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

Abstract

The invention discloses a composition of picrasone base and irinotecan and application of the composition in preparation of a medicine for resisting colorectal cancer. The invention finds that the picrasone base has an obvious anti-colorectal cancer effect and has a prospect of being developed into the medicine for treating the colorectal cancer. The invention also finds that the picrasone base can enhance the anti-colorectal cancer effect of irinotecan; on the basis, a composition of picrasone base and irinotecan can be developed and is used for preparing a medicine for treating colorectal cancer. Moreover, the invention also finds that the picrasone base and irinotecan have a synergistic effect in a certain proportion range, and can play a role in resisting colorectal cancer when 1 + 1 is greater than 2.

Description

Technical Field

[0001] The invention belongs to the field of medicine, relates to a composition and its application in preventing colorectal cancer, and specifically relates to a composition of quassinone and irinotecan and its application in preparing an anti-colorectal cancer drug. Background Art

[0002] Colorectal cancer (CRC) is a common malignant tumor. In 2022, the global incidence of colorectal cancer was 1.9261 million, ranking third among the world's major malignant tumors. In 2022, the global death from colorectal cancer was 903,900, ranking second among the world's major malignant tumors. The development of anti-colorectal drugs is of great value in the treatment of colorectal cancer.

[0003] Irinotecan (CPT-11) is a semi-synthetic water-soluble camptothecin derivative, mainly used in the first-line or second-line treatment of metastatic colorectal cancer. Its mechanism of action is to block the repair of damaged chains in DNA replication and transcription, leading to DNA breaks and inhibiting cancer cell proliferation. It is the core drug of the standard chemotherapy regimen for colorectal cancer.

[0004] Quassin is a natural alkaloid compound isolated from plants of the Quassinaceae family. There are currently no reports on its use in the treatment of colorectal cancer, let alone its use in combination with irinotecan in the treatment of colorectal cancer. Summary of the invention

[0005] The first purpose of the present invention is to provide the use of quasirone alkaloids for preparing drugs for treating colorectal cancer, the second purpose is to provide a composition of quasirone alkaloids and irinotecan and the use of the composition for preparing drugs for treating colorectal cancer, and the third purpose is to provide the use of quasirone alkaloids for preparing drugs for enhancing the anti-colorectal cancer effect of irinotecan.

[0006] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0007] The invention discloses a use of quasinone for preparing a medicine for treating colorectal cancer, wherein the active ingredient of the medicine comprises quasinone.

[0008] Furthermore, the active ingredient also includes irinotecan.

[0009] Furthermore, the molar ratio of quassinone base to irinotecan is 0.01:1 to 1:1, preferably 0.02:1 to 0.04:1 or 0.1:1 to 1:1.

[0010] The above-mentioned medicine also contains pharmaceutically acceptable excipients and is made into pharmaceutically acceptable dosage forms.

[0011] Furthermore, the auxiliary material is solid, liquid or semi-solid.

[0012] Furthermore, the dosage forms include tablets, capsules, pills, transdermal microneedle preparations and injections.

[0013] A pharmaceutical composition consists of quassinone base and irinotecan.

[0014] Furthermore, the molar ratio of quassinone base to irinotecan is 0.01:1 to 1:1, preferably 0.02:1 to 0.04:1 or 0.1:1 to 1:1.

[0015] The above-mentioned pharmaceutical composition is used for preparing a drug for treating colorectal cancer.

[0016] The invention discloses a use of quassinone for preparing a drug for enhancing the anti-colorectal cancer effect of irinotecan.

[0017] Beneficial effects:

[0018] The present invention finds that quasinone has a significant anti-colorectal cancer effect and has the prospect of being developed into a drug for treating colorectal cancer. The present invention also finds that quasinone can enhance the anti-colorectal cancer effect of irinotecan; based on this, a composition of quasinone and irinotecan can be developed for preparing a drug for treating colorectal cancer. In addition, the present invention also finds that quasinone and irinotecan have a synergistic effect within a certain ratio range and can exert an anti-colorectal cancer effect of 1+1>2. DETAILED DESCRIPTION

[0019] The essential contents of the present invention are described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0020] 1. Experimental Materials

[0021] Reagents: DMSO (Sigma-Aldrich, USA), penicillin-streptomycin (Hyclone, USA), 0.5% trypsin-EDTA solution (Boster, Wuhan Boster Biotechnology Co., Ltd.), PBS solution (Hyclone, USA), FBS (Gibco, USA), DMEM medium (Gibco, USA), MTT (Jiangsu KeyGen Biotechnology Co., Ltd.).

[0022] Consumables: 100 mm cell culture dishes, cell cryopreservation tubes, 96-well plates (Corning, USA), 15 mL / 50 mL centrifuge tubes (Guangzhou Jiete Biofiltration Co., Ltd.).

[0023] Drugs: Quassin (CAS: 18110-86-6) was purchased from Chengdu Efa Biotechnology Co., Ltd., purity: 98%; Irinotecan (CAS: 97682-44-5) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., purity: 99%.

[0024] Cells: Human colon cancer cell HCT116 (purchased from Nanjing Kebai Biotechnology Co., Ltd.).

[0025] 2. Experimental Methods

[0026] 1. HCT116 cell culture

[0027] HCT116 cells were cultured in DMEM high glucose medium containing 10% FBS, 1% penicillin-streptomycin and 89% glucose.

[0028] (1) Cell recovery: Take HCT116 cells frozen in liquid nitrogen to a 37°C constant temperature water bath, shake to quickly warm and thaw, quickly transfer to a centrifuge tube containing 10 mL of culture medium, centrifuge at 1000 rpm for 5 min. Discard the supernatant, add 2 mL of culture medium to resuspend the cells, transfer to a 100 mm culture dish containing 8 mL of culture medium, shake in an "8" shape, and place at 37°C, 5% CO 2 Culture in a constant temperature cell culture incubator.

[0029] (2) Cell passaging: When the healing rate of HCT116 cells reached 80-90%, discard the culture medium, wash twice with PBS, add 1 mL of 0.25% trypsin solution (containing EDTA), and digest for 1 min at 37°C. When the cells began to slide down like sand, quickly add 2 mL of complete culture medium to terminate the digestion, and gently pipette to collect the cell suspension into a 15 mL centrifuge tube. Centrifuge at 1000 rpm for 5 min and perform the same operation as the above "cell recovery".

[0030] (3) Cell freezing: After digestion and centrifugation, discard the supernatant and resuspend the cells in 1 mL of cell freezing solution (100 μL DMSO + 900 μL FBS). Mix by pipetting and transfer to a cryotube. Store at 4°C for 0.5 h, -20°C for 2 h, -80°C overnight, and then transfer to liquid nitrogen for long-term storage.

[0031] 2. MTT cell proliferation assay

[0032] HCT116 cells in the logarithmic growth phase were collected after trypsin digestion and the cells were collected at 3×10 3Cells were inoculated in 96-well plates at a density of 10 cells / well. The quassinone group was given different concentrations of quassinone, and the irinotecan group was given different concentrations of irinotecan. The drug concentrations of the combination group were set as the above concentrations in pairs; the solvent was the basal medium. After the cells were inoculated in 96-well plates, 5% CO 2 After overnight culture at 37°C, the blank group and the control group were replaced with fresh culture medium, and the drug-treated group was replaced with fresh drug-containing culture medium. 2 , incubated in an incubator at 37°C for 48 h. Then, 20 μL of 5 mg / mL MTT solution was added to each well and the well was kept in a 5% CO 2 , incubate in an incubator at 37°C for 4 hours, then aspirate the solution in the wells, re-dissolve with 150 mL DMSO, and place on a microplate oscillator at 300 rpm for 10 minutes. Measure the absorbance value of each well at a wavelength of 570 nm and calculate the cell growth inhibition rate. The formula for cell growth inhibition rate is as follows:

[0033] Cell growth inhibition rate (%) = [1-(OD drug group-OD blank group) / (OD control group-OD blank group)] × 100%.

[0034] 3. Evaluation indicators of drug synergy

[0035] The drug synergistic effect evaluation method adopts the Jin Zhengjun Q value method (Jin Zhengjun. Additive in combined medication [J]. Chinese Pharmacological Acta, 1980) which is recognized in the art, that is, Q = M AB / (M A +M B -M A *M B ). The numerator in the formula represents the “measured combined effect” and the denominator represents the “expected combined effect”, where M A 、M B With M AB They represent the inhibition rate of drug A, the inhibition rate of drug B, and the inhibition rate of the combination of the two drugs at the current dose. The drug synergy index Q is defined as follows: when the Q value is less than 0.85, the two drugs are considered to have an antagonistic effect; when the Q value is between 0.85 and 1.15, the two drugs are considered to be independent of each other and have an additive effect; when the Q value is greater than 1.15, the two drugs are considered to have a synergistic effect.

[0036] 3. Experimental Results

[0037] The results of the inhibition rate and Q value calculation of quassinone and irinotecan alone or in combination with HCT116 cells are shown in Table 1. The results show that quassinone and irinotecan alone inhibit the proliferation of HCT116 cells in a dose-dependent manner; the combination of the two also inhibits the proliferation of HCT116 cells in a dose-dependent manner, and when the ratio of the amount of the two substances is within a certain range, this inhibition shows a synergistic effect.

[0038] Table 1 Inhibitory rate and Q value of quassinone, irinotecan alone or in combination with HCT116 cells

[0039]

[0040]

[0041] In summary:

[0042] The present invention finds that quasinone has a significant anti-colorectal cancer effect and has the prospect of being developed into a drug for treating colorectal cancer. The present invention also finds that quasinone can enhance the anti-colorectal cancer effect of irinotecan; based on this, a composition of quasinone and irinotecan can be developed for preparing a drug for treating colorectal cancer. In addition, the present invention also finds that quasinone and irinotecan have a synergistic effect within a certain ratio range and can exert an anti-colorectal cancer effect of 1+1>2.

[0043] The purpose of the above-mentioned embodiments is to specifically introduce the essential content of the present invention, but those skilled in the art should know that the protection scope of the present invention should not be limited to the specific embodiments.

Claims

1. The use of quasirone alkaloids for preparing a drug for treating colorectal cancer, wherein the active ingredient of the drug comprises quasirone alkaloids.

2. The use according to claim 1, characterized in that: The active ingredient also includes irinotecan.

3. The use according to claim 2, characterized in that: The molar ratio of quassinone base to irinotecan is 0.01:1 to 1:1, preferably 0.02:1 to 0.04:1 or 0.1:1 to 1:

1.

4. The use according to any one of claims 1 to 3, characterized in that: The medicine also contains pharmaceutically acceptable excipients and is prepared into pharmaceutically acceptable dosage forms.

5. The use according to claim 4, characterized in that: The auxiliary material is solid, liquid or semi-solid.

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

7. A pharmaceutical composition, characterized in that: It is composed of quassinone base and irinotecan.

8. The pharmaceutical composition according to claim 7, characterized in that: The molar ratio of quassinone base to irinotecan is 0.01:1 to 1:1, preferably 0.02:1 to 0.04:1 or 0.1:1 to 1:

1.

9. Use of the pharmaceutical composition according to claim 7 or 8 for preparing a drug for treating colorectal cancer.

10. Use of quassinone for preparing a drug for enhancing the anti-colorectal cancer effect of irinotecan.

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