Use of a derivative of azulenone C-3 against erythroleukemia

Abstract

The application discloses application of aza-3 in resisting erythroleukemia, wherein the aza-3 is a compound obtained by modifying a ring of aza-3 with 3alpha-O-(Boc-proline); the application proves that the aza-3 can obviously inhibit HEL and K562 cell proliferation, reduce splenomegaly, improve anemia indexes (RBC, HGB and HCT) and splenic and hepatic pathological infiltration, and has no obvious toxic side effects on main organs such as heart, lung and kidney through in-vitro MTT experiment and a Friend virus-induced erythroleukemia mouse model.

Description

Technical Field

[0001] This invention relates to the fields of medicinal chemistry and pharmaceutical technology, and in particular to the application of a blue calyx derivative C-3 in the treatment of erythroleukemia. Background Technology

[0002] Erythroleukemia is a special type of acute myeloid leukemia characterized by abnormal proliferation of both erythroid and myeloid lineages, with rapid clinical progression and a poor prognosis. Current clinical treatments mainly involve combination chemotherapy and hematopoietic stem cell transplantation, but these methods suffer from high relapse rates, significant treatment-related toxicity, and ineffectiveness in some patients. Anti-leukemia drugs derived from natural products have been a research hotspot. Among them, styraxin, a natural diterpenoid compound, has been reported to possess certain antitumor activity. A series of derivatives obtained through structural modification of styraxin have shown proliferative inhibitory effects in various solid tumors and hematological malignancies.

[0003] Existing research on leukemia derivatives mainly focuses on evaluating their broad-spectrum antitumor activity, lacking systematic pharmacodynamic validation for specific leukemia subtypes, particularly erythroleukemia, and even more so on studies of their in vivo therapeutic effects targeting the specific pathological features of erythroleukemia. The reported leukemia derivatives suffer from poor water solubility, low bioavailability, and unclear toxicity to normal organs in vivo, limiting their prospects for clinical translation.

[0004] Therefore, in response to the problems mentioned above, this invention proposes the application of a blue calyx derivative C-3 in the treatment of erythroleukemia. Summary of the Invention

[0005] To overcome the problems of the lack of blue calyx derivatives with specific therapeutic effects on erythroleukemia, incomplete in vivo efficacy data, and insufficient safety evaluation in the existing technology, this invention proposes the application of blue calyx derivative C-3 in the treatment of erythroleukemia, which can significantly inhibit erythroleukemia and has good safety.

[0006] The technical solution of this invention is: the application of a blue sepal derivative C-3 in the treatment of erythroleukemia, wherein the blue sepal derivative C-3 is a compound obtained by modifying the A ring of blue sepal with a 3α-O-(Boc-aminoacyl) structure, and its structure is as follows:

[0007] .

[0008] (1) The blue calyx derivative C-3 exerts its anti-erythroleukemic effect by inhibiting the proliferation of erythroleukemia cells (HEL cells and K562 cells), with an inhibition rate of 81.99% against HEL cells and 78.85% against K562 cells after 48 hours; the IC50 value against HEL cells after 48 hours is 0.499±0.041μM and the IC50 value against K562 cells after 48 hours is 0.787±0.048μM;

[0009] (2) The blue calyx derivative C-3 is used to prepare a drug to reduce splenomegaly in erythroleukemia patients;

[0010] (3) The blue calyx derivative C-3 is used to prepare a drug to improve anemia caused by erythroleukemia, wherein the anemia indicators include red blood cell count, hemoglobin and hematocrit;

[0011] (4) The blue calyx derivative C-3 is used to prepare drugs that reduce pathological infiltration of the spleen and liver in patients with erythroleukemia.

[0012] Preferably, when the blue calyx derivative C-3 is used to prepare an anti-erythroleukemia drug, the drug is administered via intraperitoneal injection.

[0013] The beneficial effects of this invention are:

[0014] 1. This invention is the first to apply the blue calyx derivative C-3 to the treatment of erythroleukemia, filling the gap in the existing technology for the application of blue calyx derivatives targeting this specific subtype of erythroleukemia, and overcoming the deficiency of existing blue calyx derivatives that only generally claim anti-tumor effects without verifying their specific efficacy against erythroleukemia.

[0015] 2. This invention confirms through in vitro MTT assay that C-3 has a potent inhibitory effect on the proliferation of HEL and K562 erythroleukemia cells, with an IC50 value of 48 hours. 50 The values ​​were as low as 0.499 μM and 0.787 μM, respectively, with inhibition rates of 81.99% and 78.85%, solving the technical problem of the lack of blue calyx derivatives that are highly effective in inhibiting erythroleukemia cells in the existing technology.

[0016] 3. This invention uses a Friend virus-induced erythroleukemia mouse model to systematically verify that C-3 can significantly reduce splenomegaly, improve anemia indicators (RBC, HGB, HCT) and reduce pathological infiltration of the spleen and liver in a dose-dependent manner, which solves the problems of incomplete in vivo efficacy data and lack of evaluation of the improvement of specific pathological features of erythroleukemia in the prior art.

[0017] 4. This invention further confirms that C-3 has no obvious toxic side effects on major organs such as the heart, lungs, and kidneys in the dosage range of 1-10 mg / kg, and has good safety. It overcomes the shortcomings of the existing blue calyx derivatives with unclear in vivo safety, and provides a reliable basis for clinical application. Attached Figure Description

[0018] Figure 1 The diagram shows the inhibitory effect of C-3 on the proliferation of HEL and K562 cells of the present invention.

[0019] Figure 2 The diagram shows a comparison of the spleen morphology and spleen weight of mice in the normal control group, model group and C-3 treatment groups at various doses according to the present invention.

[0020] Figure 3 The diagram shown is a statistical illustration of the mouse blood routine indicators of the present invention;

[0021] Figure 4 The diagram shown represents the H&E staining of mouse heart, liver, spleen, lung, and kidney according to the present invention.

[0022] Figure 5 The diagram shows a comparison of the morphological appearance of the major organs of mice in the normal control group, model group, and C-3 treatment groups at various doses according to the present invention.

[0023] Figure 6 The diagram shown is a quantitative statistical analysis of the coefficients of various organs in this invention. Detailed Implementation

[0024] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are some embodiments of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] Example 1:

[0026] This embodiment provides a specific method for synthesizing the blue calyx methyl ester derivative C-3. The C-3 of this invention is obtained by using blue calyx methyl ester as a raw material, through 3α-O-(Boc-aminoacyl) structural modification of the A ring, followed by condensation reaction, deprotection, and purification. Specific steps include:

[0027] Weigh intermediate A-26 (50.5 mg, 0.1350 mmol, 1.0 eq) and DMAP (5 mg, 0.0668 mmol, 0.3 eq) into a 50 mL dry round-bottom flask, dissolve in anhydrous dichloromethane, then add EDCI (102.5 mg, 0.5348 mmol, 4.0 eq) and stir to dissolve, followed by Boc-L-proline (2.0 eq). The reaction is carried out under nitrogen protection at room temperature with stirring, and the reaction progress is monitored by TLC. After the reaction is complete, 30 mL of water is added to quench the reaction, then alkalize with 3-5 times the volume of saturated NaHCO3 solution, extract three times with 50 mL of dichloromethane each time, combine the organic phases, wash with saturated brine, dry with anhydrous Na2SO4, concentrate under reduced pressure, and proceed directly to the next reaction step.

[0028] The concentrated product was dissolved in anhydrous dichloromethane, and 0.2 mL of trifluoroacetic acid was slowly added dropwise under ice bath and nitrogen protection. The reaction was stirred at room temperature and monitored by TLC. After 10 hours of reaction, the starting material disappeared. The pH was adjusted to alkaline with saturated NaHCO3 solution (3-5 times the volume), diluted with 30 mL of water, and extracted three times with 30 mL of dichloromethane each time. The organic phases were combined, washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography with petroleum ether-acetone (V / V = 6:1 → 5:1) as the eluent. The target fraction was collected, concentrated, and yielded 48.6 mg of white powder, with a yield of 53.9%, which was C-3.

[0029] The structure of the obtained C-3 was confirmed by nuclear magnetic resonance hydrogen spectrum, carbon spectrum and mass spectrometry. 1 H NMR (600 MHz, CDCl3) δ:6.17 (1H, s), 5.41 (1H, d, J = 3.1 Hz), 4.86 (1H, s), 4.46 (1H, dd, J = 11.7,4.4 Hz), 4.37-4.23 (2H, m), 3.55-3.36 (2H, m), 3.07 (1H, s), 2.28-2.15 (1H,m), 2.02-1.86 (5H, m), 1.83-1.69 (7H, m), 1.66-1.53 ​​(2H, m), 1.42 (9H, s),1.28-1.18 (2H, m), 1.08 (3H, d, J = 10.0 Hz), 1.02-0.97 (1H, m), 0.93 (3H, s), 0.89 (3H, s). 13C NMR (150 MHz, CDCl3) δ: 207.8, 172.8, 154.0, 147.7,118.2, 80.9, 80.1, 75.0, 61.8, 59.5, 54.0, 52.8, 46.2, 39.7, 37.9, 37.6,31.2, 30.1, 28.7, 28.6, 27.8, 24.5, 23.6, 23.3, 18.2, 17.8, 17.0. ESI-MS: m / z532 [M+H] + (C) 30 H 45 NO7). The above spectral data are consistent with the structure of the target compound, indicating that a high-purity blue calyx derivative C-3 has been successfully synthesized, and its structural formula is shown below:

[0030] .

[0031] Example 2:

[0032] This example is used to study the in vitro inhibitory effect of the blue calyx derivative C-3 on the proliferation of erythroleukemia cells.

[0033] In this embodiment, the inhibitory effect of C-3 on the proliferation of erythroleukemia cell lines HEL and K562 was determined by the MTT assay, and the half-maximal inhibitory concentration (IC50) was calculated. 50 To evaluate the in vitro anti-erythroleukemia activity of C-3.

[0034] Both HEL and K562 cells were purchased from the American Type Culture Collection (ATCC). In this case, the cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and incubated at 37 °C with 5% CO2 and saturated humidity. Cells were passaged every 2-3 days, and cells in the logarithmic growth phase were used for experiments.

[0035] HEL and K562 cells in logarithmic growth phase were cultured at 5 × 10⁶ cells per well. 3 Cells were seeded at a density of 100 μL per well in 96-well plates. The plates were pre-cultured for 24 hours to allow cell adhesion (HEL cells were suspension cells, but uniformly distributed after seeding). Different concentrations of blue calyx derivative C-3 were then added. Based on preliminary results, the final concentration gradients of C-3 were set at 0.3125 μM, 0.625 μM, 1.25 μM, 2.5 μM, and 5 μM. Five replicates were performed for each concentration. A control group (with an equal volume of DMSO added, final concentration not exceeding 0.1%) and a blank group (without cells, only culture medium added, used for zeroing) were also set up. The 96-well plates were then incubated for 24, 48, and 72 hours, respectively.

[0036] At each time point, 20 μL of MTT solution (5 mg / mL, prepared with sterile PBS) was added to each well, and the entire process was performed in the dark. The culture plate was then returned to the incubator for another 4 hours of incubation. After incubation, the supernatant was carefully aspirated from the wells, and 150 μL of DMSO was added to each well. The culture plate was then placed on a shaker and shaken at low speed for 10 minutes to fully dissolve the purple formazan crystals. Finally, the optical density (OD) value of each well was measured at 490 nm using a microplate reader. The OD values ​​of each group were recorded, and the cell proliferation inhibition rate was calculated using the following formula: Cell proliferation inhibition rate (%) = (1 - OD value of experimental group / OD value of control group) × 100%.

[0037] All experiments were independently repeated three times. Data processing was performed using GraphPad Prism software. Dose-response curves were fitted using nonlinear regression to calculate the IC50 of C-3 on HEL and K562 cells at different treatment times. 50 Values. Experimental results are expressed as mean ± standard deviation (Mean ± SD).

[0038] Please see Figure 1 , Figure 1 (A) shows the inhibitory effect curve of C-3 on HEL cell proliferation. Figure 1 (B) shows the inhibitory effect of C-3 on K562 cell proliferation. MTT assay results showed that the blue calyx derivative C-3 exhibited significant inhibitory effects on the proliferation of both HEL and K562 erythroleukemia cell lines, with the best inhibitory effect on HEL cells at 48 hours, while the effect on K562 cells was time- and dose-dependent. Specifically, at different concentrations of C-3, the survival rate of both cell lines decreased with increasing drug concentration. When the C-3 concentration was 5 μM and the treatment time was 48 hours, the inhibition rate on HEL cells reached 81.99%, and the inhibition rate on K562 cells reached 78.85%. Even at a lower concentration (0.3125 μM), C-3 still showed some inhibitory effect on both HEL and K562 cells.

[0039] Please refer to Table 1. The inhibitory effect of C-3 on HEL cells is strongest at 48 hours, IC50. 50 The concentration was as low as below 0.5 μM; the inhibitory activity against K562 cells was comparable at 48 and 72 hours, both remaining around 0.78 μM. Compared with existing anti-leukemia natural product derivatives, the C-3 of this invention exhibits potent inhibitory activity at nanomolar to micromolar levels, and its sensitivity against erythroleukemia cells is superior to that of general tumor cell lines. These results fully demonstrate that the blue calyx derivative C-3 can effectively inhibit the proliferation of erythroleukemia cells in vitro.

[0040]

[0041] Example 3:

[0042] This example is used to study the in vivo anti-erythroleukemia effect of the blue calyx derivative C-3.

[0043] This embodiment uses a Friend virus-induced mouse erythroleukemia model to systematically evaluate the therapeutic effect of the blue calyx derivative C-3 on erythroleukemia through in vivo drug administration experiments, including its improvement on splenomegaly, anemia indicators, and pathological infiltration, while also assessing the safety of C-3 on major organs.

[0044] The BALB / c suckling mice used in this experiment (within 48 hours of birth) were raised under standard conditions: temperature 22±2℃, humidity 50%±10%, 12-hour light / 12-hour dark cycle, and free access to food and water.

[0045] BALB / c suckling mice within 48 hours of birth were intraperitoneally injected with 100 μL of Friend virus suspension to induce an erythroleukemia model. The Friend virus, a Friend virus complex (FV) preserved in our laboratory, was pre-diluted to an appropriate titer with serum-free medium. After modeling, the mice continued to be housed under the same conditions. Four weeks after modeling, the general condition of the model mice was observed, including changes in activity level, coat color, and body weight. At this time, the model mice showed obvious splenomegaly and anemia (pale ears and paws), indicating successful establishment of the erythroleukemia model.

[0046] Four weeks after modeling, the successfully modeled mice were randomly divided into four groups of five mice each, as follows:

[0047] The model group was given an equal volume of physiological saline after modeling.

[0048] The C-3 low-dose group was given C-3 1 mg / kg after modeling;

[0049] The C-3 medium-dose group was given C-3 5 mg / kg after modeling;

[0050] The high-dose C-3 group was given C-3 at 10 mg / kg after modeling.

[0051] A separate normal control group was set up. Mice in this group were not subjected to modeling but were injected with an equal volume of physiological saline and were raised under the same conditions.

[0052] C-3 was dissolved in a small amount of DMSO and then diluted with physiological saline to the required concentration (final DMSO concentration ≤ 1%). Administration was by intraperitoneal injection, once every other day, for a total of 8 administrations. Mice in the model group and the normal control group were intraperitoneally injected with the same volume of physiological saline (containing the same concentration of DMSO, serving as a solvent control).

[0053] Twenty-four hours after the last administration, mice were weighed and euthanized by cervical dislocation. The spleen, liver, heart, lungs, and kidneys were quickly dissected, rinsed with pre-cooled physiological saline to remove residual blood, and blotted dry with filter paper before weighing each organ. The organ coefficient was calculated using the formula: Organ coefficient (%) = (Organ weight (g) / Body weight (g)) × 100%.

[0054] Please see Figure 2 , Figure 2 (A) Comparison of spleen morphology in mice of the normal control group, model group, and various doses of C-3 treatment groups. The spleen morphology observation results show that the spleen of mice in the normal control group is dark red, small in size, and has a smooth capsule. The spleen of mice in the model group is significantly enlarged, several times or even dozens of times larger than that of the normal group, and is dark red or dark purple in color, with multiple nodular protrusions visible on the surface. The spleen weight is also significantly increased. Compared with the model group, the spleen enlargement of mice in the medium-dose (5 mg / kg) and high-dose (10 mg / kg) C-3 groups was significantly reduced, the spleen volume decreased, the color returned to normal, and the surface nodules decreased. The spleen in the low-dose group (1 mg / kg) also showed some improvement, but not as significantly as in the medium- and high-dose groups.

[0055] Figure 2 (B) is a statistical chart of spleen weight in mice. The spleen weight statistics show that the spleen weight of the normal control group mice was 0.08±0.01. The spleen coefficient of the model group mice was significantly increased to 0.92±0.32, indicating that the erythroleukemia model mice had severe splenomegaly and extramedullary hematopoiesis. Compared with the model group, the spleen coefficient of the low-dose C-3 group was 0.7±0.06, which was slightly reduced but not statistically significant; the spleen coefficient of the medium-dose C-3 group decreased to 0.42±0.27; and the spleen coefficient of the high-dose C-3 group further decreased to 0.26±0.11. The above results show a clear dose-dependent effect, that is, the improvement effect of splenomegaly is more significant with the increase of C-3 dose.

[0056] Peripheral blood was collected in EDTA anticoagulant tubes, and whole blood samples were taken. Complete blood count indicators were measured using a fully automated blood analyzer, and the red blood cell count (RBC, ×10⁻⁶) was recorded. 12 / L), hemoglobin concentration (HGB, g / L), hematocrit (HCT, %), platelet count (PLT, ×10⁻⁶). 9 / L), white blood cell count (WBC, ×10) 9 / L).

[0057] Please see Figure 3 , Figure 3 (A) is a graph showing the number of red blood cells. Figure 3 (B) Hemoglobin statistics chart, Figure 3 (C) Hematocrit chart, Figure 3(D) Platelet count chart, Figure 3 (E) Leukemia Statistical Chart. Compared with the model group, blood routine test results showed that the RBC, HGB, and HCT levels in the model group mice were significantly lower than those in the normal control group (P < 0.01), indicating severe anemia in the model mice. Compared with the model group, the RBC, HGB, and HCT levels in the medium and high dose C-3 groups were significantly higher (P < 0.05 or P < 0.01), and the anemia indicators were significantly improved. The low dose C-3 group also showed an upward trend in all indicators. The results indicate that the blue calyx derivative C-3 can effectively improve anemia caused by erythroleukemia.

[0058] Spleen and liver tissues were harvested, along with heart, lung, and kidney tissues for safety evaluation. Tissue blocks were fixed in 10% neutral formalin solution for 48 hours, followed by routine dehydration, paraffin embedding, and sectioning (approximately 4 μm thick). After dewaxing to water, sections underwent hematoxylin-eosin (HE) staining: hematoxylin staining for 5 minutes, rinsing with running water, differentiation with hydrochloric acid alcohol, eosin staining for 2 minutes, dehydration, clearing, and mounting with neutral resin. The tissues were observed and photographed under an optical microscope to assess histopathological changes. Special attention was paid to observing the spleen structure (red and white pulp boundary), extramedullary hematopoietic foci and tumor cell infiltration in the liver, and the structural integrity and presence of significant pathological changes in the heart, lung, and kidney tissues.

[0059] Please see Figure 4 In the normal control group, the spleen tissue structure of mice was clear, with a distinct boundary between the red and white pulp. The white pulp consisted of lymphoid nodules, while the red pulp was composed of splenic cords and sinuses, with no abnormal cell infiltration observed. In the model group, the spleen tissue structure was disordered, with unclear boundaries between the red and white pulp, dilated sinuses, diffuse infiltration of numerous tumor cells (erythroleukemia cells), atrophy or disappearance of splenic bodies, and hemorrhage and necrosis in some areas. In the low-dose C-3 group, the pathological changes in the spleen were somewhat alleviated, but some degree of tumor cell infiltration and tissue disorder were still observed. In the medium-dose C-3 group, the spleen structure was significantly improved, the boundary between the red and white pulp became clearer, tumor cell infiltration was significantly reduced, and splenic bodies partially recovered. In the high-dose C-3 group, the spleen tissue was basically restored to normal, with clear boundaries between the red and white pulp, and only a few scattered tumor cells were observed, closely resembling the morphology of the normal control group.

[0060] In the normal control group, the liver lobule structure of mice was intact, the hepatocyte cords were neatly arranged, the central vein was clear, and there was no abnormal infiltration. In the model group, multiple extramedullary hematopoietic foci were observed in the liver, characterized by a large number of immature erythroid and granulocytic cells clustered within the sinusoids and around the portal tract, along with tumor cell infiltration, destruction of the hepatocyte cord structure, and edema or degeneration of some hepatocytes. In the low-dose C-3 group, the number of extramedullary hematopoietic foci in the liver decreased, and the infiltration area shrank. In the medium-dose C-3 group, extramedullary hematopoietic foci in the liver were significantly reduced, with only a few scattered infiltrating cells, and the liver lobule structure tended to be normal. In the high-dose C-3 group, the liver structure was clear, with no obvious extramedullary hematopoietic foci or tumor cell infiltration, similar to the normal control group.

[0061] Please see Figure 5 To verify the toxicity of the blue calyx derivative C-3 to major organs, this experiment observed the heart, lungs, kidneys and thymus of mice in each C-3 dose group by HE staining and compared them with the normal control group and the model group.

[0062] (1) The myocardial fibers of mice in each group were neatly arranged with clear striations, and the myocardial cells were normal in morphology. No myocardial cell degeneration, necrosis or interstitial inflammatory cell infiltration was observed. There was no significant difference in cardiac tissue between the C-3 dose groups and the normal control group.

[0063] (2) The alveolar structures of each group were intact, the alveolar septa were not thickened, no obvious inflammatory exudate or granuloma formation was observed, and the tracheal epithelium was intact. No abnormal changes were observed in the lung tissue of each group.

[0064] (3) The glomerular structure of each group was clear, the morphology of the renal tubular epithelial cells was normal, and there was no edema or inflammatory cell infiltration in the renal interstitium. There was no significant difference between the renal tissue of each C-3 dose group and the normal control group.

[0065] (4) In the normal control group, the thymic cortex and medulla structure were clear, and the cortical area was rich in lymphocytes. In the model group, the thymus was slightly atrophied due to the influence of erythroleukemia. The thymic structure in the C-3 treatment group was similar to that in the model group, and no additional damage or atrophy caused by the drug was observed.

[0066] Please see Figure 6 The statistical analysis results of the major organ coefficients shown in the figure indicate that, compared with the normal control group, the liver coefficient of the model group mice was slightly increased (due to extramedullary hematopoiesis), but there were no statistically significant differences in the liver, heart, lung, and kidney coefficients of each C-3 treatment group compared with the normal control group (P > 0.05). This suggests that C-3 does not cause swelling or atrophy of major organs within the dosage range of 1-10 mg / kg.

[0067] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments that can be applied to other fields. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. The application of a blue calyx derivative C-3 in the treatment of erythroleukemia, characterized in that, include: The structural formula of the blue calyx methyl derivative C-3 is: Among them, the blue calyx derivative is used in the treatment of Friend virus-induced erythroleukemia with anti-erythroleukemia drugs.

2. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 1, characterized in that, The blue calyx derivative C-3 exerts its anti-erythroleukemia effect by inhibiting the proliferation of erythroleukemia cells.

3. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 2, characterized in that: The erythroleukemia cells mentioned are HEL cells and K562 cells.

4. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 3, characterized in that: The blue calyx derivative C-3 showed an inhibition rate of 81.99% against HEL cells and 78.85% against K562 cells after 48 hours.

5. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 3, characterized in that: The IC50 value of the blue calyx derivative C-3 against HEL cells was 0.499±0.041 μM at 48 hours, and against K562 cells was 0.787±0.048 μM at 48 hours.

6. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 1, characterized in that: The blue calyx derivative C-3 is used to prepare a drug to reduce splenomegaly in erythroleukemia.

7. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 1, characterized in that: The blue calyx derivative C-3 is used to prepare a drug to improve anemia caused by erythroleukemia.

8. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 7, characterized in that: The anemia indicators include red blood cell count, hemoglobin, and hematocrit.

9. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 1, characterized in that: The blue calyx derivative C-3 is used to prepare drugs that reduce pathological infiltration of the spleen and liver in patients with erythroleukemia.

10. The application of the blue calyx derivative C-3 in the treatment of erythroleukemia according to claim 1, characterized in that: When the blue calyx derivative C-3 is used to prepare an anti-erythroleukemia drug, the drug is administered via intraperitoneal injection.

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