Hippocampus bone-strengthening peptide and application thereof

Abstract

The application discloses a hippocampus bone-strengthening peptide and application thereof, and belongs to the technical field of biological medicines. The hippocampus bone-strengthening peptide provided by the application is any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7 or SEQ ID NO: 10. The hippocampus bone-strengthening peptide provided by the application can effectively promote the proliferation of bone marrow mesenchymal stem cells at a micro-molar concentration level, and is expected to be used for preparing bone repair drugs.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a hippocampal bone-strengthening peptide and its applications. Background Technology

[0002] Seahorse, a traditional Chinese medicine, is believed to tonify kidney yang and strengthen bones and muscles. Modern research indicates that seahorse extract is rich in proteins and peptides, which may regulate bone metabolism by influencing the activity of bone marrow mesenchymal stem cells. However, current research on seahorse peptides largely focuses on the crude extract level, and the specific active peptides that can promote the proliferation of bone marrow mesenchymal stem cells and their sequences remain unclear. Due to the complex structure-activity relationship between the amino acid sequence and the biological activity of peptides, peptides with different sequences may exhibit drastically different functions. This makes it difficult to predict which specific peptide has a proliferative effect based solely on the activity of crude seahorse extract. Summary of the Invention

[0003] In view of this, the present invention provides a hippocampal bone-strengthening peptide and its application, to provide a single active polypeptide derived from the hippocampus that can promote the proliferation of bone marrow mesenchymal stem cells.

[0004] In a first aspect, the present invention provides a hippocampal bone-strengthening peptide, wherein the amino acid sequence of the hippocampal bone-strengthening peptide is any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7 or SEQ ID NO: 10.

[0005] In a second aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned hippocampal bone-strengthening peptide or its pharmaceutically acceptable salt.

[0006] Preferably, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0007] Furthermore, the pharmaceutically acceptable excipients include at least one of diluents, carriers, binders, lubricants, disintegrants, pH adjusters, antioxidants, solvents, or penetrants.

[0008] Preferably, the dosage form of the pharmaceutical composition is at least one of the following: injection, tablet, capsule, granule, oral liquid, lyophilized powder for injection, sustained-release agent, or controlled-release agent.

[0009] Preferably, the route of administration of the pharmaceutical composition is at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, or local administration.

[0010] Thirdly, the present invention provides the use of the above-mentioned hippocampal bone-strengthening peptide or the above-mentioned pharmaceutical composition in the preparation of products that promote the proliferation of bone marrow mesenchymal stem cells.

[0011] Preferably, the product includes at least one of a drug, a health product, or a medical device.

[0012] Fourthly, the present invention provides the use of the above-mentioned hippocampal bone-strengthening peptide or the above-mentioned pharmaceutical composition in the preparation of bone repair drugs.

[0013] Preferably, the bone repair drug is used to treat osteoporosis, fractures, bone defects, or nonunion.

[0014] Compared with the prior art, the present invention has achieved the following beneficial effects: The hippocampal bone-strengthening peptide provided by this invention has a well-defined amino acid sequence, overcoming the limitations of existing technologies that can only study peptide activity at the level of crude hippocampal extracts and cannot obtain single active peptides. Because there is a complex structure-activity relationship between the amino acid sequence and the bioactivity of peptides, peptides with different sequences exhibit significant functional differences. The peptides shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7, or SEQ ID NO: 10 provided by this invention can effectively promote the proliferation of bone marrow mesenchymal stem cells at micromolar concentrations, thus showing promising application prospects in the field of bone repair. Attached Figure Description

[0015] The accompanying drawings, which form part of this specification, are used to provide a further understanding of the invention. The illustrative embodiments and descriptions of the invention are used to explain the invention and do not constitute an undue limitation thereof. Obviously, those skilled in the art can obtain other drawings based on these drawings without any inventive effort.

[0016] Figure 1 This is a graph showing the effect of different hippocampal bone-strengthening peptides on the proliferation activity of bone marrow mesenchymal stem cells (BMSCs) in Example 3 of the present invention. Figure 2 This is a graph showing the effect of different concentrations of hippocampal bone-strengthening peptide on the proliferation activity of bone marrow mesenchymal stem cells (BMSCs) in Example 4 of the present invention; where A is H01 peptide, B is H02 peptide, C is H07 peptide, and D is H10 peptide. Figure 3 This is a growth curve diagram of four hippocampal bone-strengthening peptides intervening in bone marrow mesenchymal stem cells (BMSCs) in Example 5 of the present invention; where * indicates P < 0.05; ** indicates P < 0.01; *** indicates P < 0.001; Ctrl indicates the control group. Detailed Implementation

[0017] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0018] The terminology and experimental procedures (such as molecular biology, cell culture, and protein chemistry) used in this invention are all methods well-known and conventionally used by those skilled in the art. Unless otherwise stated, the specific implementation process of this invention is carried out with reference to the conventional steps in the relevant recognized standard technical manuals or literature.

[0019] The present invention provides a hippocampal bone-strengthening peptide, wherein the amino acid sequence of the hippocampal bone-strengthening peptide is any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7 or SEQ ID NO: 10.

[0020] The hippocampal bone-strengthening peptides of this invention are derived from the abdominal hippocampus (Hippocampus abdominalis) and obtained through liquid chromatography-mass spectrometry combined with bioinformatics screening. The four polypeptide sequences shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7, and SEQ ID NO: 10 are the first single polypeptides identified from the hippocampus that exhibit activity in promoting the proliferation of bone marrow mesenchymal stem cells (BMSCs). Because there is a complex structure-activity relationship between the amino acid sequence and the biological activity of polypeptides, polypeptides with different sequences may exhibit drastically different functions. This invention, through systematic screening, discovered that the aforementioned specific sequences can significantly promote BMSC proliferation in a concentration-dependent manner, thus providing a new active material basis for bone repair.

[0021] In this invention, the hippocampal bone-strengthening peptide can be prepared using peptide chemical synthesis methods known in the art, such as solid-phase synthesis or liquid-phase synthesis. Solid-phase synthesis typically uses Fmoc (9-fluorenylmethoxycarbonyl) or Boc (tert-butyloxycarbonyl) as protecting groups, sequentially coupling amino acid residues from the C-terminus to the N-terminus on a resin support. The protecting groups are then removed using a lysis buffer, and the peptide is cleaved. After purification (e.g., reversed-phase high-performance liquid chromatography), the target peptide is obtained. The molecular weight of the synthesized peptide can be confirmed by mass spectrometry, and its purity (typically ≥95%) can be detected by high-performance liquid chromatography. Those skilled in the art can choose appropriate synthesis strategies according to specific needs; this invention does not limit such choices.

[0022] The present invention also provides a pharmaceutical composition comprising the above-mentioned hippocampal bone-strengthening peptide or its pharmaceutical salt.

[0023] In this invention, "pharmaceutical salt" refers to a pharmaceutically acceptable salt formed by seahorse bone-strengthening peptides with an acid or base. Since polypeptide molecules typically contain free amino or carboxyl groups, they can form salts with inorganic acids (such as hydrochloric acid, sulfuric acid, and phosphoric acid), organic acids (such as acetic acid, citric acid, tartaric acid, and methanesulfonic acid), or bases (such as hydroxides of sodium, potassium, calcium, and magnesium). Salt formation improves the water solubility, stability, and bioavailability of the polypeptide. Those skilled in the art can prepare pharmaceutical salts using conventional methods, such as dissolving the polypeptide in a suitable solvent, adding an equimolar or appropriate excess of acid or base, and then separating it by freeze-drying or crystallization.

[0024] In a preferred embodiment, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0025] Pharmaceutically acceptable excipients refer to all additives in pharmaceutical preparations other than the active ingredient, which should possess sufficient safety and not affect the efficacy of the active ingredient. The main functions of excipients include: imparting an appropriate form to the preparation, improving stability, regulating release rate, and improving taste or compliance. Those skilled in the art can rationally select excipients according to the route of administration and dosage form requirements.

[0026] Furthermore, the pharmaceutically acceptable excipients include at least one of diluents, carriers, binders, lubricants, disintegrants, pH adjusters, antioxidants, solvents, or penetrants.

[0027] Diluents (also known as fillers) are used to increase the weight or volume of solid dosage forms such as tablets and capsules. Common examples include lactose, microcrystalline cellulose, starch, and mannitol. Carriers are substances that can carry active ingredients and assist in their delivery to the target site, such as nanoparticles, liposomes, and cyclodextrins. Binders are used to bind powders into granules or tablets, such as polyvinylpyrrolidone, hydroxypropyl methylcellulose, and starch paste. Lubricants are used to reduce friction during tableting or filling, such as magnesium stearate, talc, and polyethylene glycol. Disintegrants promote the rapid disintegration of solid dosage forms in the digestive tract, such as sodium carboxymethyl starch, crospovidone, and low-substituted hydroxypropyl cellulose. pH adjusters are used to maintain a suitable pH value for the formulation or administration environment, such as citric acid, phosphate buffer, and tris(hydroxymethyl)aminomethane. Antioxidants are used to prevent the oxidative degradation of peptides, such as vitamin C, vitamin E, butylated hydroxyanisole, ethylenediaminetetraacetic acid, and its salts. Solvents are used to dissolve or disperse active ingredients, such as water, ethanol, propylene glycol, and polyethylene glycol. Penetrants are used to promote the permeation of active ingredients through biological membranes, such as lauryl ether and dimethyl sulfoxide. These excipients can be used alone or in combination, and their dosage can be determined according to conventional formulation processes. For example, the dosage of diluents can be 10% to 90% of the total weight of the formulation, and the dosage of lubricants can be 0.5% to 5%.

[0028] In another preferred embodiment, the dosage form of the pharmaceutical composition is at least one of the following: injection, tablet, capsule, granule, oral liquid, lyophilized powder for injection, sustained-release agent, or controlled-release agent.

[0029] Different dosage forms are suitable for different therapeutic needs and compliance requirements. Injectables (including solutions, suspensions, and emulsions) are suitable for situations requiring rapid onset of action or where oral administration is not possible; they typically require aseptic manufacturing processes and control of particulates and pyrogens. Tablets and capsules are the mainstream dosage forms for oral solid dosage forms, convenient to carry and for dosage control; coating technology can mask unpleasant tastes or achieve enteric release. Granules and oral solutions are suitable for children or patients with swallowing difficulties. Lyophilized powder injections are suitable for long-term storage of peptide drugs; moisture is removed through freeze-drying, and the powder is reconstituted with water for injection before use. Sustained-release and controlled-release formulations can prolong the duration of drug action and reduce the frequency of administration; they can be achieved using matrix materials (such as hydroxypropyl methylcellulose and ethyl cellulose) or coating technology. The preparation methods for each dosage form are conventional techniques in the field, such as wet granulation and tableting, dry granulation, fluidized bed granulation, and freeze-drying.

[0030] In yet another preferred embodiment, the route of administration of the pharmaceutical composition is at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, or local administration.

[0031] The choice of route of administration depends on the properties of the drug, the treatment goal, and the patient's condition. Intravenous injection ensures 100% drug entry into the bloodstream, has the fastest onset of action, and is suitable for the treatment of acute or critical illnesses. Intraperitoneal injection is commonly used in animal experiments, allowing for rapid absorption into the portal venous circulation. Intramuscular and subcutaneous injections are suitable for intermediate- or long-acting administration; absorption is slower but more stable than intravenous injection. Oral administration is the most convenient, but peptide drugs are susceptible to degradation by gastrointestinal enzymes and the first-pass effect; absorption can be improved through formulation technologies (such as enteric coating and nanoparticle encapsulation). Local administration (such as local injection or application to bone defects) can increase drug concentration at the lesion site and reduce systemic exposure. This invention does not limit the route of administration; those skilled in the art can choose an appropriate route based on the specific disease and formulation characteristics.

[0032] The present invention also provides the use of the above-mentioned hippocampal bone-strengthening peptide or the above-mentioned pharmaceutical composition in the preparation of products that promote the proliferation of bone marrow mesenchymal stem cells.

[0033] Bone marrow mesenchymal stem cells (BMSCs) are a type of adult stem cell with self-renewal and multi-lineage differentiation potential, playing a crucial role in bone repair and regeneration. BMSC proliferation is the initial step in bone formation and healing; only when a sufficient number of BMSCs are recruited or expanded can they further differentiate into osteoblasts and secrete bone matrix. This invention verified using the Cell Counting Kit-8 (CCK-8) method that the hippocampal bone-strengthening peptide significantly promotes BMSC proliferation in a concentration-dependent manner. Therefore, the hippocampal bone-strengthening peptide or pharmaceutical compositions containing it can be used to prepare products that promote BMSC proliferation. These products include at least one of pharmaceuticals, health supplements, or medical devices. Pharmaceuticals are used for therapeutic interventions, health supplements are used for daily bone health maintenance, and medical devices (such as bone repair scaffolds, sustained-release patches, and injectable gels) can serve as carriers of the active ingredients, enabling local delivery and sustained release.

[0034] The present invention also provides the use of the above-mentioned hippocampal bone-strengthening peptide or the above-mentioned pharmaceutical composition in the preparation of bone repair drugs.

[0035] Bone repair is a complex physiological process involving multiple stages such as the proliferation, migration, osteogenic differentiation, angiogenesis, and mineralization of bone mesenchymal stem cells (BMSCs). Since the hippocampal bone-strengthening peptide provided in this invention can effectively promote BMSC proliferation, and BMSC proliferation is a crucial initiation and continuation of bone repair, the hippocampal bone-strengthening peptide and pharmaceutical compositions containing it can be used to prepare bone repair drugs. These bone repair drugs are suitable for various bone diseases or injuries.

[0036] In a preferred embodiment, the bone repair drug is used to treat osteoporosis, fractures, bone defects, or nonunion. Osteoporosis is characterized by reduced bone mass and destruction of bone microstructure, and delayed bone healing or nonunion after fractures is a common clinical challenge. The hippocampal bone-strengthening peptide provided by this invention can increase the number of osteogenic progenitor cells at bone formation sites by promoting the proliferation of bone mesenchymal stem cells (BMSCs), thereby accelerating the bone repair process. In specific applications, the peptide can be prepared into systemic administration formulations (such as injections or oral preparations) or local implants (such as peptide-loaded bone cement, hydrogels, or 3D-printed scaffolds), and the dosage can be adjusted according to the patient's weight, disease severity, and dosage form.

[0037] The technical solution of the present invention will be further described below with reference to specific embodiments. The present invention does not impose any special restrictions on the source of reagents used in the following embodiments; commercially available products well known to those skilled in the art can be used.

[0038] In the following examples, 0.1% formic acid aqueous solution refers to an aqueous solution of formic acid with a volume fraction of 0.1%; 40% acetonitrile refers to an aqueous solution of acetonitrile with a volume fraction of 40%.

[0039] Example 1 This embodiment provides the preparation of hippocampal polypeptide extract.

[0040] (1) Take the abdominal seahorse (Hippocampus abdominalis) and grind it in liquid nitrogen. Add the lysis buffer and homogenize it. The lysis buffer contains 1 wt% sodium deoxycholate (SDC), 10 mM tris(2-carboxyethyl)phosphonic acid hydrochloride (TCEP), 40 mM chloroacetamide (CAA) and 100 mM triethylammonium bicarbonate (TEAB), with a pH of 8.5. After sonication for 10 minutes, centrifuge.

[0041] (2) Take the supernatant and add 4 times the volume of methanol to precipitate the protein. Centrifuge again and discard the supernatant. Extract the obtained solid with an extractant solution, which is a mixture of acetonitrile and dilute hydrochloric acid with a concentration of 10 wt% and a volume ratio of acetonitrile to dilute hydrochloric acid of 97:3. After lyophilization, dissolve in 0.1% formic acid aqueous solution.

[0042] (3) The above solution was desalted by a C18 desalting column: the desalting column was activated with acetonitrile, the desalting column was equilibrated with 0.1% formic acid aqueous solution, the sample was loaded, and then the impurities were removed by washing with 0.1% formic acid aqueous solution. Finally, the solution was eluted with 40% acetonitrile, the eluent was collected, and the solution was freeze-dried again to obtain the hippocampal polypeptide extract.

[0043] Example 2 This embodiment provides liquid chromatography-mass spectrometry (LC-MS) detection and candidate peptide screening.

[0044] (1) The hippocampal polypeptide extract obtained in Example 1 was dissolved in 0.1 wt% formic acid aqueous solution, and the supernatant was collected by centrifugation for liquid chromatography-mass spectrometry (LC-MS). LC conditions: The mobile phase consisted of solution A and solution B. Solution A was a 0.1% formic acid aqueous solution, and solution B was a mixture of acetonitrile and 0.1 wt% formic acid aqueous solution, with a volume ratio of acetonitrile to 0.1% formic acid aqueous solution of 80:20. The elution program was: 0–8 min 6% B, 8–15 min 6%–12% B, 15–63 min 12%–30% B, 63–73 min 30%–40% B, 73–74 min 40%–95% B, 74–84 min 95% B, and 84–85 min 95%–6% B. Mass spectrometry conditions: FAIMS Pro was used. TM Interface mass spectrometer, with a compensation voltage (CV) switching every 1 second between -45 and 65 V, Nanospray Flex TM (NSI) ion source, ion spray voltage 2.0 kV, ion transmission tube temperature 320℃; data-dependent acquisition mode, full scan range of single-stage mass spectrometry m / z 350~1500, resolution 120000 (m / z 200), AGC 4×10⁻⁶. 5The maximum C-trap injection time is 50 ms; the secondary mass spectrometer uses the "Top Speed" mode with a resolution of 15000 (m / z 200) and an AGC of 5×10⁻⁶. 4 The maximum injection time is 22 ms, and the peptide fragmentation collision energy is 33%.

[0045] (2) Data analysis was performed using Peaks 8 search software and the De Novo analysis method. Peptide sequences were obtained using the screening criteria of -10LgP ≥ 90 and mass deviation ≤ 2 ppm. Free peptides were obtained by comparison with the NCBI database, and peptide toxicity was predicted using the online tool ToxinPred. Ten candidate peptides were screened and named H01, H02, H03, H04, H05, H06, H07, H08, H09, and H10, with their amino acid sequences shown in Table 1. The above candidate peptides were synthesized using solid-phase synthesis. Mass spectrometry confirmed that the molecular weight was consistent with the theoretical value; high-performance liquid chromatography (HPLC) showed that the purity was greater than 99%, which was used for subsequent experiments.

[0046] Table 1. Amino acid sequences of candidate peptides

[0047] Example 3 This embodiment provides a screening method for the proliferative activity of hippocampal bone-strengthening peptides on bone marrow mesenchymal stem cells (BMSCs).

[0048] (1) Culture and passage of BMSCs: BMSCs cell lines were placed in α-minimum essential medium (α-MEM medium) containing 10% fetal bovine serum and penicillin (1:100, i.e., 100 U / mL penicillin and 100 μg / mL streptomycin) and cultured in a cell culture incubator at 37°C and 5% CO2. The medium was changed every 3 days. When the cells reached about 80% confluence, they were passaged, and cells in the logarithmic growth phase were used for experiments.

[0049] (2) Cell seeding and drug administration: Take BMSCs in the logarithmic growth phase, digest and resuspend them, and administer 2×10⁻⁶ cells per cell line. 3 Seeds were planted at a density of 100 μL / 100 μL in 96-well plates, with 100 μL per well. For the experimental groups, candidate peptide solutions (H01~H10) were added to a final concentration of 10 μM. The candidate peptide solutions were dissolved in phosphate-buffered saline (PBS), and each peptide was used in triplicate. An equal volume of PBS was added to the control group. The plates were incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0050] (3) CCK-8 assay: Add 10 μL of CCK-8 solution to each well and incubate for 1 hour. Measure the absorbance (OD) at 450 nm using a microplate reader. 450(Value). The cell proliferation fold of the control group was set to 1. The cell proliferation fold of the candidate polypeptide solution was calculated as follows: Cell proliferation fold = Experimental group OD. 450 / Control group OD 450 Among them, the experimental group OD 450 OD of each candidate peptide solution 450 Value, control group OD 450 OD of the control group solution 450 value.

[0051] (4) Results: such as Figure 1 As shown, different candidate peptides had significantly different effects on BMSCs activity. Compared with the control group, H06, H08, and H09 inhibited BMSCs activity, with cell proliferation rates lower than the control group; H03 and H05 had only slight promoting effects, but no statistical difference (cell proliferation rates of 1.11 and 1.06, respectively); while H01, H02, H04, H07, and H10 showed significant promoting effects on BMSCs proliferation, with statistical differences (cell proliferation rates of 1.22, 1.69, 1.16, 1.25, and 1.40, respectively). The four peptides with the most significant activity, H01, H02, H07, and H10, were selected for subsequent experiments.

[0052] Example 4 This embodiment provides the effect of different concentrations of hippocampal bone-strengthening peptide on the proliferation of BMSCs.

[0053] (1) Culture and passage of BMSCs: Same as step (1) in Example 3.

[0054] (2) Cell seeding and drug administration at different concentrations: BMSCs in the logarithmic growth phase were digested and then administered at 2×10⁻⁶ concentrations. 3 Hippocampal osteotrophic peptides H01, H02, H07, and H10 (dissolved in PBS) were seeded at final concentrations of 2 μM, 4 μM, 8 μM, and 10 μM, respectively, with three replicates for each concentration; the control group was incubated with an equal volume of PBS. The plates were incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0055] (3) CCK-8 detection and calculation: Same as step (3) in Example 3.

[0056] (4) Results: such as Figure 2As shown, the four hippocampal bone-strengthening peptides exhibited concentration-dependent proliferative effects on BMSCs. Specifically, H01 showed the strongest proliferative effect at 10 μM (cell proliferation fold of 1.34), H02 at 8 μM (cell proliferation fold of 1.90), H07 at 10 μM (cell proliferation fold of 1.34), and H10 at 10 μM (cell proliferation fold of 1.48). This indicates that the peptides of the present invention possess significant proliferative activity on BMSCs at micromolar concentration levels.

[0057] Example 5 This embodiment provides the effect of hippocampal bone-strengthening peptides on the growth curve of BMSCs.

[0058] (1) Culture and passage of BMSCs: Same as step (1) in Example 3.

[0059] (2) Cell seeding and drug administration: Take BMSCs in the logarithmic growth phase, digest and resuspend them to form a cell suspension, and administer 2×10⁶ cells per cell. 3 Hippocampal osteopeptides were seeded at 100 μL / well in 96-well plates. The experimental groups were treated with the optimal concentrations of H01 (10 μM), H02 (8 μM), H07 (10 μM), and H10 (10 μM), respectively; the control group (Ctrl) was treated with an equal volume of PBS. All samples were incubated at 37°C in a 5% CO2 incubator.

[0060] (3) Continuous detection: At the same time on day 1, day 2 and day 3 of culture, 10 μL of CCK-8 solution was added to each well, and after incubation for 0.5 hours, the OD was measured using a microplate reader. 450 Value. Three duplicate holes are set for each group at each time point.

[0061] (4) Results: such as Figure 3 As shown, with the extension of culture time, the OD of all experimental groups... 450 The values ​​were all higher than those of the control group. At 48 and 72 hours, the OD values ​​of the H01, H02, H07, and H10 treatment groups were significantly higher. 450 The values ​​were all significantly higher than those of the control group (P < 0.01). This indicates that the four hippocampal bone-strengthening peptides can continuously promote the proliferation of BMSCs, with H02 showing the most significant effect.

[0062] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A hippocampal bone-strengthening peptide, characterized in that, The amino acid sequence of the hippocampal bone-strengthening peptide is any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7 or SEQ ID NO:

10.

2. A pharmaceutical composition, characterized in that, Includes the hippocampal bone-strengthening peptide as described in claim 1 or its pharmaceutical salt.

3. The pharmaceutical composition according to claim 2, characterized in that, The pharmaceutical composition also contains pharmaceutically acceptable excipients.

4. The pharmaceutical composition according to claim 3, characterized in that, The pharmaceutically acceptable excipients include at least one of diluents, carriers, binders, lubricants, disintegrants, pH adjusters, antioxidants, solvents, or penetrants.

5. The pharmaceutical composition according to claim 2, characterized in that, The dosage form of the pharmaceutical composition is at least one of the following: injection, tablet, capsule, granule, oral liquid, lyophilized powder for injection, sustained-release agent, or controlled-release agent.

6. The pharmaceutical composition according to claim 2, characterized in that, The drug composition is administered via at least one of the following routes: intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, or local administration.

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