Application of cycloastragenol in preparation of medicine for treating lung adenocarcinoma with anxiety depression

Abstract

The invention relates to the field of biological medicine, and discloses application of cycloastragenol in preparation of a medicine for treating lung adenocarcinoma combined anxiety depression. According to the application, cycloastragenol is found to relieve anxiety and depression symptoms by adjusting the level of corticosterone and other stress hormones; the anti-tumor immune response is enhanced by inhibiting the depletion of CD8 + T cells; and the anti-tumor capability of the CD8 + T cells is recovered by inhibiting the expression of Fabp5, so that a new idea and a potential drug choice are provided for clinical treatment of lung adenocarcinoma combined anxiety depression.

Description

Technical Field

[0001] This application relates to the field of biomedicine, and more specifically, to the use of cycloastragaloyl alcohol in the preparation of a medicament for the treatment of lung adenocarcinoma complicated with anxiety and depression. Background Technology

[0002] Lung cancer is the most commonly diagnosed malignant cancer worldwide and a leading cause of cancer death. As the main histological subtype of lung cancer, lung adenocarcinoma has a higher incidence than squamous cell carcinoma and is still increasing. It has been reported that the prognosis of LUAD (lumbar angina pectoris) is associated with multiple risk factors, especially depression. On the one hand, a diagnosis of depression is more likely to develop into cancer. On the other hand, cancer diagnoses often involve depression and anxiety disorders, especially death anxiety, which severely impacts the quality of life and treatment outcomes for cancer patients, including chemotherapy, radiotherapy, and anti-PD-1 immunotherapy. However, current clinical practice often uses a combination of antitumor drugs and anti-anxiety / depressant drugs for these patients, which presents problems such as high risk of drug interactions, numerous side effects, and poor patient compliance.

[0003] Cycloastragalol is a monomeric component extracted from the traditional Chinese medicine Astragalus membranaceus. Studies have shown that it has immunomodulatory and anti-aging effects, but its dual role in the treatment of lung adenocarcinoma complicated with mood disorders has not yet been reported.

[0004] There is a lack of single drugs or treatments in the current technology that can both fight tumors and improve mood disorders. Therefore, developing a drug with dual efficacy is of great clinical significance and market demand. Summary of the Invention

[0005] The present invention aims to solve the problems of existing technologies, such as the need for combination therapy, large side effects, and unsatisfactory treatment effects for patients with lung adenocarcinoma and anxiety depression, and to provide a single drug component with dual therapeutic effects.

[0006] In the first aspect, this application provides the use of cycloastragalool in the preparation of a medicament for treating lung adenocarcinoma complicated with anxiety and depression. Secondly, this application provides a medicament for treating lung adenocarcinoma complicated with anxiety and depression, the medicament comprising a therapeutically effective amount of cycloastragalol, and a pharmaceutically acceptable carrier or excipient.

[0007] Furthermore, the dosage of the drug is 10-100 mg / kg per day.

[0008] Furthermore, the dosage of the drug is 20-50 mg / kg per day.

[0009] Furthermore, the dosage form of the drug is a pharmaceutically acceptable dosage form.

[0010] Furthermore, the dosage forms of the drug include oral preparations, injections, tablets, capsules, and granules.

[0011] Furthermore, the purity of cycloastragalool in the drug is not less than 95%.

[0012] Thirdly, this application provides a pharmaceutical composition comprising any of the pharmaceuticals described in the second aspect, and one or more additional antitumor pharmaceuticals.

[0013] Furthermore, this includes any of the drugs described in the second aspect, as well as PD-1 inhibitors.

[0014] In summary, this application has the following beneficial effects: 1. Dual therapeutic effects: A single drug simultaneously possesses anti-tumor and anti-anxiety / depressant functions, simplifying the treatment plan; 2. High safety: Derived from natural products, with few side effects and good patient compliance; 3. Mechanism is clear: by inhibiting CD8 + It works through two pathways, namely T cell depletion and the secretion of hormones that regulate the HPA axis, and is supported by sufficient scientific evidence. 4. Broad clinical application prospects: It can be used as adjuvant or first-line treatment for lung adenocarcinoma complicated with mood disorders. For example, cycloastragalool can be used in combination with other anti-tumor drugs (anti-PD-1, etc.). Attached Figure Description

[0015] Figure 1 Chemical structure and flowchart. A: Chemical structure diagram of cycloastragaloyl alcohol; B: Flowchart of animal experiments.

[0016] Figure 2 Diagrams of the open field and black-and-white box experiments. AB: Mouse open field experiment trajectory diagram and results statistics; CD: Mouse black-and-white box experiment trajectory diagram and results statistics; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0017] Figure 3 Changes in the level of the stress hormone corticosterone in the peripheral blood of mice. A: Mouse body weight; B: Concentration of corticosterone in the peripheral blood of mice as detected by ELISA; *p<0.05, **p<0.01, ***p<0.001.

[0018] Figure 4 The antitumor effect of cycloastragaloside was verified using a mouse lung adenocarcinoma model combined with a chronic restraint stress model. A: Mouse tumor volume; B: Mouse tumor weight; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0019] Figure 5Cycloastragalol can restore the anti-PD-1 anti-tumor effect in a stressed mouse model. A: Mouse tumor volume; B: Mouse tumor volume; ns: p ≥ 0.05, **p < 0.01, ****p < 0.0001.

[0020] Figure 6 Flow cytometry detection of CD3 in mouse subcutaneous tumors + and CD8 + The proportion of T cells (A), and the depleted CD8 + T cells (B) and cytotoxic CD8 + The proportion of T cells (C); *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0021] Figure 7 : Exploring the inhibition of CD8 by cycloastragaloside using Smart transcriptome sequencing + Mechanisms of T cell exhaustion. A: Smart transcriptome sequencing workflow; B: Volcano plot of differentially expressed genes in stress group vs. control group; C: Single-cell sequencing database and CD8+ in stress group vs. control group mice. + T-cell differential gene intersection diagram; D: CD8 in three groups of mice + T-cell differential gene heatmap; E: QRT-PCR verification of CD8 in three groups of mice + The expression level of Fap5 in T cells.

[0022] Figure 8 Flow cytometry detection of CD8 + The percentage of T cells exhausted; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0023] Figure 9 Flow cytometry detection of CD8 + The percentage of T cells exhausted; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0024] Figure 10 HE staining of mouse liver and kidney, scale bar: 100 μm. Detailed Implementation

[0025] The technical solutions and effects of this application will be further described in detail below with reference to the embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely for explaining the invention and are not intended to limit the invention.

[0026] This invention provides the application of cycloastragalool in the preparation of a drug for treating lung adenocarcinoma complicated with anxiety and depression, the specific technical solution of which includes: 1. Active pharmaceutical ingredient: Cycloastragenol (CAG), its chemical structure is as follows: Figure 1 As shown in A in the figure. The purity of cycloastragaloyl alcohol used in the following examples is not less than 95%.

[0027] 2. Pharmaceutical formulations: It can be prepared into pharmaceutically acceptable dosage forms such as oral preparations, injections, tablets, capsules, and granules.

[0028] 3. Administration method and dosage: The daily dosage is 10-100 mg / kg, preferably 20-50 mg / kg, which can be adjusted according to the patient's weight and condition.

[0029] 4. Mechanism of action: It alleviates anxiety and depressive symptoms by regulating the levels of stress hormones such as cortisol; By inhibiting CD8 + T cell depletion enhances anti-tumor immune responses; CD8 was restored by inhibiting Fap5 expression. + The anti-tumor ability of T cells.

[0030] Example (1) Verification of the antidepressant effect of cycloastragalool in a mouse lung adenocarcinoma model combined with a chronic restraint stress model After one week of acclimatization, each mouse was subcutaneously injected with 1×10 6 A mouse lung adenocarcinoma model was established using Lewis lung adenocarcinoma cells. Subcutaneous tumor size was measured every 3 days using calipers; tumor volume = (length × width × width) / 2. Starting from the second day after subcutaneous tumor inoculation, mice were restrained with a confinement container and deprived of food and water for 3 hours daily for 21 consecutive days to establish a mouse model of anxiety and depression. The model was established when the average tumor volume reached 50 mm². 3 Mice were administered 100 μl of 20 and 40 mg / kg cycloastragalool and PBS solution by gavage once daily for 14 days; and 100 μl of a PD-1 inhibitor (2 μg / μl) was injected intraperitoneally once daily for 5 days. After the chronic restraint stress ended, the mice's anxiety-depression-like behaviors were assessed using the open field test and the black-and-white box test (animal experiment flowchart shown in...). Figure 1 (As shown in B in the diagram).

[0031] In the open field experiment, the total distance traveled, the distance traveled in the central region, and the number of times mice entered the central region were all significantly lower in the stress group than in the control group. After treatment with cycloastragalool, all of the above indicators were significantly improved. Figure 2Similarly, in the black-and-white box experiment, the number of times mice entered the white box and the time they stayed in the white box were significantly reduced compared to the control group. After treatment with cycloastragalool, the number of times mice entered the white box and the time they stayed in the white box were significantly increased. Figure 2 (CD in the text). Therefore, cycloastragalool can improve the anxiety and depression in mice with lung adenocarcinoma.

[0032] (2) Changes in the level of the stress hormone corticosterone in peripheral blood were detected by ELISA. In addition, the level of the stress hormone corticosterone in the peripheral blood of mice in the stress group was significantly higher than that in the control group, and the weight of mice increased after treatment with cycloastragalool. Figure 3 In A), the level of corticosterone secretion was significantly reduced ( Figure 3 (B in the middle).

[0033] (3) The antitumor effect of cycloastragalool was verified by combining a mouse lung adenocarcinoma model with a chronic restraint stress model. Compared with the control group, stress significantly promoted the growth of lung adenocarcinoma in mice. After treatment with cycloastragalol, tumor growth was significantly inhibited, and this inhibitory effect was enhanced with increasing cycloastragalol concentration. Figure 4 (AB in the middle).

[0034] (4) Cycloastragalol can restore the anti-PD-1 anti-tumor effect in a stressed mouse model. Compared with the control group, mice treated with anti-PD-1 showed a significant reduction in subcutaneous tumor volume. Figure 5 In mice subjected to stress treatment, the anti-tumor effect of anti-PD-1 was significantly weakened (A). Figure 5 (B in the text), however, the addition of cycloastragalool significantly restored the anti-PD-1 anti-tumor effect ( Figure 5 (B in the middle).

[0035] (5) Detection of CD8 in mouse tumor tissue by flow cytometry + Changes in indicators related to T cell exhaustion function Flow cytometry analysis was used to detect changes in immune cells in subcutaneous tumors of mice. The results showed that CD8+ cells were significantly reduced after stress. + The proportion of T cells was significantly reduced, and cycloastragalool could increase local CD8+ in tumors. + T cell infiltration ( Figure 6 (A) Further examination of T cell exhaustion indicators revealed that stress significantly promoted CD8. + The expression of exhaustion markers such as PD-1 and LAG3 in T cells was significantly inhibited by cycloastragalool, which significantly suppressed CD8 expression. + T depletion ( Figure 6 (B in the text); In addition, cycloastragalool can significantly increase CD107a + and TNF-α +CD8 + The proportion of T cells, i.e., those promoting CD8 + The killing ability of T cells ( Figure 6 (C in the middle).

[0036] (6) Exploring the inhibition of CD8 by cycloastragaloside using Smart transcriptome sequencing + Mechanisms of T cell exhaustion After the animal experiments were completed, CD8 groups were isolated from subcutaneous tumors in mice. + Smart transcriptome sequencing of T cells ( Figure 7 A), to obtain the upregulated and downregulated genes after stress modeling compared with the control group ( Figure 7 (B in the text); in order to screen for stress-promoting CD8); + Genes of T cell exhaustion were identified by intersecting upregulated genes in the stress group with CD8Tex upregulated genes in two lung adenocarcinoma datasets (GSE139555 and GSE176021) from the TISCH2 single-cell sequencing database. Figure 7 (C) and then cross-referenced with genes downregulated in the astragaloside treatment group compared to the stress group, revealing that Fabp5 may be a possible inhibitor of CD8 by astragaloside. + The key to T cell exhaustion ( Figure 7 (D in the D), and verified the role of FFab5 in CD8 in stressed mice by QRT-PCR. + Upregulated in T cells, while its expression was significantly downregulated after treatment with cycloastragalool. Figure 7 (E in the text).

[0037] (7) Its interaction with CD8 was verified by in vitro overexpression of Fabp5 and flow cytometry. + The relationship between T cell exhaustion Next, to verify that cycloastragalool inhibits CD8 by downregulating the expression level of Fapp5... + To address T cell exhaustion, we constructed a Fap5-overexpressing virus and used it in vitro to target primary mouse CD8 cells. + T cells were infected with a virus to promote Fap5 overexpression, and then CD8 was detected by flow cytometry. + The study investigated the level of T cell exhaustion. Results showed that, compared to the negative control group (NC), overexpression of Fabp5 (Fabp5OE) significantly reduced PD-1 cell depletion. + LAG3 + and TIM-3 + CD8 + The proportion of T cells increased significantly, and treatment with cycloastragalool (Fabp5OE+CAG) could reverse this phenomenon. Figure 8-9 ).

[0038] (8) Cycloastragalol showed no toxic effects on the liver and kidneys of mice after in vivo administration. To assess the biosafety of CAG in mice, histopathological examination of mice revealed no significant pathological changes in the liver and kidneys. Figure 10 ), surface CAG did not cause significant liver and kidney damage under physiological conditions, and has good biocompatibility.

[0039] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. Application of cycloastragalool in the preparation of drugs for treating lung adenocarcinoma complicated with anxiety and depression.

2. A drug for treating lung adenocarcinoma complicated with anxiety and depression, characterized in that, The drug comprises a therapeutically effective amount of cycloastragalol, and a pharmaceutically acceptable carrier or excipient.

3. The drug according to claim 2, characterized in that, The dosage of the drug is 10-100 mg / kg per day.

4. The drug according to claim 3, characterized in that, The dosage of the drug is 20-50 mg / kg per day.

5. The drug according to claim 2, characterized in that, The dosage form of the drug is pharmaceutically acceptable.

6. The drug according to claim 5, characterized in that, The dosage forms of the drug include oral preparations, injections, tablets, capsules, and granules.

7. The drug according to claim 2, characterized in that, The purity of cycloastragalool in the drug is not less than 95%.

8. A pharmaceutical composition, characterized in that, It includes the drug as described in any one of claims 2-7, and one or more additional antitumor drugs.

9. The pharmaceutical composition according to claim 8, characterized in that, Includes the drug as described in any one of claims 2-7, and PD-1 inhibitors.

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