Use of lactate salts in the manufacture of a medicament for promoting bone / cartilage tissue growth and repair

Abstract

The application provides application of lactate in preparation of a medicine for promoting growth and repair of bone / cartilage tissue, and the medicine has the effect of any one of 1) to 2): 1) promoting growth of bone / cartilage tissue; and 2) promoting repair and healing of bone / cartilage tissue. The lactate includes one or more of magnesium lactate, sodium lactate, potassium lactate, ferrous lactate, chromium lactate, copper lactate, manganese lactate, zinc lactate, lithium lactate, aluminum lactate, chitosan lactate, halofuginone lactate, trimethoprim lactate, 1-ethyl-3-methyl imidazole L-(+)-lactate, 2-hydroxyethyl-trimethyl ammonium L-(+)-lactate and L-lactic acid-tetrabutyl ammonium salt. The chitosan lactate and the like can promote growth and repair of bone / cartilage tissue for the first time, and a new way is provided for growth and repair of bone / cartilage tissue.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of lactic acid drugs, in particular to application of lactic acid salt in preparation of drugs for promoting growth and repair of bone / cartilage tissue. BACKGROUND

[0002] Bone tissue repair is similar to bone development process, which is mainly completed through endochondral ossification and intramembranous ossification. Most bone injury repair includes the following processes occurring continuously and overlapping each other. First, after inflammatory reaction, bone and blood vessel related stem cells gather and differentiate in the area near the fracture line, and gradually form cartilaginous callus tissue. In the periphery of this central area, at the edge of the new cartilage tissue, new blood vessels form, periosteum expands, and initial bone formation process begins. With further differentiation of chondrocytes, chondrocytes eventually undergo apoptosis, and the extracellular matrix of chondrocytes is mineralized. With the gradual reduction of cartilage tissue, secondary bone formation begins, forming bony callus. Subsequently, into the bone remodeling stage, the trabecular bone is gradually replaced by compact bone, the bone marrow space is reconstructed, and the original bone marrow structure is regenerated, and the increased blood vessels gradually decrease to normal. When the fracture ends are stable and less active, no callus tissue is formed, and the repair is completed directly through the bone formation of osteoblasts and the bone absorption of osteoclasts. Bone injury repair is a continuous process composed of constantly changing cell populations and signal transmission in the regenerated tissue, and is a pathological and physiological process in which multiple cells and factors are involved in fine regulation. Although most bone injuries can heal normally without scarring, 5% to 10% will have delayed union or nonunion of bone.

[0003] Lactic acid salt has always been considered as a metabolic waste produced by carbohydrate fermentation or anaerobic glycolysis of skeletal muscle during exercise. More than a century ago, Otto Warburg noticed that cancer cells can rapidly produce lactic acid even in the presence of oxygen, a process known as aerobic glycolysis. It has been reported that lactic acid has immunosuppressive function in glycolytic tumors. Further studies have shown that lactic acid salt can replace glucose as the main carbon fuel source for most tissues, including immune organs. Recent studies have shown that lactic acid salt can increase the stemness of CD8+ T cells to enhance anti-tumor immunity. As can be seen, lactic acid salt has certain pharmacological activity, but the role of lactic acid salt in bone / cartilage tissue growth still needs further research. SUMMARY

[0004] In view of the deficiencies in the prior art, the present application provides application of lactic acid salt in preparation of drugs for promoting growth and repair of bone / cartilage tissue. It provides a new strategy for promoting growth and repair of bone / cartilage.

[0005] In a first aspect, the present application provides use of a lactate salt in the preparation of a drug for promoting growth and repair of bone / cartilage tissue, wherein the drug has the effect of any one of 1) to 2) below:

[0006] 1) promoting growth of bone / cartilage tissue;

[0007] 2) promoting repair and healing of bone / cartilage tissue.

[0008] The lactate salt includes one or several of one or more of magnesium lactate, sodium lactate, potassium lactate, ferrous lactate, chromium lactate, manganese lactate, zinc lactate, lithium lactate, aluminum lactate, chitosan lactate, halofuginone lactate, trimethoprim lactate, 1-ethyl-3-methylimidazole L-(+)-lactate, 2-hydroxyethyl-trimethylammonium L-(+)-lactate, and L-lactic acid-tetrabutylammonium salt.

[0009] Further, the lactate salt is chitosan lactate.

[0010] Further, the concentration of L-lactic acid in the lactate salt is 2 to 500 mmol / L, preferably 100 to 500 mmol / L.

[0011] Further, the lactate salt and the first active ingredient form a pharmaceutical composition.

[0012] Further, the first active ingredient has the activity of promoting growth and repair of bone / cartilage tissue.

[0013] Further, the first active ingredient includes one or several of collagen, gelatin, osteocalcin, bone morphogenetic protein, basic fibroblast growth factor, insulin growth factor, transforming growth factor β, platelet-derived growth factor, fibrin containing concentrated growth factor, and platelet-rich fibrin.

[0014] Further, the pharmaceutical composition further comprises an excipient.

[0015] Further, the excipient includes at least one of a pharmaceutically acceptable carrier, a solubilizer, a stabilizer, a filler, a binder, and a surfactant.

[0016] Further, the pharmaceutical composition is in the form of a preparation.

[0017] Further, the preparation includes a solution, a powder, an aerosol, an injection, a patch, a capsule, a pill, a tablet, or a suppository.

[0018] Compared with the prior art, the present application has the following beneficial effects:

[0019] The present application first discovers that chitosan lactate and the like can promote the growth and repair of bone / cartilage tissue, and provides a new approach for the growth and repair of bone / cartilage tissue. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The proliferation results of MC3T3 cells in different concentrations of chitosan lactate and magnesium lactate are shown in the graph. DETAILED DESCRIPTION

[0021] The technical solutions of the present application will be further described in detail below in combination with specific examples. It should be understood that the following examples are only illustratively described and explained, and should not be interpreted as limiting the scope of protection of the present application. Any technology realized based on the above description of the present application is covered within the scope of protection intended by the present application.

[0022] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods. Chitosan lactate is purchased from Shandong Aokang Biotechnology Co., Ltd.

[0023] Example 1 Promoting effect on bone cell growth

[0024] 0, 100, 250, 500 mmol / L of chitosan lactate and 250 mmol / L of magnesium lactate were added to the culture medium, respectively.

[0025] The effect of chitosan lactate on the proliferation of mouse embryonic osteoblast cells MC3T3 was tested by MTT method. First, the cells were inoculated in a 12-well plate at a concentration of 2×10 4 cells / well, and 2 mL of culture medium containing different concentrations of chitosan lactate was added to each well. Then, the medium was changed every two days, and the number of cells in each well was calculated by taking three parallel wells. 200 μL of MTT solution was added to each well, and incubated in the dark for 4 hours. Then, the supernatant in each well was removed, and 1.5 mL of dimethyl sulfoxide was added to dissolve it into a transparent solution. The reading at 570 nm was tested by an enzyme marker, and the proliferation results of MC3T3 cells in different concentrations of chitosan lactate were finally calculated.

[0026] The test results are shown in Table 1. Figure 1 Figure 1 ​It can be seen that the number of osteoblast proliferation gradually increases with the increase of the concentration of chitosan lactate. However, when the concentration of chitosan lactate continues to increase, the effect on cell proliferation is no longer obvious, that is, under the experimental conditions of the present application, the effect is the best when the concentration of chitosan lactate is 250 mmol / L. Therefore, chitosan lactate with an appropriate concentration has a significant effect on promoting the proliferation of osteoblasts. Magnesium lactate also has a good effect on promoting the proliferation of bone cells, but under the same concentration, chitosan lactate promotes the growth of bone cells more obviously than magnesium lactate. The inventors detected the effect of chitosan lactate on chondrocytes in the same way and obtained the same effect, indicating that chitosan lactate has a significant effect on promoting the proliferation of chondrocytes.

[0027] Example 2: Promoting effect on bone tissue growth

[0028] 1. Establishment of osteoporosis model:

[0029] Forty 3-month-old non-pregnant female SD rats with a body weight of about 200 g were selected, and both ovaries were removed by castration operation, and then fed for 3 months.

[0030] 2. Implantation of chitosan lactate and detection of in vivo osteogenesis:

[0031] (1) Specimen preparation

[0032] After the rats were anesthetized, they were placed on the operation panel in a prone position. The bilateral greater trochanter was incised and exposed, and a bone defect with a diameter of 2.5 mm and a depth of 3 mm was made from the greater trochanter to the lesser trochanter by an electric drill, and then the muscle and skin were sutured layer by layer. Forty castrated rats were randomly divided into five groups, eight rats in each group. They are: control group (sham operation), normal saline group, low-dose chitosan lactate group (injected at a concentration of 100 mmol / L), medium-dose chitosan lactate group (injected at a concentration of 250 mmol / L), and high-dose chitosan lactate group (injected at a concentration of 500 mmol / L). Then, according to the grouping, different doses of chitosan lactate solution or normal saline were injected into the abdominal cavity, once a day, for 4 consecutive weeks. Three months after the operation, eight rats were sacrificed from each group, and the complete femur was taken out, the soft tissue was removed, and then wrapped with normal saline gauze and stored at -20°C.

[0033] (2) Measurement of bone mineral density in the intertrochanteric region of the femur

[0034] The femur specimen was scanned by X-ray bone densitometer to determine the bone mineral content (BMC) and bone mineral density (BMD) of the intertrochanteric region. The results are shown in Table 1. As can be seen from the data in Table 1, the bone mineral content and bone mineral density of the chitosan lactate administration group were not lower than those of the control group of normal saline group 3 months after the operation, and the bone mineral content and bone mineral density values gradually increased with the increase of the concentration of chitosan lactate (the comparison of the bone mineral content and bone mineral density of the intertrochanteric region had statistical significance). Under the experimental conditions, the bone repair effect of the high-dose administration group was the best.

[0035] Table 1 Comparison of bone mineral content and bone mineral density of the intertrochanteric region of the femur 3 months after the operation of different samples (n = 8)

[0036]

[0037] (3) Observation of the trabecular structure of the intertrochanteric region of the femur by Micro CT

[0038] The femur was disconnected from the lower edge of the small trochanter, and the proximal part was placed in the sample container along the long axis. Scanning was performed under the conditions of voltage 80 kV and resolution 6.8 μm to determine the bone morphometric indexes of the intertrochanteric region of the femur. Among them, 1) trabecular thickness (Tb.Th): used to describe the morphological structure of the trabecula, under the condition of a certain quantity, the greater the thickness, the more the bone mass; 2) percentage of trabecular area (%Tb.Ar): the percentage of trabecular area to bone tissue area, reflecting the amount of bone mass; 3) trabecular spacing (Tb.Sp): used to describe the average distance between the trabecula, the greater the separation degree, the greater the distance between the trabecula, and the more osteoporotic the bone. The results are shown in Table 2.

[0039] As can be seen from the data in Table 2, the trabecular thickness value of the chitosan lactate administration group was significantly higher than that of the control group and the normal saline group 3 months after the operation, and the trabecular thickness gradually increased with the increase of the concentration of chitosan lactate (the difference between each group had statistical significance). In addition, the percentage of trabecular area of the high-dose administration group was significantly higher than that of the control group and the normal saline group, but there was no statistical significance compared with the low-dose and medium-dose chitosan lactate administration groups. The trabecular separation degree value of the chitosan lactate administration group was lower than that of the control group and the normal saline group 3 months after the operation, and the trabecular separation degree gradually decreased with the increase of the concentration of chitosan lactate (the difference between each group had statistical significance). Under the experimental conditions, the bone repair effect of the high-dose administration group was the best.

[0040] Table 2 Comparison of the trabecular bone histomorphometry of the femoral intertrochanteric bone 3 months after operation (n=8)

[0041]

[0042] Finally, it should be noted that the above examples are merely intended to illustrate the technical solutions of the present application but not to limit the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or equivalently replaced without departing from the spirit and scope of the present application, and all modifications and equivalents thereof should be included in the scope of the claims of the present application.

Claims

1. Use of chitosan lactate for the preparation of a medicament for the treatment of osteoporosis, characterized in that: The drug has the effects of increasing bone mineral content (BMC), bone mineral density (BMD) and trabecular bone thickness (Tb.Th), reducing trabecular bone separation (Tb.Sp) and reducing bone mass loss caused by osteoporosis. The concentration of L-lactic acid in the chitosan lactate is 100-500 mmol / L.

2. Use according to claim 1, characterized in that: The lactate and the first active ingredient form a pharmaceutical composition.

3. Use according to claim 2, wherein: The first active ingredient has the activity of promoting growth and repair of bone / cartilage tissue.

4. Use according to claim 3, wherein: The first active ingredient includes one or more of collagen, gelatin, osteocalcin, bone morphogenetic protein, basic fibroblast growth factor, insulin growth factor, transforming growth factor β, platelet-derived growth factor, fibrin containing concentrated growth factor and platelet-rich fibrin.

5. The use according to claim 2, characterized in that: The pharmaceutical composition further comprises an auxiliary material.

6. The use according to claim 5, characterized in that: The auxiliary material includes at least one of a solubilizing agent, a stabilizer, a filler, a binder, a surfactant.

7. The use according to claim 2, wherein: The pharmaceutical composition exists in a preparation form.

8. Use according to claim 7, wherein: The preparation is a powder, an aerosol, an injection, a patch, a capsule, a pill, a tablet or a suppository.

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