Application of alpha-caryophyllene in preparation of medicine for treating colorectal cancer
By using a drug prepared with α-caryophyllene, the abundance of Bacteroides fragilis in the gut was regulated, and ETBF-mediated colorectal cancer cell activity and migration were inhibited. This addresses the shortcomings of existing ETBF in the treatment of colorectal cancer, provides a new treatment strategy, and has significant clinical application value.
Patent Information
- Application Number
- CN202510798818.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-16
- Publication Date
- 2025-11-21
AI Technical Summary
There is a lack of drugs in the current technology that can effectively regulate the abundance of enterotoxigenic Bacteroides fragilis (ETBF), which plays a key role in the proliferation, migration and growth of colorectal cancer tumor cells, resulting in a lack of effective treatment options.
Using α-caryophyllene as the active ingredient, various pharmaceutically acceptable dosage forms were prepared to regulate the abundance of Bacteroides fragilis in the gut and inhibit ETBF-mediated colorectal cancer cell activity, migration, and growth.
α-Caryophyllene significantly inhibits ETBF-mediated proliferation and migration of colorectal tumor cells, slows the progression of colorectal cancer, and provides a new direction for the treatment of colorectal cancer, with low side effects and clinical application value.
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Figure CN120983405A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biological medicine, and particularly relates to an application of alpha-caryophyllene in preparation of a drug for treating colorectal cancer. BACKGROUND
[0002] Colorectal cancer (CRC) is the second leading cause of cancer death worldwide, and its occurrence and development are closely related to intestinal microbiota. In recent years, it has been found that enterotoxigenic Bacteroides fragilis (ETBF) plays an important role in the proliferation of colorectal cancer tumor cells.
[0003] Bacteroides fragilis is a commensal member of human microbiota, accounting for 0.1% to 0.5% of the total number of intestinal bacteria. Most Bacteroides fragilis strains are benign, but enterotoxigenic Bacteroides fragilis (ETBF) can produce a virulence factor, BFT (Bacteroides fragilis toxin), which is a zinc-dependent metalloproteinase that can damage the intestinal mucosal barrier function and cause intestinal inflammation and infection.
[0004] ETBF binds to colon epithelial cells, activates Wnt and NF-κB signaling pathways, and leads to increased cell proliferation, release of inflammatory mediators, and DNA damage, thereby promoting the occurrence and development of tumors. In addition, ETBF may create favorable conditions for the occurrence of colorectal cancer through various mechanisms such as regulation of host gene expression, alteration of intestinal barrier function, and promotion of inflammatory response.
[0005] If a drug that effectively regulates the abundance of ETBF can be developed to inhibit ETBF-mediated colorectal tumor cell proliferation and migration, it is expected to provide new strategies and methods for the treatment of colorectal cancer.
[0006] Caryophyllene is a class of bicyclic sesquiterpenes with alpha, beta, and gamma isomers. Alpha-caryophyllene is a natural monocyclic terpene that mainly exists in the essential oils of plants such as hops and can be extracted from natural plants such as hops (Humulus lupulus) and Lindera strychnifolia. Existing research suggests that alpha-caryophyllene has anti-inflammatory effects and can effectively prevent viruses that cause respiratory diseases such as colds and pneumonia from invading the human body. Some research has also explored its new use in the preparation of skin itching-relieving external drugs.
[0007] Currently, there is no research report on the regulation of ETBF and the treatment of colorectal cancer by alpha-caryophyllene. SUMMARY
[0008] In view of the above status of the prior art, the application provides application of alpha-caryophyllene in preparation of a drug for treating colorectal cancer. Alpha-caryophyllene can regulate abundance of Bacteroides fragilis in the intestinal tract, inhibit colorectal cancer cell activity, and can be used for treating or relieving colorectal cancer progression.
[0009] The technical scheme adopted by the application is as follows:
[0010] The application of alpha-caryophyllene in preparation of a drug for treating colorectal cancer.
[0011] The molecular formula of alpha-caryophyllene is shown in the following formula A:
[0012]
[0013] The application also provides application of alpha-caryophyllene in preparation of a drug for inhibiting colorectal cancer cell activity, inhibiting tumor cell proliferation mediated by enterotoxigenic Bacteroides fragilis (ETBF), inhibiting increase in tumor cell migration rate mediated by enterotoxigenic Bacteroides fragilis, inhibiting colorectal cancer growth mediated by enterotoxigenic Bacteroides fragilis, and reducing abundance of enterotoxigenic Bacteroides fragilis (ETBF).
[0014] The application also provides a drug for treating colorectal cancer, which comprises alpha-caryophyllene.
[0015] Further, the drug for treating colorectal cancer refers to a drug comprising a pharmaceutically effective amount of alpha-caryophyllene and a pharmaceutically acceptable carrier.
[0016] The drug for treating colorectal cancer can be prepared into any one of pharmaceutically acceptable dosage forms. The any one of pharmaceutically acceptable dosage forms comprises a suspension, an emulsion, a tablet, a capsule, a granule, an oral liquid preparation, a spray, an injection, etc.
[0017] The application has the following beneficial effects:
[0018] The application first discovers that alpha-caryophyllene can regulate abundance of enterotoxigenic Bacteroides fragilis (ETBF) in the intestinal tract, inhibit tumor cell proliferation and migration ability mediated by ETBF, and first verifies that alpha-caryophyllene can slow down colorectal cancer progression by improving intestinal microbial abundance. The application discloses, from cell and animal experiments, a new use of alpha-caryophyllene in preparation of a drug for treating colorectal cancer, and provides a new application direction. Alpha-caryophyllene is a natural active ingredient, has low side effects, and is easy to be clinically accepted. The application provides new scientific research evidence for developing a drug for treating colorectal cancer, and has important clinical application value and development value. BRIEF DESCRIPTION OF DRAWINGS
[0019] In order to make the technical solutions in the embodiments of the present application or the prior art clearer, the accompanying drawings needed in the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description only constitute some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained from these drawings without creative effort.
[0020] Figure 1 Cell viability graphs of HT-29 and HCT116 cells treated with different concentrations of α-caryophyllen, in which **P represents <0.01, and *** represents P<0.001.
[0021] Figure 2 Graphs of the effects of α-caryophyllen on the proliferation of HT-29 and HCT116 cells mediated by B. fragilis ETBF, in which ++ represents P<0.01 compared with the control group, and ## represents P<0.01 compared with the ETBF group.
[0022] Figure 3 Scratch photos and migration rate data graphs of HT-29 cells in different treatment groups, in which the left graph is the scratch photo, and the right graph is the migration rate; +++ represents P<0.001 compared with the control group, and ## represents P<0.01 compared with the ETBF group.
[0023] Figure 4 Scratch photos and migration rate data graphs of HCT116 cells in different treatment groups, in which the left graph is the scratch photo, and the right graph is the migration rate; +++ represents P<0.001 compared with the control group, and ## represents P<0.01 compared with the ETBF group.
[0024] Figure 5 Tumor photos and volume and weight comparison graphs of the control group, the ETBF group and the ETBF+α-caryophyllen group, in which A is the tumor photo, B is the tumor volume comparison bar graph, and C is the tumor weight comparison bar graph; +++ represents P<0.001 compared with the control group, and ### represents P<0.001 compared with the ETBF group.
[0025] Figure 6 Comparison graphs of the abundance (Bft expression level) of ETBF in different groups, in which +++ represents P<0.001 compared with the control group, and ### represents P<0.001 compared with the ETBF group. DETAILED DESCRIPTION
[0026] The technical solutions of the present application are further described below in combination with examples, and those skilled in the art can refer to the content herein to implement the process parameters. It is particularly pointed out that all similar substitutions and changes are obvious to those skilled in the art, and they are all regarded as included in the present application. The method of the present application has been described by way of preferred embodiments, and those skilled in the art can obviously make changes or appropriate changes and combinations to the products described herein without departing from the content, spirit and scope of the present application, to implement and apply the technical solutions of the present application. The protection scope of the present application is not limited thereto.
[0027] Example 1
[0028] Effect of α-caryophyllene on the viability of colon cancer tumor cells
[0029] The experimental method is as follows:
[0030] Human colon epithelial cell line (HT-29) and human colorectal cancer cell line (HCT116) were used, and RPMI-1640 medium containing 10% fetal bovine serum was used for culture in a CO2 incubator. Cells in the logarithmic growth phase were inoculated in a 96-well plate (5x10 3 The cells were divided into 5 groups, and different concentrations of α-caryophyllene (0, 5, 10, 15, 20 μg / mL) were added for 48 h. After 48 h of culture, 10 μL CCK-8 solution was incubated, and the culture was carried out in the dark for 4 h. The absorbance at 450 nm was detected by an enzyme-labeled instrument.
[0031] The cell viability graphs of HT-29 and HCT116 cells treated with different concentrations of α-caryophyllene are shown in Figure 1 The results show that with the increase of the concentration of α-caryophyllene, the cell viability of HT-29 and HCT116 cells is significantly inhibited.
[0032] Example 2
[0033] Inhibition of α-caryophyllene on B. fragilis-mediated colorectal cancer tumor cell proliferation
[0034] The experimental method is as follows:
[0035] Human colon epithelial cell line (HT-29) and human colorectal cancer cell line (HCT116) were used, and RPMI-1640 medium containing 10% fetal bovine serum was used for culture in a CO2 incubator. Cells in the logarithmic growth phase were inoculated in a 96-well plate (5x10 3 The cells were divided into 5 groups, and different concentrations of α-caryophyllene (0, 5, 10, 15, 20 μg / mL) were added for 48 h. After 48 h of culture, 10 μL CCK-8 solution was incubated, and the culture was carried out in the dark for 4 h. The absorbance at 450 nm was detected by an enzyme-labeled instrument.
[0031] The cell viability graphs of HT-29 and HCT116 cells treated with different concentrations of α-caryophyllene are shown in Figure 1 The results show that with the increase of the concentration of α-caryophyllene, the cell viability of HT-29 and HCT116 cells is significantly inhibited.
[0032] Example 2
[0033] Inhibition of α-caryophyllene on B. fragilis-mediated colorectal cancer tumor cell proliferation
[0034] The experimental method is as follows:
[0035] Human colon epithelial cell line (HT-29) and human colorectal cancer cell line (HCT116) were used, and RPMI-1640 medium containing 10% fetal bovine serum was used for culture in a CO2 incubator. Cells in the logarithmic growth phase were inoculated in a 96-well plate (5x10 3 The cells were divided into 5 groups, and different concentrations of α-caryophyllene (0, 5, 10, 15, 20 μg / mL) were added for 48 h. After 48 h of culture, 10 μL CCK-8 solution was incubated, and the culture was carried out in the dark for 4 h. The absorbance at 450 nm was detected by an enzyme-labeled instrument.
[0036] The results of the effect of a-caryophyllen on the proliferation of colorectal cancer tumor cells HT-29 and HCT116 cells mediated by B. fragilis ETBF are shown in the graph of Figure 2 As shown in the graph of
[0037] Example 3:
[0038] a-caryophyllen inhibits the increase of tumor cell migration rate mediated by B. fragilis (ETBF)
[0039] The experimental method is as follows:
[0040] The migration level of HT-29 cells and HCT116 cells was detected by scratch test. After 48h of anaerobic co-culture of HT-29 and HCT116 cells with ETBF at a multiplicity of infection (MOI) of 100, HT-29 cells and HCT116 cells (5x105cells) were planted in a six-well plate. After fusion of 80%, the cells were washed twice with DMEM medium and phosphate buffer PBS. The fused cell monolayer was cut into a linear wound with 1 mL of sterile microtubule needle tip, and the floating cells were removed for culture of the cells in serum-free medium. The scratch photos were taken after 0 and 48h of scratch operation using an optical microscope. 5
[0041] The HT-29 and HCT116 cells of the Control group were not anaerobically co-cultured with ETBF, and the rest of the operations were the same as above.
[0042] ETBF+a-caryophyllen group: when the density of HT-29 and HCT116 cells grew to 60%-70%, ETBF at a multiplicity of infection (MOI) of 100 and 15 μg / mL of a-caryophyllen were added, and the scratch operation was performed for co-culture for 48h, and the scratch photos after 0h and 48h were recorded, respectively.
[0043] The scratch photos and migration rate data of the HT-29 cell groups with different treatment modes are shown in the graph of Figure 3 , wherein the left graph is the scratch photo, and the right graph is the migration rate data.
[0044] The scratch photos and migration rate data of the HCT116 cell groups with different treatment modes are shown in the graph of Figure 4 , wherein the left graph is the scratch photo, and the right graph is the migration rate data.
[0045] Figure 3 、 Figure 4 The results showed that co-culture of Bacteroides fragilis (ETBF) promoted the migration of tumor cells (HT-29 cells and HCT116 cells), and α-caryophyllene treatment significantly inhibited the enhanced migration of tumor cells, that is, α-caryophyllene treatment can significantly inhibit ETBF-mediated tumor cell migration.
[0046] Example 4:
[0047] α-Caryophyllene inhibits ETBF-mediated colon cancer growth in vivo.
[0048] The experimental method is as follows:
[0049] In vivo tumor-bearing experiments were conducted using male BALB / c nude mice (5 weeks old, 17±2g) housed under specific pathogen-free (SPF) conditions. In xenotransplantation studies, the mice were sized at a ratio of 1×10⁻⁶. 6 HCT116 cells were subcutaneously injected into nude mice. Mice were randomly assigned to three groups after subcutaneous injection: control group, ETBF group, and ETBF+α-caryophyllene group. The ETBF group received multiple intratumoral injections of ETBF at an MOI of 500 (MOI = 500 means 500 CFU of ETBF per tumor cell, approximately 1 × 10⁶ cells per tumor cell). 6 Each cell requires an injection of 5 × 10⁸ cells. 8 CFU (eTBF / dose), twice a week for 3 consecutive weeks. The ETBF+α-caryophyllene group received 20 mg / kg of α-caryophyllene every 2 days via intraperitoneal injection in addition to the ETBF group. The control group received only DMSO.
[0050] After 3 weeks, mice were sacrificed, and tumors were collected, weighed, and preserved. Tumor volume was calculated using the following formula: (longest diameter) × (shortest diameter) × 0.5.
[0051] Tumor images and comparisons of volume and weight between the control group, ETBF group, and ETBF+α-caryophyllene group are shown below. Figure 5 As shown, Figure A is a photograph of the tumor, Figure B is a bar chart comparing tumor volume, and Figure C is a bar chart comparing tumor weight.
[0052] Compared with the control group, Bacteroides fragilis (ETBF) significantly promoted the in vivo growth of colon cancer cells; while ETBF promoted tumor growth, α-caryophyllene significantly inhibited ETBF-mediated tumor growth of colorectal cancer in vivo.
[0053] Example 5:
[0054] The effect of α-caryophyllene on reducing the abundance of Bacteroides fragilis (ETBF) in colon cancer.
[0055] The experimental method is as follows:
[0056] Total RNA was extracted from the collected tumor tissues using TRIzol reagent. The quality of the RNA was detected using a NanoDrop ND-2000 spectrophotometer, and its integrity was determined by standard denaturing agarose gel electrophoresis. The total RNA (2 μg) was then reverse transcribed using a PrimeScript RT reagent kit. qRT-PCR was performed using a GoTaq® Green qPCR SuperMix and the 2-ΔΔCT method was used to calculate the relative gene expression of RNA. The primer sequences used in the experiment are as follows:
[0057] btf Forward: 5'-AAAGTGGCTCGGAGAAGACG-3'
[0058] bft Reverse: 5'-GTCCGACATACACAGGGACG-3'
[0059] 16S Forward: 5'-GGTGAATACGTTCCCGG-3'
[0060] 16S Reverse: 5'-tacggctaccttgttacgtt-3'
[0061] A comparison chart of the abundance (Bft expression level) of B. fragilis (ETBF) in different groups is shown in FIG. 2, and the results show that a- caryophyllene can significantly inhibit the abundance of B. fragilis (ETBF) in a tumor. Figure 6
[0062] Although the present application has been disclosed in preferred embodiments as above, it is not intended to limit the present application, and any person skilled in the art can make various modifications and modifications without departing from the spirit and scope of the present application, and therefore the scope of protection of the present application should be defined by the claims.
Claims
1. Use of α-caryophyllene in the manufacture of a medicament for treating colorectal cancer.
2. Use according to claim 1, characterized in that:
2. Use of α-caryophyllene in the manufacture of a medicament for inhibiting viability of colorectal cancer cells.
3. Use according to claim 1, characterized in that:
3. Use of α-caryophyllene in the manufacture of a medicament for inhibiting Bacteroides fragilis enterotoxin-producing mediated proliferation of colorectal cancer tumor cells.
4. The use according to claim 1, characterized in that:
4. Use of α-caryophyllene in the manufacture of a medicament for inhibiting Bacteroides fragilis enterotoxin-producing mediated increase in tumor cell migration rate.
5. The use according to claim 1, characterized in that:
5. Use of α-caryophyllene in the manufacture of a medicament for inhibiting Bacteroides fragilis enterotoxin-producing mediated growth of colon cancer.
6. The use according to claim 1, characterized in that:
6. Use of α-caryophyllene in the manufacture of a medicament for reducing abundance of Bacteroides fragilis enterotoxin-producing.
7. A drug for treating colorectal cancer, characterized in that:
7. The medicament for treating colorectal cancer comprises α-caryophyllene.
8. The medicament according to claim 7, characterized in that:
8. The medicament for treating colorectal cancer comprises a pharmaceutically effective amount of α-caryophyllene and a pharmaceutically acceptable carrier.