Application of IL-9 as a drug for treating thrombocytopenia

Abstract

The application of IL-9 as a treatment for thrombocytopenia is specifically described in the preparation of drugs for treating drug-induced thrombocytopenia, where the preferred drug is a chemotherapeutic agent. IL-9, even at low doses, can exceed the effects of existing high-dose cytokines in clinical practice, and its efficacy is better than TPO and IL-11.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and more specifically to the application of IL-9 as a drug for treating thrombocytopenia. Background Technology

[0002] Platelets are an important blood component involved in hemostasis and coagulation. Abnormalities in their quantity or quality can lead to various hemorrhagic diseases. Clinical symptoms range from mild gastrointestinal bleeding to severe cardiac arrest, cerebral hemorrhage, and even death. Current treatments for platelet disorders primarily involve corticosteroids, immunosuppressants, splenectomy, and platelet transfusions. However, these methods are prone to causing infections, bone marrow suppression, and the production of platelet antibodies, and are ineffective for some patients. Current treatments for thrombocytopenia caused by chemotherapy for solid tumors mainly involve subcutaneous injections of TPO and IL-11. TPO is administered 6-24 hours after the completion of chemotherapy for solid tumors, at a dose of 300 IU per kilogram of body weight, once daily for 14 consecutive days, continuing until platelet counts recover to 100 × 10⁶. 9 / L or above, or an absolute increase of ≥50×10 9 The medication is discontinued when the platelet count reaches a certain level ( / L). IL-11 is administered subcutaneously 24-48 hours after the completion of chemotherapy for solid tumors or after the onset of thrombocytopenia, at a dose of 25-50 μg per kilogram of body weight, once daily, for a typical course of 7-14 days. The medication is discontinued once platelet counts recover. These two types of cytokines are not very effective for thrombocytopenia in chemotherapy patients, and require large doses, resulting in high costs. Therefore, finding new drug targets for treating thrombocytopenia in chemotherapy patients has become an urgent clinical problem. Summary of the Invention

[0003] Clinically, solid tumors are generally treated with a combination of chemotherapy drugs. However, chemotherapy drugs often cause bone marrow suppression in most chemotherapy patients, leading to severe thrombocytopenia, sometimes even refractory thrombocytopenia. Currently, the main treatment for this type of thrombocytopenia is subcutaneous injection of TPO and IL-11. However, these two cytokines are not very effective for thrombocytopenia in chemotherapy patients, and require large doses, resulting in high costs. To overcome the problems of existing technologies, the inventors of this invention discovered that IL-9, as a cytokine that promotes the proliferation and differentiation of megakaryocytes (progenitor cells of platelets), can rapidly reverse thrombocytopenia in chemotherapy-treated mice, restoring platelet counts to pre-chemotherapy levels. Compared with TPO and IL-11, it has the advantages of lower dosage, better efficacy, and lower cost.

[0004] The IL-9 of this invention can exert a significant effect in vivo on its own, and can reverse thrombocytopenia caused by chemotherapy drugs.

[0005] One aspect of the present invention provides the use of IL-9 in the preparation of a medicament for treating thrombocytopenia.

[0006] Another aspect of the present invention provides the use of IL-9 in the preparation of medicaments for treating drug-induced thrombocytopenia.

[0007] The drugs are anti-tumor drugs.

[0008] The antitumor drug is a chemotherapy drug, preferably carboplatin, cisplatin, 5-fluorouracil, methotrexate, cyclophosphamide, paclitaxel, or etoposide. Another aspect of this invention provides the use of IL-9 in the preparation of a drug for treating bone marrow megakaryopenia.

[0009] Another aspect of the present invention provides the use of IL-9 in the preparation of a drug that reduces the proliferation of bone marrow megakaryocytes.

[0010] The drugs are anti-tumor drugs.

[0011] Among them, the antitumor drugs are chemotherapy drugs, preferably carboplatin, cisplatin, 5-fluorouracil, methotrexate, cyclophosphamide, paclitaxel, or etoposide. These drugs can achieve effects exceeding those of currently available high-dose cytokines at low doses, and their efficacy is better than TPO and IL-11. Attached Figure Description

[0012] Figure 1 To measure platelet counts using a complete blood count instrument in C57 mouse experiments.

[0013] Figure 2 To determine platelet counts using a complete blood count instrument in BALB / c mouse experiments.

[0014] Figure 3 The results of Swiss-Gymsa staining of megakaryocytes in bone marrow smears in C57 mouse experiments.

[0015] Figure 4 The results of Swiss-Gymsa staining of megakaryocytes in bone marrow smears in BALB / c mice experiments.

[0016] Figure 5 The results of megakaryocyte proliferation percentage were obtained by flow cytometry in C57 mouse experiments.

[0017] Figure 6 The results of megakaryocyte proliferation percentage were obtained by flow cytometry in BALB / c mouse experiments. Detailed Implementation

[0018] The recombinant IL-9 cytokine used in the embodiments of this invention is endotoxin-free, has a purity of over 99%, and was purchased from Peprotech, Inc., USA.

[0019] Example 1: C57 and BALB / c mouse experiments

[0020] Eight-week-old female C57 and BALB / c mice were purchased from the Experimental Animal Center of Southern Medical University. After 7 days of injection with carboplatin (100 mg / kg, purchased from Sigma-Aldrich, USA), the mice experienced a significant decrease in platelet count, typically ranging from 40 to 200 × 10⁻⁶ in routine blood tests. 9 / L (normal platelet count in 8-week-old female C57 and BALB / c mice is 800-1200 × 10⁹ / L) 9 The levels were between 1 / L and 2.5 μg / kg. Starting on day 8, mice were intraperitoneally injected with IL-9 (2.5 μg / kg), TPO (100 μg / kg, purchased from Peprotech, USA), and IL-11 (100 μg / kg, purchased from Peprotech, USA), for 8 consecutive days. Changes in various indicators were assessed on day 17. Blood was collected from the heart of mice under general anesthesia, and whole blood was placed in anticoagulant tubes for platelet count analysis using a complete blood count (CBC) machine. Bone marrow smears from one leg were stained with Swiss-Gymsa and megakaryocytes were counted under a microscope; bone marrow cells from the other leg were double-labeled with CD41 (a specific megakaryocyte surface marker) and Edu (a specific cell proliferation marker) antibodies, and the percentage of megakaryocyte proliferation was analyzed by flow cytometry. The effects of the three cytokines on platelet count, megakaryocyte count, and megakaryocyte proliferation percentage were compared.

[0021] For detailed test results, please refer to the appendix. Figure 1-6 And Table 1-2 (* indicates statistical significance compared to the control group):

[0022] In the C57 mouse experiment (see...) Figure 1 According to Table 1, IL-9 (2.5 μg / kg), TPO (100 μg / kg), and IL-11 (100 μg / kg) can reverse carboplatin-induced thrombocytopenia to varying degrees. Among them, IL-9 has the best efficacy and can reverse platelet count to the pre-chemotherapy level.

[0023] In BALB / c mouse experiments (see...) Figure 2 According to Table 2, IL-9 (2.5 μg / kg), TPO (100 μg / kg), and IL-11 (100 μg / kg) can reverse carboplatin-induced thrombocytopenia to varying degrees. Among them, IL-9 still has the best efficacy and can reverse platelet count to the pre-chemotherapy level.

[0024] In C57 mouse experiments (see...) Figure 3According to Table 1, IL-9 (2.5 μg / kg), TPO (100 μg / kg), and IL-11 (100 μg / kg) can reverse the reduction of bone marrow megakaryocytes induced by carboplatin to varying degrees. Among them, IL-9 has the best therapeutic effect, and the number of bone marrow megakaryocytes is the highest among the three cytokines.

[0025] In BALB / c mouse experiments (see...) Figure 4 According to Table 2, IL-9 (2.5 μg / kg), TPO (100 μg / kg), and IL-11 (100 μg / kg) can reverse the reduction of bone marrow megakaryocytes induced by carboplatin to varying degrees. Among them, IL-9 still has the best therapeutic effect, and the number of bone marrow megakaryocytes is the highest among the three cytokines.

[0026] In C57 mouse experiments (see...) Figure 5 IL-9 (2.5 μg / kg), TPO (100 μg / kg), and IL-11 (100 μg / kg) could reverse the reduction in bone marrow megakaryocyte proliferation induced by carboplatin to varying degrees, with IL-9 promoting the highest percentage of megakaryocyte proliferation among the five groups.

[0027] In BALB / c mouse experiments (see...) Figure 6 IL-9 (2.5 μg / kg), TPO (100 μg / kg), and IL-11 (100 μg / kg) could reverse the reduction in bone marrow megakaryocyte proliferation induced by carboplatin to varying degrees. Among them, the percentage of megakaryocyte proliferation promoted by IL-9 was the highest among the five groups.

[0028] Table 1. Experimental results in C57 mice

[0029]

[0030] Table 2 Results of BALB / c mouse experiments

[0031]

[0032] In conclusion, IL-9 is more effective than both TPO and IL-11 in treating this condition.

Claims

1. The use of IL-9 as the sole active ingredient in the preparation of drugs for treating thrombocytopenia caused by pharmaceuticals; wherein the pharmaceuticals are carboplatin, cisplatin, 5-fluorouracil, methotrexate, cyclophosphamide, paclitaxel, or etoposide.

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