Application of AS-605240 in treatment of renal vascular smooth muscle lipoma related to nodular sclerosis
By using AS-605240 to promote apoptosis in tuberous sclerosis-associated renal angiomyolipoma cells and regulate related protein expression and signaling pathways, the limitations of existing mTOR inhibitors in efficacy and side effects have been overcome, achieving safe and effective tumor treatment.
Patent Information
- Application Number
- CN202511245715.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-02
- Publication Date
- 2025-10-21
AI Technical Summary
Existing mTOR inhibitors have limited efficacy in treating tuberous sclerosis-related renal angiomyolipoma, are expensive, have significant side effects, cannot completely eliminate lesions, and pose a risk of fatal complications.
AS-605240 was used as a therapeutic agent to promote apoptosis of tuberous sclerosis-associated renal angiomyolipoma cells, regulate the expression of pro-apoptotic protein BAX and anti-apoptotic protein BCL-2, and reduce the phosphorylation levels of AKT and PI3K. The drug was prepared into various dosage forms to achieve effective treatment.
AS-605240 can effectively inhibit tumor growth, promote cell apoptosis, reduce tumor volume, reduce the risk of complications, lower treatment costs, and provide a safe and effective treatment method.
Smart Images
Figure CN120815084A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of AS-605240 in treating tuberous sclerosis complex-related renal angiomyolipoma. Background Art
[0002] Tuberous sclerosis complex (TSC) is an autosomal dominant genetic disease that affects approximately 1 in 5,800 newborns and over 1 million people worldwide. The clinical manifestations of TSC are diverse, including skin lesions, neurological lesions, and renal involvement. For adult patients, TSC-related renal angiomyolipomas are of particular concern. Epidemiological studies have shown that 60%-80% of TSC cases will develop renal angiomyolipomas in adulthood. Although benign, tuberous sclerosis complex-related renal angiomyolipomas (TSC-RAMLs) are often large and rapidly growing, making them highly susceptible to life-threatening complications such as rupture and bleeding, renal insufficiency, and even uremia. They are the leading cause of death in adult TSC patients.
[0003] The treatment of TSC-RAML faces many difficult clinical problems. Surgery can delay the progression of renal insufficiency and reduce the risk of bleeding. However, due to the lack of knowledge of TSC-RAML among some surgeons, rashly performing surgery on patients may result in total nephrectomy or worsening of renal insufficiency. Radiofrequency ablation, arterial embolization and other methods have great limitations and are only suitable for specific cases. In recent years, several clinical trials, including EXIST, have shown that mammalian target of rapamycin (mTOR) inhibitors such as everolimus can effectively control the progression of TSC-RAML. mTOR inhibitors can inhibit the overactivation of mTORC1 caused by TSC gene mutations. mTOR inhibitors have become the first-line treatment for the disease. However, the current first-line treatments used for inhibitors targeting a single mTOR target, such as rapamycin and everolimus, still have many problems: Limited efficacy: mTOR inhibitors lack cytotoxicity, resulting in only partial reduction of renal lesions, not complete elimination. Later, fatal complications such as bleeding and renal failure can occur. Expensive: The high cost of these drugs necessitates lifelong medication (disease relapses upon discontinuation), placing a heavy financial burden on patients. Significant side effects: mTOR receptors are widely expressed in the human body, supporting many critical functions, including cell proliferation and metabolism. Taking mTOR inhibitors is a double-edged sword for TSC patients, often causing numerous intolerable side effects that force many patients to discontinue treatment.
[0004] In summary, there is an urgent need to find a more effective, safe and non-mTOR target drug for the treatment of tuberous sclerosis complex-associated renal angiomyolipoma. Summary of the Invention
[0005] The purpose of the present invention is to provide the use of AS-605240 in the treatment of tuberous sclerosis complex-related renal angiomyolipoma, providing an effective and safe treatment method for clinical application.
[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides the use of AS-605240 in preparing a medicine for treating tuberous sclerosis complex-related renal angiomyolipoma.
[0007] The present invention also provides the use of AS-605240 in preparing an apoptosis promoter for renal angiomyolipoma cells associated with tuberous sclerosis.
[0008] The present invention also provides the use of AS-605240 in preparing a reagent for regulating the expression of a pro-apoptotic protein BAX and / or a reagent for regulating the expression of an anti-apoptotic protein BCL-2.
[0009] The present invention also provides the use of AS-605240 in preparing a reagent for regulating the phosphorylation level of AKT and / or PI3K in tuberous sclerosis complex-associated renal angiomyolipoma cells.
[0010] The present invention also provides use of a composition in preparing a drug for treating tuberous sclerosis complex-related renal angiomyolipoma, wherein the composition comprises AS-605240 and a pharmaceutically acceptable excipient.
[0011] Preferably, the pharmaceutically acceptable excipient is selected from 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 or a lubricant.
[0012] The present invention also provides an application of a composition in preparing a drug for treating tuberous sclerosis complex-related renal angiomyolipoma, wherein the composition comprises AS-605240 and other therapeutically active ingredients.
[0013] Preferably, the other therapeutically active ingredients include everolimus, sirolimus or FGFR inhibitors.
[0014] Preferably, the composition is in the form of an oral preparation, a dressing preparation, an injection preparation or an inhalation preparation.
[0015] Preferably, the chemical structure of AS-605240 is as shown in Formula I:
[0016] Formula I.
[0017] Beneficial effects of the present invention: The present invention found that AS-605240 can directly inhibit the growth of tuberous sclerosis complex-associated renal angiomyolipoma cells. AS-605240 can be used to prepare anti-tuberous sclerosis complex-associated renal angiomyolipoma drugs, providing a new potential drug for the treatment of tuberous sclerosis complex-associated renal angiomyolipoma.
[0018] The present invention discovered that AS-605240 increases the protein levels of the pro-apoptotic gene BAX and decreases the protein levels of the anti-apoptotic protein BCL-2 in tuberous sclerosis complex-associated renal angiomyolipoma cells. It also reduces the phosphorylation levels of AKT and PI3K in these cells. This suggests that AS-605240 can promote apoptosis in tuberous sclerosis complex-associated renal angiomyolipoma cells through PI3K / AKT, a non-mTOR target, thereby treating tuberous sclerosis complex-associated renal angiomyolipoma. AS-605240 has a significant therapeutic and ameliorative effect on tuberous sclerosis complex-associated renal angiomyolipoma.
[0019] AS-605240 is abundant in source. Its application in the treatment of tuberous sclerosis complex-associated renal angiomyolipoma can reduce the cost of the drug, making the drug have both good efficacy and low cost. It has important clinical significance and social value for improving and treating tuberous sclerosis complex-associated renal angiomyolipoma. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 This is a line graph showing the inhibitory effect of AS-605240 on tuberous sclerosis complex-associated renal angiomyolipoma cells; Figure 2The figure shows the test results of the effect of AS-605240 on apoptosis markers; Figure 3 This is a test result chart showing the effect of AS-605240 on cycle markers. DETAILED DESCRIPTION
[0021] The present invention provides the use of AS-605240 in the preparation of a medicament for treating tuberous sclerosis complex-associated renal angiomyolipoma. The uses described herein encompass all therapeutic agents prepared using AS-605240 for treating tuberous sclerosis complex-associated renal angiomyolipoma. In this invention, "treatment" encompasses intervention at all disease stages and with different therapeutic targets, including but not limited to: early disease stages (e.g., intervention for small-volume AML), progressive disease stages (stabilization or shrinkage of enlarging AML), late disease stages (e.g., management of larger AML with risk of bleeding or rupture, or in cases of pre-existing renal impairment), adjuvant therapy after surgery or interventional therapy (preventing recurrence or delaying regrowth), and preventive measures (e.g., delaying the development or early progression of AML in high-risk patients). The "therapeutic" activity is specifically manifested in various beneficial effects, including relief of tumor-related symptoms (e.g., pain, hematuria), inhibition of tumor growth, stabilization of tumor size, promotion of tumor regression, reduction of tumor risk, prevention of complications (e.g., spontaneous rupture and bleeding), and improvement of tumor-related renal impairment.
[0022] The present invention also provides the use of AS-605240 in the preparation of an agent that promotes apoptosis in tuberous sclerosis complex-associated renal angiomyolipoma cells. This application specifically relates to the use of AS-605240 to prepare a formulation that can specifically induce programmed cell death (i.e., apoptosis) in tuberous sclerosis complex-associated renal angiomyolipoma (AML) cells. The agent is intended to increase the apoptosis rate of various abnormal cell types within tumor tissues, including but not limited to smooth muscle-like cells, adipocytes, abnormal vascular endothelial cells, and their precursors within lesions. This apoptosis-promoting effect encompasses all effects that enhance the initiation and execution of apoptosis, such as activating key nodes in the apoptotic signaling pathway, disrupting the tumor cell's intrinsic survival signaling network, and eliminating factors that inhibit apoptosis. Ultimately, this results in the selective elimination of tumor cells, thereby inhibiting tumor growth, reducing tumor size, and even inducing tumor regression.
[0023] The present invention also provides the use of AS-605240 in the preparation of reagents for regulating the expression of the pro-apoptotic protein BAX and / or the anti-apoptotic protein BCL-2. This application encompasses the use of AS-605240 to prepare reagents capable of effectively regulating the expression levels or functional states of the key apoptosis-regulating proteins BAX and / or BCL-2 in tuberous sclerosis complex-associated renal angiomyolipoma cells. "Regulation" specifically refers to upregulating the expression level or pro-apoptotic activity of the pro-apoptotic protein BAX and / or downregulating the expression level or anti-apoptotic function of the anti-apoptotic protein BCL-2. Regulation can occur at multiple levels, including modulating the transcriptional activity of related genes, affecting mRNA stability and translation efficiency, altering protein stability and degradation rates, affecting protein subcellular localization, or regulating post-translational modification (e.g., phosphorylation). This regulation of the BAX / BCL-2 balance (increasing the ratio of pro-apoptotic to anti-apoptotic proteins) is one of the key molecular mechanisms for inducing tumor cell apoptosis and achieving therapeutic effects.
[0024] The present invention also provides the use of AS-605240 in preparing a reagent for regulating the phosphorylation levels of AKT and / or PI3K in tuberous sclerosis complex-associated renal angiomyolipoma cells. The present invention relates to the use of AS-605240 to prepare a regulatory reagent capable of effectively intervening in the activity of the phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT) signaling pathway in tuberous sclerosis complex-associated renal angiomyolipoma (AML) cells.
[0025] The present invention also provides a use of a composition comprising AS-605240 and a pharmaceutically acceptable excipient in the preparation of a medicament for treating renal angiomyolipoma associated with tuberous sclerosis complex. The composition comprises AS-605240 and a pharmaceutically acceptable excipient. The use of the composition comprises a therapeutically effective amount of AS-605240 and one or more pharmaceutically acceptable excipients to prepare a medicament for treating renal angiomyolipoma associated with tuberous sclerosis complex. The functions of the pharmaceutically acceptable excipients include excipient, stabilization, solubilization, controlled release, improved absorption or facilitation of formulation molding, etc., and can be widely selected and combined according to the desired pharmaceutical dosage form and administration route. Its categories include, but are not limited to: sustained-release agents (such as poly(lactic-co-glycolic acid) copolymer PLGA, ethyl cellulose), excipients / fillers (such as lactose, microcrystalline cellulose, starch, calcium phosphate), solubilizers (such as Tween surfactants, cyclodextrins), stabilizers (such as antioxidants such as ascorbic acid, EDTA chelating agents), binders (such as hydroxypropyl methylcellulose HPMC, povidone PVP), wetting agents (such as sodium lauryl sulfate), disintegrants (such as cross-linked carboxymethyl cellulose sodium croscarmellosesodium), absorption enhancers (such as bile acid derivatives), surfactants (such as lecithin), lubricants (such as magnesium stearate, talc), and other possible functional excipients such as colorants, flavoring agents or preservatives.
[0026] The present invention also provides a composition for use in preparing a drug for treating tuberous sclerosis complex-associated renal angiomyolipoma, comprising AS-605240 and other therapeutically active ingredients. This application involves using a composition comprising a therapeutically effective amount of AS-605240 and at least one other therapeutically active pharmaceutical ingredient (i.e., a combination drug) to prepare a drug for treating tuberous sclerosis complex-associated renal angiomyolipoma. The "other therapeutically active ingredients" are intended to achieve synergistic, additive, or sequential therapeutic benefits and preferably include mTOR pathway inhibitors (such as everolimus and sirolimus) and FGFR (fibroblast growth factor receptor) signaling pathway inhibitors. Furthermore, the combination strategy may also include (but is not limited to) other signaling pathway inhibitors (such as MEK inhibitors, Hedgehog pathway inhibitors), angiogenesis inhibitors (such as VEGF / VEGFR inhibitors), immunomodulators (such as immune checkpoint inhibitors), or other pharmaceutical ingredients related to TSC / AML pathogenesis or with synergistic potential (such as other classes of PI3K inhibitors or HSP90 inhibitors). This combination approach aims to enhance efficacy, overcome potential drug resistance, reduce the dose of each single drug to reduce side effects, or achieve synergistic blockade of multiple disease mechanism pathways.
[0027] Preferably, the composition is in the form of an oral preparation, dressing, injection, or inhalation formulation. The aforementioned composition of the present invention (comprising AS-605240 and excipients or combination drugs) can be formulated into a variety of dosage forms suitable for different routes of administration, preferably including: oral preparations (e.g., tablets, including immediate-release tablets, sustained-release tablets, controlled-release tablets, and orally disintegrating tablets; capsules, including hard capsules and soft capsules; granules, suspensions, and solutions); injections (e.g., sterile injections, including aqueous injections and solutions containing solubilizers / cosolvents; suspension injections; lyophilized powder injections for pre-treatment preparation; and large-volume infusions for intravenous infusion); topical dressings (e.g., gels, ointments, creams for topical application or via catheter / interventional techniques, as well as transdermal patches, or implants suitable for local sustained-release administration); and inhalation formulations (e.g., solutions or suspensions for nebulized inhalation, metered-dose inhalers, and dry powder inhalers). The choice of the specific dosage form depends on the drug's properties, therapeutic needs (e.g., systemic or localized), patient factors, and administration convenience.
[0028] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0029] Example Primary TSC-RAML (tuberous sclerosis complex-associated renal angiomyolipoma) cells (self-collected in the laboratory) were seeded into culture dishes and divided into a control group and an AS-605240-treated group. HEK293T normal kidney cells were obtained from the cell bank of Peking Union Medical College and seeded into culture dishes. Cells were maintained under standard culture conditions (37°C, 5% CO2, humidified atmosphere).
[0030] The following operations were performed on the blank group (no AS-605240 added) and the AS-605240-treated group: After the cells grew well and the growth density was about 60%, the AS-605240-treated group and the blank group were given the same volume of 1640 culture medium containing 10% serum and treated for 24 hours at the same time. The AS-605240-treated group was divided into 7 groups. After 24 hours of treatment, AS-605240 solutions with concentrations of 0.1 μmol / mL, 0.3 μmol / mL, 0.5 μmol / mL, 1 μmol / mL, 3 μmol / mL, 5 μmol / mL, and 10 μmol / mL were added to the 7 groups of the AS-605240-treated group. The cell culture medium was aspirated, 100 μL of CCK8 solution was added to each well, and the cells were incubated for another 2 hours. The test results of the inhibitory effect of the blank group and the AS-605240-treated group on tuberous sclerosis-related renal angiomyolipoma cells are shown in the figure. Figure 1 shown.
[0031] from Figure 1As can be seen in the data, RAML cells were significantly more sensitive to AS-605240 compared to HEK293T controls (IC50: 7.8 μM vs. 11.3 μM, p<0.001), indicating selective cytotoxicity against RAML cells. Furthermore, the inhibitory effect of AS-605240 on the proliferation of TSC-associated renal angiomyolipoma cells gradually increased with increasing concentrations, reaching its strongest inhibitory effect at 10 μmol / mL. These results indicate that AS-605240 can inhibit the growth and proliferation of TSC-associated renal angiomyolipoma cells, with the strongest inhibitory effect at 10 μmol / mL.
[0032] The blank group and AS-605240-treated groups (5 μmol / mL and 10 μmol / mL) were lysed with protein lysis buffer to extract protein, or RNA extraction reagent was used to extract protein or mRNA. Western blotting or quantitative PCR were used to detect the gene expression and protein content of the pro-apoptotic gene BAX and the anti-apoptotic marker BCL-2 in each group. A549 cells were used to extract proteins with 8M urea, and protease inhibitors (purchased from Biyuntian Biotechnology Co., Ltd.) were added to the lysate. The BCA method was used for quantification, and equal amounts were loaded for polyacrylamide electrophoresis. The internal reference protein actin was used as a correction. After electrophoresis, the membrane was transferred and blocked with 5% skim milk powder at room temperature for one hour. The antibody was purchased from Wuhan Aibotek Biotechnology Co., Ltd. at a dilution concentration of 1:1000. 1% skim milk powder was used overnight. The cells were washed three times the next day, each time with 5% PBST5 for 5 minutes. Then, HRP-labeled secondary antibodies were incubated with 1% skim milk powder at room temperature for one hour, washed three times, each time with 5% PBST5 for 5 minutes. ECL reagent was used for luminescence, and photos were taken and saved for analysis. The detection results of apoptosis markers are as follows. Figure 2 As shown in the figure, the detection results of cycle markers are as follows Figure 3 shown.
[0033] from Figure 2As can be seen, compared with the blank group, the AS-605240 group showed higher Bax protein expression levels, and this expression gradually increased with increasing AS-605240 concentration. This suggests that AS-605240 can increase the protein expression levels of pro-apoptotic markers. Compared with the blank group, the AS-605240 group showed lower BCL-2 protein expression levels, and this expression gradually decreased with increasing AS-605240 concentration. This suggests that AS-605240 can reduce the protein expression levels of anti-apoptotic markers. This further suggests that AS-605240 can promote apoptosis in tuberous sclerosis complex-associated renal angiomyolipoma cells.
[0034] from Figure 3 It can be seen that compared with the blank group, the phosphorylation levels of AKT and PI3K in the AS-605240 group were reduced, and AS-605240 can reduce the phosphorylation levels of AKT and PI3K in tuberous sclerosis-related renal angiomyolipoma cells.
[0035] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. Application of AS-605240 in the preparation of a drug for the treatment of tuberous sclerosis complex-associated renal angiomyolipoma.
2. Application of AS-605240 in the preparation of apoptosis promoter for renal angiomyolipoma cells associated with tuberous sclerosis complex.
3. Application of AS-605240 in the preparation of a reagent for regulating the expression of the pro-apoptotic protein BAX and / or a reagent for regulating the expression of the anti-apoptotic protein BCL-2.
4. Application of AS-605240 in the preparation of reagents for regulating the phosphorylation levels of AKT and / or PI3K in tuberous sclerosis complex-associated renal angiomyolipoma cells.
5. Use of a composition in the preparation of a drug for treating renal angiomyolipoma associated with tuberous sclerosis, characterized in that: The composition comprises AS-605240 and pharmaceutically acceptable excipients.
6. The use according to claim 5, characterized in that The pharmaceutically acceptable excipient is selected from 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 promoter, a surfactant or a lubricant.
7. Use of a composition in the preparation of a drug for treating renal angiomyolipoma associated with tuberous sclerosis, characterized in that: The composition comprises AS-605240 and other therapeutically active ingredients.
8. The use according to claim 7, characterized in that The other therapeutically active ingredients include everolimus, sirolimus or FGFR inhibitors.
9. The use according to any one of claims 5 to 8, characterized in that The dosage form of the composition is an oral preparation, a dressing preparation, an injection preparation or an inhalation preparation.
10. The use according to any one of claims 1 to 4, characterized in that The chemical structure of AS-605240 is shown in Formula I: Formula I.