Compositions containing azvudine and their application in the prevention and treatment of colorectal cancer

Abstract

This invention relates to the field of biomedical technology, and in particular to compositions containing azvudine and their application in the prevention and treatment of colorectal cancer. Studies have shown that the combined use of azvudine and berberine (or its derivatives) has a significant preventive and therapeutic effect on colorectal cancer, which is significantly better than the use of azvudine and berberine alone, and has good application prospects in the clinical prevention and treatment of colorectal cancer.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to compositions containing azvudine and their application in the prevention and treatment of colorectal cancer. Background Technology

[0002] Colorectal cancer is the third most common and second leading cause of cancer death worldwide. Currently, treatment for colorectal cancer primarily involves surgical resection, supplemented by adjuvant chemotherapy (oxaliplatin, irinotecan, capecitabine, etc.), targeted therapy (regorafenib, bevacizumab, cetuximab, fruquintinib), and immunotherapy (ipilimumab, nivolumab, and pembrolizumab). The main challenge in treatment is postoperative recurrence, specifically due to: 1. the relatively low efficacy of existing drugs against colorectal cancer; 2. the presence of certain toxic side effects making long-term use unsuitable; and 3. the high incidence of drug resistance in chemotherapy patients. Therefore, developing a novel drug with minimal side effects for the prevention and treatment of colorectal cancer has significant clinical value and importance. Drug combination therapy, which combines drugs with different mechanisms of action to reduce the dosage of single-agent drugs, is an important avenue for new drug discovery.

[0003] Azvudine (2'-deoxy-2'-fluoro-4'-azidocytidine, FNC) is a nucleoside reverse transcriptase inhibitor with independent intellectual property rights developed by Henan Zhenshi Biotechnology Co., Ltd. It was approved in 2021 for the treatment of HIV infection, with a dosage of 3 mg / day. At this dosage, FNC has few side effects and is suitable for long-term use. Currently, there are no reports of azvudine being used for the prevention and treatment of colorectal cancer.

[0004] Berberine (BBR) is a monomeric compound extracted from the traditional Chinese medicines Coptis chinensis and Phellodendron chinense. It is a non-prescription drug used clinically for the long-term treatment of bacterial diarrhea. Our previous research found that BBR has a therapeutic effect on hyperlipidemia, and the relevant findings were published in Nature Medicine (PMID: 15531889). This research won the second prize of the National Natural Science Award in 2012 (titled "Molecular Mechanism, Chemical Basis, and Clinical Characteristics of Berberine in Correcting Hyperlipidemia"), initiating research into the repurposing of BBR. In recent years, various pharmacological effects of BBR have been discovered, including hypoglycemic, lipid-lowering, anti-inflammatory, and anti-tumor effects, with a wide range of mechanisms of action (Berberine, Science Press, 2021). A 2020 article published in a Lancet sub-journal showed that BBR significantly reduced the recurrence of colorectal adenomas (precancerous lesions of colorectal cancer) (PMID: 31926918), but the mechanism remains unclear. Our previous studies have shown that the anticancer effect of BBR is related to its antagonism of lysophospholipid-related signaling pathways (PMID: 36090480). Studies by other research groups have confirmed that BBR also reduces lysophospholipid levels (PMID: 37151876). Currently, there are no reports of combining FNC and BBR, two drugs with drastically different structures and mechanisms of action, for the prevention and treatment of colorectal cancer. Summary of the Invention

[0005] In view of this, the present invention provides compositions containing azvudine and their use in the preparation of medicaments for the prevention and treatment of colorectal cancer.

[0006] This invention provides the following solution:

[0007] In a first aspect, the present invention provides a composition for preventing and treating colorectal cancer, comprising azvudine and alkaloids; said alkaloids include berberine and / or berberine derivatives.

[0008] In the composition of the present invention, the berberine derivative includes at least one of IMB-Y53, berberine, fenthrodine, demethylene berberine, and berberine.

[0009] Azvudine, abbreviated as FNC, has the structural formula shown in Formula I:

[0010]

[0011] Berberine, abbreviated as BBR, has the structure shown in Formula II:

[0012]

[0013] In this invention, the berberine derivatives include at least one of IMB-Y53, berberine, fenthrine, demethylene berberine, and berberine.

[0014] Specifically, IMB-Y53, berberine, fendrine, demethylene berberine, and berberine successively possess the structures shown in Formulas III to VII:

[0015]

[0016] Studies have shown that, compared to using FNC and berberine alone, the combination of the two drugs can more significantly reduce tumor burden. Figure 1 D. Compared with the 0.46 mg / kg group (P<0.05), the combined treatment group showed a significant improvement in the prevention and treatment of colorectal cancer. Simultaneously, the dose of FNC was reduced to 0.46 mg / kg (the human equivalent dose is 3 mg / day). The safety of this dose has been approved by the National Medical Products Administration and tested in long-term use by HIV patients, making it more suitable for the long-term prevention and treatment of colorectal cancer.

[0017] In the composition of the present invention, the mass ratio of azvudine to alkaloids is (1-10) to (100-1500), preferably (1-7):(330-660), and more preferably (3-7):(330-660). In some specific embodiments, the mass ratio is specifically 3:330, 3:660, 5:330, 5:660, 7:330, or 7:660, or any other value within the above range.

[0018] In some specific embodiments, the composition consists of azvudine and berberine in a mass ratio of 3:330, 3:660, 5:330, 5:660, 7:330, or 7:660.

[0019] In this invention, "composition" is not limited to the physical mixing of active ingredients in the composition, but also includes the combined use of active ingredients at specific time intervals in any order. Specifically, in this invention, composition refers to the combined use of azvudine and berberine (or berberine derivatives) in the treatment of diseases; the two can exist in a mixed form or independently.

[0020] Secondly, the present invention also provides the use of any of the following in the preparation of a medicament for the prevention and treatment of colorectal cancer:

[0021] (1) The composition of the present invention;

[0022] (2) Berberine or derivatives of formulas III to VII, used in combination with azvudine.

[0023] In this invention, the intestinal cancer includes colorectal cancer and / or small bowel cancer.

[0024] This invention is the first to combine BBR, a clinical treatment for enteritis, with FNC, a nucleoside reverse transcriptase inhibitor—two drugs with drastically different structures and pharmacological activities—for the prevention and treatment of colorectal cancer. The efficacy of this combined use of azvudine (3 mg / day) and berberine (330 mg / day) in preventing and treating colorectal cancer was verified using this combination as an example. The results showed that compared to azvudine or berberine alone, the combination group with half the dose of azvudine and berberine showed the most significant reduction in tumor burden. Simultaneously, the FNC dose was reduced to 3 mg / day, an approved dose for treating HIV infection, demonstrating long-term clinical use and making it more suitable for the long-term prevention and treatment of colorectal cancer. Therefore, compared to monotherapy, the combination of azvudine and berberine has significant advantages in preventing and treating colorectal cancer.

[0025] In this invention, the prevention and treatment include both prevention and treatment. Specifically, the prevention and treatment include inhibiting tumor immune escape, reducing tumor number, reducing tumor burden, and / or antagonizing lysophospholipid-related signaling pathways.

[0026] Furthermore, the inhibition of tumor immune escape includes any one of the following:

[0027] (a) Increase the proportion of CD4+ T cells in the blood;

[0028] (b) Reduce the proportion of Treg cells in tumor tissue;

[0029] In this invention, the combined use of azvudine and the alkaloids includes administering the two drugs simultaneously or in any order.

[0030] In this invention, when berberine and azvudine are used in combination, the dosage of azvudine in a 60kg human body is 1–10 mg / day, specifically 1 mg / day, 2 mg / day, 3 mg / day, 4 mg / day, 5 mg / day, 6 mg / day, 7 mg / day, 8 mg / day, 9 mg / day, 10 mg / day, or any specific value between any two of the above values. The clinically equivalent dose of berberine in a 60kg human body is 100–1500 mg / day, specifically 300 mg / day, 330 mg / day, 500 mg / day, 660 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, or 1500 mg / day, or any specific value within the above range.

[0031] The above dosage is not the only dosage for azvudine and berberine. Clinicians can make reasonable adjustments based on the patient's physical condition and illness.

[0032] Thirdly, the present invention also provides a medicament for the prevention and treatment of colorectal cancer, comprising the composition as described above and pharmaceutically acceptable excipients.

[0033] In this invention, the dosage forms of the drug include injections and oral preparations; the injections include, but are not limited to, liquid injections, powder injections, or emulsion injections; the oral preparations include, but are not limited to, tablets, granules, pills, capsules, and oral liquid preparations.

[0034] In the drug for preventing and treating colorectal cancer described in this invention, the two active ingredients, FNC and BBR, can exist in a mixed form, that is, the two drugs are formulated into the same dosage form. Alternatively, the two drugs can exist independently, that is, they are formulated into separate preparations. When formulated into separate dosage forms, the prevention and treatment of colorectal cancer includes both simultaneous and separate administration. When administered separately, it also includes any sequential use.

[0035] Fourthly, the present invention also provides a method for preventing and treating colorectal cancer, comprising: administering the composition described in the present invention or the drug for preventing and treating colorectal cancer described in the present invention. FNC and BBR can be administered simultaneously or sequentially in any order. The specific dosage and administration order can be determined based on the patient's specific condition and the physician's clinical experience.

[0036] This invention provides compositions containing azvudine and their application in the prevention and treatment of colorectal cancer. Studies have shown that azvudine can significantly increase the proportion of CD4+ T cells in the blood, help reduce Treg cells in tumor tissue, inhibit tumor immune escape, and thus exert an effective preventive and therapeutic effect against colorectal cancer. Furthermore, this invention has found that the combined use of azvudine and berberine is more effective in preventing and treating colorectal cancer than single-agent administration. Attached Figure Description

[0037] Figure 1 This study demonstrates how FNC reduces tumor number and tumor burden in the AOM / DSS model. A: Experimental protocol. B: Typical colorectal image. CD: Tumor number (C) and tumor burden statistics (D) (n=12). EF: Flow cytometry analysis of the proportion of CD4+ cells in lymphocytes in PBMCs (E) (n=9); and CD25+ cells in tumor tissue. - FOXP3 + The percentage of cells in CD4+ T cells (F) (n=9). Compared with the AOM / DSS model group, #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001. Compared with the FNC and BBR combination group, *P<0.05, ***P<0.001. Detailed Implementation

[0038] This invention provides compositions containing azvudine and their application in the prevention and treatment of colorectal cancer. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments, and those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0039] The test materials used in this invention are all common commercial products and can be purchased on the market.

[0040] The present invention will be further illustrated below with reference to the embodiments:

[0041] Example 1: Combined use of FNC and BBR reduces the incidence of colon cancer in mice.

[0042] The AOM / DSS model is a classic model of inflammation-induced colorectal cancer. Male C57BL / 6N mice were intraperitoneally injected with 10 mg / kg azoxymethane (AOM) to induce the model. Additionally, 1.5% dextran sulfate sodium salt (DSS, dissolved in drinking water) was administered at weeks 2, 4, and 6 to aid in model induction. From day 1 of the experiment, mice were administered FNC at doses of 0.46 mg / kg / day and 1.06 mg / kg / day via gavage until the experimental endpoint. These doses are equivalent to 3 mg / day and 7 mg / day for clinical use, respectively. The BBR group received a dose of 100 mg / kg / day, while the combination group received both 0.46 mg / kg / day of FNC and 50 mg / kg / day of BBR. The experimental procedure is described below. Figure 1 Group A, see Table 1 for grouping details.

[0043] Table 1 Grouping of mouse colon cancer experiments

[0044]

[0045]

[0046] Experimental results showed that FNC significantly reduced the number of tumors and tumor burden (sum of the diameters of all tumors in one mouse) in a mouse intestinal cancer model. Figure 1 C and Figure 1As shown in Figure D, the mean number of tumors in the AOM / DSS model group was 13, while the mean number in the FNC-L and FNC-H treatment groups decreased to 9 and 7, respectively; the FNC-H group showed a statistically significant difference compared to the model group. The tumor burden in the AOM / DSS model group reached 36 mm, while the mean tumor burden in the FNC-L and FNC-H groups decreased to 23 mm and 14 mm, respectively, both showing statistically significant differences compared to the model group.

[0047] BBR reduced the number of tumors and the tumor burden to 7.5 and 17 mm, respectively, which was slightly less effective than the FNC-H group.

[0048] The combination of a low-dose FNC (0.46 mg / kg / day) and a halved dose of BBR (50 mg / kg / day) achieved optimal efficacy in each group, reducing the number of tumors to 6 and the tumor burden to 12 mm, both of which were significantly different from the model group. Figure 1 C and Figure 1 D). The combination therapy group showed a significantly better effect in reducing tumor burden than the FNC group using 0.46 mg / kg / day alone, reaching a statistically significant difference. Figure 1 (D) This indicates that the two have a synergistic effect when used together. It also suggests that the mechanism of action of FNC differs from that of BBR.

[0049] To further elucidate the mechanism of action of FNCs, we used flow cytometry to detect the proportion of immune cells in mouse peripheral blood mononuclear cells (PBMCs) and tumor tissues. The results showed that FNCs significantly increased the proportion of CD4+ cells in PBMCs. Figure 1 E), and reduce CD25 in tumor tissue. - FOXP3 + (Treg) cell ratio ( Figure 1 F), thus inhibiting tumor immune escape and the occurrence of colorectal cancer.

[0050] It is worth noting that BBR affects CD4+ cells in PBMCs and CD25 in tumor tissue. - FOXP3 + The ratio of (Treg) cells had no significant effect. This is consistent with the efficacy results mentioned above. Figure 1 C and Figure 1 D), meaning that the mechanism of action of FNC differs from that of BBR. Although BBR cannot increase CD4+ cells in the blood, its combination with FNC significantly enhances the effect of FNC, resulting in a significantly better CD4+ cell increase than either FNC or BBR alone, achieving a statistically significant difference. Figure 1 E).

[0051] Furthermore, this invention uses the Jin Zhengjun Q-value method to calculate the combined effect, with the formula: Q = E A+B / E A +E B -E A ·E B E A E represents the effect of drug A when administered alone. B E represents the effect of drug B when administered alone. A+B This represents the combined effect of drugs A and B, and is the expected value of the combined effect. When Q < 0.85, the combined effect is antagonistic; when 0.85 ≤ Q < 1.15, the combined effect is additive; and when Q ≥ 1.15, the combined effect is synergistic.

[0052] Single-use groups: FNC group (0.46 mg / kg / day) and BBR group (100 mg / kg / day);

[0053] Combination therapy: 0.46 mg / kg / day of FNC and 50 mg / kg / day of BBR.

[0054] The effect of the two drugs in increasing blood CD4+ cells was calculated using the above formula. The calculated Q value was 3.17, indicating that the combination of the two drugs has a highly significant synergistic effect.

[0055] In summary, the combined use of azvudine and berberine for the prevention and treatment of colorectal cancer achieved a synergistic effect (1+1>2), demonstrating a significant synergistic effect. Furthermore, it allowed for a reduction in the azvudine dosage to the lifelong dose of 3 mg / day recommended for HIV-infected individuals, making it more suitable for long-term prevention and treatment of colorectal cancer. The novel mechanisms of action of azvudine and berberine, unlike existing drugs, offer new hope for the prevention and treatment of colorectal cancer.

[0056] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition for preventing and treating colorectal cancer, characterized in that, Including azvudine and alkaloids; The alkaloids include berberine and / or berberine derivatives.

2. The composition according to claim 1, characterized in that, The berberine derivatives include at least one of IMB-Y53, berberine, fensleyine, demethylene berberine, and berberine.

3. The composition according to claim 1, characterized in that, The mass ratio of azvudine to alkaloids is (1-10):(100-1500).

4. The composition according to claim 3, characterized in that, The mass ratio of azvudine to alkaloids is 1:330, 1:660, 3:330, 3:660, 5:330, 5:660, 7:330, or 7:

660.

5. Any of the following applications in the preparation of drugs for the prevention and treatment of colorectal cancer: (1) The composition according to any one of claims 1 to 4; (2) Berberine or its derivatives, used in combination with azvudine.

6. The application according to claim 5, characterized in that, The colorectal cancers include colorectal cancer and / or small bowel cancer.

7. The application according to claim 5 or 6, characterized in that, The prevention and treatment include inhibiting tumor immune escape, reducing tumor number, reducing tumor burden, and / or antagonizing lysophospholipid-related signaling pathways.

8. The application according to claim 7, characterized in that, The inhibition of tumor immune escape includes any of the following: (a) Increase the proportion of CD4+ T cells in the blood; (b) Reduce the proportion of Treg cells in tumor tissue.

9. The application according to any one of claims 5 to 8, characterized in that, The azvudine is administered simultaneously with berberine or its derivatives, or in any order thereof.

10. A drug for the prevention and treatment of colorectal cancer, characterized in that, It includes the composition according to any one of claims 1 to 4 and pharmaceutically acceptable excipients.

11. The medicament according to claim 10, characterized in that, The dosage forms of the drug include injections and oral preparations; the injections include liquid injections, powder injections, or emulsion injections; the oral preparations include tablets, granules, pills, capsules, and oral liquid formulations.

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