Use of lactate in preparation of drug for promoting growth and repair of bone / cartilage tissue

Abstract

Provided in the present invention is the use of lactate in the preparation of a drug for promoting the growth and repair of bone / cartilage tissue. The drug has the following effect of either 1) or 2): 1) promoting the growth of bone / cartilage tissue; or 2) promoting the repair and healing of bone / cartilage tissue. The lactate comprises 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-methylimidazolium L-(+)-lactate, 2-hydroxyethyl-trimethylammonium L-(+)-lactate and L-lactic acid-tetrabutylammonium salt. It is found in the present invention for the first time that chitosan lactate, etc. can promote the growth and repair of bone / cartilage tissue, thereby providing a new way to grow and repair bone / cartilage tissue.

Description

Application of lactate in the preparation of drugs for promoting bone / cartilage tissue growth and repair Technical Field

[0001] The present 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, similar to skeletal development, occurs primarily through two pathways: endochondral ossification and intramembranous ossification. Most bone injury repair involves the following sequential and overlapping processes. First, following an inflammatory response, skeletal and vascular stem cells gather and differentiate near the fracture line, gradually forming cartilaginous callus tissue. Peripherally to this central region, at the edges of the new cartilage, new blood vessels form, the periosteum expands, and initial bone formation begins. As chondrocytes further differentiate, they eventually undergo apoptosis, and the cartilage extracellular matrix mineralizes. With the gradual reduction of cartilage tissue, secondary bone formation begins, forming a bony callus. Subsequently, bone remodeling occurs, with compact bone gradually replacing trabecular bone. The bone marrow space is reconstructed, the original bone marrow structure regenerates, and the increased vascularity gradually decreases to normal. When the fracture ends are stable and inactive, callus tissue does not form, and repair is accomplished directly through bone formation by osteoblasts and bone resorption by osteoclasts. Bone injury repair is a continuous process involving constantly changing cell populations and signaling within the regenerating tissue, a pathophysiological process meticulously regulated by multiple cells and factors. 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 produced by carbohydrate fermentation, or anaerobic glycolysis, in skeletal muscle during exercise. Over a century ago, Otto Warburg noted that cancer cells rapidly produce lactate even in the presence of oxygen, a process known as aerobic glycolysis. Literature has reported that lactate has immunosuppressive functions in glycolytic tumors. Further research has shown that lactate can replace glucose as the primary carbon fuel source for most tissues, including immune organs. Recent studies have demonstrated that lactate can increase the stemness of CD8+ T cells, thereby enhancing anti-tumor immunity. While lactate has certain pharmacological activities, its role in bone and cartilage tissue growth remains to be further investigated.

[0004] Summary of the Invention

[0005] In view of the shortcomings of the prior art, the present invention provides the use of lactate in the preparation of a drug for promoting bone / cartilage tissue growth and repair, thereby providing a new strategy for promoting bone / cartilage growth and repair.

[0006] In a first aspect, the present invention provides the use of lactate in the preparation of a drug for promoting bone / cartilage tissue growth and repair, wherein the drug has any of the following effects 1)-2):

[0007] 1) Promote the growth of bone / cartilage tissue;

[0008] 2) Promote bone / cartilage tissue repair and healing.

[0009] 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.

[0010] Furthermore, the lactate is chitosan lactate.

[0011] Furthermore, the concentration of L-lactic acid in the lactate is 2 to 500 mmol / L, preferably 100 to 500 mmol / L.

[0012] Furthermore, the lactate and the first active ingredient form a pharmaceutical composition.

[0013] Furthermore, the first active ingredient has the activity of promoting the growth and repair of bone / cartilage tissue.

[0014] 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 factors, and platelet-rich fibrin.

[0015] Furthermore, the pharmaceutical composition further comprises excipients.

[0016] Furthermore, the excipients include at least one of a pharmaceutically acceptable carrier, a solubilizer, a stabilizer, a filler, a binder, and a surfactant.

[0017] Furthermore, the pharmaceutical composition exists in the form of a preparation.

[0018] Furthermore, the preparation includes solution, powder, aerosol, injection, patch, capsule, pill, tablet or suppository.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] 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

[0021] FIG1 is a graph showing the proliferation results of MC3T3 cells in different concentrations of chitosan lactate and magnesium lactate. DETAILED DESCRIPTION

[0022] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative 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 encompassed within the scope of protection that the present invention is intended to protect.

[0023] 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.

[0024] Example 1: Promoting effect on bone cell growth

[0025] 0, 100, 250, 500 mmol / L chitosan lactate and 250 mmol / L magnesium lactate were added to the culture medium respectively.

[0026] The MTT method was used to test the effect of chitosan lactate on the proliferation of mouse embryonic osteoblasts MC3T3. 4 The number of cells per well was inoculated into 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 the number of cells in each well was calculated by taking three parallel wells. 200 μL of MTT solution was added to each well. After incubation in the dark for 4 hours, the supernatant in each well was removed and then dissolved in 1.5 mL of dimethyl sulfoxide until a transparent solution was obtained. The readings at 570 nm were measured by a microplate reader, and the proliferation results of MC3T3 cells in different concentrations of chitosan lactate were finally calculated.

[0027] The test results are shown in Figure 1. As can be seen from Figure 1, as the concentration of chitosan lactate increases, the number of osteoblast proliferation also gradually increases. 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 invention, the effect is best when the concentration of chitosan lactate is 250mmol / L. Therefore, chitosan lactate at an appropriate concentration has a significant effect on promoting osteoblast proliferation. Magnesium lactate also has a good effect on promoting bone cell proliferation, but at the same concentration, the effect of chitosan lactate on promoting bone cell growth is more obvious than magnesium lactate. The inventor tested 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 chondrocyte proliferation.

[0028] Example 2 Promoting effect on bone tissue growth

[0029] 1. Establishment of osteoporosis model:

[0030] Forty 3-month-old non-pregnant female SD rats weighing about 200 g were selected and underwent bilateral ovariectomy. The rats were then fed for 3 months.

[0031] 2. Implantation of chitosan lactate and in vivo osteogenesis detection:

[0032] (1) Specimen preparation

[0033] Rats were anesthetized and placed prone on a surgical panel. The greater trochanters of both femurs were incised and exposed. Bone defects of 2.5 mm in diameter and 3 mm in depth were created using an electric drill from the greater trochanter toward the lesser trochanter. The muscles and skin were then sutured layer by layer. Forty castrated rats were randomly divided into five groups, each consisting of eight rats: a control group (sham surgery), a saline group, a low-dose chitosan lactate group (injection concentration of 100 mmol / L), a medium-dose chitosan lactate group (injection concentration of 250 mmol / L), and a high-dose chitosan lactate group (injection concentration of 500 mmol / L). Subsequently, chitosan lactate solution or saline solution were injected intraperitoneally at different doses daily for four weeks, depending on group status. Three months after surgery, eight rats from each group were sacrificed, and intact femurs were removed, soft tissue removed, wrapped in saline gauze, and stored at -20°C.

[0034] (2) Measurement of bone density in the intertrochanteric region of the femur

[0035] Femoral specimens were scanned using an X-ray absorptiometry instrument to measure bone mineral content (BMC) and bone mineral density (BMD) in the intertrochanteric region. The results are shown in Table 1. As can be seen from the data in Table 1, three months after surgery, the BMC and BMD in the chitosan lactate group were comparable to those in the saline control group. Furthermore, with increasing chitosan lactate concentration, the BMC and BMD values ​​gradually increased (both differences in the intertrochanteric region were statistically significant). Under these experimental conditions, the high-dose group achieved the best bone repair effect.

[0036] Table 1 Comparison of intertrochanteric bone mineral content and bone density of different samples 3 months after surgery (n=8)

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

[0038] Femora were dissected from the inferior margin of the lesser trochanter, and the proximal portion was placed along its long axis in a sample container. Scanning was performed at 80 kV and a resolution of 6.8 μm to measure bone histomorphometric parameters in the intertrochanteric region. These parameters included: 1) trabecular thickness (Tb.Th): describes the morphology of trabecular bone. Given a given amount of trabeculae, greater thickness indicates greater bone mass; 2) trabecular area percentage (%Tb.Ar): measures the percentage of trabecular area to bone tissue area, reflecting bone mass; and 3) trabecular spacing (Tb.Sp): describes the average distance between trabeculae. Greater spacing indicates greater trabecular spacing and more porous 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-treated group were significantly higher than those of the control group and the saline group 3 months after surgery, and as the chitosan lactate concentration increased, the trabecular thickness gradually increased (the differences between the groups were statistically significant). In addition, the trabecular area percentage of the high-dose group was significantly higher than that of the control group and the saline group, but there was no statistically significant difference compared with the low-dose and medium-dose chitosan lactate-treated groups. The trabecular separation values ​​of the chitosan lactate-treated group were lower than those of the control group and the saline group 3 months after surgery, and as the chitosan lactate concentration increased, the trabecular separation gradually decreased (the differences between the groups were statistically significant). Under the conditions of this experiment, the high-dose 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] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. 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 solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should all 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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