Application of clofazimine in preparation of medicine for treating cancer

Abstract

The invention provides an application of clofazimine in preparation of a medicine for treating cancers. Animal experiments show that clofazimine can regulate the tumor immune environment and enhance the anti-tumor immune effect; the number of tumors is reduced, and tumor bodies are reduced; the survival time of tumor animals is prolonged.

Description

Field of the Invention

[0001] The present application belongs to the field of tumor treatment. Specifically, the present application provides the use of clofazimine in the preparation of drugs for treating cancer. Background Art

[0002] Clofazimine has significant inhibitory activity against various mycobacteria such as Mycobacterium leprae and Mycobacterium tuberculosis. It is mainly used in the treatment of leprosy in clinical practice. In addition, it has anti-inflammatory and lysosome stabilizing effects, and has certain effects on certain types of lupus erythematosus and certain special skin diseases. There are currently no confirmed reports on the use of clofazimine in the treatment of tumors.

[0003] Colon cancer is one of the most common tumor types in clinical practice. Currently, there are many chemical, biological, and traditional Chinese medicine drugs for treatment, but each drug has its limitations. There is still a continuous demand for new drugs to treat colon cancer in clinical practice. Summary of the invention

[0004] In one aspect, the present application provides the use of clofazimine in the preparation of a drug for treating cancer.

[0005] Furthermore, the cancer is colon cancer.

[0006] Furthermore, the drug reduces the number of tumors, reduces the size of tumors and / or prolongs survival time.

[0007] Furthermore, the drug regulates the tumor immune environment and enhances anti-tumor immune effects.

[0008] Furthermore, the drug is in oral or injectable form.

[0009] Furthermore, the drug contains pharmaceutically acceptable excipients.

[0010] On the other hand, the present application provides the use of clofazimine in the preparation of a drug for improving anti-tumor immunity.

[0011] Furthermore, the drug increases PD1-positive CD3+CD8+T cells and Treg cells in peripheral blood.

[0012] Furthermore, the drug is in oral or injectable form.

[0013] Furthermore, the drug contains pharmaceutically acceptable excipients.

[0014] The dosage forms available in this application include but are not limited to: tablets, capsules, oral liquids, water injections, powder injections, etc. According to the dosage form prepared / administered, various pharmaceutically acceptable excipients can be selected in the preparation, including but not limited to coating materials, solvents, solubilizers, fillers, disintegrants, binders, stabilizers, antioxidants, pH regulators, osmotic pressure regulators, flavoring agents, taste masking agents, etc. These excipient components can be selected by those skilled in the art according to common knowledge of pharmacy.

[0015] The drug of the present application does not exclude co-administration with other anti-tumor drugs, and these drugs can be co-administered with the drug of the present application or prepared in the same package or preparation.

[0016] The present application found that clofazimine has a significant therapeutic effect on tumors, especially colon cancer, which expands the use of clofazimine and provides a new class of drug options for tumor treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The results are for counting PD1+Treg cells in the peripheral blood of mice;

[0018] Figure 2 The results are for the CD8+PD1+T cell counts in the peripheral blood of mice;

[0019] Figure 3 The number of mouse tumors and the maximum tumor diameter are shown;

[0020] Figure 4 Shown are the overall survival results for mice with colon cancer. DETAILED DESCRIPTION

[0021] Example 1 Basic Materials and Methods

[0022] Animal Materials:

[0023] Animals: C57 mice (Speff); Apcmin / + mice (Jicui Yaokang); Gender: male; Age: 6-8 weeks.

[0024] Mouse peripheral blood flow cytometry:

[0025] The peripheral blood immune cells of mice were detected by flow cytometry, and the detection was performed on a BD Fortessa flow cytometer, and the data were analyzed using Diva analysis software. The materials used include:

[0026] Table 1 All major reagents for flow cytometry

[0027]

[0028]

[0029] Mouse peripheral blood immune cell detection:

[0030] Take 200ul of mouse peripheral blood and place it in a test tube. Add CD3 / CD4 / CD8 / CD45 / PD-1 / CD25 according to the recommended amount of antibodies, vortex and mix well, and incubate in the dark for 15 minutes.

[0031] Add 1 mL of 1x fixative permeabilization reagent, vortex to mix, and incubate in the dark for 15 min;

[0032] Centrifuge at 1500 rpm for 5 min and discard the supernatant to enrich the cells;

[0033] Add 1 mL 1X Buffer, centrifuge at 1500 rpm for 5 min, and discard the supernatant;

[0034] Add FOXP3 according to the recommended amount of antibody, vortex to mix, and incubate in the dark for 30 minutes;

[0035] Add 1 mL of Buffer, centrifuge at 1500 rpm for 5 min, and discard the supernatant. Repeat once;

[0036] Add 200uL 1X Buffer to resuspend the sample and test on the instrument.

[0037] Experimental animals:

[0038] Within 48 hours before dissection, mouse feces were collected and stored at -80 degrees. Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate, with an injection dose of 0.3 ml / 100 g. The mouse chest cavity was cut open, and blood was collected from the abdominal aorta. Part of the whole blood was taken to separate serum and stored at -80 degrees. The colon tissue was taken out for 4% paraformaldehyde fixation, glutaraldehyde fixation and -80 degrees cryostasis.

[0039] Example 2 Experimental process

[0040] Apcmin / + mice:

[0041] After 1 week of adaptive feeding, 20 male mice were divided into two groups, 10 mice in each group. The control group was fed normally without any intervention or treatment for 90 days; the drug group was fed normally and given clofazimine (45.5 mg / kg) by gavage once a day for 90 days.

[0042] C57 mice:

[0043] After 1 week of adaptive feeding, 20 male mice were divided into two groups, 10 mice in each group.

[0044] Control group: mice were given a single intraperitoneal injection of 10 mg / kg azoxymethane (AOM). One week after the injection of AOM, the mice were given drinking water containing 2% dextran sodium sulfate (DSS) for 5 days, and then normal drinking water was restored for 16 days, and the cycle was repeated 3 times until the 90th day of the experiment.

[0045] Treatment group: mice were given a single intraperitoneal injection of azoxymethane (AOM) at a dose of 10 mg / kg. One week after the injection of AOM, the mice were given drinking water containing 2% dextran sodium sulfate (DSS) for 5 days, and then normal drinking water was restored for 16 days, with a cycle of 3 times; and at the beginning of the experiment, clofazimine (45.5 mg / kg) was gavaged once a day for 90 days.

[0046] Observe whether clofazimine affects the overall survival time of mice:

[0047] Mice were selected from 20 Apcmin / + mice and 20 C57 mice (AOM / DSS), 10 of which were control group and 10 of which were drug-treated group. The modeling and drug-treatment procedures were the same as above. The death experiment was recorded until the mice died and the survival curve was drawn.

[0048] Example 3 Experimental results

[0049] Figure 1 and Figure 2 The results showed that clofazimine treatment can increase the PD1-positive CD3+CD8+T cells and Treg cells in the peripheral blood of mice, proving that clofazimine can regulate the tumor immune environment and enhance anti-tumor immunity. After clofazimine treatment, CD3+CD8+PD1+T cells in the peripheral blood of mice increased (3.6% vs 7.1%, P<0.05); at the same time, PD1-positive Treg cells in the peripheral blood also increased (15% vs 31%, P<0.05).

[0050] Figure 3 The results showed that clofazimine treatment can significantly reduce colon malignant tumors and reduce the number of colon malignant tumor metastases.

[0051] Figure 4 It showed that clofazimine treatment could significantly prolong the overall survival of colon cancer mice.

Claims

1. The use of clofazimine in the preparation of drugs for treating cancer.

2. The use according to claim 1, wherein the cancer is colon cancer.

3. The use according to claim 1 or 2, wherein the drug reduces the number of tumors, reduces the tumor volume and / or prolongs the survival time.

4. The use according to claim 1 or 2, wherein the drug regulates the tumor immune environment and enhances anti-tumor immune effects.

5. The use according to any one of claims 1 to 4, wherein the drug is in oral or injectable dosage form.

6. The use according to claim 5, wherein the medicine contains pharmaceutically acceptable excipients.

7. Application of clofazimine in the preparation of drugs for improving anti-tumor immunity.

8. The use according to claim 7, wherein the drug increases PD1-positive CD3+CD8+T cells and Treg cells in peripheral blood.

9. The use according to claim 7 or 8, wherein the drug is in oral or injectable form.

10. The use according to claim 9, wherein the drug contains pharmaceutically acceptable excipients.

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