Application of a short peptide from rice bran in prevention and treatment of bone-related diseases

Abstract

The application discloses application of a rice bran-derived short peptide in a drug for preventing and treating bone-related diseases, wherein the amino acid sequence of the rice bran-derived short peptide is Lys-His-Asn-Arg-Gly-Asp-Glu-Phe (from the N terminal), or an amino acid sequence with equivalent functions formed by replacing, deleting or adding one or more amino acids in the sequence, which is used for preventing and treating osteoporosis, promoting bone tissue repair and regeneration or any combination thereof. The rice bran-derived short peptide in the application can be synthesized by using existing conventional methods. The application also provides the short peptide after functional enhancement modification (such as connection of nucleic acid aptamer and / or antibody, wrapping of a lipid membrane and the like), and application of the short peptide in preventing and treating bone diseases related to bone mass loss, bone defects and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to an application of a short peptide from rice bran in a drug for preventing and treating bone-related diseases. BACKGROUND

[0002] Osteoporosis is a common metabolic bone disease in the elderly, which is characterized by bone loss and bone microstructure destruction, and further leads to increased bone fragility and easy fracture. With the aging of the population, osteoporosis has become a serious social problem that threatens human health. According to different causes, osteoporosis can be divided into primary and secondary categories. Primary osteoporosis is further divided into postmenopausal, senile and idiopathic osteoporosis, while secondary osteoporosis is mostly caused by diseases or other factors, including disuse osteoporosis. The occurrence of osteoporosis and its resulting fractures not only affects the survival and quality of life of patients, but also brings huge economic pressure to society. Therefore, actively developing research on the prevention and treatment of osteoporosis has great social and economic significance. Current anti-osteoporosis drugs have some effect on bone regeneration, but usually have long treatment cycles, high costs and side effects. Therefore, it is urgent and necessary to explore new strategies for preventing or treating osteoporosis that are safer, more efficient and cheaper.

[0003] Rice bran is the byproduct of rice processing, the first largest grain in China, and is rich in plant protein. Studies have found that the amino acid sequence of rice bran protein is basically the same as that of soybean protein, both of which are important protein sources recommended by the Food and Agriculture Organization of the United Nations and the World Health Organization, and are rich in content, easy to be digested and absorbed by the human body, and safe and non-toxic. Rice bran short peptides are a mixture of short peptides obtained by enzymatic hydrolysis of rice bran protein. Several studies have reported that rice bran short peptides have antioxidant, anti-fatigue, immune-enhancing and anti-tumor effects. Our team used a MicroTOF-QII liquid chromatography-mass spectrometry high-resolution mass spectrometer to isolate a short peptide composed of Lys-His-Asn-Arg-Gly-Asp-Glu-Phe from rice bran for the first time, and reported that it can protect endothelial cells from damage induced by hydrogen peroxide by inhibiting the activation of the NF-κb pathway. There is no report on the application of this short peptide in the prevention and treatment of osteoporosis. SUMMARY

[0004] The present application provides an application of a short peptide from rice bran in a drug for preventing and treating bone-related diseases, which aims to solve the above-mentioned problems in the prior art. The short peptide from rice bran in the drug for preventing and treating bone-related diseases can prevent and treat osteoporosis.

[0005] In order to achieve the above object, the present application provides an application of a rice bran-derived short peptide in a drug for preventing and treating bone-related diseases, wherein the amino acid sequence of the rice bran-derived short peptide is Lys-His-Asn-Arg-Gly-Asp-Glu-Phe, and the amino acid sequence starts from the N terminal; wherein Lys represents lysine, His represents histidine, Asn represents asparagine, Arg represents arginine, Gly represents glycine, Asp represents aspartic acid, Glu represents glutamic acid, and Phe represents phenylalanine.

[0006] It should be noted that the amino acid sequence of the rice bran-derived short peptide of the present application is shown in SEQ ID: 1.

[0007] It should be noted that Table 1 below shows the English name, three-letter abbreviation and single letter of the exemplary amino acid.

[0008] Table 1 English name, three-letter abbreviation and single letter of amino acid

[0009]

[0010] According to an aspect of the present application, the molecular weight of the rice bran-derived short peptide is 1002.6 Da.

[0011] According to an aspect of the present application, the rice bran-derived short peptide can be artificially synthesized by any one of methods including chemical synthesis and polypeptide synthesizer synthesis.

[0012] It should be noted that the above synthesis method of the rice bran-derived short peptide of the present application is a conventional method, and the present application does not describe it in more detail.

[0013] According to an aspect of the present application, the drug for preventing and treating bone-related diseases further comprises a functionally enhanced modifier.

[0014] According to an aspect of the present application, the functionally enhanced modifier is one or more of a nucleic acid aptamer or an antibody, and a lipid membrane.

[0015] According to an aspect of the present application, the nucleic acid aptamer or antibody is a nucleic acid aptamer or antibody having targeting property.

[0016] According to an aspect of the present application, the nucleic acid is mRNA.

[0017] According to an aspect of the present application, the drug for preventing and treating bone-related diseases further comprises one or more pharmaceutically acceptable carriers.

[0018] According to an aspect of the present application, the carrier is one or more of an excipient, a filler, a binder, a humectant, a sustained-release agent, an absorption enhancer, a surfactant, and a lubricant.

[0019] According to one aspect of the present application, the drug for preventing and treating bone-related diseases comprises tablets, pills, capsules, injections.

[0020] Advantages of the present application:

[0021] The present application provides a use of a rice bran-derived short peptide in a drug for preventing and treating bone-related diseases (such as osteoporosis), and the amino acid sequence of the rice bran-derived short peptide is Lys-His-Asn-Arg-Gly-Asp-Glu-Phe (from the N-terminal). The natural bioactive short peptide, i.e., the rice bran-derived short peptide, can be synthesized by a conventional method. The advantages of the rice bran-derived short peptide are as follows: compared with small-molecule compound drugs, the rice bran-derived short peptide has less toxic and side effects; compared with protein macromolecular drugs, the rice bran-derived short peptide is cheaper and has lower immunogenicity; compared with traditional Chinese medicines, the rice bran-derived short peptide has a more definite efficacy and mechanism. The rice bran-derived short peptide in the present application can increase bone mass, improve bone quality, and promote bone regeneration. Therefore, the rice bran-derived short peptide can be used for preparing a drug for preventing and treating bone-related diseases, and provides a new safe and effective biological treatment method for preventing and treating bone-related diseases. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 A comparison chart of femoral bone microstructure data after two months of intervention by the method in Example 1 for the six groups of mice in Example 1 after one week of sham operation or ovariectomy;

[0023] Figure 2 A three-dimensional imaging chart of the distal femur after two months of intervention by the method in Example 1 for the six groups of mice in Example 1 after one week of sham operation or ovariectomy;

[0024] Figure 3 A comparison chart of femoral bone microstructure data after two months of intervention by the method in Example 2 for the four groups of mice in Example 2 after two months of sham operation or ovariectomy;

[0025] Figure 4 A three-dimensional imaging chart of the distal femur after two months of intervention by the method in Example 2 for the four groups of mice in Example 2 after two months of sham operation or ovariectomy;

[0026] Figure 5 A comparison chart of femoral bone microstructure data after three weeks of intervention by the method in Example 3 for the four groups of mice in Example 3, i.e., a normal group and tail suspension;

[0027] Figure 6 A three-dimensional imaging chart of the distal femur after three weeks of intervention by the method in Example 3 for the four groups of mice in Example 3, i.e., a normal group and tail suspension;

[0028] Figure 7This is a comparison of femoral bone microstructure data of three groups of elderly mice in Example 4 of the present application after being treated with the method in Example 4 for two months;

[0029] Figure 8 These are three-dimensional images of the distal femurs of the three groups of elderly mice in Example 4 of the present application after two months of intervention using the method in Example 4;

[0030] Figure 9 This is a comparison of femoral bone microstructure data of the three groups of mice in Example 5 of the present application after being treated with the method in Example 5 for two months;

[0031] Figure 10 These are three-dimensional images of the distal femur of the three groups of mice in Example 5 of the present application after two months of intervention using the method in Example 5;

[0032] Figure 11 This is a comparison of femoral bone microstructure data of the six groups of mice in Example 6 of the present application after being treated with the method in Example 6 for two months;

[0033] Figure 12 These are three-dimensional images of the distal femur of the six groups of mice in Example 6 of the present application, respectively, after being intervened with the method in Example 6 for two months. DETAILED DESCRIPTION

[0034] To make the present invention easier to understand, the present invention is further described below with reference to specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Unless otherwise defined, the professional terms used below are consistent with the meanings understood by professional and technical personnel in this field; unless otherwise specified, the raw materials and reagents involved in this article can be purchased from the market or prepared by known methods.

[0035] Definitions of specific terms are described in more detail below.This disclosure is not intended to be limited in any way by the exemplary list of substitutes described herein.

[0036] The terms "bone disease" and "bone-related disease" are used interchangeably and generally refer to a disease of the skeleton, non-limiting examples of conditions affecting the skeleton include achondroplasia, acromegaly, callus, demineralization of bone, fracture, marrow disease, marrow tumor, dyskeratosis congenita, leukemia (e.g., hairy cell leukemia, lymphocytic leukemia, myeloid leukemia, Philadelphia chromosome-positive leukemia, plasma cell leukemia, stem cell leukemia), systemic mastocytosis, myelodysplastic syndrome, paroxysmal nocturnal hemoglobinuria, myeloid sarcoma, myelofibrotic disease, multiple myeloma, polycythemia vera, pearson marrow-pancreas syndrome, bone tumor, marrow tumor, Ewing sarcoma, osteochondroma, osteoclastoma, osteosarcoma, phalangeal dysplasia, progressive diaphyseal dysplasia syndrome, craniosynostosis, Crouzon craniofacial dysostosis, dwarfism, achondroplasia, Bloom syndrome, Cockayne syndrome, Elhers-Danlos syndrome, Seckel syndrome, spondyloepiphyseal dysplasia, spondyloepiphyseal dysplasia congenita, Werner syndrome, hyperostosis, osteophytosis, cleidocranial dysostosis, Marfan syndrome, multiple fibrous dysplasia, osteitis, osteoarthritis, osteochondritis, Morquio's disease, Kasabach disease, Leri-Weill dyschondroplasia, osteochondrodystrophy, osteodystrophy, osteogenesis imperfecta, osteolysis, Gorham-Stout syndrome, osteomalacia, osteomyelitis, osteonecrosis, osteopenia, osteosclerosis, osteoporosis, ossification of the skeleton, oto-spondylo-epiphyseal dysplasia, pachydermoperiostosis, Paget's disease of bone, polydactyly, McEwen syndrome, rickets, Rothmund-Thomson syndrome, Sotos syndrome, spondyloepiphyseal dysplasia, spondyloepiphyseal dysplasia congenita, syndactyly, Apert syndrome, type II syndactyly, or Werner syndrome. In certain embodiments, the bone disease is a cartilage disease, such as chondroma, osteochondritis, Morquio's disease, Kasabach disease, or Leri-Weill dyschondroplasia. In certain embodiments, the bone disease is a herniation, such as a lumbar disc herniation. In certain embodiments, the bone disease is a joint disease, such as joint pain, arthritis (e.g., gout (e.g., Chikungunya syndrome, Lesniak syndrome)), Lyme disease, osteoarthritis, psoriatic arthritis, reactive arthritis, rheumatic fever, rheumatoid arthritis, Felty syndrome, synovial membrane disease, Blau syndrome, nail patella syndrome, spondyloarthropathy, reactive arthritis, Still's syndrome, synovial membrane disease, or Blau syndrome. In certain embodiments, the bone disease is Langerhan's cell histiocytosis.In certain embodiments, the skeletal disease is a muscle disease, such as Barth syndrome, mitochondrial encephalomyopathy, MELAS syndrome, MERRF syndrome, MNGIE syndrome, mitochondrial myopathy, Kearns-Sayre syndrome, myalgia, fibromyalgia, polymyalgia rheumatica, myoma, myositis, dermatomyositis, neuromuscular disease, Kearns-Sayre syndrome, muscular dystrophy, myasthenia, myasthenia congenita, Lambert-Eaton myasthenic syndrome, myasthenia gravis, myotonia, myotonia congenita, spinal muscular atrophy, tetany, external ophthalmoplegia, or rhabdomyolysis. In certain embodiments, the skeletal disease is Proteas syndrome. In certain embodiments, the skeletal disease is a rheumatic disease, such as arthritis (e.g., gout (e.g., Keyser syndrome, Lessner syndrome, Lyme disease)), osteoarthritis, psoriatic arthritis, reactive arthritis, rheumatic fever, rheumatoid arthritis, Felty syndrome, synovitis, Blau syndrome, gout (e.g., Keyser syndrome, Lessner syndrome), polymyalgia rheumatica, rheumatic fever, rheumatic heart disease, or Sjögren's syndrome. In certain embodiments, the skeletal disease is Sturgeon-Janzheimer syndrome. In certain embodiments, the musculoskeletal disease is a skeletal disease, such as Leri-Weill chondrodysplasia, skeletal malformations, Meissler-Nies syndrome, pachydermatosis periostealis, Rieger syndrome, spinal disease, herniated disc, scoliosis, spina bifida, spondylitis, ankylosing spondylitis, spondyloarthropathies, reactive arthritis, spondyloepiphyseal dysplasia, congenital spondyloepiphyseal dysplasia, or cervical spondylosis. In some embodiments, the disease is a musculoskeletal disease.

[0037] The term "bone-related disease" refers to any disease that can affect bones.

[0038] The present invention first uses a MicroTOF-QII liquid chromatography-mass spectrometer with high resolution to separate a short peptide consisting of Lys-His-Asn-Arg-Gly-Asp-Glu-Phe from rice bran. After obtaining the short peptide sequence, the present invention synthesizes the rice bran-derived short peptide by chemical synthesis to facilitate subsequent experiments.

[0039] The inventors conducted animal experiments using short peptides derived from rice bran. By intervening in a postmenopausal osteoporosis mouse model, a disuse osteoporosis mouse model, and an elderly osteoporosis mouse model, they observed that the short peptides derived from rice bran can significantly promote bone regeneration in the above-mentioned various mouse models.

[0040] It should be noted that this application Figure 1 、 3 , 5, 7, 9, 11 * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001; Figure 2 、4 White scale in 6, 8, 10, 12 is 1 mm (top) and 500 pm (bottom).

[0041] Example 1

[0042] Rice bran-derived short peptides can prevent bone loss in postmenopausal osteoporosis mouse models:

[0043] Mouse model: 10-week-old C57BL / 6 female SPF mice weighing 20-22 g were selected and purchased from Hunan Slaik Jingda Animal Experiment Co., Ltd. Each group had 7-10 mice, and they were free to eat and drink, maintaining the circadian rhythm.

[0044] Grouping experiment: C57BL / 6 female SPF mice were randomly divided into 6 groups, including Sham+Veh group, Sham+RBAP (i.g.) group, Sham+RBAP (i.v.) group, OVX+Veh group, OVX+RBAP (i.g.) group, and OVX+RBAP (i.v.) group; each group had 7-10 mice.

[0045] Modeling: One week after sham operation or ovariectomy, the mice in each group were intervened.

[0046] Administration method:

[0047] Sham+Veh group: One week after the mice were sham-operated, phosphate buffer was administered by gavage and injected into the tail vein, with a volume of 100 μL for both gavage and tail vein injection, twice a week, and samples were collected after two months of intervention.

[0048] Sham+RBAP (i.g.) group: One week after the mice were sham-operated, rice bran-derived short peptide (RBAP) solution (dissolved in phosphate buffer) was administered by gavage, with an intervention concentration of 30 mg / kg, and phosphate buffer was injected into the tail vein at the same time, with a volume of 100 μL for both gavage and tail vein injection, twice a week, and samples were collected after two months of intervention.

[0049] Sham+RBAP (i.v.) group: One week after the mice were sham-operated, rice bran-derived short peptide solution was injected into the tail vein, with an intervention concentration of 30 mg / kg, and phosphate buffer was administered by gavage at the same time, with a volume of 100 μL for both gavage and tail vein injection, twice a week, and samples were collected after two months of intervention.

[0050] OVX+Veh group: One week after the mice were ovariectomized, phosphate buffer was administered by gavage and injected into the tail vein at the same time, with a volume of 100 μL for both gavage and tail vein injection, twice a week, and samples were collected after two months of intervention.

[0051] OVX+RBAP(ig) group: One week after the mice underwent ovariectomy, they were gavaged with a rice bran-derived short peptide solution at an intervention concentration of 30 mg / kg. At the same time, phosphate buffer was injected into the tail vein. The volume of gavage and tail vein injection was 100 μL twice a week. Samples were collected after two months of intervention.

[0052] OVX+RBAP(iv) group: One week after the mice underwent ovariectomy, they were injected with a rice bran-derived short peptide solution through the tail vein at an intervention concentration of 30 mg / kg. At the same time, they were gavaged with phosphate buffer. The volume of gavage and tail vein injection was 100 μL twice a week. Samples were collected after two months of intervention.

[0053] The femurs of the six groups of mice collected two months after the intervention were subjected to three-dimensional imaging and bone microstructure analysis. The results are as follows: Figures 1-2 shown.

[0054] like Figures 1-2 As shown in the results, intravenous injection of rice bran-derived short peptides can significantly increase the cancellous bone volume fraction (Tb.BV / TV) and cancellous bone number (Tb.N) of Sham and OVX mice, and oral gavage intervention of rice bran-derived short peptides can significantly increase Tb.BV / TV and trabecular thickness (Tb.Th) of OVX (ovariectomized) mice, indicating that both oral gavage and tail vein injection of rice bran-derived short peptides can prevent bone loss in the postmenopausal osteoporosis mouse model.

[0055] Example 2

[0056] Rice bran-derived short peptides can treat bone loss in a postmenopausal osteoporosis mouse model:

[0057] Mouse model: 10-week-old female C57BL / 6 SPF mice weighing 20-22 g were purchased from Hunan Slake Jingda Animal Experiment Co., Ltd. Each group consisted of 7-10 mice with free access to food and water, and maintained a circadian rhythm.

[0058] Grouping experiment: C57BL / 6 female SPF mice were randomly divided into 4 groups, including Sham+Veh group, OVX+Veh group, OVX+RBAP(ig) group, and OVX+RBAP(iv) group; there were 7 to 10 mice in each group.

[0059] Modeling: Each group of mice underwent sham surgery or ovariectomy and then began to be intervened two months later.

[0060] Dosage:

[0061] Sham+Veh group: Two months after the mice underwent sham surgery, they were gavaged with phosphate buffer and injected into the tail vein at the same time. The volume of gavage and tail vein injection was 100 μL twice a week. Samples were collected two months after the intervention.

[0062] OVX+Veh group: Two months after ovariectomy, the mice were given phosphate buffer by gavage and tail vein injection, with a volume of 100 μL each, twice a week, and samples were collected after two months of intervention.

[0063] OVX+RBAP(i.g.) group: Two months after ovariectomy, the mice were given a rice bran-derived short peptide solution (dissolved in phosphate buffer) by gavage, with an intervention concentration of 30 mg / kg, and phosphate buffer was given by tail vein injection, with a volume of 100 μL each, twice a week, and samples were collected after two months of intervention.

[0064] OVX+RBAP(i.v.) group: Two months after ovariectomy, the mice were given a rice bran-derived short peptide solution by tail vein injection, with an intervention concentration of 30 mg / kg, and phosphate buffer was given by gavage, with a volume of 100 μL each, twice a week, and samples were collected after two months of intervention.

[0065] The femurs of the 4 groups of mice collected after two months of intervention were subjected to three-dimensional imaging and bone microstructure analysis, and the results are shown in Figures 3-4 .

[0066] As shown in Figures 3-4 , intravenous injection of rice bran-derived short peptides can significantly increase Tb.BV / TV and Tb.N in OP mice, indicating that rice bran-derived short peptides can treat bone loss in postmenopausal osteoporosis mouse models.

[0067] Example 3

[0068] Rice bran-derived short peptides can improve bone mass in disuse osteoporosis mouse models:

[0069] Mouse model: 10-week-old C57BL / 6 male SPF mice weighing 20-22 g were selected, all purchased from Hunan Slike Jingda Animal Experiment Co., Ltd. There were 7-10 mice in each group, and they were free to eat and drink, and the circadian rhythm was maintained.

[0070] Grouping experiment: C57BL / 6 male SPF mice were randomly divided into 4 groups, including Veh group, TS+Veh group, TS+RBAP(i.g.) group, and TS+RBAP(i.v.) group; there were 7-10 mice in each group.

[0071] Modeling: The mice in each group were subjected to tail suspension while starting the intervention.

[0072] Dosage form:

[0073] Veh group: the mice were normally fed, and phosphate buffer was gavaged and injected into the tail vein, the volume of gavage and tail vein injection was 100 μL, twice a week, and the samples were collected after three weeks of intervention.

[0074] TS+Veh group: the mice were tail-suspended, and phosphate buffer was gavaged and injected into the tail vein, the volume of gavage and tail vein injection was 100 μL, twice a week, and the samples were collected after three weeks of intervention.

[0075] TS+RBAP(i.g.) group: the mice were tail-suspended, and rice bran-derived short peptide solution was gavaged and phosphate buffer was injected into the tail vein, the intervention concentration of rice bran-derived short peptide was 30 mg / kg, the volume of gavage and tail vein injection was 100 μL, twice a week, and the samples were collected after three weeks of intervention.

[0076] TS+RBAP(i.v.) group: the mice were tail-suspended, and rice bran-derived short peptide solution was injected into the tail vein and phosphate buffer was gavaged, the intervention concentration of rice bran-derived short peptide was 30 mg / kg, the volume of gavage and tail vein injection was 100 μL, twice a week, and the samples were collected after three weeks of intervention.

[0077] The femurs of the 4 groups of mice collected after three weeks of intervention were subjected to three-dimensional imaging and bone microstructure analysis, and the results are shown in Figures 5-6

[0078] As shown in Figures 5-6 , intravenous injection of rice bran-derived short peptide can significantly increase Tb.BV / TV and Tb.N of tail-suspended mice, indicating that rice bran-derived short peptide can increase bone mass in disuse osteoporosis mouse models.

[0079] Example 4

[0080] Rice bran-derived short peptide can increase bone mass in elderly osteoporosis mouse models:

[0081] Mouse model: 18-month-old C57BL / 6 male SPF mice weighing 28-35 g were selected, all purchased from Hunan Slike Jingda Animal Experiment Co., Ltd. Each group had 7-10 mice, and they were free to eat and drink, and the circadian rhythm was maintained.

[0082] Grouping experiment: C57BL / 6 male SPF mice were randomly divided into 3 groups, including Veh group, RBAP(i.g.) group, and RBAP(i.v.) group; each group had 7-10 mice.

[0083] Modeling: the elderly mice in each group were intervened at the same time.

[0084] Dosage form:

[0085] ​Veh group: phosphate buffer solution tail vein injection and gavage were performed on old mice at the same time, the volume of gavage and tail vein injection was 100 μL, twice a week, samples were collected after two months of intervention.

[0086] RBAP(i.g.) group: old mice were given rice bran-derived short peptide gavage at the same time with phosphate buffer solution tail vein injection, the intervention concentration of rice bran-derived short peptide was 30 mg / kg, the volume of gavage and tail vein injection was 100 μL, twice a week, samples were collected after two months of intervention.

[0087] RBAP(i.v.) group: old mice were given rice bran-derived short peptide tail vein injection at the same time with phosphate buffer solution gavage, the intervention concentration of rice bran-derived short peptide was 30 mg / kg, the volume of gavage and tail vein injection was 100 μL, twice a week, samples were collected after two months of intervention.

[0088] The femurs of the 3 groups of mice collected after two months of intervention were subjected to three-dimensional imaging and bone microstructure analysis, and the results are shown in Figures 7-8 .

[0089] As shown in Figures 7-8 , intravenous injection of rice bran-derived short peptide can significantly increase Tb.BV / TV and Tb.N of old mice, indicating that rice bran-derived short peptide can improve osteoporosis in old mice.

[0090] Example 5

[0091] Rice bran-derived short peptide can accelerate the repair of osteoporotic mouse fracture:

[0092] Mouse model: 10-week-old C57BL / 6 male SPF mice weighing 28-35 g were selected, all purchased from Hunan Slike Jingda Animal Experiment Co., Ltd. Each group had 7-10 mice, free access to food and water, and maintained circadian rhythm.

[0093] Grouping experiment: C57BL / 6 female SPF mice were randomly divided into 3 groups, including OPF+Veh group, OPF+RBAP(i.g.) group, and OPF+RBAP(i.v.) group; each group had 7-10 mice.

[0094] Modeling: After two months of ovariectomy, the mice in each group were subjected to femur fracture modeling, and intervention began one week later.

[0095] Dosing method:

[0096] OPF group: phosphate buffer solution tail vein injection and gavage were performed on the fractured mice at the same time, the volume of gavage and tail vein injection was 100 μL, twice a week, samples were collected after two months of intervention.

[0097] OPF+RBAP(i.g.) group: the fractured mice were given rice bran-derived short peptide by gavage at the same time, and phosphate buffer was injected into the tail vein. The intervention concentration of rice bran-derived short peptide was 30 mg / kg, and the volume of gavage and tail vein injection was 100 μL. The intervention was performed twice a week, and the samples were collected after two months of intervention.

[0098] OPF+RBAP(i.v.) group: the fractured mice were given rice bran-derived short peptide by tail vein injection at the same time, and phosphate buffer was given by gavage. The intervention concentration of rice bran-derived short peptide was 30 mg / kg, and the volume of gavage and tail vein injection was 100 μL. The intervention was performed twice a week, and the samples were collected after two months of intervention.

[0099] The femurs of the three groups of mice collected after two months of intervention were subjected to three-dimensional imaging and bone microstructure analysis, and the results are shown in Figures 9-10

[0100] As shown in Figures 9-10 , intravenous injection of rice bran-derived short peptide can significantly increase the bone mass of the callus at the fracture end of osteoporotic mice, indicating that rice bran-derived short peptide can accelerate the repair of osteoporotic mouse fracture.

[0101] Example 6

[0102] Functional rice bran-derived short peptide after connecting aptamer targeting BMSC (RBAP) has a stronger anti-osteoporosis effect:

[0103] Mouse model: 10-week-old C57BL / 6 male SPF mice weighing 28-35 g were selected, all purchased from Hunan Slike Jingda Animal Experiment Co., Ltd. There were 7-10 mice in each group, and they were free to eat and drink, and the circadian rhythm was maintained.

[0104] Grouping experiment: C57BL / 6 female SPF mice were randomly divided into 6 groups, including Sham+Veh group, OVX+Veh group, OVX+Apt group, OVX+RBAP group, OVX+Rd-RBAP group, and OVX+Apt-RBAP group; each group had 7-10 mice.

[0105] Modeling: the mice in each group were subjected to sham operation or ovariectomy one week before intervention.

[0106] Dosage form:

[0107] Sham+Veh group: the sham-operated mice were given phosphate buffer by tail vein injection, and the volume of tail vein injection was 100 μL. The intervention was performed twice a week, and the samples were collected after two months of intervention.

[0108] OVX+Veh group: the ovariectomized mice were given phosphate buffer by tail vein injection, and the volume of tail vein injection was 100 μL. The intervention was performed twice a week, and the samples were collected after two months of intervention.​

[0109] OVX+Apt group: OVX mice were injected with simple nucleic acid aptamer (Aptamer targeting BMSCs) via tail vein, the volume of tail vein injection was 100 μL, twice a week, and samples were collected after two months of intervention.

[0110] OVX+RBAP group: OVX mice were injected with rice bran-derived short peptide via tail vein, the intervention concentration of rice bran-derived short peptide was 30 mg / kg, the volume of tail vein injection was 100 μL, twice a week, and samples were collected after two months of intervention.

[0111] OVX+Rd-RBAP group: OVX mice were injected with rice bran-derived short peptide connected with random sequence (complex) via tail vein, the intervention concentration of rice bran-derived short peptide in rice bran-derived short peptide connected with random sequence was 30 mg / kg, the volume of tail vein injection was 100 μL, twice a week, and samples were collected after two months of intervention;

[0112] wherein the random sequence is mRNA, as shown in SEQ ID NO: 2, specifically:

[0113] 5'-CGCGAATTAGACAGACTCATTGACTCGGATTACTCAAAGA-3'.

[0114] OVX+Apt-RBAP group: OVX mice were injected with rice bran-derived short peptide connected with nucleic acid aptamer (Aptamer targeting BMSCs) via tail vein, the intervention concentration of rice bran-derived short peptide in rice bran-derived short peptide connected with nucleic acid aptamer was 30 mg / kg, the volume of tail vein injection was 100 μL, twice a week, and samples were collected after two months of intervention;

[0115] wherein the nucleic acid aptamer (Aptamer targeting BMSCs) is mRNA, as shown in SEQ ID NO: 3, specifically:

[0116] 5'-ACGACGGTGATATGTCAAGGTCGTATGCACGAGTCAGAGG-3'.

[0117] The femurs of the 6 groups of mice collected after two months of intervention were subjected to three-dimensional imaging and bone microstructure analysis, and the results are shown in Figures 11-12 .

[0118] As shown in Figures 11-12As shown, simple nucleic acid aptamer injection has no obvious effect on the bone mass of OVX mice, rice bran-derived short peptides significantly increase the bone mass of OVX mice; the rice bran-derived short peptides connected with random sequences significantly increase the bone mass of OVX mice, but there is no obvious difference compared with simple rice bran-derived short peptides; the rice bran-derived short peptides connected with the aptamer targeting BMSCs significantly increase the bone mass of OVX mice, and compared with simple rice bran-derived short peptides and rice bran-derived short peptides connected with random sequences, the bone mass is also significantly increased, indicating that the functionalized RBAP after connecting the aptamer targeting BMSCs has a stronger anti-osteoporosis effect.

[0119] The above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto, any person skilled in the art can easily think of changes or replacements within the scope of the present application, which should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. Use of a rice bran-derived short peptide in the preparation of a drug for preventing and treating bone-related diseases, characterized in that: The bone-related disease is any one of postmenopausal osteoporosis, disuse osteoporosis, senile osteoporosis, and osteoporotic fractures; the amino acid sequence of the rice bran-derived short peptide is Lys-His-Asn-Arg-Gly-Asp-Glu-Phe, and the above amino acid sequence starts from the N-terminus; wherein Lys represents lysine, His represents histidine, Asn represents asparagine, Arg represents arginine, Gly represents glycine, Asp represents aspartic acid, Glu represents glutamic acid, and Phe represents phenylalanine.

2. The use of the rice bran-derived short peptide according to claim 1 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The molecular weight of the rice bran-derived short peptide is 1002.6 Da.

3. The use of the rice bran-derived short peptide according to claim 1 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The rice bran-derived short peptide is artificially synthesized by a chemical synthesis method.

4. The use of the rice bran-derived short peptide according to claim 1 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The rice bran-derived short peptide is artificially synthesized by using a peptide synthesizer.

5. The use of the rice bran-derived short peptide according to claim 1 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The drug for preventing and treating bone-related diseases also includes a modified form with enhanced function.

6. The use of the rice bran-derived short peptide according to claim 5 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The function-enhancing modified body is one or more of a nucleic acid aptamer, an antibody, and a lipid membrane package.

7. The use of the rice bran-derived short peptide according to claim 6 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The nucleic acid aptamer or antibody is a targeting nucleic acid aptamer or antibody.

8. The use of the rice bran-derived short peptide according to claim 6 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The nucleic acid is mRNA.

9. The use of the rice bran-derived short peptide according to claim 1 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The drug for preventing and treating bone-related diseases also contains one or more pharmaceutically acceptable carriers.

10. Use of the rice bran-derived short peptide according to claim 9 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The carrier is one or more of a filler, a binder, a wetting agent, a sustained-release agent, an absorption promoter, a surfactant, and a lubricant.

11. Use of the rice bran-derived short peptide according to claim 9 in preparing a drug for preventing and treating bone-related diseases, characterized in that: The medicine for preventing and treating bone-related diseases includes tablets, pills, capsules and injections.

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