Organic molecular self-assembly serving as carrier-free system, and preparation method therefor and use thereof

Abstract

The present invention belongs to the field of biomedicine. Provided are an organic molecular self-assembly serving as a carrier-free system, and a preparation method therefor and the use thereof. Provided is a self-assembly, which is composed of steroidal molecules, and by means of optimizing the molecular structures of these steroids and critical aggregation concentrations thereof, a carrier-free system formed via self-assembly of the steroidal molecules is prepared. The carrier-free system can be loaded with a drug to prepare a drug nanocomposite. In the nanocomposite, the efficacy of the drug is improved, and the effects of reducing toxicity and improving efficacy can be achieved. If the steroid molecules used are themselves biologically active, the pharmacological activity of the steroid molecules remains unaffected after being prepared into the drug nanocomposite. The carrier-free system has the advantages of both a carrier-free nano-preparation and a nano-preparation using a conventional material as a carrier.

Description

An organic molecular self-assembled system as a material-free drug delivery system, its preparation method and applications Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to an organic molecular self-assembled system as a material-free drug delivery system, its preparation method, and its uses. Background Technology

[0002] Whether a drug molecule can fully exert its therapeutic effect depends not only on the physiological activity brought about by its single-molecule chemical structure, but also on its formulation. The latter determines how the drug enters the human body, how it is absorbed, distributed, metabolized, and excreted, thus determining its actual efficacy and potential side effects. With the continuous integration of nanotechnology and biomedical technology, nano-formulations have become an extremely important development direction in the field of modern drug research and development (Biomed Phys Eng Express. 2023, 9(5). doi:10.1088 / 2057-1976 / acedb2). Preparing drugs into nano-formulations can bring advantages that single-molecule drug forms cannot possess in terms of targeting, sustained release, and improved bioavailability, thereby achieving the goal of reducing toxicity and increasing efficacy (Molecules. 2024, 29(9):2073. doi:10.3390 / molecules29092073.). For example, nano-sized anti-tumor drug formulations can target tumor tissues through enhanced penetration and retention (EPR) effects, thereby enhancing tumor killing while reducing toxicity to normal tissues (Front Pharmacol.2024,15:1363346.doi:10.3389 / fphar.2024.1363346.); preparing water-soluble local anesthetics into nano-formulations can achieve long-acting sustained release, thus meeting the need for prolonged postoperative analgesia (Drug Des Devel Ther.2023,17:2639-2655.doi:10.2147 / DDDT.S417051.).

[0003] Currently, strategies for preparing nanoparticles are mainly divided into two categories: carrier-free and carrier-based. The carrier-free strategy involves preparing the drug itself into nanoscale crystals through bottom-up crystallization or top-down physical fragmentation and dispersion (Recent Pat Nanotechnol. 2023, 17(4):307-326. doi:10.2174 / 1872210516666220523120313.). However, this strategy is only suitable for drugs with poor water solubility and has many problems in terms of the uniformity and controllability of crystal morphology and size, as well as the dispersibility and stability of the formulation, limiting its widespread application. The carrier-based strategy uses various nanobiomaterials as carriers to form nanoparticles through adsorption and loading of drugs (Pharmaceutics, 2024, 16(10):1339. doi:10.3390 / pharmaceutics16101339.). While the use of carrier materials makes the preparation of nanoparticles more precise and controllable, enabling the production of nanoparticles with uniform size and good dispersibility, the introduction of additional materials also brings new problems. For example, due to considerations of efficacy and production cost, there are high requirements for drug loading and encapsulation efficiency; most carrier materials are non-natural components with large molecular weights, and their biodegradability and biocompatibility bring new safety issues; additional materials and the more complex formulation production processes they bring inevitably lead to increased costs.

[0004] In addition to the two categories of nano-formulations mentioned above—carrier-free and carrier-based (using traditional materials as carriers)—recent studies have proposed a new category: carrier-free nanosystems. Unlike the classification of nano-formulations based on whether or not a carrier is included, carrier-free nanosystems can be considered a category between the two. They refer to small molecules with physiological activity that can be used as therapeutic drugs, which can self-assemble into nanostructures under certain conditions. Among these, carrier-free nanosystems based on drug molecule self-assembly have begun to attract attention in recent years (Med Res Rev. Sep 2020;40(5):1754-1775.doi:10.1002 / med.21669, Emerging carrier-free nanosystems based on molecular self-assembly of pure drugs for cancer therapy). They do not rely on exogenous carrier materials, reducing the immune response caused by the carrier; they have high drug loading capacity; they are simple to prepare; and they themselves can be used as drugs for disease treatment. Material-free drug-loaded nanosystems can also serve as drug carriers to load other drugs, thereby obtaining dual- or multi-active drug formulations. These dual- or multi-active drug formulations not only combine the advantages of the two types of nanoformulations (carrier-free and carrier-based), but the combination of two or more drug molecules may also have a synergistic effect of "reducing toxicity and increasing efficacy." Therefore, the development of novel material-free drug-loaded nanosystems is of great significance.

[0005] Therefore, exploring novel material-free drug delivery systems provides more options for the development of novel nanomedicine formulations and will greatly promote the development of nanomedicine formulations. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides a self-assembled steroidal organic molecule system, with the aim of preparing novel material-free drug delivery systems, thus offering more options for the development of novel nanomedicine formulations.

[0007] This invention provides an organic molecular self-assembled structure, which is obtained by the self-assembly of steroid organic molecules; wherein the steroid organic molecules are selected from at least one of steroid organic molecules with a pregnane backbone structure and an estradiol backbone structure.

[0008] When the steroid organic molecule is selected from at least one of the steroid organic molecules with a pregnane skeleton structure, the concentration of the steroid organic molecule is ≥1 mg / mL;

[0009] When the steroid organic molecule is selected from at least one of steroid organic molecules with an estradiol skeleton structure, the concentration of the steroid organic molecule is ≥0.4 mg / mL.

[0010] Preferably, when the steroid organic molecule is selected from at least one of the steroid organic molecules with a pregnane skeleton structure, the concentration of the steroid organic molecule is ≥2 mg / mL; when the steroid organic molecule is selected from at least one of the steroid organic molecules with an estradiol skeleton structure, the concentration of the steroid organic molecule is ≥0.6 mg / mL.

[0011] Preferably, the steroidal organic molecule with the pregnane skeleton structure is selected from at least one of hydrocortisone, betamethasone, dexamethasone, methylprednisolone, prednisolone, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable ester thereof;

[0012] And / or, the steroidal organic molecule with the estradiol skeleton structure is selected from at least one of estradiol, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable ester thereof.

[0013] Preferably, the steroid organic molecule is selected from at least one of hydrocortisone sodium succinate, betamethasone sodium phosphate, dexamethasone sodium phosphate, methylprednisolone sodium succinate, prednisolone sodium phosphate, and beta-estradiol 17-hemisuccinate.

[0014] Preferably, the specific conditions for self-assembly are: stirring for 4-6 hours at a temperature of 20-25℃ and a stirring speed of 600-1000 rpm, followed by standing for 12-18 hours at a temperature of 2-20℃.

[0015] This invention provides a method for preparing the organic molecular self-assembly according to any one of the above claims, comprising: obtaining it by self-assembly of steroid organic molecules; wherein the steroid organic molecules are selected from at least one of steroid organic molecules with a pregnane backbone structure and an estradiol backbone structure;

[0016] When the steroid organic molecule is selected from at least one of the steroid organic molecules with a pregnane skeleton structure, the concentration of the steroid organic molecule is ≥1 mg / mL;

[0017] When the steroid organic molecule is selected from at least one of the steroid organic molecules with an estradiol skeleton structure, the concentration of the steroid organic molecule is ≥0.4 mg / mL.

[0018] The present invention provides the use of the organic molecular self-assemblies described in any of the above claims as drug carriers and / or active pharmaceutical ingredients.

[0019] The present invention provides a pharmaceutical composition comprising an organic molecular self-assembly of any one of the above-described embodiments loaded with a drug; the drug is selected from at least one of a compound having pharmaceutical activity containing a tertiary amine group and a hydrophobic aromatic ring structure, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable ester thereof;

[0020] The molar ratio of the steroid organic molecules used to form the organic molecule self-assembly to the drug is 1:2.5-40.

[0021] Preferably, the pharmaceutical composition is prepared by mixing the pharmaceutical composition with a steroidal organic molecule that constitutes the organic molecule self-assembly.

[0022] Preferably, the specific preparation process is as follows: at a temperature of 20-25°C, the steroid organic molecules are added dropwise to the drug and mixed under stirring at a speed of 600-1000 rpm for 4-6 hours, and then allowed to stand at a temperature of 2-20°C for 12-18 hours.

[0023] And / or, the steroid organic molecules are adjusted to pH ≤ 7.0 with a pH adjuster before being mixed with the drug.

[0024] The term "organic molecule self-assembly" refers to a nanostructure formed by the self-assembly of organic molecules, which can be either drug-active or non-drug-active organic molecules.

[0025] This invention provides a self-assembled system composed of steroid molecules. By optimizing the molecular structure and critical aggregation concentration of these steroids, a material-free drug-loaded system formed by the self-assembly of steroid molecules is prepared. This material-free drug-loaded system can load drugs as drug carriers to prepare drug nanocomposites. In this nanocomposite, the efficacy of the drug is enhanced, achieving a "reduced toxicity and enhanced efficacy" effect. Furthermore, if the steroid molecules used are inherently bioactive, their pharmacological activity is not affected after being made into the drug nanocomposite of this invention. The material-free drug-loaded system prepared by this invention possesses the advantages of both carrier-free and traditional material-based nanoparticle formulations, showing promising prospects in the development of novel nanomedicine formulations.

[0026] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0027] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0028] Figure 1 is a transmission electron microscope image of the nanospheres formed by HSS self-assembly;

[0029] Figure 2 shows the changes in zeta potential after HSS loading with different proportions of DHM.

[0030] Figure 3 shows transmission electron microscopy images of nanosphere formulations formed by loading HSS with different proportions of DHM.

[0031] Figure 4 shows the changes in zeta potential after HSS loading with different proportions of LB.

[0032] Figure 5 shows transmission electron microscopy images of HSS-loaded nanospheres with different proportions of LB.

[0033] Figure 6 shows a transmission electron microscope image of nanospheres formed by BSP self-assembly.

[0034] Figure 7 shows the changes in zeta potential after loading BSP with different proportions of ATL.

[0035] Figure 8 shows transmission electron microscopy images of nanosphere formulations formed by loading BSP with different proportions of ATL.

[0036] Figure 9 shows the changes in zeta potential after loading BSP with different proportions of ROP.

[0037] Figure 10 shows the changes in zeta potential after BSP loading with different proportions of PMZ.

[0038] Figure 11 shows transmission electron microscopy images of nanosphere formulations formed by loading BSP with different proportions of PMZ.

[0039] Figure 12 is a transmission electron microscope image of nanospheres formed by the self-assembly of DMSP;

[0040] Figure 13 shows the changes in zeta potential after loading DMSP with different proportions of BUP.

[0041] Figure 14 shows transmission electron microscopy images of nanosphere formulations formed by loading different proportions of BUP onto DMSP.

[0042] Figure 15 shows the changes in zeta potential after loading DMSP with different proportions of OCD.

[0043] Figure 16 shows transmission electron microscopy images of nanosphere formulations formed by loading DMSP with different proportions of OCD.

[0044] Figure 17 is a transmission electron microscope image of MPSS self-assembly forming nanospheres;

[0045] Figure 18 shows an atomic force electron microscope image of MPSS self-assembly forming nanospheres;

[0046] Figure 19 shows the changes in zeta potential after MPSS loading with different proportions of TDZ;

[0047] Figure 20 shows transmission electron microscopy images of nanosphere formulations formed by loading MPSS with different proportions of TDZ;

[0048] Figure 21 shows the changes in zeta potential after MPSS is loaded with different proportions of LB;

[0049] Figure 22 shows transmission electron microscopy images of nanosphere formulations formed by loading MPSS with different proportions of LB.

[0050] Figure 23 shows the changes in zeta potential after MPSS is loaded with different proportions of MP;

[0051] Figure 24 shows an atomic force electron microscope image of MPSS loaded with ROP;

[0052] Figure 25 shows a transmission electron microscope image of nanospheres formed by the self-assembly of PNSP.

[0053] Figure 26 shows the changes in zeta potential after PNSP loading with different proportions of ROP.

[0054] Figure 27 shows transmission electron microscopy images of nanosphere formulations formed by loading different proportions of ROP onto PNSP.

[0055] Figure 28 shows a transmission electron microscope image of nanospheres formed by BEH self-assembly;

[0056] Figure 29 shows the changes in zeta potential after BEH loading with different proportions of DHM.

[0057] Figure 30 shows transmission electron microscopy images of nanosphere formulations formed by loading BEH with different proportions of DHM.

[0058] Figure 31 shows the results of the determination of the critical aggregation concentration of HSS;

[0059] Figure 32 shows the results of the determination of the critical aggregation concentration of BSP;

[0060] Figure 33 shows the results of the determination of the critical aggregation concentration of DMSP;

[0061] Figure 34 shows the results of the determination of the critical aggregation concentration of MPSS;

[0062] Figure 35 shows the results of the determination of the critical aggregation concentration of PNSP;

[0063] Figure 36 shows the results of the determination of the critical aggregation concentration of BEH;

[0064] Figure 37 shows the systemic inflammatory factor levels in rats after administration of ROP and ROP-MPSS (*: p<0.05);

[0065] Figure 38 shows the survival rate of A549 cells under different drug concentrations of LB and LB-MPSS (*: p<0.05, ***: p<0.001);

[0066] Figure 39 shows the survival rate of A549 cells under different drug concentrations of BUP and BUP-DMSP (*: p<0.05, **: p<0.01);

[0067] Figure 40 shows the survival rate of A549 cells under different drug concentrations of ROP and ROP-PNSP (*: p<0.05, **: p<0.01). Detailed Implementation

[0068] Unless otherwise specified, all reagents and materials used in the following examples and experimental cases are commercially available.

[0069] Example 1: Preparation of Sodium Hydrocortisone Succinate Nanospheres

[0070] I. Preparation of hydrocortisone sodium succinate nanospheres

[0071] Weigh a measured amount of hydrocortisone sodium succinate (HSS, hereinafter referred to as HSS), add it to ultrapure water to prepare a 10 mg / mL HSS stock solution (i.e., a 1% HSS stock solution by mass), and sonicate in a water bath for 10 minutes to ensure complete dissolution. Stir magnetically at 600 rpm for 4 hours at room temperature, and then incubate overnight at 4°C to allow for complete self-assembly.

[0072] II. Characterization of Hydrocortisone Sodium Succinate Nanospheres

[0073] The nanostructures formed by the self-assembly of HSS were observed using transmission electron microscopy (TEM). As shown in Figure 1, HSS forms nanospheres with a diameter of 10-50 nm in aqueous solution. This demonstrates that the method of this embodiment successfully prepared hydrocortisone sodium succinate nanospheres.

[0074] Example 2: Nanoformulation of diphenhydramine hydrochloride (DHM-HSS) loaded onto hydrocortisone sodium succinate nanospheres

[0075] I. Preparation of DHM-HSS Nanoparticles

[0076] The concentration of HSS was fixed at 5 mg / mL, and carrier-free nano-formulations were prepared according to the molar ratios of HSS to diphenhydramine hydrochloride (DHM, molecular weight 291.82) of 1:5 and 1:10 (Table 1).

[0077] Table 1. Raw material ratio in DHM-HSS nano-formulation

[0078] The specific preparation process is as follows:

[0079] Weigh a measured amount of DHM, add an appropriate amount of ultrapure water, and sonicate in a water bath for 15 minutes to completely dissolve it. At room temperature, magnetically stir the DHM solution at 600 rpm, and slowly and uniformly add 0.4 mL of 1% HSS stock solution dropwise while stirring to prepare the formulations shown in Table 2. After stirring for 4 hours, let it stand overnight to allow the system to fully load the drug molecules onto the self-assembled nanospheres.

[0080] II. Characterization of DHM-HSS Nanoformulations

[0081] The zeta potential of the formulation was measured using a Malvern nanoparticle potentiometer (Figure 2). Each sample was measured three times, and the average value was taken. The results showed that the HSS nanospheres were negatively charged, with a zeta potential of -28.4 mV. After loading with DHM, the negative charge on the nanospheres decreased, and the decrease in negative charge increased with increasing drug loading. Furthermore, TEM revealed that the microstructure of the formulation consisted of nanospheres with a size of 50-100 nm as the proportion of DHM increased (Figure 3). This demonstrates that the method described in this embodiment successfully prepared DHM-HSS nanoformulations with different molar ratios.

[0082] Example 3: Nanoformulation of levobupivacaine hydrochloride (LB-HSS) loaded onto hydrocortisone sodium succinate nanospheres.

[0083] I. Preparation of LB-HSS Nanoparticles

[0084] The concentration of HSS was fixed at 2 mg / mL, and carrier-free nano-formulations were prepared according to the molar ratios of HSS to levobupivacaine hydrochloride (LB) of 1:2.5, 1:10, and 1:40 (Table 2).

[0085] Table 2. Raw material ratios in LB-HSS nano-formulations

[0086] The specific preparation process is as follows:

[0087] Weigh a measured amount of LB, add an appropriate amount of ultrapure water, and sonicate in a water bath for 30 minutes to completely dissolve it. At room temperature, magnetically stir the LB solution at 600 rpm, and slowly and uniformly add 0.4 mL of 1% HSS stock solution dropwise while stirring to prepare the formulations shown in Table 3. After stirring for 4 hours, let it stand overnight to allow the system to fully load the drug molecules onto the self-assembled nanospheres.

[0088] II. Characterization of LB-HSS Nanoformulations

[0089] The zeta potential of the formulation was measured using a Malvern nanoparticle potentiometer (Figure 4). The results showed that after loading LB onto HSS nanospheres, the negative charge of the HSS nanospheres was significantly reduced, and this reduction continued with increasing total LB content. Furthermore, TEM revealed that the microstructure of the formulation consisted of nanospheres ranging from 30 to 150 nm with increasing LB ratio (Figure 5). This demonstrates that the method described in this embodiment successfully prepared LB-HSS nanoformulations, and that a higher LB ratio resulted in a larger particle size of the LB-HSS nanoformulations.

[0090] Example 4: Preparation of Betamethasone Sodium Phosphate Nanospheres

[0091] I. Preparation of Betamethasone Sodium Phosphate Nanospheres

[0092] Weigh a measured amount of betamethasone sodium phosphate (BSP) and add it to ultrapure water to prepare a 10 mg / mL BSP stock solution (i.e., a 1% BSP stock solution by mass). Sonicate in a water bath for 20 minutes to ensure complete dissolution. Stir magnetically at 600 rpm for 4 hours at room temperature, and then incubate overnight at 4°C to allow for complete self-assembly.

[0093] II. Characterization of Betamethasone Sodium Phosphate Nanospheres

[0094] The nanostructures formed by the self-assembly of BSP were observed using TEM. As shown in Figure 6, BSP formed nanospheres with a diameter of 10-50 nm in aqueous solution. This demonstrates that the method of this embodiment successfully prepared betamethasone sodium phosphate nanospheres.

[0095] Example 5: Nanoformulation of amitriptyline hydrochloride (ALT-BSP) loaded onto betamethasone sodium phosphate nanospheres.

[0096] I. Preparation of ALT-BSP Nanoparticles

[0097] Amitriptyline hydrochloride (ATL) was loaded into a 5 mg / mL BSP solution, and carrier-free nanoformulations were prepared at BSP to ATL molar ratios of 1:5 and 1:10 (Table 3). The preparation method was as follows: first, the pH of the BSP was adjusted to 6.0 with 1M hydrochloric acid, and then the formulation was prepared according to the method described in Example 2, except that the formulations were prepared in the corresponding proportions shown in Table 3.

[0098] Table 3. Raw material ratios in ALT-BSP nano-formulation

[0099] II. Characterization of ALT-BSP Nanoformulation

[0100] The zeta potential of the formulation was measured using a Malvern nanoparticle potentiometer. The BSP nanospheres were negatively charged, with a zeta potential of -24.9 mV. After loading with ATL, the negative charge on the nanospheres decreased significantly, becoming positively charged (Figure 7). Furthermore, TEM revealed that with increasing ATL ratio, the microstructure of the formulation consisted of nanospheres with a size of 40-80 nm (Figure 8). This demonstrates that the method described in this embodiment successfully prepared the ALT-BSP nanosphere formulation.

[0101] Example 6: Nanoformulation of betamethasone sodium phosphate nanospheres loaded with ropivacaine hydrochloride (ROP-BSP)

[0102] I. Preparation of ROP-BSP Nanoparticle Formulation

[0103] Ropivacaine hydrochloride (ROP) was loaded with a BSP solution at a molar concentration of 2.5 times and 10 times that of BSP, and ROP was loaded with a BSP solution at a molar concentration of 1.333 mg / mL at a molar concentration of 30 times. Specific parameters and ratios are shown in Table 4. The preparation conditions were the same as in Example 5, except that the formulations were prepared in the corresponding proportions shown in Table 4.

[0104] Table 4. Raw material ratios in ROP-BSP nano-formulations

[0105] II. Characterization of ROP-BSP Nanoformulation

[0106] When BSP is loaded with ROP, the negative charge of the nanospheres decreases, and this decrease is significant with increasing total ROP content (Figure 9). This demonstrates that the method of this embodiment successfully prepared ROP-BSP nanoformulations.

[0107] Example 7: Nanoformulation of betamethasone sodium phosphate nanospheres loaded with promethazine hydrochloride (PMZ-BSP)

[0108] I. Preparation of PMZ-BSP Nanoparticles

[0109] Promethazine hydrochloride (PMZ) was loaded into a 5 mg / mL BSP solution, and carrier-free nano-formulations were prepared at BSP to PMZ molar ratios of 1:5 and 1:10 (Table 5). A measured amount of PMZ was weighed and added to an appropriate amount of ultrapure water, and the mixture was sonicated in a water bath for 20 minutes to ensure complete dissolution. At room temperature and with magnetic stirring at 600 rpm, 0.4 mL of 1% BSP stock solution (pH 6.0) was slowly added dropwise (20 μL / s), and stirring continued for 4 hours to obtain the nano-formulations in the corresponding proportions shown in Table 5. The system was then incubated overnight at 4°C to stabilize the system.

[0110] Table 5. Raw material ratio in PMZ-BSP nano-formulation

[0111] II. Characterization of PMZ-BSP Nanoformulation

[0112] After loading with PMZ, the negative charge on the BSP nanospheres was significantly reduced, and the PMZ-BSP formulation nanospheres became positively charged (Figure 10). Furthermore, TEM revealed that with increasing PMZ content, the formulation's microstructure consisted of nanospheres ranging from 50 to 150 nm (Figure 11). This demonstrates that the method described in this embodiment successfully prepared the PMZ-BSP nanoformulation.

[0113] Example 8: Preparation of dexamethasone sodium phosphate nanospheres

[0114] I. Preparation of dexamethasone sodium phosphate nanospheres

[0115] Weigh a measured amount of dexamethasone sodium phosphate (DMSP), add it to ultrapure water to prepare a 10 mg / mL stock solution, and sonicate in a water bath for 20 minutes to ensure complete dissolution. Perform self-assembly according to the conditions of Example 1.

[0116] II. Characterization of Dexamethasone Sodium Phosphate Nanospheres

[0117] Under TEM, it can be observed that DMSP self-assembles into relatively uniform nanospheres with a diameter of 20-40 nm in aqueous solution (Figure 12). This demonstrates that the method of this embodiment successfully prepared dexamethasone sodium phosphate nanospheres.

[0118] Example 9: Nanoformulation of dexamethasone sodium phosphate nanospheres loaded with bupivacaine hydrochloride (BUP-DMSP)

[0119] I. Preparation of BUP-DMSP Nanoparticles

[0120] The DMSP was loaded with bupivacaine hydrochloride (BUP) at a molar concentration of 2 mg / mL, which was 2.5 and 10 times that of DMSP, respectively. The DMSP was loaded with BUP at a molar concentration of 1.333 mg / mL, which was 30 times that of DMSP. Specific parameters and ratios are shown in Table 6. The preparation method was as follows: first, the pH of the DMSP was adjusted to 6.0 with 1M hydrochloric acid, and then the preparation was carried out according to the method described in Example 2, except that the formulation was prepared in the corresponding proportions shown in Table 6.

[0121] Table 6. Raw material ratios in BUP-DMSP nanoformulations

[0122] II. Characterization of BUP-DMSP Nanoformulation

[0123] DMSP nanospheres are negatively charged with a zeta potential of -26.3 mV. After loading with BUP, the negative charge of the DMSP nanospheres decreases, and this decrease further with increasing total BUP content (Figure 13). TEM observation shows that with increasing BUP proportion, the BUP-DMSP formulation exhibits a nanosphere structure with a diameter of 60-100 nm (Figure 14). This demonstrates that the method of this embodiment successfully prepared the BUP-DMSP nanoformulation.

[0124] Example 10: Nanoformulation of dexamethasone sodium phosphate nanospheres loaded with oxycodone hydrochloride (OCD-DMSP)

[0125] I. Preparation of OCD-DMSP Nanoparticles

[0126] Carrier-free nanoformulations were prepared using 1 mg / mL DMSP loaded with oxycodone hydrochloride (OCD) at DMSP to OCD molar ratios of 1:2.5 and 1:10 (Table 7). The preparation conditions were the same as in Example 9, except that the formulations were prepared in the corresponding proportions shown in Table 7.

[0127] Table 7. Raw material ratio in OCD-DMSP nano-formulation

[0128] II. Characterization of OCD-DMSP Nanoformulation

[0129] After loading with OCD, the negative charge of the nanosphere formulation was significantly reduced, and this reduction further increased with the increase of the total amount of OCD loaded (Figure 15). TEM observation showed that, with the increase of the OCD ratio, the OCD-DMSP formulation exhibited a nanosphere structure with a diameter of 30-120 nm (Figure 16). This demonstrates that the method of this embodiment successfully prepared the OCD-DMSP nanosphere formulation.

[0130] Example 11 Preparation of Methylprednisolone Sodium Succinate Nanospheres

[0131] I. Preparation of Methylprednisolone Sodium Succinate Nanospheres

[0132] A measured amount of methylprednisolone sodium succinate (MPSS) was weighed and added to ultrapure water to prepare a 10 mg / mL stock solution. The solution was then sonicated in a water bath for 20 minutes to ensure complete dissolution. Following the conditions of Example 1, the solution was allowed to self-assemble.

[0133] II. Characterization of Methylprednisolone Sodium Succinate Nanospheres

[0134] TEM revealed that MPSS self-assembled in aqueous solution into relatively uniform nanospheres with a diameter of 30-50 nm (Figure 17). This demonstrates that the method described in this embodiment successfully prepared methylprednisolone sodium succinate nanospheres.

[0135] Example 12 Preparation of Methylprednisolone Sodium Succinate Nanospheres

[0136] I. Preparation of Methylprednisolone Sodium Succinate Nanospheres

[0137] Methylprednisolone sodium succinate nanospheres were prepared according to the preparation method of Example 11, except that the concentration of MPSS was 5 mg / mL.

[0138] II. Characterization of Methylprednisolone Sodium Succinate Nanospheres

[0139] Atomic force microscopy revealed that MPSS self-assembled into relatively uniform nanospheres in aqueous solution (Figure 18). This demonstrates the successful preparation of methylprednisolone sodium succinate nanospheres using the method described in this embodiment.

[0140] Example 13: Nanoformulation of methylprednisolone sodium succinate nanospheres loaded with thioridazine hydrochloride (TDZ-MPSS)

[0141] I. Preparation of TDZ-MPSS Nanoparticle Formulation

[0142] Carrier-free nanoformulations were prepared by loading thioridazine hydrochloride (TDZ) at a concentration of 5 mg / mL MPSS and at molar ratios of 1:5 and 1:10 to TDZ (Table 8). The preparation method was the same as in Example 2.

[0143] Table 8. Raw material ratio in TDZ-MPSS nano-formulation

[0144] II. Characterization of TDZ-MPSS Nanoformulation

[0145] MPSS nanospheres are negatively charged with a zeta potential of -41.5 mV. After loading with TDZ, the negative charge disappears, and the nanospheres become positively charged (Figure 19). TEM observation shows that as the proportion of TDZ increases, the TDZ-MPSS formulation exhibits a nanosphere structure with a diameter of 30-120 nm (Figure 20). This demonstrates that the method of this embodiment successfully prepared the TDZ-MPSS nanosphere formulation.

[0146] Example 14: Nanoformulation of methylprednisolone sodium succinate nanospheres loaded with levobupivacaine hydrochloride (LB-MPSS)

[0147] I. Preparation of LB-MPSS Nanoparticles

[0148] LB was loaded with 2 mg / mL MPSS, and carrier-free nanoformulations were prepared at MPSS to LB molar ratios of 1:2.5, 1:10, and 1:40 (Table 9). The preparation method was the same as in Example 3, except that the formulations were prepared in the corresponding proportions shown in Table 9.

[0149] Table 9. Raw material ratios in LB-MPSS nanoformulations

[0150] II. Characterization of LB-MPSS Nanoformulations

[0151] After loading with LB, the negative charge of the MPSS nanospheres decreased, and this decrease was significant with increasing total LB loading (Figure 21). TEM revealed that the LB-MPSS formulation exhibited nanosphere structures with diameters of 30-150 nm with increasing LB proportions (Figure 22). This demonstrates that the method of this embodiment successfully prepared the LB-MPSS nanosphere formulation.

[0152] Example 15: Nanoformulation of methylprednisolone sodium succinate nanospheres loaded with morphine hydrochloride (MP-MPSS)

[0153] I. Preparation of MP-MPSS Nanoparticle Formulation

[0154] Using 1 mg / mL MPSS loaded with morphine hydrochloride (MP), carrier-free nanoformulations were prepared at MPSS to MP molar ratios of 1:2.5 and 1:10 (Table 10). The preparation method was the same as in Example 2, except that the formulations were prepared in the corresponding proportions shown in Table 10.

[0155] Table 10. Raw material ratios in MP-MPSS nanoformulations

[0156] II. Characterization of MP-MPSS Nanoformulations

[0157] After MP was loaded onto MP nanospheres, the negative charge was significantly reduced, and this reduction gradually increased with the total amount of MP loaded (Figure 23). This demonstrates that the method of this embodiment successfully prepared MP-MPSS nanoformulations.

[0158] Example 16: Nanoformulation of methylprednisolone sodium succinate nanospheres loaded with ropivacaine hydrochloride (ROP-MPSS)

[0159] I. Preparation of ROP-MPSS Nanoparticle Formulation

[0160] Ropivacaine hydrochloride was prepared using a 5 mg / mL MPSS loading molar concentration 15 times that of ropivacaine hydrochloride, in the same manner as in Example 2.

[0161] II. Characterization of ROP-MPSS Nanoformulation

[0162] As shown in Figure 24, atomic force microscopy reveals that ROP-MPSS exists as nanospheres in aqueous solution, with a particle size slightly larger than MPSS nanospheres prepared at the same MPSS concentration (Figure 18). This demonstrates that the method of this embodiment successfully prepared the ROP-MPSS nanosphere formulation.

[0163] Example 17 Preparation of Prednisolone Sodium Phosphate Nanospheres

[0164] I. Preparation of Prednisolone Sodium Phosphate Nanospheres

[0165] Weigh a measured amount of prednisolone sodium phosphate (PNSP), add it to ultrapure water to prepare a 10 mg / mL stock solution, and sonicate in a water bath for 20 minutes to ensure complete dissolution. Perform self-assembly according to the conditions of Example 1.

[0166] II. Characterization of Prednisolone Sodium Phosphate Nanospheres

[0167] Under TEM, it can be observed that PNSP self-assembles into relatively uniform nanospheres with a diameter of 50-100 nm in aqueous solution (Figure 25). This demonstrates that the method of this embodiment successfully prepared prednisolone sodium phosphate nanospheres.

[0168] Example 18: Nanoformulation of prednisolone sodium phosphate nanospheres loaded with ropivacaine hydrochloride (ROP-PNSP)

[0169] I. Preparation of ROP-PNSP Nanoparticle Formulation

[0170] Ropivacaine hydrochloride (ROP) with a PNSP loading molar concentration of 2 mg / mL was used at 2.5 and 10 times its concentration, and ROP with a PNSP loading molar concentration of 1.333 mg / mL was used at 30 times its concentration. Specific parameters and ratios are shown in Table 11. The preparation method is the same as in Example 3, except that the formulations were prepared in the corresponding proportions shown in Table 11.

[0171] Table 11. Raw material ratios in ROP-PNSP nano-formulations

[0172] II. Characterization of ROP-PNSP Nanoformulation

[0173] PNSP nanospheres are negatively charged with a zeta potential of -20.2 mV. After loading with ROP, the negative charge of the nanospheres decreases, and this decrease is significant with increasing total ROP content (Figure 26). TEM observation shows that with increasing ROP proportion, the ROP-PNSP formulation exhibits a nanosphere structure with a diameter of 50-150 nm (Figure 27). This demonstrates that the method of this embodiment successfully prepared the ROP-PNSP nanoformulation.

[0174] Example 19 Preparation of beta-estradiol 17-hemisuccinate nanospheres

[0175] I. Preparation of beta-estradiol 17-hemisuccinate nanospheres

[0176] Weigh a measured amount of beta-estradiol 17-hemisuccinate (BEH), add it to ultrapure water, and then slowly add 1M sodium hydroxide dropwise until the pH reaches 7.0. Sonicate in a water bath for 20 minutes, supplemented with shaking, to ensure thorough dispersion in water, preparing a 10 mg / mL stock solution. Perform self-assembly according to the conditions of Example 1.

[0177] II. Characterization of beta-estradiol 17-hemisuccinate nanospheres

[0178] Under TEM, it can be observed that BEH self-assembles into relatively uniform nanospheres with a diameter of approximately 50 nm in aqueous solution (Figure 28). This demonstrates that the method of this embodiment successfully prepared beta-estradiol 17-hemisuccinate nanospheres.

[0179] Example 20: Nanoformulation of beta-estradiol 17-hemisuccinate loaded with diphenhydramine hydrochloride (DHM-BEH)

[0180] I. Preparation of DHM-BEH Nanoparticles

[0181] Diphenhydramine hydrochloride (DHM) with a BEH loading molar concentration of 2.5 mg / mL was used, at 5 and 10 times that concentration, as shown in Table 12. The preparation method was the same as in Example 2, except that the formulation was prepared in the corresponding proportions shown in Table 12.

[0182] Table 12. Raw material ratios in DHM-BEH nano-formulations

[0183] II. Characterization of DHM-BEH Nanoparticle Formulation

[0184] BEH nanospheres are negatively charged with a zeta potential of -30.04 mV. After loading with DHM, the negative charge of the nanospheres decreases, and this decrease is significant with increasing total DHM content (Figure 29). TEM observation shows that with increasing DHM proportion, the DHM-BEH formulation exhibits a nanosphere structure with a diameter of 100-150 nm (Figure 30). This demonstrates that the method of this embodiment successfully prepared the DHM-BEH nanoformulation.

[0185] The technical solution of the present invention will be further illustrated by the following experiments. The ROP-MPSS nanoparticle formulations tested in the following experimental examples were prepared by the method of Example 16.

[0186] Experimental Example 1: Determination of the critical aggregation concentration (HSS) of self-assembled molecules

[0187] I. Experimental Methods

[0188] Take 499 μL of HSS of different concentrations (see Table 13 for specific concentrations) or ultrapure water (control), add 1 μL of pyrene stock solution (2 mM, prepared with dimethyl sulfoxide), incubate at room temperature for 2 minutes, add to a quartz cuvette, and use a fluorescence spectrophotometer to detect the emission spectrum of pyrene in HSS or water in the range of 360-440 nm under 336 nm excitation light.

[0189] Table 13. I1 / I3 ratio of pyrene in HSS solutions of different concentrations

[0190] II. Experimental Results

[0191] The experimental results are shown in Table 13 and Figure 31. Five characteristic peaks were detected in the free pyrene within the 360-440 nm range. When pyrene was in a hydrophobic environment, the ratio of the first peak (I1) to the third peak (I3) decreased. When pyrene was in highly polar water, its I1 / I3 ratio was 1.60. Compared to the ratio in water, when the HSS concentration was higher than 0.8 mg / mL, the I1 / I3 ratio of pyrene in the HSS solution began to decrease sharply, significantly lower than the ratio in water. This result indicates that when the HSS concentration is higher than 0.8 mg / mL, a large number of hydrophobic regions begin to appear in the solution environment, indicating that HSS possesses self-assembly characteristics. As shown in Figure 31, the critical aggregation concentration (CAC) of HSS is 1-2 mg / mL.

[0192] The above results indicate that when the concentration of HSS solution is higher than 1-2 mg / mL, HSS possesses self-assembly properties and can be used to self-assemble nanomaterials.

[0193] Experimental Example 2: Determination of the critical aggregation concentration of self-assembled molecules (BSP)

[0194] I. Experimental Methods

[0195] The critical aggregation concentration of the self-assembled molecule BSP was determined according to the method in Experimental Example 1. The difference is that the concentration of BSP is shown in Table 14.

[0196] Table 14. I1 / I3 ratio of pyrene in BSP solutions of different concentrations

[0197] II. Experimental Results

[0198] The experimental results are shown in Table 14 and Figure 32. When the BSP concentration is higher than 0.8 mg / mL, the I1 / I3 ratio of pyrene in it is significantly lower than that in water (1.60). As shown in Figure 32, the CAC for BSP self-assembly is 1-2 mg / mL.

[0199] The above results indicate that when the concentration of BSP solution is higher than 1-2 mg / mL, BSP possesses self-assembly properties and can be used to self-assemble nanomaterials.

[0200] Experimental Example 3: Determination of the critical aggregation concentration of self-assembled molecules DMSP

[0201] I. Experimental Methods

[0202] The critical aggregation concentration of the self-assembled molecule DMSP was determined according to the method in Experimental Example 1. The difference is that the concentration of DMSP is shown in Table 15.

[0203] Table 15. I1 / I3 ratio of pyrene in DMSP solutions of different concentrations

[0204] II. Experimental Results

[0205] The experimental results are shown in Table 15 and Figure 33. With increasing DMSP concentration, the I1 / I3 ratio decreased, especially when the concentration was greater than 0.4 mg / mL, where the I1 / I3 ratio of pyrene was significantly lower than that in water (1.60). Figure 33 shows that the CAC for DMSP self-assembly was 1 mg / mL.

[0206] The above results indicate that when the concentration of DMSP solution is higher than 1 mg / mL, DMSP possesses self-assembly properties and can be used to self-assemble nanomaterials.

[0207] Experimental Example 4: Determination of the critical aggregation concentration of self-assembled molecules MPSS

[0208] I. Experimental Methods

[0209] The critical aggregation concentration of self-assembled molecules MPSS was determined according to the method in Experimental Example 1. The difference is that the concentration of MPSS is shown in Table 16.

[0210] Table 16. I1 / I3 ratio of pyrene in MPSS solutions of different concentrations

[0211] II. Experimental Results

[0212] The experimental results are shown in Table 16 and Figure 34. When the MPSS concentration is greater than 0.5 mg / mL, the I1 / I3 ratio decreases with increasing MPSS concentration, especially when the concentration is greater than 1 mg / mL, the I1 / I3 ratio of pyrene in MPSS is significantly lower than that in water (1.60). As shown in Figure 34, the CAC for MPSS self-assembly is 1-2 mg / mL.

[0213] The above results indicate that when the concentration of MPSS solution is higher than 1-2 mg / mL, MPSS possesses self-assembly properties and can be used to self-assemble nanomaterials.

[0214] Experimental Example 5: Determination of the critical aggregation concentration of self-assembled molecules PNSP

[0215] I. Experimental Methods

[0216] The critical aggregation concentration of self-assembled PNSP molecules was determined according to the method in Experimental Example 1. The difference is that the concentration of PNSP is shown in Table 17.

[0217] Table 17. I1 / I3 ratio of pyrene in PNSP solutions of different concentrations

[0218] II. Experimental Results

[0219] The experimental results are shown in Table 17 and Figure 35. With increasing PNSP concentration, the I1 / I3 ratio decreased, especially when the concentration was greater than 0.8 mg / mL, where the I1 / I3 ratio of pyrene was significantly lower than that in water (1.60). Figure 35 shows that the CAC for PNSP self-assembly was 1-2 mg / mL.

[0220] The above results indicate that when the concentration of PNSP solution is higher than 1-2 mg / mL, PNSP possesses self-assembly properties and can be used to self-assemble nanomaterials.

[0221] Experimental Example 6: Determination of the critical aggregation concentration (BEH) of self-assembled molecules

[0222] I. Experimental Methods

[0223] The critical aggregation concentration of the self-assembled molecule BEH was determined according to the method in Experimental Example 1. The difference is that the concentration of BEH is shown in Table 18.

[0224] Table 18. I1 / I3 ratio of pyrene in BEH solutions of different concentrations

[0225] II. Experimental Results

[0226] The experimental results are shown in Table 18 and Figure 36. With increasing BEH concentration, the I1 / I3 ratio decreased, especially when the concentration was greater than 0.1 mg / mL, where the I1 / I3 ratio of pyrene was significantly lower than that in water (1.60). Figure 36 shows that the CAC for BEH self-assembly was 0.4–0.6 mg / mL.

[0227] The above results indicate that when the concentration of BEH solution is higher than 0.4-0.6 mg / mL, BEH possesses self-assembly properties and can be used to self-assemble nanomaterials.

[0228] Anti-inflammatory effect of ROP-MPSS in Experiment 7

[0229] The ROP-MPSS nanoparticle formulation tested in this experimental example was prepared using the method described in Example 16.

[0230] I. Experimental Methods

[0231] 1. Experimental Grouping

[0232] This experiment was divided into three groups: a control group, a ROP group, and a ROP-MPSS group. The control group received physiological saline, the ROP group received ROP solution, and the ROP-MPSS group received ROP-MPSS solution. The preparation methods for each group's drug solution are as follows:

[0233] ROP group: ROP was dissolved in ultrapure water to form a 10 mg / mL ROP solution.

[0234] ROP-MPSS group: The pH of the ROP-MPSS nano-formulation was adjusted to 7.5-8.5 with 1M sodium hydroxide solution, lyophilized and resuspended to obtain a ROP-MPSS solution with a ROP concentration of 10 mg / mL.

[0235] 2. Animal experiments

[0236] Eight rats were injected with 0.2 mL of physiological saline, ROP solution (10 mg / mL), and ROP-MPSS solution (ROP concentration 10 mg / mL) into the lateral thigh muscle of each rat group. Tissue samples were collected from the injection sites at 12 and 24 hours post-administration. Tissue homogenates were prepared, centrifuged, and the supernatant was stored at -80°C for analysis. The concentrations of TNF-α, IL-1β, and IL-6 were determined using an ELISA kit (Thermo Fisher Scientific Inc., USA) according to the manufacturer's instructions.

[0237] II. Experimental Results

[0238] The results are shown in Figure 37. Compared with the saline control group, injection of 10 mg / mL ROP induced significant local inflammation, manifested by increased expression levels of TNF-α, IL-1β, and IL-6. In rats injected with ROP-MPSS, the expression levels of inflammatory factors were similar to those in the saline group, but compared with the ROP group, the expression of TNF-α (12h:p=0.0079, 24h:p=0.011), IL-1β (12h:p=0.0079, 24h:p=0.015), and IL-6 (12h:p=0.0119, 24h:p=0.015) was significantly reduced.

[0239] The above results indicate that MPSS in ROP-MPSS can not only act as a carrier to load ROP to form nano-formulations, but also exert its original anti-inflammatory effect, achieving a "toxicity reduction" effect.

[0240] Experimental Example 8: Cytotoxicity of Nanocomposites

[0241] I. Experimental Methods

[0242] 1. Cells and their culture conditions

[0243] This experiment used human non-small cell lung cancer cells A549. The cells were cultured in DMEM medium (Hyclone) supplemented with 10% fetal bovine serum (Gibco), 100 U / ml penicillin, and 100 μg / ml streptomycin at 37°C under 5% carbon dioxide conditions.

[0244] 2. Experimental Grouping

[0245] The experiment was divided into 7 groups: LB group, LB-MPSS group, BUP group, BUP-DMSP group, ROP group, ROP-PNSP group, and control group. The drug in the LB group was free LB; the drug in the LB-MPSS group was the LB-MPSS nanoformulation prepared in Example 14 with a molar concentration ratio of 1:40; the drug in the BUP group was free BUP; the drug in the BUP-DMSP group was the BUP-DMSP nanoformulation prepared in Example 9 with a molar concentration ratio of 1:30; the drug in the ROP group was free ROP; the drug in the ROP-PNSP group was the ROP-PNSP nanoformulation prepared in Example 18 with a molar concentration ratio of 1:40; and the control group used fresh culture medium without the drug.

[0246] 3. Cell processing and viability determination

[0247] (1) A549 cells were used at a density of 1×10⁶ cells per well. 4 The cells were seeded at a density of 100 cells / well in 96-well plates and incubated for 16 hours (to ensure cell adhesion and a stable state).

[0248] (2) Replace each culture medium with 100 μL of fresh medium and add different concentrations of drug according to the grouping. The final drug concentrations in the culture medium are 0.1 mM, 0.3 mM, and 1.0 mM, respectively. The concentration of LB-MPSS nanoparticles is based on the concentration of LB, the concentration of BUP-DMSP nanoparticles is based on the concentration of BUP, and the concentration of ROP-PNSP nanoparticles is based on the concentration of ROP.

[0249] (3) After incubation for 4 hours, remove the culture medium and replace it with 100 μL of fresh culture medium. Continue culturing for 24 hours, add 1×CCK8 (APExBIO) reagent, and then use a microplate reader (BioTek) to detect the absorbance (OD) at a wavelength of 450 nm. The cell viability is calculated as follows: Cell viability (%) = (OD) / (APExBIO) / (100 μL) 样品 –OD 空白 ) / (OD 对照 –OD 空白 )×100%

[0250] The result is the average of three trials.

[0251] II. Experimental Results

[0252] As shown in Figure 38, both LB and LB-MPSS showed cytotoxic effects on A549 cells at drug concentrations of 0.1-1 mM. In particular, at the same LB concentration, the cytotoxic effect of LB-MPSS was significantly higher than that of free LB.

[0253] As shown in Figure 39, both BUP and BUP-DMSP exhibited cytotoxic effects on A549 cells at drug concentrations of 0.1–1 mM. In particular, at the same BUP concentration, the cytotoxic effect of BUP-DMSP was significantly higher than that of free BUP.

[0254] As shown in Figure 40, both ROP and ROP-PNSP exhibited cytotoxic effects on A549 cells at drug concentrations of 0.1-1 mM. In particular, at the same ROP concentration, the cytotoxic effect of ROP-PNSP was significantly higher than that of free ROP.

[0255] The above experimental results show that loading LB, BUP and ROP onto the material-free drug carrier of the present invention can significantly enhance the killing effect of drugs on tumor cells.

[0256] The self-assembled molecules of this invention are themselves drugs. Hydrocortisone sodium succinate (HSS) is a corticosteroid with various pharmacological effects including anti-inflammatory, anti-allergic, and immunosuppressive properties. It can be used for toxic infections, anaphylactic shock, severe adrenocortical insufficiency, connective tissue diseases, severe bronchial asthma, and other allergic diseases, as well as for the prevention and treatment of acute graft rejection. It is also used for rheumatoid arthritis, osteoarthritis, tenosynovitis, and tendon strain. Betamethasone sodium phosphate (BSP) is a corticosteroid with various pharmacological effects including anti-inflammatory, anti-allergic, and immunosuppressive properties. Dexamethasone sodium phosphate (DM...) Prednisolone sodium phosphate (SP) is a type of adrenocortical hormone with various pharmacological effects, including anti-inflammatory, anti-allergic, anti-rheumatic, and immunosuppressive effects. It is mainly used for allergic and autoimmune inflammatory diseases. Methylprednisolone sodium succinate (MPSS) is a synthetic glucocorticoid with various pharmacological effects, including anti-inflammatory, immunosuppressive, and anti-allergic effects. Prednisolone sodium phosphate (PNSP) is a synthetic intermediate-acting glucocorticoid with various pharmacological effects, including anti-inflammatory, anti-allergic, and immunosuppressive effects. Beta-estradiol 17-hemisuccinate (BEH) is an estrogen drug mainly used to supplement estrogen deficiency or regulate estrogen levels.

[0257] In Experiments 1-6 above, the critical aggregation concentrations of self-assembled molecules HSS, BSP, DMSP, MPSS, PNSP, and BEH were screened. Exceeding these critical aggregation concentrations, the self-assembled molecules exhibited self-assembly characteristics. Examples 1, 4, 8, 11, 12, 17, and 19 above demonstrated that under the experimental conditions of this invention, self-assembled molecules HSS, BSP, DMSP, MPSS, PNSP, and BEH can self-assemble to form nanostructures. Examples 2, 3, 5-7, 9, 10, 13-16, 18, and 20 above demonstrated that the nanostructures formed by self-assembled molecules HSS, BSP, DMSP, MPSS, PNSP, and BEH can be used as drug carriers to load drugs. Experiment 7 above demonstrated that self-assembled molecules can still exert their original pharmacological effects after self-assembly, alleviating drug-induced inflammatory responses and achieving a "detoxification" effect. Experiment 8 above demonstrated that the drug carrier of this invention, without materials, can significantly enhance drug efficacy after further loading.

[0258] As can be seen from the above embodiments and experimental examples, the present invention provides a self-assembled system composed of steroid series molecules. By optimizing the molecular structure and critical aggregation concentration of these steroids, a material-free drug-loaded system formed by the self-assembly of steroid series molecules is prepared. This material-free drug-loaded system can load drugs as drug carriers to prepare drug nanocomposites. In this nanocomposite, the efficacy of the drug is enhanced, achieving a "reduced toxicity and enhanced efficacy" effect. Furthermore, if the steroid molecules used themselves are biologically active, their pharmacological activity is not affected after being made into the drug nanocomposites of the present invention. The material-free drug-loaded system prepared by the present invention possesses the advantages of both carrier-free and traditional material-based nanoparticle formulations, showing promising prospects in the development of novel nanomedicine formulations.

Claims

1. An organic molecular self-assembled structure, characterized in that: It is prepared by the self-assembly of steroid organic molecules; the steroid organic molecules are selected from at least one of steroid organic molecules with a pregnane skeleton structure and an estradiol skeleton structure. When the steroid organic molecule is selected from at least one of the steroid organic molecules with a pregnane skeleton structure, the concentration of the steroid organic molecule is ≥1 mg / mL; When the steroid organic molecule is selected from at least one of steroid organic molecules with an estradiol skeleton structure, the concentration of the steroid organic molecule is ≥0.4 mg / mL.

2. The organic molecular self-assembled assembly according to claim 1, characterized in that: When the steroid organic molecule is selected from at least one of the steroid organic molecules with a pregnane skeleton structure, the concentration of the steroid organic molecule is ≥2 mg / mL; when the steroid organic molecule is selected from at least one of the steroid organic molecules with an estradiol skeleton structure, the concentration of the steroid organic molecule is ≥0.6 mg / mL.

3. The organic molecular self-assembled assembly according to claim 1, characterized in that: The steroidal organic molecule with the pregnane skeleton structure is selected from at least one of hydrocortisone, betamethasone, dexamethasone, methylprednisolone, prednisolone or a pharmaceutically acceptable salt or ester thereof; And / or, the steroidal organic molecule with the estradiol skeleton structure is selected from at least one of estradiol, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable ester thereof.

4. The organic molecular self-assembled assembly according to claim 3, characterized in that: The steroid organic molecule is selected from at least one of hydrocortisone sodium succinate, betamethasone sodium phosphate, dexamethasone sodium phosphate, methylprednisolone sodium succinate, prednisolone sodium phosphate, and beta-estradiol 17-hemisuccinate.

5. The organic molecular self-assembled body according to any one of claims 1-4, characterized in that, The specific conditions for self-assembly are: stirring for 4-6 hours at 20-25℃ with a stirring speed of 600-1000 rpm, and then standing for 12-18 hours at 2-20℃.

6. The method for preparing the organic molecular self-assembled body according to any one of claims 1-5, characterized in that, It comprises: prepared by self-assembly of steroidal organic molecules; wherein the steroidal organic molecules are selected from at least one of steroidal organic molecules with a pregnane backbone structure and an estradiol backbone structure; When the steroid organic molecule is selected from at least one of the steroid organic molecules with a pregnane skeleton structure, the concentration of the steroid organic molecule is ≥1 mg / mL; When the steroid organic molecule is selected from at least one of steroid organic molecules with an estradiol skeleton structure, the concentration of the steroid organic molecule is ≥0.4 mg / mL.

7. Use of the organic molecular self-assembly according to any one of claims 1-5 as a drug carrier and / or active pharmaceutical ingredient.

8. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises an organic molecular self-assembly of any one of claims 1-5 loaded with a drug; the drug is selected from at least one of a pharmaceutically active compound containing a tertiary amine group and a hydrophobic aromatic ring structure, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable ester thereof. The molar ratio of the steroid organic molecules used to form the organic molecule self-assembly to the drug is 1:2.5-40.

9. The pharmaceutical composition according to claim 8, characterized in that, The pharmaceutical composition is prepared by mixing the drug with a steroid organic molecule used to form the organic molecule self-assembly.

10. The pharmaceutical composition according to claim 9, characterized in that, The specific preparation process is as follows: at a temperature of 20-25℃, the steroid organic molecules are added dropwise to the drug and mixed under stirring at a speed of 600-1000 rpm for 4-6 hours, and then allowed to stand at a temperature of 2-20℃ for 12-18 hours. And / or, the steroid organic molecules are adjusted to pH ≤ 7.0 with a pH adjuster before being mixed with the drug.

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