Application of MG53 recombinant protein

Abstract

The invention relates to an application of an MG53 recombinant protein, which is applied to treatment medicines for age-related, glucocorticoid-induced or disuse sarcopenia, and can play a role in increasing muscle mass and muscle bundle cross-sectional area, thereby inhibiting muscle atrophy and improving skeletal muscle functions.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to an application of MG53 recombinant protein. Background Art

[0002] Sarcopenia refers to the decline in skeletal muscle mass, muscle strength or physical function associated with aging, mainly seen in the elderly. Its clinical characteristics are progressive and widespread muscle function loss, with an insidious onset but far-reaching effects. It not only threatens the patient's health, but also puts tremendous pressure on family medical burdens and social public health expenditures.

[0003] At present, the treatment of sarcopenia mainly relies on skeletal muscle rehabilitation training and nutritional support. Drug treatment methods are extremely limited, and a considerable number of patients still fail to benefit from existing treatments. Therefore, the development of new therapeutic drugs for sarcopenia has become an urgent problem to be solved. Summary of the invention

[0004] In view of the above analysis, the embodiments of the present invention aim to provide an application of MG53 recombinant protein to solve the problem of limited existing therapeutic drugs for sarcopenia.

[0005] The present invention provides an application of MG53 recombinant protein, which is applied in therapeutic drugs for age-related, glucocorticoid-induced or disuse sarcopenia.

[0006] Furthermore, the amino acid sequence encoding the MG53 recombinant protein was referenced to the Uniprot database with an ID number of Q6ZMU5.

[0007] Furthermore, the application includes using the MG53 recombinant protein to prepare drugs for treating age-related, glucocorticoid-induced or disuse sarcopenia.

[0008] Furthermore, the purity of the MG53 recombinant protein is not less than 90%.

[0009] Further, the MG53 recombinant protein is dissolved in water or protein buffer;

[0010] The protein buffer is PBS buffer or TrisS basic buffer.

[0011] Furthermore, the PBS buffer solution includes phosphate and NaCl, the concentration of phosphate is 10 mmol / L, the concentration of NaCl is 150 mmol / L, and the solvent is deionized water.

[0012] Furthermore, the TrisS basic buffer comprises tris-HCl and NaCl, the concentration of tris-HCl is 20 mmol / L, the concentration of NaCl is 0.5 mol / L, and the solvent is deionized water.

[0013] Furthermore, the dosage of the MG53 recombinant protein in the mouse age-related, glucocorticoid-induced or disuse sarcopenia model is 10 mg / Kg.

[0014] Furthermore, for patients with age-related, glucocorticoid-induced or disuse sarcopenia, the daily dosage of MG53 recombinant protein for adults is 1.10 mg / Kg body weight.

[0015] Furthermore, the administration routes of the drug include intraperitoneal injection and intravenous infusion.

[0016] Compared with the prior art, the present invention can achieve at least one of the following beneficial effects: The present invention finds that the MG53 recombinant protein can be used in therapeutic drugs for age-related, glucocorticoid-induced or disuse sarcopenia, and can increase muscle mass and muscle bundle cross-sectional area, thereby inhibiting muscle atrophy and improving skeletal muscle function.

[0017] In the present invention, the above-mentioned technical solutions can also be combined with each other to achieve more preferred combination solutions. Other features and advantages of the present invention will be described in the subsequent description, and some advantages can become obvious from the description, or can be understood by practicing the present invention. The purpose and other advantages of the present invention can be realized and obtained through the contents particularly pointed out in the description and the drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The drawings are only for the purpose of illustrating specific embodiments and are not to be considered limiting of the present invention. Like reference symbols denote like components throughout the drawings.

[0019] Figure 1 A schematic diagram of establishing a mouse glucocorticoid-induced muscle atrophy model in a specific embodiment;

[0020] Figure 2 It is a graph showing the results of H&E staining of mouse skeletal muscle sections in the mouse glucocorticoid-induced muscle atrophy model control group, treatment group, model group and model treatment group in a specific embodiment;

[0021] Figure 3 It is a statistical graph of the skeletal muscle mass and the cross-sectional area of ​​the muscle bundles of mice in the control group, treatment group, model group and model treatment group of the mouse glucocorticoid-induced muscle atrophy model in a specific embodiment;

[0022] Figure 4It is a statistical graph of the limb grasping force of mice in the mouse glucocorticoid-induced muscle atrophy model control group, treatment group, model group and model treatment group in a specific embodiment;

[0023] Figure 5 A schematic diagram of establishing a mouse disuse muscle atrophy model in a specific embodiment;

[0024] Figure 6 It is a diagram showing the results of H&E staining of mouse skeletal muscle sections in a mouse disuse muscle atrophy model control group, treatment group, model group, and model treatment group in a specific embodiment;

[0025] Figure 7 It is a statistical graph of the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the mouse disuse muscle atrophy model control group, treatment group, model group and model treatment group in a specific embodiment;

[0026] Figure 8 It is a statistical graph of the limb grasping force of mice in the mouse disuse muscular atrophy model control group, treatment group, model group and model treatment group in a specific embodiment;

[0027] Fig. 9 A schematic diagram of establishing an age-related muscle atrophy model in mice in a specific embodiment;

[0028] Fig.10 It is a diagram showing the results of H&E staining of mouse skeletal muscle sections in the control group, treatment group, model group and model treatment group of the mouse age-related muscular atrophy model in a specific embodiment;

[0029] Fig.11 It is a statistical graph of the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the control group, treatment group, model group and model treatment group of the mouse age-related muscular atrophy model in a specific embodiment;

[0030] Fig.12 It is a statistical graph of the limb grasping strength of mice in the control group, treatment group, model group and model treatment group of the mouse age-related muscular atrophy model in a specific embodiment. DETAILED DESCRIPTION

[0031] The preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings, wherein the accompanying drawings constitute a part of the present invention and are used together with the embodiments of the present invention to illustrate the principles of the present invention, but are not used to limit the scope of the present invention.

[0032] In the description of the embodiments of the present invention, it should be noted that, unless otherwise clearly specified and limited, the term "connected" should be understood in a broad sense, for example, it can be a fixed connection, or a detachable connection, or an integral connection, which can be a mechanical connection, or an electrical connection, which can be a direct connection, or can be indirectly connected through an intermediate medium. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0033] The terms "top", "bottom", "above", "lower", and "on" used throughout the description are relative to the relative positions of components of a device, such as the relative positions of the top and bottom substrates within a device. It is understood that devices are multifunctional regardless of their orientation in space.

[0034] The common working surface of the present invention can be a plane or a curved surface, can be inclined, or can be horizontal. For the convenience of description, the embodiment of the present invention is placed on a horizontal plane and used on the horizontal plane, and "high and low" and "up and down" are defined in this way.

[0035] A specific embodiment of the present invention discloses an application of MG53 recombinant protein, which provides a new drug treatment strategy for age-related, glucocorticoid-induced or disuse sarcopenia, that is, MG53 recombinant protein can be used in therapeutic drugs for age-related, glucocorticoid-induced or disuse sarcopenia.

[0036] The amino acid sequence encoding the MG53 (Mitsugumin 53) recombinant protein is referenced to the Uniprot database, and the ID number is Q6ZMU5.

[0037] The present invention provides an application of MG53 recombinant protein, which includes using the MG53 recombinant protein to prepare a drug for treating age-related, glucocorticoid-induced or disuse sarcopenia.

[0038] When the MG53 recombinant protein is used to prepare a drug for treating age-related, glucocorticoid-induced, or disuse sarcopenia, the purity of the MG53 recombinant protein is not less than 90%.

[0039] When the MG53 recombinant protein is used to prepare a drug for treating age-related, glucocorticoid-induced or disuse sarcopenia, the MG53 recombinant protein is dissolved in water or a protein buffer, wherein the protein buffer is a PBS buffer or a TrisS basic buffer, wherein the PBS buffer solution includes 10 mM phosphate and 150 mM NaCl, that is, the concentration of phosphate is 10 mmol / L, the concentration of NaCl is 150 mmol / L, and the solvent is deionized water; the TrisS basic buffer includes 20 mM tris-HCl (trishydroxymethylaminomethane hydrochloride or Tris hydrochloride), 0.5 M NaCl, that is, the concentration of tris-HCl is 20 mmol / L, the concentration of NaCl is 0.5 mol / L, and the solvent is deionized water.

[0040] The dosage of the MG53 recombinant protein in the age-related, glucocorticoid-induced or disuse sarcopenia mouse model is 10 mg / Kg, which is equivalent to about 1.10 mg / Kg body weight per day for adults. That is, the dosage of the MG53 recombinant protein in the age-related, glucocorticoid-induced or disuse sarcopenia mouse model is 10 mg / Kg, and for patients with age-related, glucocorticoid-induced or disuse sarcopenia, the daily dosage of the MG53 recombinant protein for adults is 1.10 mg / Kg body weight.

[0041] When the MG53 recombinant protein is used to prepare a drug for treating age-related, glucocorticoid-induced or disuse sarcopenia, the administration routes of the drug include intraperitoneal injection and intravenous infusion.

[0042] The application of MG53 recombinant protein in the treatment of age-related, glucocorticoid-induced or disuse sarcopenia can effectively increase muscle mass and muscle bundle cross-sectional area, thereby inhibiting muscle atrophy and improving skeletal muscle function.

[0043] like Figure 1 As shown in the figure, in order to construct a mouse glucocorticoid-induced sarcopenia model, male C57BL / 6 mice aged 6-8 weeks were randomly divided into a control group (Control PBS), a treatment group (Control rhMG53), a model group (DEX PBS) and a model treatment group (DEX rhMG53). The mice in the model group and the model treatment group were intraperitoneally injected with dexamethasone at a dose of 23 μg / day for 14 days. The mice in the treatment group and the model treatment group were intraperitoneally injected with rhMG53 recombinant protein solution from the 4th day, once every 4 days, with a dose of 10 mg / kg body weight each time. After the experiment, the mice in each group were sampled and H&E tested to evaluate the effectiveness of the treatment. The evaluation results are shown in the figure. Figures 2 to 4 shown. Specifically: Figure 4To construct the H&E staining results of skeletal muscle sections of mice in the control group, treatment group, model group and model treatment group in the mouse glucocorticoid-induced sarcopenia model, the results showed that the cross-section of the skeletal muscle bundles of mice in the model group was significantly smaller than that of mice in the control group; there was no significant difference in the skeletal muscle pathology of mice in the treatment group compared with the control group after simple rhMG53 recombinant protein treatment. After treatment with rhMG53 recombinant protein, the cross-section size of the skeletal muscle bundles of mice in the model treatment group was significantly restored compared with that of mice in the model group, indicating that rhMG53 recombinant protein significantly improved glucocorticoid-induced muscle atrophy. Figure 5 To construct the statistical graphs of skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the control group, treatment group, model group and model treatment group in the mouse glucocorticoid-induced sarcopenia model, the results showed that the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the model group were significantly reduced compared with those in the control group; there was no significant difference in skeletal muscle mass and muscle bundle cross-sectional area between the mice in the treatment group and those in the control group after simple administration of rhMG53 recombinant protein. After treatment with rhMG53 recombinant protein, the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the model treatment group were significantly restored compared with those in the smoked group, indicating that rhMG53 recombinant protein significantly improved the reduction of skeletal muscle mass and muscle atrophy induced by glucocorticoid. Figure 6 To construct a statistical graph of the limb grasping strength of mice in the control group, treatment group, model group and model treatment group in the mouse glucocorticoid-induced sarcopenia model, the results showed that the skeletal muscle strength (i.e., limb grasping strength) of the model group mice was significantly reduced compared with the control group mice; there was no significant difference in the skeletal muscle strength of the treatment group mice compared with the control group after simple administration of rhMG53 recombinant protein. After treatment with rhMG53 recombinant protein, the skeletal muscle strength of the model treatment group mice was significantly improved compared with the model group mice, indicating that rhMG53 recombinant protein significantly improved the decline in skeletal muscle strength induced by glucocorticoids.

[0044] like Figure 5 As shown in the figure, in order to construct a mouse disuse sarcopenia model, male C57BL / 6 mice aged 6-8 weeks were randomly divided into a control group (Control PBS), a treatment group (Control rhMG53), a model group (DEN PBS) and a model treatment group (DEN rhMG53). The sciatic nerve of the right hind limb of the mice in the model group and the model treatment group was surgically removed. The control group and the treatment group were not treated. The mice in the treatment group and the model treatment group were intraperitoneally injected with rhMG53 recombinant protein solution from the 4th day after surgery, once every 4 days, with a dosage of 10 mg / Kg body weight each time. After 2 weeks, samples were collected and H&E tests were performed on each group of mice to evaluate the effectiveness of the treatment. The evaluation results are shown in the figure. Figure 6 to Figure 8 shown. Specifically, Figure 6To construct the H&E staining results of skeletal muscle sections of mice in the control group, treatment group, model group and model treatment group in the mouse disuse sarcopenia model, the results showed that the cross-section of skeletal muscle bundles in the model group mice was significantly smaller than that in the control group mice; there was no significant difference in skeletal muscle pathology between the treatment group mice and the control group mice when rhMG53 recombinant protein was given alone. After treatment with rhMG53 recombinant protein, the cross-section size of skeletal muscle bundles in the model treatment group mice was significantly restored compared with that in the model group mice, indicating that rhMG53 recombinant protein significantly improves disuse muscle atrophy. Figure 7 To construct the statistical graphs of skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the control group, treatment group, model group and model treatment group in the mouse disuse sarcopenia model, the results showed that the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the model group were significantly reduced compared with those in the control group; there was no significant difference in skeletal muscle mass and muscle bundle cross-sectional area between the mice in the treatment group and those in the control group after simple administration of rhMG53 recombinant protein. After treatment with rhMG53 recombinant protein, the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the model treatment group were significantly restored compared with those in the smoked group, indicating that rhMG53 recombinant protein significantly improves disuse-related skeletal muscle mass reduction and muscle atrophy. Figure 8 To construct a statistical graph of the limb grasping strength of mice in the control group, treatment group, model group and model treatment group in the mouse disuse sarcopenia model, the results showed that the skeletal muscle strength (i.e., limb grasping strength) of the model group mice was significantly reduced compared with the control group mice; there was no significant difference in the skeletal muscle strength of the treatment group mice compared with the control group after simple administration of rhMG53 recombinant protein. After treatment with rhMG53 recombinant protein, the skeletal muscle strength of the model treatment group mice was significantly improved compared with the model group mice, indicating that rhMG53 recombinant protein significantly improves the disuse-related skeletal muscle strength decline.

[0045] like Fig. 9 As shown, in order to construct an age-related sarcopenia model in mice, male C57BL / 6 mice aged 6-8 weeks were randomly divided into a control group (Control PBS), a treatment group (Control rhMG53), an elderly group (AGED PBS) and an elderly treatment group (AGED rhMG53). The mice in the model group and the model treatment group grew naturally to 52 weeks. The control group and the treatment group grew naturally to 8 weeks. The mice in the treatment group were intraperitoneally injected with rhMG53 recombinant protein solution from the 8th week, and the mice in the elderly treatment group were intraperitoneally injected with rhMG53 recombinant protein solution from the 52nd week, twice a week, with each dose of 10 mg / Kg body weight. After 15 days, samples were collected from each group of mice and H&E tests were performed to evaluate the effectiveness of the treatment. The evaluation results are shown in the following table. Figure 10 to Figure 12 shown. Specifically, Fig.10To construct the results of H&E staining of skeletal muscle sections of mice in the control group, treatment group, elderly group and elderly treatment group in the mouse age-related sarcopenia model, the results showed that the cross-section of skeletal muscle bundles in the elderly group was significantly smaller than that in the control group; there was no significant difference in skeletal muscle pathology between the treated group and the control group after simple administration of rhMG53 recombinant protein. After treatment with rhMG53 recombinant protein, the cross-section size of skeletal muscle bundles in the elderly treatment group was significantly restored compared with that in the elderly group, indicating that rhMG53 recombinant protein significantly improves age-related muscle atrophy. Fig.11 To construct a statistical chart of skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the control group, treatment group, elderly group and elderly treatment group in the mouse age-related sarcopenia model, the results showed that the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the elderly group were significantly reduced compared with those in the control group; there was no significant difference in skeletal muscle mass and muscle bundle cross-sectional area between the mice in the treatment group and those in the control group after simple administration of rhMG53 recombinant protein. After treatment with rhMG53 recombinant protein, the skeletal muscle mass and muscle bundle cross-sectional area of ​​mice in the elderly treatment group were significantly restored compared with those in the smoked group, indicating that rhMG53 recombinant protein significantly improved age-related skeletal muscle mass reduction and muscle atrophy. Fig.12 To construct a statistical graph of the limb grasping strength of mice in the control group, treatment group, elderly group and elderly treatment group in the mouse age-related sarcopenia model, the results showed that the skeletal muscle strength (i.e., limb grasping strength) of the elderly group mice was significantly reduced compared with the control group mice; there was no significant difference in the skeletal muscle strength of the treatment group mice compared with the control group after simple rhMG53 recombinant protein treatment. After rhMG53 recombinant protein treatment, the skeletal muscle strength of the elderly treatment group mice was significantly improved compared with the elderly group mice, indicating that rhMG53 recombinant protein significantly improves the age-related decline in skeletal muscle strength.

[0046] The MG53 recombinant protein provided by the present invention can be used in drugs for treating age-related, glucocorticoid-induced or disuse sarcopenia. After treatment with the MG53 recombinant protein, the muscle mass and muscle bundle cross-sectional area of ​​age-related, glucocorticoid-induced or disuse sarcopenia model mice can be increased; at the same time, the MG53 recombinant protein can improve the skeletal muscle function (such as limb grip strength) of age-related, glucocorticoid-induced or disuse sarcopenia model mice.

[0047] The MG53 recombinant protein provided by the present invention is not toxic to important organs, and mice treated with the MG53 recombinant protein do not show significant toxic and side effects.

[0048] The MG53 recombinant protein provided by the present invention is dissolved in PBS and then directly injected systemically by intraperitoneal injection, which is feasible for clinical application and provides ideas and directions for the subsequent treatment and prevention of age-related, glucocorticoid-induced or disuse sarcopenia.

[0049] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by any technician familiar with the technical field within the technical scope disclosed by the present invention should be covered within the protection scope of the present invention.

Claims

1. An application of MG53 recombinant protein, characterized in that: Application in the treatment of age-related, glucocorticoid-induced or disuse sarcopenia.

2. The use according to claim 1, characterized in that: The amino acid sequence encoding the MG53 recombinant protein was referenced to the Uniprot database with the ID number Q6ZMU5.

3. The use according to claim 1, characterized in that: Including MG53 recombinant protein for preparing drugs for treating age-related, glucocorticoid-induced or disuse sarcopenia.

4. The use according to claim 3, characterized in that: The purity of the MG53 recombinant protein is not less than 90%.

5. The use according to claim 3, characterized in that: The MG53 recombinant protein is dissolved in water or protein buffer; The protein buffer is PBS buffer or TrisS basic buffer.

6. The use according to claim 5, characterized in that: The PBS buffer solution includes phosphate and NaCl, the concentration of phosphate is 10 mmol / L, the concentration of NaCl is 150 mmol / L, and the solvent is deionized water.

7. The use according to claim 5, characterized in that: The TrisS basic buffer comprises tris-HCl and NaCl, the concentration of tris-HCl is 20 mmol / L, the concentration of NaCl is 0.5 mol / L, and the solvent is deionized water.

8. The use according to claim 3, characterized in that: The dosage of the MG53 recombinant protein in the mouse age-related, glucocorticoid-induced or disuse sarcopenia model is 10 mg / Kg.

9. The use according to claim 3, characterized in that: For patients with age-related, glucocorticoid-induced or disuse sarcopenia, the daily dosage of MG53 recombinant protein for adults is 1.10 mg / Kg body weight.

10. The use according to claim 3, characterized in that: The administration routes of the drug include intraperitoneal injection and intravenous infusion.

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