Application of deaminotyrosine in preparation of medicine for preventing and / or treating osteoporosis

Abstract

The invention discloses application of deaminotyrosine in preparation of a medicine for preventing and / or treating osteoporosis, and belongs to the technical field of medicines. In-vitro cell experiments and in-vivo animal model experiments prove that the deaminotyrosine can remarkably promote proliferation and differentiation of osteoblasts and inhibit generation and activity of osteoclasts, so that the bone density is effectively increased, the bone microstructure is improved, and the bone biomechanical strength is improved. The invention provides a brand-new, safe and effective candidate medicine for preventing and treating osteoporosis.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to application of desaminotyrosine in preparation of a drug for preventing and / or treating osteoporosis. BACKGROUND

[0002] Osteoporosis is a systemic bone disease characterized by low bone mass, damage to bone microstructure, increased bone fragility, and easy fracture. It is a common disease that seriously endangers the health of the elderly, especially postmenopausal women. At present, the drugs for treating osteoporosis in the clinic are mainly divided into two categories: bone resorption inhibitors (such as bisphosphonates, estrogen, RANKL inhibitors, etc.) and bone formation promoters (such as parathyroid hormone analogs). However, the existing drugs have many limitations, for example: bisphosphonates can cause mandibular osteonecrosis or atypical femoral fractures; long-term use of estrogen increases the risk of breast cancer and thrombosis; parathyroid hormone analogs are expensive and need to be injected daily, and the patient's compliance is poor. Therefore, developing a new type of safe, effective and multi-mechanism anti-osteoporosis drug is still the focus and difficulty of current research.

[0003] Desaminotyrosine (DAT) is a tyrosine metabolite naturally present in the human body and also found in the fermentation products of some probiotics. Studies have shown that DAT has anti-inflammatory and immunoregulatory activity, such as providing protection in an influenza infection model. However, to date, the skilled person in the art has never been suggested or disclosed in any public literature or patent that DAT has the use of regulating bone metabolism, particularly for treating osteoporosis. SUMMARY

[0004] In order to overcome the above-mentioned defects in the prior art, the present application provides the application of desaminotyrosine in preparation of a drug for preventing and / or treating osteoporosis. Studies have shown that DAT has the dual effect of promoting bone formation and inhibiting bone resorption, which provides a brand-new and extremely potential solution for the treatment of osteoporosis.

[0005] In order to achieve the above-mentioned application purposes, the present application provides the following technical solutions: In one aspect, the present application provides the application of desaminotyrosine in preparation of a drug for preventing and / or treating osteoporosis.

[0006] Optionally, the desaminotyrosine exerts effects by promoting osteoblast activity and / or inhibiting osteoclast activity.

[0007] Optionally, the application dose of the desaminotyrosine is 10-100 mg / kg. Here, "mg / kg" is relative to the body such as a mouse or a human, and is administered per kilogram of body weight.

[0008] Optionally, the osteoporosis comprises at least one of primary osteoporosis, secondary osteoporosis, postmenopausal osteoporosis, senile osteoporosis, and glucocorticoid-induced osteoporosis.

[0009] Optionally, the dosage form of the drug comprises an oral preparation or an injection preparation.

[0010] Optionally, the oral preparation comprises a tablet, a capsule, a granule, or an oral liquid. The injection preparation comprises an injection liquid or a freeze-dried powder injection.

[0011] Optionally, the drug further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

[0012] In a second aspect, the present application provides a pharmaceutical composition for preventing and / or treating osteoporosis, wherein the pharmaceutical composition takes deaminotyrosine or a pharmaceutically acceptable salt thereof as an active ingredient.

[0013] Optionally, the pharmaceutical composition further comprises at least one of a calcium agent, vitamin D, a bisphosphonate, a RANKL inhibitor, a parathyroid hormone analogue, and a selective estrogen receptor modulator.

[0014] Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

[0015] Compared with the prior art, the present application has the following beneficial effects: The present application discloses, for the first time, the use of DAT in the preparation of a drug for preventing and / or treating osteoporosis, which opens up a new application field of the compound. The present application researches and finds that DAT not only can promote the activity of osteoblasts and accelerate bone formation, but also can inhibit the generation and function of osteoclasts and slow down bone resorption, thus achieving a two-way regulation of bone metabolism in a “opening source and saving” manner, which has a significant advantage compared with most existing drugs of a single mechanism. Moreover, DAT is an endogenous small molecule substance in the human body, and is expected to have good biocompatibility and low toxic side effects. In addition, DAT can be used in combination with existing anti-osteoporosis drugs, which can produce a synergistic effect, reduce the dosage and side effects of the existing drugs, and provide a new choice for combination medication. BRIEF DESCRIPTION OF DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments. It should be understood that the following drawings only show some embodiments of the present application, and therefore should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0017] Figure 1 Effect of DAT on bone mineral density (BMD) of ovariectomized (OVX) mouse model and effect of DAT on tibia microstructure of ovariectomized (OVX) mouse model (Micro-CT three-dimensional reconstruction map); Figure 2 Effect of DAT on bone mineral density (BMD) of ovariectomized (OVX) mouse model and effect of DAT on tibia microstructure of ovariectomized (OVX) mouse model (Micro-CT three-dimensional reconstruction map); Figure 3 Effect of DAT on bone tissue structure (HE and MASSON staining) of ovariectomized (OVX) mouse model; Figure 4 Effect of DAT on bone absorption and expression of osteogenesis-related proteins of ovariectomized (OVX) mouse model (Note: A represents TRAP staining picture; B represents TRAP staining statistical column chart in A; C represents NFATC1 immunohistochemical staining picture; D represents statistical column chart of staining results in C; E represents CTSK immunohistochemical staining picture; F represents statistical column chart of staining results in E; G represents RUNX2 immunohistochemical staining picture; H represents statistical column chart of staining results in G; I represents OPN immunohistochemical staining picture; J represents statistical column chart of staining results in I; ** represents p<0.01; *** represents p<0.001; **** represents p<0.0001); Figure 5 Effect of DAT on bone mineral density (BMD) of ovariectomized (OVX) mouse model and effect of DAT on tibia microstructure of ovariectomized (OVX) mouse model (Micro-CT three-dimensional reconstruction map); Figure 6 Effect of DAT on bone tissue structure (HE and MASSON staining) of ovariectomized (OVX) mouse model; DETAILED DESCRIPTION

[0018] The present application will be further described below in conjunction with specific examples. The following description is merely some embodiments of the present application, and does not limit the present application in any form. Although the present application is disclosed as follows with preferred embodiments, it is not intended to limit the present application. Any person skilled in the art can make some changes or modifications to the disclosed technical contents without departing from the scope of the technical solutions of the present application, and such equivalent embodiments are also included in the scope of the technical solutions.

[0019] Unless otherwise specified, the raw materials in the examples of the present application are purchased through commercial channels and directly used without any special treatment.

[0020] Unless otherwise specified, the analysis methods in the examples all use the conventional settings of instruments or equipment and conventional analysis methods. Example

[0021] Effect of DAT on osteoblast differentiation As Figure 1 shown, mouse bone marrow mesenchymal stem cells were inoculated in 24-well plates, cultured in osteogenic induction differentiation medium, and DAT was added at 0 μM, 0.5 μM, 1 μM, 3 μM, and 5 μM, respectively. The medium was changed every 3 days. After 7 days of culture, alkaline phosphatase (ALP) staining was performed. The results are shown in B of Figure 1 , compared with the control group, the ALP staining and alizarin red staining of the DAT treatment group were significantly deepened, indicating that the osteogenic activity was significantly enhanced, suggesting that DAT effectively promoted the differentiation and maturation of osteoblasts. Example

[0022] Therapeutic effect of DAT on ovariectomy (OVX) mouse osteoporosis model 8-week-old female C57 mice were selected and randomly divided into sham operation group (Sham), model group (OVX), DAT low-dose group (100 mM), DAT high-dose group (150 mM), and positive drug control group (alendronate sodium, 1 mg / kg / w). One week after the operation to establish the OVX model, gavage administration was started and lasted for 8 weeks. After the last administration, the femur of the mouse was taken for bone mineral density (BMD) detection and Micro-CT scanning.

[0023] Micro-CT results ( Figure 2 ): The femur BMD of the OVX group of mice was significantly lower than that of the Sham group, while the DAT high and low dose groups could dose-dependently reverse the decrease in BMD, with an effect comparable to that of the positive drug. The bone trabeculae of the OVX group were sparse and broken, while the bone trabeculae of the DAT treatment group, especially the high-dose group, were significantly improved in number, thickness, and connectivity.

[0024] HE staining was used to observe the bone trabeculae around the distal femoral epiphysis ( Figure 3 ). The bone trabeculae structure of the control group mice was dense, mainly in a plate-like structure; the bone trabeculae of the OVX mice were thin and scattered, the number of bone trabeculae was small, and the gap between the bone trabeculae was wide, mainly in a rod-like structure. After intervention with DAT, the number of bone trabeculae increased, the gap decreased, and the structure of the bone trabeculae changed from rod-like to plate-like. MASSON staining was used to observe the collagen content and structural continuity of the bone trabeculae and the surrounding bone marrow / soft tissue, and whether the surface of the bone trabeculae was covered with a continuous blue collagen layer. It was found that the OVX group had thin and interrupted collagen, while the high-dose DAT group had significantly thickened collagen. The width of the red-stained area (osteoid / unmineralized bone) on the edge of the bone trabeculae was examined, and it was found that the OVX group had widened (poor mineralization), while the high-dose DAT group had narrowed, indicating improved mineralization.

[0025] The immunohistochemical results showed that Figure 4OVX group mice femur bone resorption related indicators TRAP, NFATC1 and CTSK were higher than Sham group, while DAT high and low dose groups could dose-dependently reverse the increase of bone resorption related indicators, and the effect was comparable to the positive drug. The femur bone formation related indicators RUNX2 and OPN of OVX group mice were lower than Sham group, while DAT high and low dose groups could dose-dependently reverse the decrease of bone formation related indicators, and the effect was comparable to the positive drug. Embodiments

[0026] Therapeutic effect of DAT on senile mouse osteoporosis model Six-month-old male SAMP8 mice were selected and randomly divided into control group (SAMR1 anti-aging mice), model group (SAMP8), DAT low dose group (100 mM), DAT high dose group (150 mM) and positive drug control group (alendronate sodium, 1 mg / kg / w). Each group of mice started to drink water for 12 weeks. After the last administration, the femur of the mouse was taken for immunohistochemical detection and Micro-CT scanning.

[0027] Micro-CT results Figure 5 ): The trabeculae of SAMP8 group were sparse and broken, while the trabeculae number, thickness and connectivity of DAT treatment group, especially high dose group, were significantly improved.

[0028] HE staining observed the trabeculae around the distal femoral epiphysis Figure 6 The trabeculae structure of SAMR1 group mice was dense, mainly plate structure; the trabeculae of SAMP8 mice were thin and scattered, the trabeculae number was few, and the trabeculae gap was widened, mainly rod structure. After using DAT intervention, the trabeculae number increased, the gap decreased, and the trabeculae structure recovered from rod to plate structure. MASSON staining observed the collagen content and structural continuity of trabeculae and surrounding bone marrow / soft tissue, whether the trabeculae surface was covered by continuous blue collagen layer, and it was found that the SAMP8 group could observe the thin and interrupted collagen, and the high dose of DAT could be seen obvious thickening. The width of the red stained area (osteoid / unmineralized bone) of the trabeculae edge was checked, and the SAMP8 was widened (poor mineralization), and the high dose of DAT group was narrowed, indicating that the mineralization was improved.

[0029] The above is only a few embodiments of the present application, and does not limit the present application in any form. Although the preferred embodiments are disclosed as above, they are not intended to limit the present application. Any skilled person in the art can make some changes or modifications to the above disclosed technical content without departing from the scope of the technical solution of the present application, which are equivalent to equivalent embodiments, and belong to the scope of the technical solution.

Claims

1. Use of deaminotyrosine in the preparation of a medicament for preventing and / or treating osteoporosis.

2. Use according to claim 1, characterized in that, The deaminotyrosine exerts an effect by promoting osteoblast activity and / or inhibiting osteoclast activity.

3. Use according to claim 1, characterized in that, The deaminotyrosine is used in a dosage of 10 to 100 mg / kg.

4. Use according to claim 1, characterized in that, The osteoporosis includes at least one of primary osteoporosis, secondary osteoporosis, postmenopausal osteoporosis, senile osteoporosis, and glucocorticoid-induced osteoporosis.

5. The use according to claim 1, characterized in that, The medicament is in a dosage form including an oral preparation or an injection preparation.

6. Use according to claim 5, characterized in that, The oral preparation includes a tablet, a capsule, a granule, or an oral solution. The injection preparation includes an injection solution or a freeze-dried powder injection.

7. The use according to claim 1, characterized in that, The medicament further includes a pharmaceutically acceptable carrier, diluent, or excipient.

8. A pharmaceutical composition for preventing and / or treating osteoporosis, characterized by, The pharmaceutical composition includes deaminotyrosine or a pharmaceutically acceptable salt thereof as an active ingredient.

9. The pharmaceutical composition for preventing and / or treating osteoporosis according to claim 8, wherein The pharmaceutical composition further includes at least one of a calcium agent, vitamin D, a bisphosphonate, a RANKL inhibitor, a parathyroid hormone analogue, and a selective estrogen receptor modulator.

10. The pharmaceutical composition for preventing and / or treating osteoporosis according to claim 8, wherein The pharmaceutical composition further includes a pharmaceutically acceptable carrier, diluent, or excipient.

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