Application of thyroid hormone T3 in the preparation of drugs to delay or treat intervertebral disc degeneration

Abstract

This invention relates to the application of thyroid hormone T3 in the preparation of drugs for delaying or treating intervertebral disc degeneration. T3 can inhibit the glycolytic capacity of degenerated nucleus pulposus cells, reduce lactic acid accumulation in the intervertebral disc, thereby alleviating nucleus pulposus cell aging and apoptosis, and delaying or treating intervertebral disc degeneration. Compared with existing technologies, this invention utilizes thyroid hormone T3 to delay or treat intervertebral disc degeneration, providing a new use for thyroid hormone T3, greatly enhancing its application value, expanding its application scope, and opening up new fields of application, with broad application prospects and commercial value.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the use of a thyroid hormone T3 in the preparation of a drug for delaying or treating intervertebral disc degeneration. Background Technology

[0002] Low back pain is a common ailment with a high incidence and wide prevalence in modern populations. With social development and changes in lifestyle, the incidence of low back pain is rising year by year, and patients are becoming increasingly younger. Numerous studies have demonstrated that intervertebral disc degeneration is one of the main causes of low back pain. Currently, the efficacy of non-surgical treatments is unclear, and most patients still require discectomy in the later stages. However, surgery is invasive; therefore, how to slow down intervertebral disc degeneration has become an important issue in the field of spinal surgery.

[0003] Patent CN108606969A discloses the application of aspirin in the preparation of drugs for treating intervertebral disc degeneration. This invention constructs an intervertebral disc degeneration model by using LPS to induce oxidative stress, observes the therapeutic effect of aspirin on intervertebral disc degeneration, evaluates the therapeutic effect of aspirin on intervertebral disc degeneration by observing various indicators of oxidative stress and intervertebral disc degeneration, and demonstrates the brief mechanism by which aspirin reduces oxidative stress and alleviates intervertebral disc degeneration through Western blot. Oxidative stress in the nucleus pulposus of the intervertebral disc causes intervertebral disc degeneration, and aspirin delays intervertebral disc degeneration by inhibiting the oxidative stress of the nucleus pulposus and reducing the degradation of the cellular matrix and type II collagen.

[0004] Patent CN108635349A discloses the application of melatonin in the preparation of drugs for treating intervertebral disc degeneration. This invention establishes a rat caudal intervertebral disc degeneration model by using needle puncture, observes the therapeutic effect of melatonin on intervertebral disc degeneration, evaluates the therapeutic effect of melatonin on intervertebral disc degeneration by observing various indicators of intervertebral disc degeneration, and elucidates the brief mechanism by which melatonin delays intervertebral disc degeneration by observing the expression of Mel-type receptors and RANKI on the cartilaginous endplate of the rat caudal intervertebral disc through immunohistochemical staining and TRAP staining. The destruction of the cartilaginous endplate of the intervertebral disc and the increase in subchondral bone loss may be important mechanisms causing intervertebral disc degeneration. Meltonin delays intervertebral disc degeneration by inhibiting the expression of RANKI by endplate chondrocytes, thereby reducing osteoclast formation in the cartilaginous endplate region.

[0005] Patent CN115068463B discloses the application of a specific metabolic kinase PFKFB3 inhibitor (KAN) in products that improve intervertebral disc degeneration. KAN can partially restore the anaerobic glycolytic energy metabolism phenotype of nucleus pulposus cells that are overactivated under inflammatory stimulation, thereby reducing the excretion of lactic acid into the extracellular matrix, improving the acidification of the extracellular environment and its effects on nucleus pulposus cells and the extracellular matrix, and improving intervertebral disc degeneration.

[0006] The intervertebral disc is the largest avascular tissue in the human body, with blood vessels mainly located at its periphery. Nucleus pulposus cells primarily obtain oxygen and nutrients from surrounding tissues through diffusion. The oxygen tension in the nucleus pulposus region is extremely low (<1%), thus the cells are chronically in a hypoxic environment. This environment forces them to rely primarily on glycolysis for energy. Lactic acid, the main metabolic product of glycolysis, is usually transported extracellularly, leading to an acidic microenvironment in the nucleus pulposus. The glycolysis-dependent nature of the nucleus pulposus cells leads to lactic acid accumulation, further exacerbating the local acidic environment, which is detrimental to cell function and may accelerate degeneration. Therefore, improving the nucleus pulposus cell microenvironment (e.g., reducing lactic acid accumulation or alleviating the acidic environment) helps slow down intervertebral disc degeneration.

[0007] Thyroid hormones (THs), namely 3,5,3'-triiodo-L-tyrosine (T3) and 3,5,3',5'-tetraiodo-L-tyrosine (T4), are widely recognized for their involvement in various physiological and pathological processes, including cell growth, cell metabolism, and tumorigenesis and development. T4 is the primary hormone secreted by the thyroid gland, accounting for over 90% of total thyroid hormone secretion. T4 is converted into the more active form, T3, in the body; therefore, T4 is often considered a precursor hormone. T3 is the active form of thyroid hormone, and its physiological effects are 4-5 times stronger than those of T4.

[0008] Currently, most inventions related to thyroid hormones focus on thyroid hormone receptor agonists, such as a polycyclic thyroid hormone β receptor agonist and its uses (CN202210999229.5), and the application of thyroid hormone β receptor agonists in the preparation of drugs to improve gut microbiota (CN202410212000.1), etc.

[0009] Thyroid hormones possess a wide range of pharmacological activities, with their core functions being the regulation of metabolism, promotion of growth and development, and maintenance of nervous and cardiovascular function. They play a crucial role in the treatment of endocrine disorders (such as hypothyroidism and hyperthyroidism) and also demonstrate significant clinical value in metabolic diseases, neurological disorders, and reproductive disorders. For example, patent CN111419834A discloses the application of thyroid hormones and their analogues in the preparation of treatments for sickle cell disease; patent CN111388459A discloses the application of thyroid hormones and their analogues in the preparation of treatments for α* thalassemia.

[0010] Yang Chuncheng et al. conducted a study on the mechanism by which thyroid hormones inhibit the progression of liver cancer by regulating thyroid hormone-reactive protein, and found that thyroid hormone T3 can enhance the anti-tumor effect of the multi-kinase inhibitor lenvatinib.

[0011] There are currently no applications of thyroid hormones to delay or treat intervertebral disc degeneration. Summary of the Invention

[0012] The purpose of this invention is to overcome the defects of the prior art and provide an application of thyroid hormone T3 in the preparation of drugs for delaying or treating intervertebral disc degeneration. By utilizing thyroid hormone T3 to inhibit the glycolytic ability of senescent nucleus pulposus cells and reduce lactic acid accumulation in the intervertebral disc, the senescence and apoptosis of nucleus pulposus cells are alleviated, thereby using thyroid hormone T3 to delay or treat intervertebral disc degeneration.

[0013] The objective of this invention can be achieved through the following technical solutions:

[0014] The first objective of this invention is to protect the use of a thyroid hormone T3 in the preparation of a medicament for delaying or treating intervertebral disc degeneration, with thyroid hormone T3 as the component for exerting the therapeutic effect.

[0015] Furthermore, the application of thyroid hormone T3 in the preparation of drugs that delay intervertebral disc degeneration.

[0016] Furthermore, the application of thyroid hormone T3 in the preparation of drugs for treating intervertebral disc degeneration.

[0017] Furthermore, the intervertebral disc referred to is the lumbar intervertebral disc.

[0018] Furthermore, the intervertebral disc degeneration includes one or more of the following: degeneration of the annulus fibrosus, degeneration of the nucleus pulposus, degeneration of the cartilaginous endplate, and osteophyte formation; more preferably, the intervertebral disc degeneration is degeneration of the nucleus pulposus.

[0019] Furthermore, the thyroid hormone T3 is used in the preparation of drugs that inhibit the glycolytic capacity of aging nucleus pulposus cells, reduce lactic acid accumulation in intervertebral discs, and alleviate nucleus pulposus cell aging and apoptosis.

[0020] Furthermore, the thyroid hormone T3 is used in the preparation of drugs to delay or treat intervertebral disc degeneration caused by strong glycolytic activity of aging nucleus pulposus cells, lactic acid accumulation, and apoptosis of nucleus pulposus cells.

[0021] Furthermore, the drug is an oral or injectable formulation.

[0022] More preferably, the drug is an injectable formulation.

[0023] Furthermore, the drug comprises a therapeutically effective amount of thyroid hormone T3 and pharmaceutically acceptable excipients.

[0024] More preferably, the pharmaceutically acceptable excipients include one or more of solvents, carriers, and excipients.

[0025] More preferably, the pharmaceutically acceptable excipients specifically include one or more of ethanol, glycerol, exosomes, etc.

[0026] Furthermore, the dosage of the drug to exert its therapeutic effect is 1–100 nM, calculated as thyroid hormone T3.

[0027] More preferably, the dosage of the drug to exert its therapeutic effect is 10 nM, calculated as thyroid hormone T3.

[0028] Furthermore, the thyroid hormone T3 is triiodothyronine, with the following structural formula:

[0029]

[0030] A second objective of this invention is to protect a medicament for delaying or treating intervertebral disc degeneration, said medicament comprising the aforementioned thyroid hormone T3.

[0031] Furthermore, the drug uses thyroid hormone T3 (abbreviated as T3) as the sole component that exerts its therapeutic effect.

[0032] Furthermore, the dosage of the drug to exert its therapeutic effect is 1–100 nM (nM represents nanomoles per liter, 1 nM = 10) based on thyroid hormone T3. -9 (mol / L).

[0033] Furthermore, the dosage of the drug to exert its therapeutic effect is 1–10 nM, calculated as thyroid hormone T3.

[0034] More preferably, the dosage of the drug to exert its therapeutic effect is 10 nM, calculated as thyroid hormone T3.

[0035] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0036] More preferably, the pharmaceutically acceptable excipients include one or more of solvents, carriers, and excipients.

[0037] More preferably, the pharmaceutically acceptable excipients specifically include one of ethanol, glycerol, exosomes, etc.

[0038] Furthermore, the specific dosage of thyroid hormone T3 in the drug is 10 mg / kg body weight.

[0039] This invention, through research, has discovered that thyroid hormone T3, as the active pharmaceutical ingredient, can delay or treat intervertebral disc degeneration. Specifically, this invention has found that, in cell experiments, thyroid hormone T3 inhibits the glycolytic activity of senescent nucleus pulposus cells, reduces lactic acid accumulation in the intervertebral disc, and thus alleviates the aging and apoptosis of nucleus pulposus cells. Furthermore, this invention has also found through animal experiments that intradiscal injection of thyroid hormone T3 into the tail intervertebral disc of rats can effectively delay or treat intervertebral disc degeneration.

[0040] Compared with the prior art, the present invention has the following advantages:

[0041] 1) The drug prepared in this invention uses thyroid hormone T3 as the component that exerts its pharmacological effect. It can inhibit the glycolytic ability of senescent nucleus pulposus cells, reduce the accumulation of lactic acid in the intervertebral disc, thereby alleviating the aging and apoptosis of nucleus pulposus cells and delaying or treating intervertebral disc degeneration.

[0042] 2) This invention utilizes the technology of thyroid hormone T3 to delay or treat intervertebral disc degeneration, providing a new use for thyroid hormone T3, greatly improving its application value, expanding the application scope of thyroid hormone T3, and opening up new fields for the application of thyroid hormone T3, which has broad application prospects and commercial value. Attached Figure Description

[0043] Figure 1 This is a graph showing the results of detecting the viability of nucleus pulposus cells treated with different concentrations of thyroid hormone T3 using the CCK8 kit in Example 1 of the present invention.

[0044] Figure 2 This is a Western blotting result of the inhibition of glycolysis-related proteins pyruvate dehydrogenase kinase 1 (PDK1) and lactate dehydrogenase isoenzyme 5 (LDH5) by thyroid hormone T3 in senescent nucleus pulposus cells, as shown in Example 1 of the present invention.

[0045] Figure 3 This is a diagram showing the results of reducing lactate accumulation in senescent nucleus pulposus cells using thyroid hormone T3 in Example 1 of the present invention.

[0046] Figure 4 This is a diagram showing the SA-β-gal staining results of thyroid hormone T3 reducing nucleus pulposus cell senescence in Example 1 of the present invention.

[0047] Figure 5 This is a flow cytometry result of the reduction of nucleus pulposus cell apoptosis by thyroid hormone T3 in Example 1 of the present invention.

[0048] Figure 6 The image shows the HE staining and histological scoring results of the rat tail vertebrae in Example 2 of this invention, which showed that thyroid hormone T3 delayed or treated intervertebral disc degeneration in vivo.

[0049] Figure 7 This diagram illustrates the mechanism of action of thyroid hormone T3 in delaying or treating intervertebral disc degeneration in this invention. Detailed Implementation

[0050] The present invention will now be described in detail with reference to specific accompanying drawings and embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several modifications and improvements without departing from the concept of the present invention. These modifications and improvements are all protected by the present invention.

[0051] Any preparation methods, materials, structures, or composition ratios not explicitly described in this technical solution are considered common technical features disclosed in the prior art.

[0052] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0053] In this embodiment, thyroid hormone T3 was purchased from Yisheng Biotechnology (Shanghai) Co., Ltd. (Catalog No.: 54574ES). Normal human nucleus pulposus cells were purchased from Wuhan Pronosei Life Sciences Co., Ltd. (Catalog No.: CP-H097), cultured in DMEM / F12 medium containing 10% fetal bovine serum and 1% penicillin antibiotics, and placed in a 37℃, 5% CO2 cell culture incubator. The medium was changed every other day, and cell growth was observed under a microscope. Healthy human nucleus pulposus cells used in the experiment were passaged to the 2nd-3rd generation (P2-3), and aged human nucleus pulposus cells were obtained from the 6th-8th generation (P6-8). SD rats were purchased from the Experimental Animal Center of Fudan University.

[0054] Example 1

[0055] This embodiment provides an experimental process for thyroid hormone T3 to inhibit the glycolytic capacity of nucleus pulposus cells in vitro, reduce lactate accumulation, and alleviate nucleus pulposus cell senescence and apoptosis. This embodiment sets up three experimental groups: a control group (P2-3 nucleus pulposus cells without any treatment, i.e., normal cells), an aged group (P6-8 nucleus pulposus cells without any treatment, i.e., senescent cells), and an aged + T3 group (T3 group, aged human nucleus pulposus cells were seeded in 6cm culture dishes and cultured for 24 hours until cell adhesion. After washing the cells once with serum-free medium, medium containing 10nM T3 was added, and the cells were cultured for another 48 hours, i.e., senescent cells + T3).

[0056] The glycolytic capacity, lactate accumulation, cellular senescence, and apoptosis of nucleus pulposus cells in the three experimental groups were measured respectively.

[0057] The CCK8 kit was used to detect the viability of nucleus pulposus cells treated with different concentrations of T3. Senescent nucleus pulposus cells showed significantly decreased viability, and cell viability exhibited a dose-dependent relationship with T3 concentration. When the T3 concentration exceeded 10 nM, the increase in cell viability was minimal; therefore, a concentration of 10 nM was chosen for the experiment. Figure 1 ).

[0058] Western blotting was used to detect glycolysis-related proteins in three groups of cells. Pyruvate dehydrogenase kinase 1 (PDK1) inhibits glucose entry into the mitochondrial tricarboxylic acid cycle, thereby enhancing anaerobic glycolysis; lactate dehydrogenase isoenzyme 5 (LDH5) preferentially converts pyruvate to lactate. Results showed that the expression levels of PDK1 and LDH5 in senescent nucleus pulposus cells were higher than in normal cells, indicating a higher level of glycolysis. T3 treatment decreased the expression levels of both, indicating inhibition of glycolysis. Figure 2 ).

[0059] The lactate assay kit was used to detect lactate accumulation in culture media. Results showed that, compared to normal cells in the control group, degenerated nucleus pulposus cells in the senescent group accumulated lactate, while, compared to the senescent group, lactate production decreased in the T3 group after using T3. Figure 3 ).

[0060] SA-β-gal staining was used to detect cellular senescence. Results showed that, compared to normal cells in the control group, the degenerated nucleus pulposus cells in the senescent group were significantly aged. However, compared to the senescent group, T3 treatment in the T3 group reduced the senescence level of nucleus pulposus cells. Figure 4 ).

[0061] Flow cytometry was used to detect cell apoptosis. Results showed that, compared to normal cells in the control group, degenerated nucleus pulposus cells in the aging group had a higher apoptosis rate, while T3 treatment reduced the apoptosis level of nucleus pulposus cells compared to the aging group. Figure 5 ).

[0062] This embodiment reveals that thyroid hormones can inhibit the glycolytic activity of senescent nucleus pulposus cells, reduce lactic acid accumulation in the intervertebral disc, thereby alleviating the aging and apoptosis of nucleus pulposus cells and delaying or treating intervertebral disc degeneration.

[0063] Example 2

[0064] This embodiment provides an experimental procedure for delaying or treating intervertebral disc degeneration in vivo using thyroid hormones. Thirty 8-week-old SD rats were used. In this embodiment, three experimental groups were set up: control group (no intervention), degeneration group (IVDD, intervertebral disc degeneration induced by acupuncture), and degeneration + T3 group (T3, T3 group injected with 10 nM T3 after acupuncture modeling).

[0065] The specific experiments of the above experimental group are as follows:

[0066] After general anesthesia, rats were placed in a prone position. Under fluoroscopic guidance, a 20-gauge needle was used to puncture the intervertebral disc at segment 8-9 (Co8 / 9). The needle was inserted through the center of the disc to the contralateral side, rotated 180°, and held for 10 seconds. In the degeneration + T3 group, 2 μL of 10 nM T3 was injected immediately after modeling using a 33-gauge needle. After modeling, the wound was covered with gauze, and standard postoperative management was performed. Postoperatively, the rats' health was monitored daily, and they were allowed free movement, normal food and water intake.

[0067] Eight weeks after modeling, all rats were sacrificed, and their tail vertebrae were collected for HE staining and histological scoring. The degeneration group had the highest intervertebral disc histological score, showing disordered nucleus pulposus structure on the sections; while the degeneration + T3 group, injected with T3, was able to better preserve the structure of the intervertebral disc. Figure 6 ).

[0068] This embodiment demonstrates that intradiscal injection of thyroid hormone into the caudal intervertebral disc of degenerated rats can effectively delay or treat intervertebral disc degeneration. The mechanism of action of thyroid hormone in delaying or treating intervertebral disc degeneration is as follows: Figure 7 As shown.

[0069] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. The use of a thyroid hormone T3 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, The drug uses thyroid hormone T3 as the component that exerts its therapeutic effect. By using thyroid hormone T3 to inhibit the glycolytic capacity of senescent nucleus pulposus cells and reduce lactic acid accumulation in the intervertebral disc, the aging and apoptosis of nucleus pulposus cells can be alleviated, thus allowing thyroid hormone T3 to be used to delay or treat intervertebral disc degeneration.

2. The application of the thyroid hormone T3 according to claim 1 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, The application of the thyroid hormone T3 in the preparation of drugs that inhibit the glycolytic capacity of senescent nucleus pulposus cells, reduce lactic acid accumulation in intervertebral discs, and alleviate nucleus pulposus cell senescence and apoptosis.

3. The application of the thyroid hormone T3 according to claim 1 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, The drug is an oral or injectable formulation.

4. The application of the thyroid hormone T3 according to claim 1 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, The drug comprises a therapeutically effective amount of thyroid hormone T3 and pharmaceutically acceptable excipients.

5. The application of the thyroid hormone T3 according to claim 4 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, Pharmaceutically acceptable excipients include solvents.

6. The application of the thyroid hormone T3 according to claim 1 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, The dosage of the drug to exert its therapeutic effect is 1–100 nM, calculated as thyroid hormone T3.

7. The use of thyroid hormone T3 according to claim 6 in the preparation of a drug for delaying or treating intervertebral disc degeneration, characterized in that, The dosage at which the drug exerts its therapeutic effect is 10 nM, calculated as thyroid hormone T3.

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