Application of artemisinin compounds in the preparation of drugs for treating renal angiomyolipoma associated with tuberous sclerosis

Abstract

The present invention belongs to the field of biomedicine technology and specifically relates to the use of artemisinin compounds in the preparation of drugs for treating tuberous sclerosis complex-associated renal angiomyolipoma. The artemisinin compounds can inhibit the growth of tuberous sclerosis complex-associated renal angiomyolipoma cells and have therapeutic efficacy in treating tuberous sclerosis complex-associated renal angiomyolipoma. Furthermore, artemisinin compounds are relatively abundant in source and relatively low in cost. Artemisinin compounds can be used to prepare drugs for treating tuberous sclerosis complex-associated renal angiomyolipoma, providing a new potential drug for the treatment of tuberous sclerosis complex-associated renal angiomyolipoma.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of artemisinin compounds in the preparation of drugs for treating tuberous sclerosis complex-related renal angiomyolipoma. Background Art

[0002] Tuberous sclerosis complex-associated renal angiomyolipoma (TSC-RAML) is a benign renal tumor composed of blood vessels, smooth muscle, and adipose tissue. Its etiology is closely linked to genetic factors. Genetic studies have shown that patients with TSC-RAML harbor mutations or abnormal expression of specific genes, TSC1 / 2. This activates the m-Tor pathway, affecting cell growth, differentiation, and proliferation. This leads to abnormal intermixing and proliferation of blood vessels, smooth muscle, and adipose tissue in the kidney, ultimately forming a hamartoma. Furthermore, endocrine disorders and imbalanced cytokine networks are also believed to play a role in the development and progression of TSC-RAML. For example, changes in estrogen levels may affect the local renal microenvironment, stimulating abnormal cell growth. Furthermore, the complex interactions between cytokines may disrupt normal cell signaling pathways, leading to an imbalance between renal cell proliferation and apoptosis.

[0003] Treatment options for tuberous sclerosis complex-associated renal angiomyolipoma depend primarily on tumor size, presenting symptoms, and the patient's overall physical condition. For small, asymptomatic tuberous sclerosis complex-associated renal angiomyolipomas, regular follow-up is recommended, with imaging studies such as ultrasound, CT, or MRI closely monitoring tumor growth. Once the tumor diameter exceeds a certain range or significant compromising symptoms develop, such as low back pain, hematuria, or compression of surrounding tissues and organs by the abdominal mass, treatment options primarily include surgery and medical therapy. mTOR inhibitors are currently key treatments for tuberous sclerosis complex-associated renal angiomyolipomas, with sirolimus and its derivative, everolimus, being the most commonly used in clinical practice. These drugs bind to the intracellular FKBP12 protein, forming a complex that directly inhibits mTOR activity, thereby blocking the mTOR signaling pathway and suppressing tumor cell proliferation, inducing apoptosis, and suppressing angiogenesis.

[0004] Although existing treatments have achieved certain results in the treatment of tuberous sclerosis complex-associated renal angiomyolipoma, many problems still exist.

[0005] Although surgical treatment can directly remove the tumor, it is highly invasive and can lead to complications such as bleeding, infection, and renal impairment. Furthermore, resection of tumors in specific locations or multiple tumors can be technically challenging, leading to incomplete resection and an increased risk of recurrence. The response to everolimus treatment varies greatly between patients. Some patients experience tumor shrinkage and symptom relief, while others are less responsive, with continued tumor growth or no improvement. This makes radical cure difficult, and discontinuation of treatment can lead to recurrence. Long-term use of everolimus not only contributes to tumor resistance and reduced efficacy, but also places a heavy financial burden on patients' families due to its high price, making adherence difficult for those with limited financial resources and impacting treatment continuity. Furthermore, it is harmful to the fetus and is contraindicated in pregnant and lactating women, limiting treatment options for specific populations. Furthermore, the drug is metabolized by the liver and kidneys, increasing the burden on the liver and kidneys and the risk of adverse reactions in patients with impaired liver and kidney function. Caution or dose adjustment is therefore necessary. In summary, everolimus has numerous shortcomings in the treatment of renal angiomyolipoma associated with tuberous sclerosis complex, and improvements or alternatives are urgently needed.

[0006] In summary, finding a more effective, safe, comprehensive and targeted treatment for tuberous sclerosis complex-related renal angiomyolipoma has important clinical significance and social value. Summary of the Invention

[0007] Based on the above technical background, the main purpose of the present invention is to provide an application of artemisinin compounds in the preparation of drugs for treating tuberous sclerosis complex-related renal angiomyolipoma, so as to overcome the deficiencies in the prior art.

[0008] To achieve the aforementioned object of the invention, the technical solutions adopted by the present invention include:

[0009] The first aspect of the present invention is to provide the use of artemisinin compounds in the preparation of drugs for treating renal angiomyolipoma associated with tuberous sclerosis.

[0010] Preferably, the artemisinin compound is artemisinin and its derivatives.

[0011] More preferably, the artemisinin compounds include artemisinin, artemether, artesunate, and dihydroartemisinin.

[0012] Preferably, the artemisinin-based compound can inhibit the growth of tuberous sclerosis complex-associated renal angiomyolipoma cells.

[0013] Preferably, the dosage form of the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma includes an oral preparation, a dressing preparation, an injection preparation or an inhalation preparation.

[0014] Preferably, the active ingredient of the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma is an artemisinin-based compound, and the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma further comprises a pharmaceutically acceptable excipient.

[0015] More preferably, the excipients include one or more of a sustained-release agent, an excipient, a solubilizer, a stabilizer, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, a surfactant, and a lubricant.

[0016] The second aspect of the present invention is to provide a drug for treating tuberous sclerosis complex-related renal angiomyolipoma, wherein the active ingredient of the drug for treating tuberous sclerosis complex-related renal angiomyolipoma is an artemisinin compound.

[0017] The beneficial effects of the present invention are:

[0018] (1) The present invention discovered that artemisinin compounds can inhibit the growth of tuberous sclerosis complex-associated renal angiomyolipoma cells. The artemisinin compounds can be used to prepare drugs for treating tuberous sclerosis complex-associated renal angiomyolipoma, providing new potential drugs for the treatment of tuberous sclerosis complex-associated renal angiomyolipoma.

[0019] (2) The artemisinin-based compounds have the efficacy of treating tuberous sclerosis complex-related renal angiomyolipoma by inhibiting the growth of tuberous sclerosis complex-related renal angiomyolipoma cells. They can not only directly inhibit the growth of tuberous sclerosis complex-related renal angiomyolipoma tumor cells, but also, as a drug that has been widely used in the treatment of malaria and has a relatively abundant source, have a low cost. Their application in the treatment of tuberous sclerosis complex-related renal angiomyolipoma will have a good cost-effectiveness advantage, bring better treatment effects and prognosis to the majority of patients, and have important clinical significance and social value. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Shows the inhibitory effect of artemisinin on tuberous sclerosis complex-associated renal angiomyolipoma cells (including control drugs);

[0021] Figure 2 The results showed that dihydroartemisinin has an inhibitory effect on renal angiomyolipoma cells associated with tuberous sclerosis complex;

[0022] Figure 3 The results showed that artemether has an inhibitory effect on renal angiomyolipoma cells associated with tuberous sclerosis complex;

[0023] Figure 4 The results show that artesunate has an inhibitory effect on tuberous sclerosis complex-associated renal angiomyolipoma cells. DETAILED DESCRIPTION

[0024] The present invention will be described in detail below, and the characteristics and advantages of the present invention will become clearer and more distinct with the following description.

[0025] The first aspect of the present invention is to provide the use of artemisinin compounds in the preparation of drugs for treating renal angiomyolipoma associated with tuberous sclerosis.

[0026] The artemisinin compounds are artemisinin and its derivatives.

[0027] Preferably, the artemisinin compounds include artemisinin, artemether, artesunate, and dihydroartemisinin.

[0028] The chemical structure of artemisinin is shown in formula (1):

[0029] Formula (1).

[0030] The chemical structural formula of artemether is shown in formula (2):

[0031] Formula (2).

[0032] The chemical structure of artesunate is shown in formula (3):

[0033] Formula (3).

[0034] The chemical structure of dihydroartemisinin is shown in formula (4):

[0035] Formula (4).

[0036] The artemisinin compound can inhibit the growth of tuberous sclerosis complex-related renal angiomyolipoma cells. That is, the artemisinin compound treats tuberous sclerosis complex-related renal angiomyolipoma by inhibiting the growth of tuberous sclerosis complex-related renal angiomyolipoma cells.

[0037] The dosage form of the drug for treating tuberous sclerosis complex-related renal angiomyolipoma includes oral preparations, dressing preparations, injection preparations or inhalation preparations.

[0038] The active ingredient of the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma is an artemisinin compound, and the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma further comprises pharmaceutically acceptable excipients.

[0039] Preferably, the excipients include one or more of a sustained-release agent, an excipient, a solubilizer, a stabilizer, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, a surfactant and a lubricant.

[0040] The second aspect of the present invention is to provide a drug for treating tuberous sclerosis complex-related renal angiomyolipoma, wherein the active ingredient of the drug for treating tuberous sclerosis complex-related renal angiomyolipoma is an artemisinin compound.

[0041] Example

[0042] The present invention is further described below through specific examples. These examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0043] Example 1 Cell culture

[0044] Quickly remove the frozen TSC-RAML (tuberous sclerosis complex-associated renal angiomyolipoma) cells from the liquid nitrogen tank, place them in a constant temperature water bath at 37°C, and shake quickly to thaw the cell suspension as quickly as possible. The shaking should be completed within 1 to 2 minutes. After thawing, immediately transfer the cell suspension to a centrifuge tube containing an appropriate amount of culture medium. The culture medium is DMEM commonly used for cell culture. 10% to 15% fetal bovine serum and double antibodies are added to the culture medium. Subsequently, centrifuge at a speed of 1000 to 1500 rpm for 3 to 5 minutes, discard the supernatant, and resuspend the precipitated cells with fresh DMEM culture medium.

[0045] TSC-RAML cells were cultured at an appropriate density (generally 1 × 10 per cm2). 5 Cells (about 100 cells) are inoculated into culture flasks or culture dishes using a specialized cell culture medium (RPMI 1640 medium supplemented with 10% fetal bovine serum, glutamine, growth factors, etc.). The cells are cultured in an incubator at 37°C, 5% carbon dioxide, and saturated humidity. The cell growth status is observed daily, and the culture medium is replaced in a timely manner according to the color change of the culture medium and the cell density to maintain normal cell growth and proliferation.

[0046] Example 2 Cell activity detection

[0047] The CCK-8 assay is primarily used to detect cell proliferation and cytotoxicity. First, AML cells in the logarithmic growth phase are trypsinized to create a single-cell suspension, and the cell concentration is adjusted by cell counting. Next, an appropriate number of cells are seeded into a 96-well plate, typically 10,000 cells per well (the specific number depends on the cell type and experimental purpose), with multiple replicates. The 96-well plate is then placed in an incubator at 37°C and 5% CO2 for a specified period of time to allow the cells to adhere. The plates were divided into artemisinin, artemether, artesunate, and dihydroartemisinin groups.

[0048] The artemisinin group was divided into 7 parallel groups, one of which did not add any substance and served as the blank group. 100nM artemisinin, 200nM artemisinin, 400nM artemisinin, 800nM artemisinin, 1600nM artemisinin, and 3200nM artemisinin were added to the remaining 6 groups, respectively. At a specific time point before the end of the culture, CCK-8 reagent was added to each well, generally about 10μL per well, gently mixed and incubated at 37°C for 1 to 4 hours. Finally, the absorbance of each well was detected at a wavelength of 450nm (reference wavelength 630nm) using an enzyme reader, and the proliferation of tuberous sclerosis-related renal angiomyolipoma cells or the degree of toxicity of tuberous sclerosis-related renal angiomyolipoma cells was analyzed by comparing the absorbance values. The test results are as follows. Figure 1 shown.

[0049] from Figure 1 It can be seen that compared with the blank group, artemisinin has an inhibitory effect on the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells, and with the increase of artemisinin concentration, its inhibitory effect on the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells gradually increases, indicating that artemisinin can inhibit the growth and proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells.

[0050] Example 3

[0051] The artemether group in Example 2 was divided into 7 parallel groups, one of which did not add any substance and served as a blank group. 100nM artemether, 200nM artemether, 400nM artemether, 800nM artemether, 1600nM artemether, and 3200nM artemether were added to the remaining 6 groups, respectively. At a specific time point before the end of the culture, CCK-8 reagent was added to each well, generally about 10μL per well, and gently mixed and incubated at 37°C for 1 to 4 hours. Finally, the absorbance of each well was detected at a wavelength of 450nm (reference wavelength 630nm) using a microplate reader, and the proliferation of tuberous sclerosis-related renal angiomyolipoma cells or the degree of toxicity of tuberous sclerosis-related renal angiomyolipoma cells was analyzed by comparing the absorbance values. The test results are as follows: Figure 2 shown.

[0052] from Figure 2 It can be seen that compared with the blank group, artemisinin has an inhibitory effect on the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells, and with the increase of artemisinin concentration, its inhibitory effect on the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells gradually increases, indicating that artemisinin can inhibit the growth and proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells.

[0053] Example 4

[0054] The artesunate group in Example 2 was divided into 7 parallel groups, one of which did not add any substance and served as a blank group. 100 nM artesunate, 200 nM artesunate, 400 nM artesunate, 800 nM artesunate, 1600 nM artesunate, and 3200 nM artesunate were added to the remaining 6 groups, respectively. At a specific time point before the end of the culture, CCK-8 reagent was added to each well, generally about 10 μL per well, and the mixture was gently mixed and incubated at 37°C for 1 to 4 hours. Finally, the absorbance of each well was detected at a wavelength of 450 nm (reference wavelength 630 nm) using an enzyme reader, and the proliferation of tuberous sclerosis-related renal angiomyolipoma cells or the degree of cytotoxicity of tuberous sclerosis-related renal angiomyolipoma cells was analyzed by comparing the absorbance values. The test results are as follows: Figure 3 shown.

[0055] from Figure 3 It can be seen that compared with the blank group, artesunate can inhibit the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells, and with the increase of artesunate concentration, its inhibitory effect on the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells gradually increases, indicating that artesunate can inhibit the growth and proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells.

[0056] Example 5

[0057] The dihydroartemisinin group in Example 2 was divided into 7 parallel groups, one of which did not add any substance and served as a blank group. 100 nM dihydroartemisinin, 200 nM dihydroartemisinin, 400 nM dihydroartemisinin, 800 nM dihydroartemisinin, 1600 nM dihydroartemisinin, and 3200 nM dihydroartemisinin were added to the remaining 6 groups, respectively. At a specific time point before the end of the culture, CCK-8 reagent was added to each well, generally about 10 μL per well, and gently mixed and incubated at 37°C for 1 to 4 hours. Finally, the absorbance of each well was detected at a wavelength of 450 nm (reference wavelength 630 nm) using an enzyme reader, and the proliferation of tuberous sclerosis-related renal angiomyolipoma cells or the degree of cytotoxicity of tuberous sclerosis-related renal angiomyolipoma cells was analyzed by comparing the absorbance values. The test results are as follows: Figure 4 shown.

[0058] Figure 4In the experiment, compared with the blank group, dihydroartemisinin could inhibit the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells, and its inhibitory effect on the proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells gradually increased with the increase of dihydroartemisinin concentration, indicating that dihydroartemisinin could inhibit the growth and proliferation of tuberous sclerosis complex-associated renal angiomyolipoma cells.

[0059] The present invention has been described in detail above with reference to specific embodiments and exemplary examples. However, these descriptions should not be construed as limiting the present invention. Those skilled in the art will appreciate that various equivalent substitutions, modifications, or improvements may be made to the technical solutions and implementations of the present invention without departing from the spirit and scope of the present invention, all of which fall within the scope of the present invention. The scope of protection of the present invention shall be determined by the appended claims.

Claims

1. The use of artemisinin compounds in the preparation of drugs for the treatment of renal angiomyolipoma associated with tuberous sclerosis complex; The artemisinin compounds include artemisinin, artemether, artesunate and dihydroartemisinin.

2. The use according to claim 1, characterized in that The artemisinin compound inhibits the growth of renal angiomyolipoma cells associated with tuberous sclerosis.

3. The use according to claim 1, characterized in that The dosage form of the drug for treating tuberous sclerosis complex-related renal angiomyolipoma includes oral preparations, dressing preparations, injection preparations or inhalation preparations.

4. The use according to claim 1, characterized in that The active ingredient of the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma is an artemisinin compound, and the drug for treating tuberous sclerosis complex-associated renal angiomyolipoma further comprises pharmaceutically acceptable excipients.

5. The use according to claim 4, characterized in that The auxiliary materials include one or more of sustained-release agents, excipients, solubilizers, stabilizers, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants and lubricants.

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