A self-assembly material for preventing and treating osteoporosis and its preparation method and application

Abstract

The present invention relates to a self-assembling material for preventing and treating osteoporosis, and its preparation method and application. The self-assembling material for preventing and treating osteoporosis comprises a targeting unit R3, a self-assembling unit R2, and a hydrophobic unit R1, which are sequentially connected by amide bonds. R2 is derived from a multi-hydrogen bond polypeptide sequence; R3 is derived from a RANK targeting peptide and / or a RANKL targeting peptide; and R1 is derived from a C10-C20 alkyl carboxylic acid. The self-assembling material self-assembles into nanoparticles in vivo. After binding to the receptor RANK protein or RANKL protein, the nanoparticles capture the receptor RANKL or RANK molecules through ligand-receptor binding and molecular deformation into a fibrous network structure, competitively inhibiting the binding of RANKL to RANK, reducing osteoclast differentiation, and thus achieving the effect of preventing and treating osteoporosis.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and relates to a self-assembling material, a preparation method and an application thereof, and in particular to a self-assembling material for preventing and treating osteoporosis, a preparation method and an application thereof. Background Art

[0002] Osteoporosis (OP) is the most common systemic bone disease among the elderly. With the accelerating aging of the population, its prevention and treatment have become a hot topic and a challenge in modern medical research. OP is characterized by decreased bone formation and enhanced bone resorption. Osteoclasts are multinucleated giant cells that act as bone resorbing cells during bone remodeling. They are formed by the fusion of multiple mononuclear cells. When osteolysis is active, they attach to bone, forming resorption gaps. They secrete proteases and hydrochloric acid to induce mineral dissolution and bone degradation. Under normal physiological conditions, bone resorption by osteoclasts and bone formation by osteoblasts are in a dynamic equilibrium. When bone resorption exceeds bone formation, osteoporosis develops. In vivo, the binding of RANKL to RANK promotes osteoclast differentiation and leads to bone resorption. OPG, secreted by osteoblasts, inhibits osteoclastogenesis by binding to RANKL, thereby hindering the interaction between RANKL and RANK. Given the central role of RANKL and RANK in the development and progression of osteoporosis, inhibiting the binding process between RANKL and RANK has become an important approach for the treatment of osteoporosis.

[0003] In recent years, the rapid development of nanomaterials has led to their widespread attention and a growing range of applications. CN114984235A discloses a bone-targeted nanomaterial and its preparation method. The material comprises calcium-aluminum double hydroxide layered nanosheets surface-modified with bone-targeting groups, wherein the calcium-aluminum double hydroxide layered nanosheets contain calcium and aluminum. This bone-targeted nanomaterial is used in the treatment of osteoporosis to inhibit osteoclast activity and promote the repair of damaged bones. The degradation products, calcium and aluminum ions, pose no risk of drug accumulation.

[0004] CN109289054A discloses a PLGA-PEG-ZOL drug-loaded nanomaterial that specifically targets bone tissue. Zoledronic acid is coupled to the surface of the PLGA shell. The material not only has the ability to actively target bone tissue in vivo but also has the potential to treat osteoporosis and osteolysis. The material can serve as a drug carrier shell, encapsulating therapeutic drugs. This can delay the initial burst dose of the drug, extend the drug's half-life, increase drug stability and solubility, and impart a certain sustained-release effect and passive targeting to the nanoparticles. Specifically, the PLGA-PEG-ZOL drug-loaded nanomaterial can be loaded with different drugs according to different treatment needs, targeting these drugs to bone tissue and increasing the drug concentration in bone tissue. Therefore, the material has the potential to be applied to a variety of skeletal diseases.

[0005] However, currently, there are relatively few strategies in the existing technology for treating osteoporosis by utilizing the combination of RANKL and RANK. Therefore, it is very important to develop a new product with significant efficacy for treating osteoporosis. Summary of the Invention

[0006] In view of the deficiencies in the prior art, the present invention aims to provide a self-assembling material and a preparation method and application thereof, and in particular to provide a self-assembling material for preventing and treating osteoporosis and a preparation method and application thereof.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a self-assembling material for preventing and treating osteoporosis, wherein the self-assembling material for preventing and treating osteoporosis comprises a targeting unit R3, a self-assembling unit R2, and a hydrophobic unit R1 connected in sequence by an amide bond, and its chemical structure is shown in formula (I):

[0009] R1-R2-R3 formula (Ⅰ);

[0010] Among them, R2 comes from a multiple hydrogen bond polypeptide sequence; R3 comes from a RANK targeting peptide and / or a RANKL targeting peptide; R1 comes from a C10-C20 alkyl carboxylic acid, and the number of carbon atoms in the alkyl group can be, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

[0011] Taking advantage of the fact that nanoparticles can be converted into nanofibers through fiber peptides with multiple hydrogen bonds within the molecule, we designed and prepared a self-assembling material that can bind to RANK protein or RANKL protein. The self-assembling material has three structural units: a targeting unit, a self-assembling unit, and a hydrophobic unit. It self-assembles into nanoparticles in the body. After binding to the receptor RANK protein or RANKL protein, it captures the receptor RANKL or RANK molecules through the ligand-receptor binding force and molecular deformation into a fibrous network structure, competitively inhibiting the binding of RANKL to RANK and reducing the differentiation of osteoclasts, thereby preventing and treating osteoporosis. This provides new ideas and new methods for improving or treating osteoporosis.

[0012] Preferably, the carbon number of the alkyl chain of the alkyl carboxylic acid is 10-15.

[0013] Preferably, the alkyl chain of the alkyl carboxylic acid is a straight chain.

[0014] Preferably, the R1 is derived from lauric acid or n-hexadecanoic acid.

[0015] Preferably, the multiple hydrogen-bonding polypeptide sequence comprises FFVLK or DFFPL.

[0016] Preferably, the RANK targeting peptide includes NVLKLCSGE, and the RANKL targeting peptide includes YLEIEFSLKHR.

[0017] Preferably, R1 is derived from lauric acid, R2 is derived from FFVLK, and R3 is derived from NVLKLCSGE, and its chemical structure is CH3(CH2) 10 CO-FFVLK-NVLKLCSGE.

[0018] Preferably, R1 is derived from n-hexadecanoic acid, R2 is derived from FFVLK, and R3 is derived from YLEIEFSLKHR, and its chemical structure is CH3(CH2) 14 CO-FFVLK-YLEIEFSLKHR.

[0019] In a second aspect, the present invention provides a method for preparing the self-assembly material for preventing and treating osteoporosis according to the first aspect, the preparation method comprising:

[0020] The self-assembly material for preventing and treating osteoporosis is synthesized by solid phase synthesis using amino acids with protected terminal amino groups and side chain amino groups and C10-C20 alkyl carboxylic acids as raw materials.

[0021] The preparation method of the self-assembling material for improving atherosclerosis involved in the present invention is synthesized according to the standard solid phase polypeptide synthesis method (SPPS). For example, the preparation method can be carried out according to the following steps:

[0022] (1) swelling the carrier resin;

[0023] (2) Using amino acids with Fmoc protection at the terminal amino group and Boc protection at the side chain amino group and a hydrophobic material as raw materials, first, according to the amino acid sequence of R3, the first amino acid of R3 is added to the carrier resin, coupled with the carrier resin and connected; the Fmoc protecting group on the first amino acid of R3 is removed, and the second amino acid of R3 is coupled with the first amino acid of R3 and connected; until the condensation of all amino acids in R3 is completed;

[0024] (3) removing the Fmoc protecting group of the last amino acid of R3, and subjecting the first amino acid of R2 to a coupling reaction with the last amino acid of R3 according to the amino acid sequence of R2, and then connecting them; removing the Fmoc protecting group of the first amino acid of R2, and subjecting the second amino acid of R2 to a coupling reaction with the first amino acid of R2, and then connecting them; until the condensation of all amino acids in R2 is completed;

[0025] (4) removing the Fmoc protecting group of the last amino acid of R2, and coupling the carboxyl end of the hydrophobic molecule with the last amino acid of R2 and connecting them;

[0026] (5) removing the product obtained in step (4) from the carrier resin to obtain the self-assembly material.

[0027] In a third aspect, the present invention provides use of the self-assembly material for preventing and treating osteoporosis according to the first aspect in the preparation of a drug for preventing and treating osteoporosis.

[0028] In a fourth aspect, the present invention provides a drug for preventing and treating osteoporosis, wherein the active ingredients of the drug include a first self-assembling material and a second self-assembling material; the first self-assembling material is lauric acid, a polypeptide sequence FFVLK and a polypeptide sequence NVLKLCSGE(CH3(CH2) 10 CO-FFVLK-NVLKLCSGE); the second self-assembly material is n-hexadecanoic acid, a polypeptide sequence FFVLK and a polypeptide sequence YLEIEFSLKHR(CH3(CH2) 14 CO-FFVLK-YLEIEFSLKHR).

[0029] Preferably, the dosage form of the drug is any pharmaceutically acceptable dosage form.

[0030] Preferably, the drug further comprises pharmaceutically acceptable excipients, which include any one or a combination of at least two of diluents, excipients, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, coating materials, colorants, pH regulators, antioxidants, antibacterial agents or buffers.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] Taking advantage of the fact that nanoparticles can be converted into nanofibers through fiber peptides with multiple hydrogen bonds within the molecule, we designed and prepared a self-assembling material that can bind to RANK protein or RANKL protein. The self-assembling material has three structural units: a targeting unit, a self-assembling unit, and a hydrophobic unit. It self-assembles into nanoparticles in the body. After binding to the receptor RANK protein or RANKL protein, it captures the receptor RANKL or RANK molecules through the ligand-receptor binding force and molecular deformation into a fibrous network structure, competitively inhibiting the binding of RANKL to RANK and reducing the differentiation of osteoclasts, thereby preventing and treating osteoporosis. This provides new ideas and new methods for improving or treating osteoporosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is a mass spectrometry characterization of the self-assembly material BIFP for preventing and treating osteoporosis prepared in Example 1;

[0034] Figure 2 This is a mass spectrometry characterization of the self-assembly material BIFY for preventing and treating osteoporosis prepared in Example 2;

[0035] Figure 3 These are transmission electron microscopy images of BIFP or BIFY co-incubated with RANK protein or RANKL protein, respectively;

[0036] Figure 4 Circular dichroism spectra of BIFP or BIFY after co-incubation with RANK protein or RANKL protein, respectively;

[0037] Figure 5 These are the laser confocal microscopy images of BIFP or BIFY labeled with Cy5 and binding to the corresponding target cells (RAW264.7 cells and MC3T3-E1 cells);

[0038] Figure 6 are scanning electron microscopic images of BIFP or BIFY after co-incubation with corresponding target cells (RAW264.7 cells and MC3T3-E1 cells);

[0039] Figure 7 This is the result of the inhibitory effect of BIFP or BIFY on osteoclast differentiation;

[0040] Figure 8 This is a diagram showing the formation of absorption pits on the surface of bone slices after mononuclear cells seeded on the bone slices were treated with BIFP or BIFY;

[0041] Figure 9 This is a statistical graph showing the absorption pit area on the surface of bone slices after mononuclear cells seeded on the bone slices were treated with BIFP or BIFY;

[0042] Figure 10 This is a diagram of the F-actin ring structure of cells observed using a laser confocal microscope after coincubation of BIFP or BIFY with monocytes;

[0043] Figure 11 is the result of Western blot analysis of pAKT and NFAT after co-incubation of BIFP or BIFY with monocytes;

[0044] Figure 12 Micro CT images of the tibia after BIFP or BIFY treatment in mice. DETAILED DESCRIPTION

[0045] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely for the purpose of helping to understand the present invention and should not be regarded as specific limitations of the present invention.

[0046] Example 1

[0047] This embodiment constructs a self-assembling material for preventing and treating osteoporosis (hereinafter referred to as BIFP). The chemical structure of the self-assembling material for preventing and treating osteoporosis is:

[0048] CH3(CH2) 10 CO-FFVLK-NVLKLCSGE

[0049] The preparation method comprises the following steps: using lauric acid, each free amino acid in the polypeptide sequence FFVLK and each free amino acid in the polypeptide sequence NVLKLCSGE as raw materials, and synthesizing the peptide according to a standard polypeptide solid phase synthesis method (SPPS).

[0050] The prepared self-assembled material was characterized by mass spectrometry. The mass spectrometry characterization results are as follows: Figure 1 As shown by Figure 1 It can be seen from the main peak of the mass spectrum that the molecular weight of the polypeptide material is basically consistent with that of the designed one, thereby inferring that the target molecule has been synthesized, indicating that the self-assembly material with the above structure has been successfully synthesized.

[0051] Example 2

[0052] This embodiment constructs a self-assembling material for preventing and treating osteoporosis (hereinafter referred to as BIFY). The chemical structure of the self-assembling material for preventing and treating osteoporosis is:

[0053] CH3(CH2) 14 CO-FFVLK-YLEIEFSLKHR

[0054] The preparation method comprises the following steps: using n-hexadecanoic acid, each free amino acid in the polypeptide sequence FFVLK and each free amino acid in the polypeptide sequence YLEIEFSLKHR as raw materials, and synthesizing the product according to a standard polypeptide solid phase synthesis method (SPPS).

[0055] The prepared self-assembled material was characterized by mass spectrometry. The mass spectrometry characterization results are as follows: Figure 2 As shown by Figure 2 It can be seen from the main peak of the mass spectrum that the molecular weight of the polypeptide material is basically consistent with that of the designed one, thereby inferring that the target molecule has been synthesized, indicating that the self-assembly material with the above structure has been successfully synthesized.

[0056] Example 3

[0057] In this example, the self-assembled nanoparticle solution and nanofiber dispersion were prepared by the following method:

[0058] The self-assembly materials prepared in Example 1 and Example 2 were dissolved in DMSO solvent (the concentration of the self-assembly material was 3×10 -3 M), 10 μL of the above solution was placed in a centrifuge tube, and 990 μL of deionized water was slowly added into the centrifuge tube to prepare a mixed solution with a water content of 98%.

[0059] RANK protein (Example 1) or RANKL protein (Example 2) was added to the self-assembled nanoparticle solution obtained above to make the protein concentration 1×10 -7 M, the morphology transformation of the self-assembled particles was characterized by transmission electron microscopy at 0h, 4h, 8h, 12h, and 24h, respectively. Figure 3 As shown by Figure 3 It can be seen that the self-assembled polypeptide material has become short fibers. The circular diffraction detection was used to characterize the material at 0h, 4h, 8h, 12h, and 24h. The results are as follows: Figure 4 shown.

[0060] Example 4

[0061] This example explores the binding of self-assembled materials to corresponding target cells:

[0062] BIFP prepared in Example 1 and BIFY prepared in Example 2 were labeled with Cy5. RAW264.7 cells and MC3T3-E1 cells expressing RANK or RANKL were seeded into Confocol dishes and allowed to adhere overnight. They were then co-incubated with Cy5-labeled BIFP and Cy5-labeled BIFY, respectively. Laser confocal microscopy was performed at 4, 8, and 24 hours of co-incubation to observe the co-localization of the two materials and cells at different time points. The results are shown in Figure 2. Figure 5 As shown (a is a laser confocal 3D reconstruction image of BIFP co-incubated with RAW264.7 for 4 hours, b is a laser confocal image of BIFP co-incubated with RAW264.7 for 4 hours, c is a laser confocal 3D reconstruction image of BIFY co-incubated with MC3T3-E1 cells for 4 hours, d is a laser confocal image of BIFY co-incubated with MC3T3-E1 cells for 4 hours, and e is a statistical graph of fluorescence intensity at three time points: 4, 8, and 24 hours after the two materials were co-incubated with cells). It can be seen from the figure that the self-assembling polypeptide material can target and bind to BIFP / BIFY.

[0063] Example 5

[0064] This example explores the surface morphology of the self-assembled material after binding to the corresponding target cells:

[0065] RAW264.7 cells and MC3T3-E1 cells expressing RANK or RANKL were seeded on silicon wafers and attached overnight, and then incubated with BIFP prepared in Example 1 and BIFY prepared in Example 2 for 4 hours. They were fixed with 4% paraformaldehyde and dehydrated in a gradient of different concentrations of alcohol (10, 30, 50, 70, 90 and 100%) for 10 minutes in each step. The samples were sprayed with gold for 40 seconds, and the surface morphology of the cells was observed by scanning electron microscopy. The results are shown in Figure 2. Figure 6 As shown in the figure, there are significant differences in the surface morphology between untreated cells and BIFP / BIFY-treated cells. Furthermore, a distinct fibrous network covers BIFP / BIFY-treated RAW264.7 or MC3T3-E1 cells, respectively. These results confirm that BIFP / BIFY binds to the target protein and forms a fibrous network on the target cell surface.

[0066] Example 6

[0067] This example explores the inhibitory effect of self-assembled materials on osteoclast differentiation:

[0068] Mouse BMM cells were cultured in DMEM medium containing 10.0% fetal bovine serum, 100.0U / mL penicillin, 100.0μg / mL streptomycin, and 50ng / mL macrophage colony stimulating factor (M-CSF). The culture temperature was 37.0°C, and the concentration of CO2 in the humidified air was 5.0%. 100ng / mL RANKL protein was used to induce the cells to differentiate into osteoclasts. At the same time, BIFP nanoparticles and BIFY nanoparticles were added at a concentration of 5μM or 50μM, respectively, and 5μM or 50μM P (hydrophilic targeting unit peptide of BIFP) or Y (hydrophilic targeting unit peptide of BIFY) was used as a control. The cells were cultured for 6 days, and the culture medium was changed every other day. The cells were stained for osteoclast-specific tartrate-resistant acid phosphatase (TRAP). Figure 7 As shown in Figure a, BMM cells fused to form red-stained multinuclear giant cells under the stimulation of RANKL, indicating that osteoclasts were formed in the +RANKL group. However, different concentrations of BIFP and BIFY significantly reduced TRAP-positive multinuclear cells to varying degrees. Figure 7 As shown in Figure b, compared with the +RANKL group, the TRAP MNCs in the 50 μM BIFP / BIFY peptide group decreased by 68.1% and 61.2%, respectively. Figure 7 As shown in Figure c, the nuclear fusion index was inhibited by 54.0% and 49.6% respectively, and the tartrate-resistant acid phosphatase activity detection of the cell supernatant was detected. TRAP excretion into the extracellular space is a more specific behavior of osteoclasts. TRAP determination of the cell supernatant can reflect the number of osteoclast differentiation on the side. Figure 7 As shown in middle d, high concentrations of peptides (BIFP and BIFY) can reduce TRAP concentrations by 55.9% and 54.0%, respectively.

[0069] Depend on Figure 7 The results showed that in the osteoclast differentiation inhibition experiment, the high-peptide group was more effective than the low-peptide group (p < 0.001). More importantly, the self-assembling material of the present invention was more effective in inhibiting osteoclast differentiation than the simple targeting peptides (P and Y), and the in vitro experimental results verified the effectiveness of the material.

[0070] Example 7

[0071] This example explores the inhibitory effect of self-assembled materials on osteoclast function:

[0072] Mouse BMM cells were cultured in DMEM medium containing 10.0% fetal bovine serum, 100.0 U / mL penicillin, 100.0 μg / mL streptomycin, and 50 ng / mL macrophage colony-stimulating factor (M-CSF). The culture temperature was 37.0°C and the CO2 concentration in the humidified air was 5.0%.

[0073] Pit formation experiment: Mouse BMMs cells (5000 cells, 200 μL / well) were cultured on bovine cortical bone slices in 96-well culture plates. Overnight culture was performed to allow the cells to be planted on the surface of the bone slices. The next day, BIFP nanoparticles or BIFY nanoparticles (5 or 50 μM) were added to fresh DMEM culture medium containing M-CSF (50 ng / mL) and RANKL (100 ng / mL). The culture medium was replaced every other day (including deformable material treatment). At the end of the 10th day of culture, the bone slices were ultrasonically cleaned to remove attached osteoclasts and macrophages, and then the absorption pits were observed under a microscope, as shown in FIG. Figure 8 As shown, the pit area is counted and the statistical results are shown as follows Figure 9 As shown. Figure 8 and Figure 9 It can be seen that the high concentration BIFP group (17.4±9.3μm 2 ) was significantly reduced in bone resorption pit area compared with the control + RANKL group (92.4 ± 7.6 μm 2 ) was reduced to 75.0%. The high-concentration BIIFY group (30.7±7.0μm 2 ) was reduced to 61.6%. The difference between the experimental group and the control group was statistically significant (p<0.001). The results show that the self-assembly material of the present invention can reduce the bone resorption function of osteoclasts.

[0074] Osteoclast actin ring staining: BMMs cells were seeded in 24-well plates and the culture medium was changed once a day. Cells cultured with BIFP nanoparticles or BIFY nanoparticles (5 or 50 μM) for 6 days were fixed with 4% paraformaldehyde for 15 minutes and permeabilized with 0.2% Triton X-100 for 3 minutes. Cells were stained with rhodamine-phalloidin for 30 minutes and the nuclei were counterstained with Hoechst 33258. The F-actin ring structure of the cells was observed using laser confocal microscopy. The results are shown in Figure 2. Figure 10 As shown in the figure, the F-actin ring is an important structure of osteoclasts in bone resorption, which reflects the bone resorption function of cells to a certain extent. The formation of a complete F-actin ring was clearly detected in cells stimulated by RANKL, and aggregated cell nuclei were seen in the ring. Figure 10 The results showed that the RANKL-induced differentiation control group formed multinucleated, relatively large giant cells. In contrast, in the high-concentration BIFP and BIFY treatment group, only small red rings were observed, and no obvious nuclear aggregation was detected. In the low-concentration treatment group, although no complete large ring structures were detected, nuclear aggregation was observed. These results indicate that BIFP and BIFY significantly inhibit the bone resorption capacity of osteoclasts.

[0075] Example 8

[0076] This example explores the effects of self-assembly materials on cell expression:

[0077] Mouse BMM cells were starved for 2 hours and then treated with self-assembly materials BIFP or BIFY (50 μM) and co-incubated with or without RANKL (100 ng / mL). After 6 days of culture, total cell protein was obtained. It was co-incubated with antibodies to Akt, p-Akt, NFAT, and β-actin, and β-actin was used as an internal control protein. The effects of treatment with self-assembly materials BIFP or BIFY on cell protein expression were detected. The results of pAKT and NFAT Western blotting experiments are shown in Figure 2. Figure 11 As shown in a, the statistical results of p-Akt and NFAT are as follows: Figure 11 As shown in b and c. Figure 11 It can be seen that the self-assembled materials BIFP or BIFY inhibited the expression of pAKT and NFAT signals in BMM cells induced by RANKL, indicating that BIFP and BIFY may inhibit osteoclast differentiation by downregulating the expression of downstream signals such as RANK-RANKL.

[0078] Example 9

[0079] This example explores the effect of self-assembled materials on an osteoporosis mouse model:

[0080] Eight-week-old female C57BL / 6 mice underwent ovariectomy to establish an ovariectomized osteoporosis model in mice. A sham-operated group was also established. Starting one week after surgery, mice were intraperitoneally injected with PBS (5 mg / kg), BIFP (5 mg / kg), BIFY (5 mg / kg), or a combination of BIFP and BIFY (2.5 mg / kg + 2.5 mg / kg), depending on body weight and group. After four weeks of treatment, mice were sacrificed using CO2 anesthesia. Tibiae were harvested and preserved in 4% paraformaldehyde solution.

[0081] Micro-CT scanning was performed using a micro-CT scanner. The micro CT images of the tibia of mice in each group after treatment are shown in the figure below. Figure 12 As shown in the figure, the BIFP and BIFY groups showed excellent osteoporosis prevention and treatment effects, and the BIFP group had significantly better bone microstructure, including trabecular bone, than the other groups. The statistical results of bone density and bone histological parameters are shown in Table 1 below.

[0082] Table 1

[0083]

[0084] As shown in Table 1, both the BIFP and BIFY groups demonstrated excellent therapeutic effects, with the BIFP group showing the most significant effect. The BMD value reached 2.55 times that of the PBS group, and the BV / TV ratio reached 3.99 times. Furthermore, the Tb.Sp value in the BIFP group was 3.65 times lower than that in the PBS group, indicating that the BIFP group exhibited a stronger bone anabolic effect. The BIFY group also showed significant differences from the PBS group. The proximal tibial structural parameters of the half-dose BIFP & BIFY combination group were intermediate between the BIFP and BIFY groups. These results indicate that BIFP is more effective than BIFY in treating osteoporosis.

[0085] The applicant declares that the present invention uses the above-mentioned embodiments to illustrate the self-assembling material for preventing and treating osteoporosis, its preparation method, and its application, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent replacements for various raw materials in the product of the present invention, addition of auxiliary ingredients, and selection of specific methods, etc., are all within the scope of protection and disclosure of the present invention.

[0086] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0087] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A self-assembly material for preventing and treating osteoporosis, characterized in that: The self-assembly material for preventing and treating osteoporosis comprises a targeting unit R3, a self-assembly unit R2 and a hydrophobic unit R1 connected in sequence via amide bonds, and its chemical structure is shown in formula (I): R1-R2-R3 formula (Ⅰ); Among them, R2 comes from a multiple hydrogen-bonding polypeptide sequence, and the multiple hydrogen-bonding polypeptide sequence is FFVLK; R3 comes from a RANK targeting peptide and / or a RANKL targeting peptide, and the RANK targeting peptide is NVLKLCSGE, and the RANKL targeting peptide is YLEIEFSLKHR; R1 comes from a C10-C20 alkyl carboxylic acid, and the alkyl chain of the alkyl carboxylic acid is a straight chain.

2. The self-assembly material for preventing and treating osteoporosis according to claim 1, characterized in that: The carbon number of the alkyl chain of the alkyl carboxylic acid is 10-15.

3. The self-assembly material for preventing and treating osteoporosis according to claim 1, characterized in that: The R1 is derived from lauric acid or n-hexadecanoic acid.

4. The self-assembly material for preventing and treating osteoporosis according to claim 1, characterized in that: The R1 is derived from lauric acid, R2 is derived from FFVLK, and R3 is derived from NVLKLCSGE.

5. The self-assembly material for preventing and treating osteoporosis according to claim 1, characterized in that: The R1 is derived from n-hexadecanoic acid, R2 is derived from FFVLK, and R3 is derived from YLEIEFSLKHR.

6. The method for preparing a self-assembly material for preventing and treating osteoporosis according to any one of claims 1 to 5, characterized in that: The preparation method comprises: The self-assembly material for preventing and treating osteoporosis is synthesized by solid phase synthesis using amino acids with protected terminal amino groups and side chain amino groups and C10-C20 alkyl carboxylic acids as raw materials.

7. Use of the self-assembly material for preventing and treating osteoporosis according to any one of claims 1 to 5 in the preparation of a medicament for preventing and treating osteoporosis.

8. A drug for preventing and treating osteoporosis, characterized in that: The active ingredients of the drug include a first self-assembly material and a second self-assembly material; the first self-assembly material is lauric acid, a polypeptide sequence FFVLK, and a polypeptide sequence NVLKLCSGE connected in sequence through amide bonds; the second self-assembly material is n-hexadecanoic acid, a polypeptide sequence FFVLK, and a polypeptide sequence YLEIEFSLKHR connected in sequence through amide bonds.

9. The drug according to claim 8, characterized in that The dosage form of the drug is any pharmaceutically acceptable dosage form.

10. The drug according to claim 8, characterized in that The drug also includes pharmaceutically acceptable pharmaceutical excipients, which include any one or a combination of at least two of diluents, excipients, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, coating materials, colorants, pH regulators, antioxidants, antibacterial agents or buffers.

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