Use of daphnetin in preparation of a drug for treating thyroid nodules

Abstract

The application discloses a use of daphnetin in preparation of a drug for treating thyroid nodules, relates to the field of medicines, and particularly relates to a new use of a drug, and can be used for treating thyroid nodules and has a wide application prospect.

Description

[0001] This disclosure claims priority to Chinese Patent Application No. 2025107913358, filed on June 13, 2025, entitled "Use of Daphne in a Drug for the Treatment of Thyroid Nodules", the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of medicine, particularly to the field of new uses of pharmaceuticals, and especially to the use of daphne in the preparation of thyroid nodule treatment drugs. Background Technology

[0003] Thyroid nodules refer to scattered lesions caused by abnormal local growth of thyroid cells. Most patients with thyroid nodules are asymptomatic. However, when combined with thyroid dysfunction, corresponding clinical manifestations may appear. Some patients experience compression symptoms such as hoarseness, a feeling of pressure in the air, and difficulty breathing / swallowing due to the nodules compressing surrounding tissues. Thyroid nodules are usually discovered during a physical examination, followed by evaluation to determine their benign or malignant nature. According to the 2012 Guidelines for the Diagnosis and Treatment of Thyroid Nodules and Differentiated Thyroid Cancer, the clinical evaluation of thyroid nodules in adults includes taking a medical history, physical examination, TSH measurement, and ultrasound examination. Patients with thyroid nodules and serum thyroid-stimulating hormone (TSH) levels below normal have a lower proportion of malignant nodules compared to those with normal or elevated TSH levels.

[0004] Thyroid nodules can progress to thyroid cancer (TC) if they become malignant. Of TC, 90% are differentiated thyroid cancer (DTC), which originates from thyroid follicular epithelial cells and mainly includes approximately 80% papillary thyroid carcinoma (PTC) and approximately 20% follicular thyroid carcinoma (FTC). DTC generally has a better prognosis and longer survival. Treatment methods for DTC mainly include surgical resection, radioactive iodine therapy, and TSH suppression therapy. Thyroid cancer also includes approximately 4% medullary thyroid carcinoma (MTC) and undifferentiated thyroid carcinoma (ATC). ATC grows rapidly, is prone to early metastasis, and has a poor response to treatment, resulting in a very poor prognosis; patients often survive no more than six months. The malignancy of MTC is between that of DTC and ATC.

[0005] Malignant progression of thyroid nodules can also lead to problems such as enlargement of the thyroid mass, vocal cord paralysis, endocrine disorders, and metastasis to nearby and distant organs and tissues.

[0006] To prevent thyroid nodules from becoming malignant, more medications are still needed to treat them.

[0007] Daphne, chemically named 7,8-dihydroxycoumarin, also known as Daphne acetonide, with the chemical formula C9H6O4, is a natural coumarin derivative. It is currently mainly used as an adjunct treatment for thromboangiitis obliterans and other occlusive vascular diseases and coronary heart disease. In addition, daphne is also used to treat rheumatoid arthritis. Summary of the Invention

[0008] The main objective of this invention is to provide new pharmaceutical uses for daphne.

[0009] To achieve the above objectives, this invention proposes the use of daphne in the preparation of thyroid nodule treatment drugs.

[0010] Daphne is a natural coumarin-type active ingredient, mainly derived from plants in the Thymelaeaceae family, such as Daphne genkwa and Edgeworthia chrysantha, as well as other plants in the Thymelaeaceae family. Its extraction typically involves several steps: raw material pretreatment, extraction, separation and purification, and component identification, ultimately yielding daphne as the active pharmaceutical ingredient.

[0011] Previous studies, such as the paper "Daphne triggers ROS-induced ovarian cancer cell death and protective autophagy by regulating the AMPK / Akt / mTOR pathway" published in the journal Phytomedicine, described how daphne exerts its anti-tumor effect by inducing ROS-dependent apoptosis in ovarian cancer cells, and how N-acetylcysteine ​​(NAC) can reverse this effect. The AMPK / Akt / mTOR pathway is involved in DAPH-mediated protective autophagy. When DAPH-mediated AMPK expression and autophagy are blocked, cell proliferation is strongly inhibited and apoptosis is induced. The journal *Biomedicine & pharmacotherapy* also reported the inhibitory effect of daphne in breast cancer treatment in its paper "Daphne alleviates 7,12-dimethylbenzo[a]phenanthrene-induced breast cancer via NRF-2-Keap1 and NF-κB pathways." Daphne's anticancer mechanism and multi-target anti-inflammatory effects make it valuable for drug development. Currently, there are no reports on the effects of daphne on thyroid nodules. This invention, through rat experiments, found that daphne exhibits good therapeutic effects on thyroid nodules in rats and can be applied as a promising drug for the treatment of thyroid nodules.

[0012] Preferably, the structural formula of daphne is: .

[0013] The present invention also provides a drug for treating thyroid nodules.

[0014] Specifically, medications for treating thyroid nodules contain thymol and pharmaceutically acceptable salts.

[0015] Or, to be more specific, medications for treating thyroid nodules include daphne and pharmaceutically acceptable excipients.

[0016] More preferably, the thyroid nodule treatment drug is an in vivo treatment drug, and the effective dose of the active ingredient daphne contained in the thyroid nodule treatment drug is 150~1350 mg / day.

[0017] More preferably, the thyroid nodule treatment drug is an in vivo treatment drug, and the dosage form of the thyroid nodule treatment drug is selected from: oral dosage form or injectable dosage form. It is used to deliver thymol, as the active ingredient, into the body. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 Ultrasound imaging of the thyroid gland in each group of rats (A. transverse section B. longitudinal section); Figure 2 Quantitative graphs of left and right thyroid gland volumes in rats from different treatment groups (in the figure, **** indicates p<0.0001). Figure 3 HE staining results of thyroid glands in rats from different treatment groups (green arrows indicate thyroid follicles, and red arrows indicate thyroid follicular epithelial cells). Figure 4 A visual image of the thyroid glands of rats in each group; Figure 5 The graph shows the relative mass of the thyroid gland in rats under different treatment groups (in the graph, **** indicates p<0.0001). Figure 6 Figures showing the changes in body weight of rats in different treatment groups before and after treatment. Figure 7 Quantitative graphs of FT3, FT4 and TSH concentrations in the serum of rats in different treatment groups (in the figure, * indicates p<0.05, **** indicates p<0.0001). Figure 8Quantitative graphs of serum TG, TRH, and TPO concentrations in rats under different treatment groups (in the graph, * indicates p<0.05, ** indicates p<0.01, and **** indicates p<0.0001). Figure 9 A quantitative graph showing the concentration of serum inflammatory factors in rats under different treatment groups (in the graph, * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001, and **** indicates p<0.0001).

[0020] The realization of the purpose, functional features and advantages of this application will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0021] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the 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. In addition, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

[0022] This application provides a drug for treating thyroid nodules.

[0023] In a specific embodiment, the thyroid nodule treatment drug includes daphne and pharmaceutically acceptable excipients, typically including: diluents, wetting agents, binders, disintegrants, lubricants, antioxidants, solubilizers, fillers, buffers, stabilizers, solubilizers, flavoring agents, coloring agents, etc. Different excipients are selected and formulated according to different dosage forms, and appropriate proportions are adjusted during production. The dosage form of the thyroid nodule treatment drug is usually the same as that of an in vivo therapeutic drug, and the specific dosage form selection is not limited.

[0024] In a specific embodiment, the thyroid nodule treatment drug includes daphne and a pharmaceutically acceptable salt, such as using metal hydroxides such as sodium or potassium or alkaline substances such as organic bases to neutralize the acidic groups of daphne to form the corresponding salt, so as to improve the physicochemical properties or pharmacokinetic characteristics.

[0025] In a preferred embodiment, the thyroid nodule treatment drug is an in vivo treatment drug, and the dosage form of the thyroid nodule treatment drug is selected from: oral dosage form or injectable dosage form.

[0026] In a specific embodiment, the oral dosage form is selected from any one of the following: tablets, capsules, pills, granules, powders, drops, oral films, and oral liquid preparations.

[0027] In a preferred embodiment, the thyroid nodule treatment drug is an in vivo treatment drug, and the effective dose range of the active ingredient daphne contained in the thyroid nodule treatment drug is 150~1350 mg / day. In specific embodiments, the effective dose can be 150 mg / day, 175 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 400 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, or 1200 mg / day.

[0028] This application provides an example of the use of daphne in the preparation of a thyroid nodule treatment drug.

[0029] In the following specific embodiments, the daphne used was purchased from Jilin Xidian Pharmaceutical Technology Development Co., Ltd. (CAS No.: 486-35-1). The structural formula of the daphne used in this application embodiment is: .

[0030] This experiment used SPF-grade SD rats as the research subject to study the therapeutic effect of daphnetin at different doses on a rat model of nodular goiter. The therapeutic efficacy of daphnetin on nodular goiter was evaluated by observing changes in thyroid pathology and blood biochemical indicators after treatment with different doses.

[0031] This experiment mainly focused on three stages: the construction of a rat model of nodular goiter, the treatment with different doses of daphne and control drugs, and the evaluation of the efficacy of daphne on rats with nodular goiter.

[0032] Experimental methods 1. Establishment and treatment of experimental animal models: 72 SD rats were randomly divided into 6 groups (n=12 per group). The groups included a blank control group, a model group, a high-dose daphne group, a medium-dose daphne group, a low-dose daphne group, and a levothyroxine sodium positive drug (Euthyrox) group. During the modeling phase, except for the blank control group rats which were administered an equal volume of physiological saline by gavage, the other groups were administered 1 mL·100 g of saline solution. -1 ·d -1The dosage of the drug administered via gavage was 0.1% PTU solution, and all groups were administered the drug continuously for 8 weeks. From week 9, drug treatment began. Rats in each group were administered different doses of powder suspensions via gavage according to their body weight. The daphne powder was fully dissolved in polyethylene glycol 400 (PEG 400) and vortexed to prepare a clear and homogeneous drug solution. The specific dosages were set as follows: high-dose daphne group (powder dose) 100 mg / kg, medium-dose daphne group (powder dose) 50 mg / kg, and low-dose daphne group (powder dose) 25 mg / kg, once daily. Levothyroxine sodium tablets (clinical standard preparation) were ground and fully dissolved in 0.9% sodium chloride injection (physiological saline) and vortexed to prepare a clear and homogeneous drug solution. The specific dosage was set as follows: levothyroxine sodium tablets positive control group (powder dose) 15 μg / kg, once daily. The model group rats were administered the same volume of physiological saline via gavage. Four weeks after treatment, the rats were euthanized and samples were collected for subsequent experimental index testing.

[0033] Table 1. Dosage of drugs administered to rats in each group 2. Experimental instruments and reagents Electronic balance; Multifunctional ELISA reader; Low-temperature high-speed centrifuge; Physiological sodium chloride solution (Sichuan Kelun Pharmaceutical Co., Ltd.); Propylthiouracil tablets (Shanghai Zhaohui Pharmaceutical Co., Ltd.); Euthyrox (Merck Pharmaceuticals (Jiangsu) Co., Ltd.); HE staining kit (Shanghai Beyotime Biotechnology Co., Ltd.); Free triiodothyronine (FT3) ELISA kit (Nanjing Boyan Biotechnology Co., Ltd.); Free thyroxine (FT4) ELISA kit (Nanjing Boyan Biotechnology Co., Ltd.); Thyroid-stimulating hormone (TSH) ELISA kit (Nanjing Boyan Biotechnology Co., Ltd.); Thyroglobulin (TG) ELISA kit (Nanjing Boyan Biotechnology Co., Ltd.); Thyrotropin-releasing hormone (TRH) ELISA kit (Nanjing Boyan Biotechnology Co., Ltd.); Thyroid peroxidase (TPO) ELISA kit (Nanjing Boyan Biotechnology Co., Ltd.); Rat IL-1 beta ELISA Kit (Youke Life Science Technology (Hangzhou) Co., Ltd.); Rat IL-6 ELISA Kit (Youke Life Science Technology (Hangzhou) Co., Ltd.); Rat TNF alpha ELISA Kit (Youke Life Science Technology (Hangzhou) Co., Ltd.); 4% Paraformaldehyde (Eubia (Shanghai) Biotechnology Co., Ltd.).

[0034] 3. Experimental detection indicators: 1) Relative Thyroid Mass: After blood collection from rats, the rats were fixed on a rat table, and the neck skin was incised. The skin was dissected layer by layer to fully expose the trachea. Red oval tissues were visible on both sides of the trachea, which were the thyroid glands. After complete separation, the surrounding tissues were removed, the rats were rinsed with cold physiological saline, blotted dry with filter paper, and the thyroid gland mass was generally observed and recorded. The relative thyroid mass was then weighed using a balance, and the relative thyroid mass was calculated. Relative thyroid mass = thyroid mass / rat body mass × 100%. A portion of the left thyroid gland was taken from each rat and immersed in 4% paraformaldehyde buffer for preparing tissue sections; the remaining thyroid tissue was stored at -80°C.

[0035] 2) Thyroid volume: Gently place the ultrasound probe in front of the rat's neck, using the trachea as an anatomical landmark, and slowly move the probe to locate the thyroid tissue. The rat thyroid gland has one lobe on each side, located on either side of the trachea. It is of medium echogenicity, regularly shaped, and surrounded by hypoechoic neck muscles, clearly distinguishing the thyroid gland from the surrounding tissues. First, perform a transverse scan to find the longest diameter of the upper and lower poles of the thyroid gland, freeze the image, and record the vertical diameter (vertical height).

[0036] The probe angle was then adjusted to perform a longitudinal section scan. The image was frozen at the largest longitudinal section of the thyroid gland, and the left-right diameter (lateral length) and anteroposterior diameter (longitudinal thickness) of the section were recorded. Each section was scanned three times, and the image with the clearest boundary and the most accurate diameter measurement was selected and saved to the instrument's built-in storage system. The sample number, detection time, and group information were also labeled.

[0037] 3) Serum TSH, FT3, and FT4 levels: After blood was collected from rats (blood was drawn from the heart during rat sampling), the samples were allowed to stand at 4°C for 2 hours, followed by centrifugation at 3000g for 5 minutes at 4°C to separate rat serum. Immediately afterward, the levels of TSH, FT3, FT4, TPO, TRH, and TG in the serum were measured strictly according to the ELISA kit instructions (using rat TSH, FT3, FT4, TPO, TRH, and TG ELISA kits respectively). The levels of IL-6, IL-1β, and TNF-α in rat serum were measured using IL-6, IL-1β, and TNF-α ELISA kits.

[0038] The procedure is as follows: (1) Add the sample to be tested to each well of the reaction plate, mix well and incubate at 37°C for 60 minutes; (2) Wash the plate 3-4 times; (3) Add the first antibody working solution, mix well and incubate at 37°C for 60 minutes; (4) Wash the plate 3-4 times; (5) Add the enzyme-labeled antibody working solution and incubate at 37°C for 30 minutes; (6) Wash the plate 3-4 times; (7) Add the substrate working solution and incubate at 37°C in the dark for 20 minutes; (8) After adding the stop solution, immediately measure the absorbance of the reaction plate at the corresponding wavelength using an enzyme-linked immunosorbent assay reader.

[0039] 4) Pathological changes in rat thyroid gland: Thyroid tissue that had been fixed in 4% paraformaldehyde buffer for 24 h was removed, repaired and smoothed in a fume hood, and prepared into tissue sections through a series of processes such as dehydration and embedding for HE staining.

[0040] The specific steps for HE staining are as follows: (1) Prepare paraffin sections; (2) Dewax in xylene for 5-10 minutes; (3) Replace with fresh xylene and dewax again for 5-10 minutes; (4) Stain with anhydrous ethanol for 5 minutes; (5) Stain with 90% ethanol for 2 minutes; (6) Stain with 80% ethanol for 2 minutes; (7) Stain with 70% ethanol for 2 minutes; (8) Stain with hematoxylin staining solution for 5-10 minutes; (9) Rinse with tap water to remove excess staining solution for about 10 minutes; (10) Wash again with distilled water; (11) Stain with eosin staining solution; (12) Stain with distilled water for 2 minutes; (13) Dehydrate, clear, and mount.

[0041] 4) Statistical analysis: Statistical analysis was performed using GraphPad Prism 10.4, and the results are expressed as mean ± standard error. One-way ANOVA was used for comparisons among multiple groups, and linear regression analysis was used for correlation. Statistical significance was defined as p-value < 0.05.

[0042] 4. Experimental Results 1) Thyroid volume In the process of establishing a rat model of nodular goiter, ultrasound technology can comprehensively evaluate the model establishment effect from multiple dimensions. In terms of morphological assessment, ultrasound can clearly display the overall outline, size, and internal echo characteristics of the rat thyroid gland, accurately identify the number, location, diameter, boundary clarity, morphological regularity, and internal echo homogeneity of nodules, and distinguish between solid, cystic, or mixed nodules, providing intuitive evidence for judging whether nodules have formed and their morphological characteristics.

[0043] Experimental results show (e.g.) Figure 1 , 2 As shown in the figure, the thyroid volume in the model group (Model) was significantly larger than that in the control group (Control), and the thyroid volume was significantly reduced after treatment with thymol.

[0044] 2) Pathological changes in the rat thyroid gland As an important endocrine organ, the integrity of the thyroid gland's tissue structure directly affects the synthesis and secretion of thyroid hormones, thereby regulating various physiological functions such as metabolism, growth, and development. Thyroid diseases are often accompanied by abnormal changes in the morphology of thyroid tissue, and tissue staining techniques are key means to directly observe tissue morphology and structure, assess the degree of lesions, and evaluate the effectiveness of drug interventions.

[0045] The results of the thyroid tissue staining experiment showed (e.g.) Figure 3 As shown in the figure, the control group rats had numerous round or oval follicles in their thyroid glands, which were of medium size and regular shape. The epithelial cells were arranged in a cuboidal or flattened pattern, and no abnormal nodular hyperplasia was observed in the loose connective tissue between the cells and follicles. In contrast, the model group rats had fewer thyroid follicles, which were irregularly arranged, and the epithelial cells showed highly nodular hyperplasia. The boundaries between the follicles were blurred, indicating the successful establishment of the model. The administration of daphnetin and levothyroxine significantly inhibited these typical histological patterns. The shape and arrangement of the follicles were more regular, and their size was more uniform. There was almost no abnormal nodular hyperplasia in the loose connective tissue between the cells and follicles, and the boundaries between the follicles were relatively clear, and the thyroid tissue structure was basically restored to normal. Compared with the model group, daily administration of daphnetin and levothyroxine for 4 weeks could reduce follicular epithelial nodular hyperplasia to varying degrees. These results indicate that daphnetin has a certain therapeutic effect on PTU-induced nodular goiter in rats.

[0046] 3) Relative thyroid mass The relative mass of the thyroid gland (thyroid weight / rat body weight) is a core biological indicator reflecting the degree of thyroid tissue hyperplasia and changes in morphology and function. Fluctuations in its value can directly reflect the regulatory effect of drugs on thyroid tissue. In experimental studies related to nodular goiter, setting up intervention groups and a control group with different doses of daphne, and dynamically monitoring and comparing the changes in the relative mass of the thyroid gland in each group, is a key step in evaluating the regulatory effect of daphne on the nodular goiter model, providing important experimental evidence for clarifying the dose-response relationship and mechanism of action of the drug.

[0047] Experimental results show (e.g.) Figure 4 , 5 As the disease progressed, the relative thyroid mass of rats in the model group (as shown) was significantly increased compared with the control group. Compared with the model group (as shown), both the Daph-High and L-T4 groups effectively reduced the relative thyroid mass of rats with the disease, with the Daph-High group (as shown) exhibiting a more significant therapeutic effect.

[0048] 4) Mouse body weight Changes in body weight are one of the core indicators for assessing drug safety and the overall physiological state of animals. In particular, in drug intervention studies of chronic disease models such as nodular goiter, it can directly reflect the effects of drugs on the metabolic function, nutrient absorption and body tolerance of rats.

[0049] Experimental results show (e.g.) Figure 6 As shown in the figure, before treatment, the model group (Model) showed weaker weight gain than the control group (Control), indicating that the modeling drug had a characteristic effect on the growth status of rats, indirectly supporting the success of model construction. After treatment, the weight gain of each treatment group was closer to that of the control group. The weight gain reflects that each treatment group had a positive regulatory effect on the physiological state of the model rats. Among them, the Daph-High group and the Levothyroxine (L-T4) group showed more significant therapeutic effects.

[0050] 5) Thyroid function related indicators (TG, TRH, FT3, FT4 and TPO) Maintaining homeostasis of thyroid function and related molecular indicators is crucial for the normal physiological function of the thyroid gland. Free triiodothyronine (FT3) and free thyroid hormone (FT4), as the active forms of thyroid hormones, directly participate in key physiological processes such as metabolism and growth and development. Thyroid-stimulating hormone (TSH) and thyrotropin-releasing hormone (TRH) constitute the core regulatory pathway of the hypothalamus-pituitary-thyroid axis (HPT axis), maintaining the balance of thyroid hormone secretion through hierarchical regulation. Thyroglobulin (TG) is a precursor protein for thyroid hormone synthesis, and its level changes reflect the synthetic function and damage status of thyroid tissue. Thyroid peroxidase (TPO) is a key enzyme in the process of thyroid hormone synthesis, and its activity or expression level directly affects the efficiency of hormone synthesis.

[0051] Experimental results show (e.g.) Figure 7 , 8As the disease progressed, compared with the control group, the model group rats showed significantly decreased FT3 and FT4 levels and significantly increased TSH levels. Compared with the model group, the Daph-Low, Daph-Med, and Daph-High groups of daphne significantly increased the FT3 and FT4 levels in rats, and there was no significant difference in therapeutic effect compared with the levothyroxine group (L-T4). The Daph-Low, Daph-Med, and Daph-High groups of daphne significantly decreased the TSH levels in rats, and there was no significant difference in therapeutic effect compared with the levothyroxine group (L-T4). As the disease progressed, compared with the control group, the model group rats showed significantly decreased TG and TPO levels and significantly increased TRH levels. Compared with the model group, the high-dose daphne group (Daph-High) significantly increased TG levels in rats, and there was no significant difference in therapeutic effect compared with the levothyroxine group (L-T4). The low-dose daphne group (Daph-Low), medium-dose daphne group (Daph-Med), and high-dose daphne group (Daph-High) all significantly reduced TRH levels in rats, and the medium-dose daphne group (Daph-Med) showed a more significant therapeutic effect than the levothyroxine group (L-T4).

[0052] 6) Effects on serum inflammatory factor levels (IL-1β, TNF-α, and IL-6) Imbalance in the inflammatory cytokine network is a core element in the pathological process of various inflammation-related diseases, including thyroid disease. Interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), as key pro-inflammatory factors, directly reflect the intensity of the body's inflammatory response and the disease progression through changes in their serum levels. IL-1β can amplify the immune response by activating downstream inflammatory signaling pathways, TNF-α can induce inflammatory cell infiltration and promote tissue damage, while IL-6 plays a dual role in the inflammatory cascade, acting as both a pro-inflammatory agent and an immunomodulatory agent. These four factors synergistically constitute the core regulatory network of the inflammatory response.

[0053] Experimental results show (e.g.) Figure 9As the disease progressed, compared with the control group, the serum levels of inflammatory factors IL-1β and TNF-α in the model group rats were significantly increased, indicating a high level of inflammation in the disease group. Compared with the model group, the Daph-Low and Daph-Med groups showed no significant difference in reducing the serum inflammatory factor IL-6 in rats. The Daph-High and L-T4 groups significantly reduced the serum levels of inflammatory factors IL-1β, IL-6, and TNF-α in rats. Among them, the Daph-Low group was superior to the L-T4 group in reducing the inflammatory factors IL-1β and TNF-α, suggesting that Daph-Low has a significant advantage in improving the inflammatory dysregulation state caused by nodular goiter.

[0054] The above test results show that: After the drug treatment phase, different doses of daphne showed therapeutic effects on rats with nodular goiter. Among them, the medium and high dose groups showed better efficacy in the treatment of rats with nodular goiter. The overall therapeutic effect of daphne was better than that of the positive control drug levothyroxine. The results of this experiment show that daphne has a good therapeutic effect on thyroid nodules in rats.

[0055] According to the internationally recognized body surface area method (referencing ICH S9, FDA, and relevant guidelines from the Chinese NMPA), the dosage was converted using the rat body surface area coefficient (Km=6.0) and the human body surface area coefficient (Km=37.0). The conversion formula is: Human equivalent dose (HED, mg / kg) = Rat dose (AD, mg / kg) × Rat Km / Human Km. Based on the gavage administration doses of daphne in rats in this study (low dose 25 mg / kg / day, medium dose 50 mg / kg / day, high dose 100 mg / kg / day, once daily), the corresponding human oral equivalent doses were calculated as follows: low dose approximately 4.05 mg / kg / day (total dose approximately 283.5 mg / day for a standard adult weight of 70 kg), medium dose approximately 8.11 mg / kg / day (total dose approximately 567.7 mg / day), and high dose approximately 16.22 mg / kg / day (total dose approximately 1135.4 mg / day). The conversion result is highly consistent with the safe and effective dosage range of 300-1000 mg per day for adults in existing clinical studies of daphne. Considering the oral bioavailability (25%-35%) and human half-life (8-10 hours) of daphne, it is recommended to use fractionated dosing (2-3 times per day) in clinical applications to maintain a stable blood drug concentration. At the same time, individual adjustments can be made according to the patient's actual weight, severity of the disease, and liver and kidney function, providing a scientific dosimetric reference for the translation of daphne from animal experiments to clinical applications.

[0056] In summary, this study demonstrates that daphne can be considered a promising candidate drug for the treatment of thyroid nodules.

[0057] The above description is merely a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural transformations made using the contents of the present invention under the inventive concept of the present invention, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present invention.

Claims

1. The use of daphne in the preparation of drugs for the treatment of thyroid nodules.

2. The use of daphne as described in claim 1 in the preparation of a thyroid nodule treatment drug, wherein the structural formula of daphne is: 。 3.The use of daphnetin in the preparation of a drug for treating thyroid nodules according to claim 1, characterized in that, The medication for treating thyroid nodules contains daphne and a pharmaceutically acceptable salt.

4. The use of daphne as described in claim 1 in the preparation of a thyroid nodule treatment drug, characterized in that, The medication for treating thyroid nodules contains daphne and pharmaceutically acceptable excipients.

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