Application of cevaltinib in preparation of medicine for treating tendinopathy and medicine

Abstract

The invention provides application of cevaltinib in preparation of a medicine for treating tendinopathy, the tendinopathy is a tendon degenerative disease caused by tendon stem cell senescence and tendon degeneration, and the medicine for treating the tendinopathy can target tendon stem cells, inhibit tyrosine kinase receptors and inhibit expression of tendon stem cell senescence markers p16 and p21. Therefore, histological pathologic progresses such as tendon degeneration and the like are inhibited. The invention further provides a medicine, and the effective component of the medicine is the cevaltinib. According to the application disclosed by the invention, the remarkable effect of the cevaltinib in the aspect of treating the tendinopathy is found for the first time, and the senescence of tendon stem cells is inhibited and the tendon degeneration is delayed by highly selectively inhibiting MET tyrosine kinase. In addition, the used cevaltinib is high in biological safety, the synthesis method is mature, intensive production has been achieved, the requirements for storage and transportation conditions are not high, and the cost of the cevaltinib serving as a medicine is greatly reduced.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of savotinib in preparing a tendinopathy therapeutic drug, and a tendinopathy therapeutic drug. Background Art

[0002] Tendinopathy is a common degenerative disorder of the tendon characterized by pain, decreased function, and impaired exercise tolerance. Its incidence is increasing annually among professional athletes and the aging population. Pathological studies have shown that tendinopathy manifests not only in structural changes such as collagen fiber disarray and imbalanced extracellular matrix metabolism, but is also accompanied by microenvironmental remodeling, including pathological neovascularization and abnormal proliferation of sensory nerve endings. Currently, conservative treatments such as nonsteroidal anti-inflammatory drugs, glucocorticoid injections, and extracorporeal shock wave therapy (ESWT) are the primary treatments used clinically. However, systematic reviews have shown that these treatments only provide short-term symptom relief and lack long-term efficacy. Although tendon loading exercises have been shown to promote collagen remodeling through mechanical stimulation, poor patient compliance limits their clinical application. For recalcitrant cases, surgical intervention can repair tendon structure, but the rate of postoperative reinjury remains high. Therefore, it is crucial to elucidate the key signaling pathways in the pathogenesis of tendinopathy and explore new treatments targeting its pathogenesis.

[0003] In recent years, the role of tendon stem / progenitor cells (TSPCs) aging in tendon degeneration has attracted considerable attention. As a core cell population that maintains tendon homeostasis, TSPCs exacerbate tendinopathy through dual mechanisms: (1) Aged TSPCs activate the NF-κB signaling pathway by secreting senescence-associated secretory phenotype (SASP) factors such as IL-6 and MMP-3, triggering local inflammatory responses and disrupting extracellular matrix homeostasis; (2) Aging significantly decreases the proliferation and differentiation capacity of TSPCs. Experimental data show that the clone formation rate of TSPCs from elderly donors is significantly lower than that from young donors, and the collagen synthesis rate is significantly reduced. Notably, in a tendon injury model, the elimination of senescent cells can improve tendon mechanical strength, suggesting that targeting cell senescence may become a new therapeutic approach.

[0004] Hepatocyte growth factor receptor (MET) is a cell surface receptor tyrosine kinase that plays an important role in a variety of physiological and pathological processes in the human body. The extracellular domain of the transmembrane receptor MET with tyrosine kinase activity, encoded by the MET proto-oncogene, binds to the intracellular domain of human growth factor (HGF) and undergoes autophosphorylation, which then leads to biological responses such as cell movement, proliferation, and morphological changes through a series of signal transductions. Studies have shown that it may be related to aging-related tissue repair disorders, stem cell function decline, and chronic inflammation regulation. Previous studies used artificial intelligence machine learning models to construct a tendon stem cell aging prediction model and combined it with the GEO public database to screen out the tendon stem cell aging-related target MET.

[0005] Savolitinib is a highly selective MET tyrosine kinase inhibitor with good biological activity and biosafety. Studies have shown that it has a good therapeutic effect on advanced non-small cell lung cancer (NSCLC) and MET-driven papillary renal cell carcinoma carrying MET exon 14 skipping mutations. It blocks tumor growth and metastasis by precisely inhibiting the MET signaling pathway (including RAS-MAPK, PI3K-AKT, etc.). Currently, there are still gaps in basic research and clinical evidence for the therapeutic potential of savolitinib in the field of tendinopathy and chronic inflammatory diseases. Its mechanism of regulating MET-mediated local inflammatory cascades and tissue repair has not yet been clarified, and still requires systematic pharmacological exploration and translational medicine verification. Summary of the Invention

[0006] In view of the problems existing in the prior art, the present invention provides the use of savotinib in the preparation of a tendinopathy therapeutic drug, and a tendinopathy therapeutic drug.

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] (1) The present invention provides a use of savotinib, which is used in the preparation of a drug for treating tendinopathy.

[0009] Furthermore, the present invention may also have the following characteristics: the tendinopathy therapeutic drug can target tendon stem cells and inhibit tyrosine kinase receptors, thereby inhibiting the expression of tendon stem cell aging markers, thereby inhibiting tendon stem aging.

[0010] Furthermore, the present invention may also have the following characteristics: the tendinopathy is caused by aging of tendon stem cells and tendon degeneration.

[0011] (2) The present invention also provides a drug for treating tendinopathy, wherein the active ingredient of the drug is savotinib.

[0012] Furthermore, the present invention may also have the following characteristics: the drug further comprises a pharmaceutically acceptable carrier or excipient.

[0013] Furthermore, the present invention may also have such a feature: the dosage form of the drug is an injection, capsule, granule or tablet.

[0014] Furthermore, the present invention may also have the following characteristics: the drug is an injection composed of savotinib dissolved in a clinical injection solution.

[0015] Furthermore, the present invention may also have the following feature: the clinical injection solution is physiological saline.

[0016] The beneficial effects of the present invention are:

[0017] (1) The present invention is the first to discover the significant effect of savotinib in treating tendinopathy. Savotinib has an anti-cellular aging effect, delaying the aging and degeneration of tendon stem cells by highly selectively inhibiting MET tyrosine kinase, inhibiting local inflammatory responses, and maintaining extracellular matrix homeostasis, thereby delaying the damage of aging to tendon stem cells and delaying tendon degeneration;

[0018] (2) The savotinib used in the present invention has been used in a variety of animal models, including advanced non-small cell lung cancer, MET-driven papillary renal cell carcinoma, and hepatocellular carcinoma, carrying MET exon 14 skipping mutations, and has high biosafety. Furthermore, the synthesis method of savotinib is mature, and intensive production has long been achieved. The storage and transportation conditions are not demanding, which greatly reduces the cost of savotinib as a drug.

[0019] (3) The present invention can use an injection preparation for intratendon injection, thereby increasing the utilization rate of savotinib in tendon tissue and maximizing its therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Represents the expression level of SA-β-Gal in the D-gal-induced tendon stem cell aging model after savotinib treatment, where Figure 1 a is a staining diagram showing the changes in SA-β-Gal content in the control group, D-gal treatment group, D-gal + Terevalefim treatment group, and D-gal + Savolitinib treatment group; Figure 1 b is the relative quantitative analysis of SA-β-Gal expression levels in the control group, D-gal-treated group, D-gal + Terevalefim-treated group, and D-gal + Savolitinib-treated group;

[0021] Figure 2 Indicates the expression levels of p16 and p21 in the D-gal-induced tendon stem cell aging model after treatment with savotinib; Figure 2 a is the immunofluorescence staining diagram of the changes in p16 and p21 levels in the control group, D-gal treatment group, D-gal + Terevalefim treatment group, and D-gal + Savolitinib treatment group; Figure 2 b is the relative quantitative analysis of p16 in each treatment group; Figure 2 c is the relative quantitative analysis of p21 in each treatment group;

[0022] Figure 3 Masson staining and H&E staining images of the sham operation group, collagenase-induced tendinopathy model (CIT) group, CIT+Terevalefim treatment group, and CIT+Savolitinib treatment group;

[0023] Figure 4 are immunofluorescence staining images of the changes in p16 and p21 levels in the sham-operated group, CIT-treated group, CIT+Terevalefim-treated group, and CIT+Savolitinib-treated group;

[0024] Figure 5 It is the relative quantitative analysis of p16 and p21 in the sham operation group, CIT treatment group, CIT + Terevalefim treatment group and CIT + Savolitinib treatment group. DETAILED DESCRIPTION

[0025] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention and the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0026] This invention proposes for the first time the use of savotinib in the preparation of a drug for treating tendinopathy, a condition caused by tendon stem cell aging and tendon degeneration. Based on this application, the invention also provides a drug for treating tendinopathy, the active ingredient of which is savotinib. Savotinib targets tendon stem cells and inhibits tyrosine kinase receptors, suppressing the expression of tendon stem cell aging markers and thereby inhibiting tendon stem cell aging.

[0027] The dosage form of the tendinopathy treatment drug is preferably an injection, comprising savotinib and normal saline.

[0028] Efficacy test:

[0029] 1. Experimental study on the effect of savotinib in the D-gal-induced tendon stem cell aging model.

[0030] Cultured tendon stem cells were treated with D-gal to induce senescence, starting at a concentration of 100 mM for seven days. A senescence model was established. Savotinib was added to the senescent cell culture system at a concentration of 10 μM. After seven days of induction, the expression levels of SA-β-Gal, a marker associated with senescence, and p16 and p21, markers of senescence, were measured.

[0031] The specific groups were: control group, D-gal treatment group, D-gal + Terevalefim treatment group and D-gal + Savolitinib treatment group.Terevalefim is a Met agonist.

[0032] Figure 1 represents the expression level of SA-β-Gal after savotinib treatment, where Figure 1 a is a staining diagram showing the changes in SA-β-Gal content in the control group, D-gal treatment group, D-gal + Terevalefim treatment group, and D-gal + Savolitinib treatment group. Figure 1 b is the relative quantitative analysis of SA-β-Gal expression levels in the control group, D-gal-treated group, D-gal + Terevalefim-treated group, and D-gal + Savolitinib-treated group.

[0033] Depend on Figure 1 a and Figure 1 b It can be seen that after using D-gal to induce tendon cell senescence, the expression of aging-related indicator SA-β-Gal was significantly increased. Compared with the D-gal treatment group and the D-gal+Terevalefim treatment group, the expression level of SA-β-Gal in the D-gal+Savolitinib treatment group was significantly decreased, indicating that Savolitinib can inhibit the expression of SA-β-Gal, a cell senescence-related indicator.

[0034] Figure 2 represents the expression levels of p16 and p21 after savotinib treatment, where Figure 2 a is the immunofluorescence staining graph of the changes in p16 and p21 levels in the control group, D-gal treatment group, D-gal + Terevalefim treatment group, and D-gal + Savolitinib treatment group. Figure 2 b Relative quantitative analysis of p16 in the control group, D-gal-treated group, D-gal + Terevalefim-treated group, and D-gal + Savolitinib-treated group. Figure 2c is the relative quantitative analysis of p21 in the control group, D-gal-treated group, D-gal + Terevalefim-treated group, and D-gal + Savolitinib-treated group.

[0035] Depend on Figure 2 As can be seen, the expression levels of p16 and p21 were significantly reduced in the D-gal + Savolitinib group compared to the D-gal and D-gal + Terevalefim groups, indicating that Savolitinib can inhibit the expression of aging-related markers p16 and p21. In other words, it can be concluded that Savolitinib treatment significantly inhibits tendon stem cell aging.

[0036] 2. Experimental study on the effect of savotinib in tendinopathy model (CIT).

[0037] Establishment of a rat Achilles tendon tendinopathy model (CIT) induced by in situ injection of type I collagenase: 8-week-old male Sprague-Dawley rats were anesthetized with isoflurane inhalation anesthesia and their hind limbs shaved. Type I collagenase (5 mg / mL, 60 μl) was injected intratendinously into the Achilles tendon using a 31-gauge insulin syringe. This was repeated three times a week for two consecutive weeks to successfully establish the model.

[0038] In situ administration of savotinib into the Achilles tendon of rats with tendinopathy model: savotinib preparation was injected into the Achilles tendon of rats with tendinopathy model. The drug dosage was 10 μM, the volume of each administration was 60 μL, and the administration frequency was 3 times a week for 4 weeks. The administration method is shown in Table 1.

[0039] Table 1 - Dosage

[0040] combination Frequency Grouping Dosage form—method of administration 1 3 times a week Sham+normal saline Injection - Achilles tendon injection 2 3 times a week CIT+normal saline Injection - Achilles tendon injection 3 3 times a week CIT+Terevalefim (10 μM, 60 μL) Injection - Achilles tendon injection 4 3 times a week CIT+Savolitinib (10 μM, 60 μL) Injection - Achilles tendon injection

[0041] Method for savotinib injection into the rat Achilles tendon: Rats were anesthetized with isoflurane gas, and the hind limbs were shaved and disinfected with 75% ethanol. The rat's right lower limb was grasped with the left hand, flexed 90°, and the needle was inserted parallel to the calcaneus using the right hand, using the posterior edge of the calcaneus as the insertion point. Savotinib was injected intratendinously into the Achilles tendon using a 31-gauge insulin syringe.

[0042] After 4 weeks of continuous administration, the experimental animals were euthanized, and Achilles tendon specimens were obtained and sliced ​​for histological staining. Immunohistochemical staining was used to evaluate the aging and degeneration of the Achilles tendon, and the expression levels of cell aging-related indicators were examined.

[0043] The specific groups were: (1) sham operation group (Sham + normal saline); (2) tendinopathy model group (CIT + normal saline); (3) tendinopathy model + Met agonist group (CIT + Terevalefim); (4) tendinopathy model + Met inhibitor group (CIT + Savolitinib).

[0044] Figure 3 The following are Masson staining and H&E staining images of the sham operation group, CIT treatment group, CIT + Terevalefim treatment group, and CIT + Savolitinib treatment group. Figure 3 It can be seen that compared with the CIT-treated group and CIT+Terevalefim group, savotinib treatment can significantly inhibit the progression of tendinopathy.

[0045] Figure 4 This is the immunofluorescence staining diagram of the changes in p16 and p21 levels in the sham operation group, CIT treatment group, CIT + Terevalefim treatment group, and CIT + Savolitinib treatment group. Figure 5 It is the relative quantitative analysis of p16 and p21 in each treatment group. Figure 4 and Figure 5 As can be seen, the expression levels of p16 and p21 were significantly decreased in the CIT + Savolitinib group compared to the CIT and CIT + Terevalefim groups, indicating that Savolitinib can inhibit the expression of p16 and p21, markers of tenocyte senescence. In other words, Savolitinib treatment significantly inhibits tenocyte senescence.

[0046] In summary, the present invention clarifies that savotinib can alleviate D-gal-induced tendon stem cell aging and the progression of CIT-induced tendinopathy animal models.

[0047] This invention demonstrates for the first time the remarkable efficacy of savotinib in treating tendinopathy. By selectively inhibiting MET tyrosine kinase, it delays aging-related damage to tendon stem cells and slows tendon degeneration. Furthermore, the savotinib used in this invention has high biosafety, mature synthesis methods, and has long been intensively produced, requiring minimal storage and transportation conditions, significantly reducing the cost of savotinib as a pharmaceutical.

[0048] The above are merely preferred embodiments of the present invention. The scope of protection of the present invention is not limited to the above embodiments. All technical solutions based on the principles of the present invention are within the scope of protection of the present invention. It should be noted that for those skilled in the art, various improvements and modifications that do not depart from the principles of the present invention should be considered within the scope of protection of the present invention.

Claims

1. The use of savotinib, characterized in that: The application is the application in preparing a drug for treating tendinopathy.

2. The use according to claim 1, characterized in that The tendinopathy therapeutic drug can target tendon stem cells and inhibit tyrosine kinase receptors, thereby inhibiting the expression of tendon stem cell aging markers and thus inhibiting tendon stem cell aging.

3. The use according to claim 1, characterized in that The tendinopathy is caused by aging of tendon stem cells and tendon degeneration.

4. A drug for treating tendinopathy, characterized in that: The active ingredient of the drug is savotinib.

5. The tendinopathy treatment drug according to claim 4, characterized in that: The medicine further includes a pharmaceutically acceptable carrier or excipient.

6. The tendinopathy treatment drug according to claim 4, characterized in that: The dosage form of the medicine is injection, capsule, granule or tablet.

7. The tendinopathy treatment drug according to claim 4, characterized in that: The drug is an injection prepared by dissolving savotinib in a clinical injection solution.

8. The tendinopathy treatment drug according to claim 7, characterized in that: The clinical injection solution is normal saline.

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