Application of lactate in preparation of medicine for promoting growth and repair of bone / cartilage tissue

Abstract

The invention provides application of lactate in preparation of a medicine for promoting growth and repair of bone / cartilage tissues. The medicine has any one of the following effects: 1) promoting growth of the bone / cartilage tissues; (2) bone / cartilage tissue repair and healing are promoted; the lactate comprises 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-methylimidazole L-(+)-lactate and 2-hydroxyethyl-trimethyl ammonium L-(+)-lactate; and L, L-lactic acid-tetrabutylammonium salt is one or more. It is found for the first time that chitosan lactate and the like can promote growth and repair of bone / cartilage tissue, and a new way is provided for growth and repair of bone / cartilage tissue.

Description

Technical Field

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

[0002] Bone tissue repair is similar to the process of bone development, and is mainly completed through two methods: endochondral ossification and intramembranous ossification. Most bone injury repair includes the following continuous and overlapping processes. First, after the inflammatory response, bone and blood vessel-related stem cells gather and differentiate in the area near the fracture line, gradually forming cartilaginous callus tissue. On the periphery of this central area, at the edge of the new cartilage tissue, new blood vessels form, the periosteum expands, and the initial bone formation process begins. With the further differentiation of chondrocytes, chondrocytes eventually undergo apoptosis, and the extracellular matrix of chondrocytes undergoes mineralization. With the gradual reduction of cartilage tissue, secondary bone formation begins, forming bone callus. Subsequently, the bone remodeling stage begins, compact bone gradually replaces trabecular bone, the bone marrow space is reconstructed, the original bone marrow structure is regenerated, and the increased blood vessels gradually decrease and return to normal. When the fracture ends are stable and less active, callus tissue is not formed, and repair is completed directly through osteoblast bone formation and osteoclast bone resorption. Bone injury repair is a continuous process composed of constantly changing cell clusters and signal transmission in regenerating tissues. It is a pathophysiological process in which multiple cells and factors participate in fine regulation. Although most bone injuries can heal normally without leaving scars, 5% to 10% will experience delayed healing or nonunion.

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

[0004] In view of the deficiencies in the prior art, the present invention provides the use of lactate in the preparation of drugs for promoting the growth and repair of bone / cartilage tissue, which provides a new strategy for promoting the growth and repair of bone / cartilage.

[0005] In a first aspect of the present invention, there is provided a use of lactate in the preparation of a drug for promoting the growth and repair of bone / cartilage tissue, wherein the drug has the effect of any one of the following 1)-2):

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

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

[0008] The lactate includes 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] Furthermore, the lactate is chitosan lactate.

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

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

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

[0013] Furthermore, 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.

[0014] Furthermore, the pharmaceutical composition also contains excipients.

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

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

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

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

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

[0020] Figure 1 This is a graph showing the proliferation results of MC3T3 cells in different concentrations of chitosan lactate and magnesium lactate. DETAILED DESCRIPTION

[0021] The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[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 was purchased from Shandong Aokang Biotechnology Co., Ltd.

[0023] Example 1 Promoting effect on bone cell growth

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

[0025] The MTT method was used to test the effect of chitosan lactate on the proliferation of mouse embryonic osteoblasts MC3T3. 4 / well were inoculated in a 12-well plate, 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 3 parallel wells were taken to calculate the number of cells in each well. 200 μL of MTT solution was added to each well, and after incubation for 4 hours in the dark, the supernatant in each well was removed and then dissolved into a transparent solution with 1.5 mL of dimethyl sulfoxide. The reading at 570 nm was tested by an ELISA instrument, and the proliferation results of MC3T3 cells in different concentrations of chitosan lactate were finally calculated.

[0026] Test results such as Figure 1 As shown. Figure 1It can be seen that with the increase of chitosan lactate concentration, the number of osteoblast proliferation also gradually increases. However, when the chitosan lactate concentration continues to increase, the influence on cell proliferation is no longer obvious, that is, under the experimental conditions of the present invention, when the chitosan lactate concentration is 250mmol / L, the effect is best. Therefore, chitosan lactate at an appropriate concentration has a significant effect on promoting osteoblast proliferation. Magnesium lactate also has a good effect on promoting osteocyte proliferation, but the effect of chitosan lactate in promoting osteocyte growth is more obvious than magnesium lactate at the same concentration. The inventor detected the effect of chitosan lactate on chondrocytes by the same method and obtained the same effect, indicating that chitosan lactate has a significant effect on promoting chondrocyte proliferation.

[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 weighing about 200 g were selected and underwent castration surgery to remove bilateral ovaries. The rats were then fed for 3 months after surgery.

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

[0031] (1) Specimen preparation

[0032] Rats were anesthetized and placed prone on the surgical panel. The greater trochanters of both femurs were cut and exposed. 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 with an electric drill, and the muscles and skin were sutured in layers. Forty ovariectomized rats were randomly divided into five groups, with 8 rats in each group. They were: control group (sham operation), normal saline group, low-dose chitosan lactate group (injection concentration of 100 mmol / L), medium-dose chitosan lactate group (injection concentration of 250 mmol / L), and high-dose chitosan lactate group (injection concentration of 500 mmol / L). Subsequently, different doses of chitosan lactate solution or normal saline were intraperitoneally injected according to the grouping, once a day for 4 consecutive weeks. Eight rats in each group were killed 3 months after surgery, and the intact femurs were removed. After the soft tissues were removed, they were wrapped with normal saline gauze and stored at -20°C.

[0033] (2) Bone density measurement of the intertrochanteric region

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

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

[0036]

[0037] (3) Micro CT observation of trabecular bone structure in the intertrochanteric region of the femur

[0038] The femur was cut off from the lower edge of the lesser trochanter, and the proximal part was placed in a sample container along the long axis. The femur was scanned at a voltage of 80 kV and a resolution of 6.8 μm to measure the bone morphometric indicators of the intertrochanteric region of the femur. Among them, 1) trabecular thickness (Tb.Th): used to describe the structural morphology of trabecular bone. Under the condition of a certain number, the greater the thickness, the more 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 trabeculae. The greater the separation, the greater the trabecular spacing, and the more porous the bone. The results are shown in Table 2.

[0039] As can be seen from the data in Table 2, the trabecular thickness values ​​of the chitosan lactate administration group were significantly higher than those of the control group and the saline group 3 months after surgery, and as the concentration of chitosan lactate increased, the trabecular thickness gradually increased (the differences between the groups were statistically significant). In addition, the percentage of trabecular area in the high-dose administration group was significantly higher than that in the control group and the saline group, but there was no statistically significant difference compared with the low-dose and medium-dose chitosan lactate administration groups. The trabecular separation values ​​of the chitosan lactate administration group were lower than those of the control group and the saline group 3 months after surgery, and as the concentration of chitosan lactate increased, the trabecular separation gradually decreased (the differences between the groups were statistically significant). Under the experimental conditions, the high-dose administration group had the best bone repair effect.

[0040] Table 2 Comparison of intertrochanteric trabecular histomorphology of different samples 3 months after surgery (n=8)

[0041]

[0042] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. The use of lactate in the preparation of drugs for promoting the growth and repair of bone / cartilage tissue, characterized in that: The drug has the effect of any one of the following 1)-2): 1) Promote the growth of bone / cartilage tissue; 2) Promote bone / cartilage tissue repair and healing. The lactate includes 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.

2. The use according to claim 1, characterized in that: The lactate is chitosan lactate.

3. The use according to claim 1, characterized in that: The concentration of L-lactic acid in the lactate is 2 to 500 mmol / L, preferably 100 to 500 mmol / L.

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

5. The use according to claim 4, characterized in that: The first active ingredient has the activity of promoting the growth and repair of bone / cartilage tissue.

6. The use according to claim 5, characterized in that: 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.

7. The use according to claim 4, characterized in that: The pharmaceutical composition further comprises excipients.

8. The use according to claim 7, characterized in that: The auxiliary materials include at least one of a pharmaceutically acceptable carrier, a solubilizer, a stabilizer, a filler, a binder, and a surfactant.

9. The use according to claim 4, characterized in that: The pharmaceutical composition is in the form of a preparation.

10. The use according to claim 9, characterized in that: The preparations include solutions, powders, aerosols, injections, patches, capsules, pills, tablets or suppositories.

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