Dipeptide capable of preventing and delaying colorectal cancer and application thereof
By using high levels of carnosine to inhibit colorectal cancer cells, a safe and effective method for preventing and delaying the progression of colorectal cancer is provided, solving the problems of high invasiveness and strong side effects of existing treatments, and achieving the effects of preventing and delaying the progression of colorectal cancer.
Patent Information
- Application Number
- CN202511102514.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-07
- Publication Date
- 2025-12-12
AI Technical Summary
Current treatments for colorectal cancer, such as surgery, radiotherapy, and chemotherapy, have problems such as being highly invasive, having strong side effects, and being difficult to detect early. There is a lack of effective methods to prevent and slow the progression of colorectal cancer.
Using high-carnosine as a dipeptide, this drug inhibits the proliferation and activity of colorectal cancer cells through different dosage forms and routes of administration. It is prepared in various forms such as suspensions and capsules, with a dosage concentration of 100~300mg/kg/day, for the prevention and delay of colorectal cancer development.
High levels of carnosine can significantly inhibit the activity and proliferation of colorectal cancer cells, reduce tumor volume, and decrease adverse reactions in patients, providing a safe and effective method for preventing and delaying the progression of colorectal cancer.
Smart Images
Figure CN121102207A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of pharmaceutical technology, specifically to a dipeptide that can prevent and delay colorectal cancer and its applications. Background Technology
[0002] Colorectal cancer is a collective term for colon cancer, rectal cancer, and anal cancer, and it is one of the most common malignant tumors. The occurrence of colorectal cancer is influenced by a variety of factors, including genetic factors, personal medical history, lifestyle, age, gender, and gut microbiota.
[0003] Currently, the main clinical treatment for colorectal cancer (CRC) is surgical resection combined with radiotherapy and chemotherapy. Surgical treatment is the core and preferred method for treating colorectal cancer. However, surgery is highly invasive, has a long recovery time, and requires a high level of surgical skill. Radiotherapy is a treatment method that uses high-energy rays to kill tumor cells. In the treatment of colorectal cancer, radiotherapy is mainly used as adjuvant therapy for mid-to-lower rectal cancer and as palliative treatment for advanced rectal cancer. However, the radiotherapy dosage is relatively difficult to control. Lower doses may not achieve the desired therapeutic effect, while higher doses may increase the risk of surgical complications such as radiation enteritis. Chemotherapy is a treatment method that uses chemical drugs to kill or inhibit the growth of tumor cells. In the treatment of colorectal cancer, chemotherapy is usually used as an adjunct to surgery to improve treatment efficacy and reduce recurrence rates. However, chemotherapy drugs have certain toxicity and side effects, which may lead to a decline in the patient's physical condition, causing adverse reactions such as nausea, vomiting, and hair loss, thereby affecting surgical tolerance. Furthermore, due to the insidious nature of colorectal cancer and the lack of obvious symptoms in its early stages, it is often mistaken for other digestive system diseases, such as indigestion, gastroenteritis, hemorrhoids, and anal fistulas. Therefore, exploring new methods for effective prevention and slowing the progression of colorectal cancer is particularly important.
[0004] Dipeptides are an important class of endogenous small-molecule metabolites in the body, widely distributed, diverse in type, and with varied functions. They play important nutritional and biological roles in the body, including anti-inflammatory, blood pressure-lowering, and blood lipid-lowering effects. Homocarnosine is a dipeptide composed of γ-aminobutyric acid (GABA) and L-histidine amino acids, possessing antioxidant, anti-inflammatory, DNA damage-preventing, and anti-advanced glycation end products (AGEs) effects. Currently, there are no reports on the use of homocarnosine for the prevention or delay of colorectal cancer. Summary of the Invention
[0005] The purpose of this invention is to provide a dipeptide that can prevent and delay colorectal cancer and its application, which offers promising prospects for the prevention of colorectal cancer.
[0006] To achieve the above objectives, the present invention provides a dipeptide for preventing and delaying the development of colorectal cancer, with the chemical formula C. 10 H 16N4O3, structural formula as follows: .
[0007] Specifically, the present invention provides the following technical solution: The first aspect of the present invention is to provide the use of a dipeptide in the preparation of a medicament for treating cancer, characterized in that the dipeptide is selected from one or more of carnosine, homocarnosine, or anserine.
[0008] Furthermore, the dipeptide is a high-carnosine peptide.
[0009] Furthermore, the effective concentration of the high carnosine is 100~300 mg / kg / day.
[0010] Furthermore, the cancer in question is colorectal cancer.
[0011] Furthermore, the dosage form of the drug includes at least one of suspension, granules, capsules, powders, tablets, pills, injections, suppositories, and drops.
[0012] Furthermore, the route of administration of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, and perfusion administration.
[0013] Furthermore, the drug is characterized in that it further includes pharmaceutically acceptable excipients, including at least one of the following: diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, or buffers.
[0014] A second aspect of the present invention is to provide a pharmaceutical preparation for treating colorectal cancer, characterized in that the pharmaceutical preparation comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is a dipeptide selected from one or more of carnosine, homocarnosine, or anserine.
[0015] Furthermore, the dosage of the active ingredient is 100~300 mg / kg / day.
[0016] Furthermore, the dipeptide is a high-carnosine peptide.
[0017] Furthermore, the drug is characterized in that the dosage form includes at least one of suspension, granules, capsules, powders, tablets, pills, injections, suppositories, and drops.
[0018] Further, the pharmaceutically acceptable excipients include at least one of the following: diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, or buffers.
[0019] A third aspect of the present invention is to provide the use of a dipeptide in the preparation of an agent that inhibits the proliferation of cancer cells, characterized in that the inhibition is in vitro inhibition, and the dipeptide is selected from one or more of carnosine, homocarnosine, or anserine.
[0020] Furthermore, the cancer cells are colorectal cancer cells, preferably HT-29 cells.
[0021] Furthermore, the concentration of the dipeptide used is 4000 μM to 8000 μM.
[0022] Furthermore, the dipeptide is a high-carnosine peptide.
[0023] The beneficial effects of this invention: This invention proposes a dipeptide that can prevent and delay colorectal cancer. It inhibits the activity and proliferation of colorectal cancer cells, thus slowing the progression of colorectal cancer. It is easily absorbed by the human body and has the ability to supplement essential amino acids and provide nutritional benefits. Therefore, it shows promising application prospects as a drug for preventing and delaying the development of colorectal cancer. Attached Figure Description
[0024] Figure 1 This figure shows the results of different carnosine effects on the formation of colonies in colorectal cancer HT-29 cells.
[0025] Figure 2 This figure shows the results of the activity of high-carnosine on colorectal cancer HT-29 cells.
[0026] Figure 3 This is a graph showing the effect of high carnosine on the proliferation of colorectal cancer HT-29 cells.
[0027] Figure 4 This is a schematic diagram of an experiment involving HT-29 tumor-bearing mice.
[0028] Figure 5 Image showing the body weight of HT-29 tumor-bearing mice.
[0029] Figure 6 This is a statistical chart showing the tumor status and volume of HT-29 tumor-bearing mice.
[0030] Figure 7 This is a pathological staining image of a tumor in a HT-29 tumor-bearing mouse. Detailed Implementation
[0031] The following detailed embodiments further illustrate the concept and technical effects of the present invention to fully understand its purpose, features, and effects. Unless otherwise specified, all methods described are conventional methods. Unless otherwise specified, all materials are available from publicly available commercial sources. The illustrative embodiments and descriptions of the present invention are used to explain the invention and do not constitute an undue limitation thereof. It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.
[0032] Example 1: Effects of high carnosine on colorectal cancer HT-29 cells Human colorectal cancer cell line HT-29 was cultured in DMEM complete medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and cultured at 37 °C in a 5% CO2 incubator until the cells adhered and grew to 70%–80% of the bottom of the dish. It was suitable for trypsin digestion and passage.
[0033] Test Example 1: The effect of different carnosines on the colony formation of colorectal cancer HT-29 cells.
[0034] Cell colony formation was detected using Giemsa staining. HT-29 cells (1×10⁻⁶) were used. 3 (Cells were seeded in 6-well plates and cultured overnight. After cell attachment, the culture medium was discarded, and the medium was replaced with complete culture medium containing different carnosine concentrations of 0 μM, 2000 μM, or sulindac concentration of 2000 μM, and cultured for 48 h. After fixation with 70% ethanol at room temperature for 10 min, Giemsa staining solution was added and incubated at room temperature for 20 min. The cells were then rinsed with distilled water to remove any residual staining solution and air-dried.
[0035] like Figure 1 As shown, compared with the control group, the size of cell clones decreased after treatment with sulindac, indicating that sulindac can inhibit the formation of colorectal cancer cell clones. Carnosine, high carnosine, and anserine can all inhibit the size of HT-29 cell clones, among which high carnosine has a better effect than carnosine and anserine.
[0036] Test Example 2: Effect of high carnosine on the activity of colorectal cancer HT-29 cells.
[0037] Cell viability was determined using the CCK-8 assay. HT-29 cells (1×10⁻⁶) were used. 3 (Each well) was seeded in a 96-well plate and cultured overnight. After cell attachment, the culture medium was discarded, and the medium was replaced with complete culture medium containing high concentrations of carnosine (0 μM, 100 μM, 500 μM, 1000 μM, 2000 μM, 4000 μM, 8000 μM) and sulindac (2000 μM), and cultured for 48 h. 20 μL of LCK-8 was added to each well and incubated for 1 h. The absorbance at 450 nm was measured using a microplate reader.
[0038] like Figure 2 As shown, the activity of HT-29 cells gradually decreased with increasing high carnosine concentration. The cell activity at high carnosine concentrations of 4000 μM and 8000 μM was similar to that of the positive control drug sulindac, indicating that high carnosine at 4000 μM and above can significantly inhibit the activity of HT-29 cells.
[0039] Test Example 3: Effect of high carnosine on the proliferation of colorectal cancer HT-29 cells.
[0040] Cell proliferation was detected using EdU staining. HT-29 cells (1×10⁻⁶) were used for cell proliferation analysis. 4 Cells were seeded in 12-well plates with built-in cell spreaders and cultured overnight. After cell attachment, the culture medium was discarded, and the medium was replaced with complete medium containing high concentrations of carnosine (0 μM, 4000 μM, and sulindac, respectively) for 48 h. Then, an equal volume of 20 μM EdU complete medium was added and incubated at 37 °C for 2 h. After labeling, the culture medium was removed, and 1 mL of 4% paraformaldehyde fixative was added for fixation at room temperature for 15 min. The fixative was removed, and the cells were washed three times with PBS for 3 min each. 1 mL of PBS containing 0.3% Triton X-100 was added to each well and incubated at room temperature for 10–15 min. The permeabilization buffer was removed, and the cells were washed once with PBS for 3 min. 0.5 mL of Click reaction solution was added to each well and incubated at room temperature in the dark for 30 min. The permeabilization buffer was removed, and the cells were washed three times with PBS for 3 min each. Cell nuclei were stained using Hoechst 33342 and incubated at room temperature in the dark for 10 min. The staining solution was then removed, and the cells were washed three times with PBS solution for 3 min each time. The cells were then observed under a fluorescence microscope.
[0041] like Figure 3 As shown, Azide 488 exhibits green fluorescence with a maximum excitation wavelength of 495 nm; Hoechst 33342 exhibits blue fluorescence with a maximum excitation wavelength of 346 nm. Compared to the control group, cell proliferation decreased after high carnosine treatment. The results indicate that the green fluorescent spot representing cell proliferation tends to decrease after high carnosine treatment, suggesting that high carnosine can inhibit HT-29 cell proliferation.
[0042] Example 2: Effects of carnosine on HT-29 tumor-bearing mice and tumors Thirty SPF-grade male BALB / c mice were housed in an SPF-grade animal laboratory at a temperature of (21±4)℃ and humidity of (50±10)%, with a natural diurnal rhythm and free access to water and food under light conditions. Experiments began after one week of acclimatization. Figure 4As shown, normally cultured HT-29 cells were subcutaneously inoculated into the right groin area of mice. One week later, small nodules were palpable, indicating successful model establishment. The mice were then randomly divided into a model group, a sulindac group, a carnosine group, a high-carnosine group, and a anserine group. The sulindac group was administered 50 mg / kg sulindac solution daily by gavage, each carnosine group was administered 200 mg / kg of a different carnosine solution daily by gavage, and the model group was administered an equal volume of physiological saline by gavage.
[0043] Test Example 4: Changes in body weight of HT-29 tumor-bearing mice.
[0044] During the experiment, the weight of the mice was monitored every 3 days.
[0045] like Figure 5 As shown, compared with the model group, mice treated with high carnosine showed a significant increase in body weight, indicating that high carnosine can reduce the adverse effect of weight loss during tumor development.
[0046] Test Example 5: Tumor status and volume in HT-29 tumor-bearing mice.
[0047] After the experiment, the mice were euthanized, the intact tumor tissue was dissected, and the longest diameter (a) and shortest diameter (b) of the tumor were measured. The tumor volume V was calculated as V = 0.5 × ab. 2 .
[0048] like Figure 6 As shown, compared with the model group, the tumor volume was significantly reduced after high carnosine treatment, indicating that high carnosine can inhibit the development of colorectal cancer.
[0049] Test Case 6: Tumor pathology in HT-29 tumor-bearing mice.
[0050] Tumor tissue was fixed in 4% paraformaldehyde solution, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE) staining kit, and examined for pathological conditions of the tumor tissue using an optical microscope.
[0051] like Figure 7 As shown, in the control group, tumor cells exhibited diffuse growth, with tight intercellular arrangement, relatively intact morphology, and large, deeply stained nuclei. Compared to the control group, the tumor cells in the sulindac group were more sparsely arranged, with obvious necrotic areas, manifested as unstructured sheet-like regions and nuclear pyknosis. Compared to carnosine and anserine, high-carnosine treatment resulted in the most significant pathological changes in tumor tissue, indicating that high-carnosine treatment has a stronger inhibitory effect on colorectal cancer than carnosine and anserine.
[0052] The embodiments described above are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.
Claims
1. The application of a dipeptide in the preparation of a drug for treating cancer, characterized in that, The dipeptide is selected from one or more of high carnosine or anserine.
2. The application according to claim 1, characterized in that, The dipeptide mentioned is high-carnosine.
3. The application according to claim 2, characterized in that, The effective concentration of the high carnosine is 100~300 mg / kg / day.
4. The application according to claim 1, characterized in that, The cancer in question is colorectal cancer.
5. The application according to claim 2, characterized in that, The dosage form of the drug includes at least one of the following: suspension, granules, capsules, powders, tablets, pills, injections, suppositories, and drops.
6. The application according to claim 1, characterized in that, The route of administration of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, and perfusion administration.
7. The application according to any one of claims 1-6, characterized in that, The drug also includes pharmaceutically acceptable excipients, including at least one of the following: diluents, binders, wetting agents, lubricants, disintegrants, solvents, emulsifiers, cosolvents, preservatives, pH adjusters, osmotic pressure adjusters, surfactants, coating materials, antioxidants, or buffers.
8. A pharmaceutical preparation for treating colorectal cancer, characterized in that, The pharmaceutical preparation comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is a dipeptide selected from one or more of carnosine or anserine, and the dosage of the active ingredient is 100-300 mg / kg / day.
9. The pharmaceutical preparation according to claim 8, characterized in that, The dipeptide mentioned is high-carnosine.
10. The pharmaceutical preparation according to claim 9, characterized in that, The dosage form of the drug includes at least one of the following: suspension, granules, capsules, powders, tablets, pills, injections, suppositories, and drops.