cell culture

HCT116 cells were cultured in high-glucose medium containing 10% FBS and 1% penicillin-streptomycin and 89% DMEM.

1.1 cell recovery: freezing HC stored in liquid nitrogenThe T116 cells are put into a constant-temperature water bath at 37 ℃, shaken to be rapidly heated and melted, and then rapidly transferred into a centrifuge tube containing 10mL of culture medium, and centrifuged at 1000rpm for 5 min. The supernatant was discarded, 2mL of medium was added to resuspend the cells, the cells were transferred to a 100mm petri dish containing 8mL of medium, the cells were shaken up "8", and the mixture was placed at 37 ℃ in 5% CO₂Culturing in a constant-temperature cell culture box.

1.2 cell passage: when the HCT116 cell healing rate was 80-90%, the medium was discarded, washed twice with PBS, and digested at 37 ℃ for 1min with the addition of 1ml of 0.25% trypsin solution (containing EDTA). When the cells are sandy and slippery, 2mL of complete culture medium is quickly added to stop digestion, the cells are gently blown, the cell suspension is collected into a 15mL centrifuge tube, and the centrifugation is carried out at 1000rpm for 5min, and then the operation is carried out in the same way as the 'cell recovery'.

1.3 cell cryopreservation: after the cells were digested and centrifuged, the supernatant was discarded, and the cells were resuspended in 1mL of cell cryopreservation solution (100. mu.L DMSO + 900. mu. LFBS), blown and mixed well, and transferred to a cryopreservation tube. Storing at 4 deg.C for 0.5 hr, storing at-20 deg.C for 2 hr, storing at-80 deg.C overnight, and transferring to liquid nitrogen for long-term storage.

2. MTT cell proliferation assay

Collecting HCT116 cells in logarithmic growth phase, digesting with pancreatin, collecting cells, and processing at 3 × 10³cells/well were seeded in 96-well plates at different cell densities, with different concentrations of wogonin given to the wogonin group alone, and different concentrations of irinotecan given to the irinotecan group alone, with the drug concentrations in the combination group set to intersect each other; the solvent is a basal culture medium. Cells were seeded in 96-well plates followed by 5% CO₂After the cells are cultured at 37 ℃ overnight for adherence, the blank group and the control group are replaced by fresh culture media, the administration group is replaced by fresh drug-containing culture media, and the cells are continuously placed in 5% CO₂Incubate at 37 ℃ for 48 hours. Thereafter, 20. mu.L of 5mg/mL MTT solution was added to each well and the mixture was placed on 5% CO₂And then incubated in an incubator at 37 ℃ for 4 hours, and then the solution in the wells was aspirated, redissolved with 150ml of DMSO, and placed on a microplate shaker at 300rpm for 10 min. The absorbance of each well was measured at a wavelength of 570 nm. Cell growth inhibition (%) (1- (OD administration group-OD blank)/(OD control-OD blank)]×100％。

3. Evaluation index of drug synergy

The evaluation method of the drug synergy adopts a Jinzhengyun Q value method (Jinzhengyun, addition in combined medication [ J ] recognized in the field]The chinese pharmacology bulletin, 1980). I.e. Q ═ M_AB/(M_A+M_B-M_A*M_B). The numerator in the formula represents the 'measured merging effect' and the denominator is the 'expected merging effect', wherein M_A、M_BAnd M_ABRespectively shows the inhibition rate of the drug A, the inhibition rate of the drug B and the inhibition rate of the combination of the two drugs under the current dose. Drug synergy index Q is defined as follows: when the Q value is less than 0.85, the two drugs are considered to have antagonistic effect; when the Q value is between 0.85 and 1.15, the two medicines are considered to be independent of each other and have additive effect; when the Q value is greater than 1.15, a synergistic effect is believed to exist between the two drugs.

Third, experimental results

The results of the inhibition rate and Q value calculation of different monomers or compositions thereof on the HCT116 cells are shown in the table. The results show that wogonin and irinotecan have obvious synergistic effect on the inhibitory activity of HCT116 cells.

In conclusion, the composition of wogonin and irinotecan has the prospect of being developed into the anti-colorectal cancer medicament.

The above-described embodiments are intended to be illustrative of the nature of the invention, but those skilled in the art will recognize that the scope of the invention is not limited to the specific embodiments.

Claims (6)

1. A composition characterized by: consists of wogonin and irinotecan.

2. The composition of claim 1, wherein: the ratio of the amounts of wogonin to irinotecan is 0.125:1 to 1.5: 1.

3. Use of the composition of claim 1 or 2 for the preparation of a medicament against colorectal cancer.

4. Use according to claim 3, characterized in that: the medicine takes the composition as an active ingredient for resisting colorectal cancer, also contains pharmaceutically acceptable carriers or auxiliary materials, and is prepared into pharmaceutically acceptable dosage forms.

5. Use according to claim 4, characterized in that: the carrier or adjuvant is solid, liquid or semisolid.

6. Use according to claim 4, characterized in that: the dosage forms include tablets, capsules, pills, transdermal microneedle preparations and injections.