Periplaneta americana extract preparation as well as preparation method and application thereof

Abstract

The invention discloses a periplaneta americana extract preparation as well as a preparation method and application thereof. The periplaneta americana extract preparation comprises a periplaneta americana extract, a nano drug loading system, an antioxidant synergistic component, a mucous membrane protective agent and pharmaceutical auxiliary materials, wherein the weight ratio of the periplaneta americana extract is 10-50%, the weight ratio of the nano drug loading system is 5-30%, the weight ratio of the antioxidant synergistic component is 3-15%, the weight ratio of the mucous membrane protective agent is 5-25%, and the balance is the pharmaceutical auxiliary materials. The preparation method comprises the following steps: 1) extracting the American cockroach extract; 2) combining the periplaneta americana extract with a nano-carrier material through an emulsification-solvent evaporation method or an ultrasonic coating method to obtain a nano-drug delivery system; and 3) forming the dosage form to obtain the periplaneta americana extract preparation. The periplaneta americana extract preparation is applied to prevention and treatment of radiation injury, comprehensive protection of radiation injury of multiple organs such as lung, intestine, hematopoietic system and the like is realized, the curative effect is improved, and the toxicity is reduced.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and relates to an American cockroach extract (PAE) preparation and a preparation method thereof and application thereof in preventing and treating radiation damage. Background Art

[0002] When ionizing radiation (such as gamma rays and X-rays) acts on the body, it causes damage through direct ionization and indirect oxidation. On the one hand, radiation can directly destroy the structure of biological macromolecules, causing DNA double-strand breaks and protein conformation changes; on the other hand, radiation can induce the decomposition of water molecules to produce a large amount of reactive oxygen species (ROS), such as OH - 、H2O2、O2 - These ROS can trigger lipid peroxidation, destroying the cell membrane phospholipid bilayer and causing organelle dysfunction; they can also attack DNA bases, causing gene mutations or cell apoptosis; at the same time, ROS can activate inflammatory signaling pathways such as NF-κB and MAPK, prompting the release of pro-inflammatory factors such as TNF-α, IL-6, and TGF-β, forming a vicious cycle of "oxidation-inflammation" and ultimately leading to multi-organ radiation damage.

[0003] Currently, amifostine, a first-line clinical free radical scavenger, has significant limitations in its use for radiation protection. From a pharmacokinetic perspective, its half-life is only 15-30 minutes, requiring frequent dosing to maintain effective concentrations. Regarding safety, approximately 30% of patients experience adverse reactions such as hypotension and nausea, significantly limiting its use in frail patients. Furthermore, amifostine lacks targeted properties; after systemic administration, only a small amount of the drug reaches the site of radiation damage, making it difficult to balance efficacy and toxicity.

[0004] Given the limitations of synthetic drugs, natural medicines have become a research hotspot in the field of radiation protection due to the synergistic effects of their multiple components. As an ancient organism, the American cockroach has extremely strong tolerance to extreme environments. Studies have found that American cockroach extract (PAE) is rich in various active ingredients such as peptides, polysaccharides, and amino acids. Among them, peptides can scavenge various free radicals and reduce inflammatory activation; polysaccharides can regulate the process of fibrosis; and nucleosides help promote tissue damage repair. Although certain results have been achieved in the research on PAE radiation protection, existing American cockroach extract preparations have problems such as insufficient targeting and low bioavailability, which limit their application in the prevention and treatment of multi-organ radiation damage. Nanodelivery systems can increase the enrichment concentration of drugs in damaged tissues and reduce off-target toxicity through enhanced permeation and retention effect (EPR) or active targeted modification, providing new ideas for solving the above problems. Summary of the Invention

[0005] To address the shortcomings of existing PAE preparations, which suffer from uneven distribution across multiple organs and low bioavailability, the present invention aims to provide an American cockroach extract preparation, its preparation method, and its application in preventing and treating radiation damage. This nanodelivery-based PAE preparation provides comprehensive protection against radiation damage to multiple organs, including the lungs, intestines, and hematopoietic system, while improving efficacy and reducing toxicity.

[0006] The preparation of the present invention obtains the active ingredients of PAE (including polypeptides, polysaccharides and nucleosides) through ultra-high pressure and low temperature extraction technology, and combines it with nano-carriers (such as polymer nanoparticles, liposomes, nanoemulsions, etc.) to form a nano-drug delivery system of nanometer scale (10-3000nm) (Table 1). The present invention significantly improves the enrichment efficiency of PAE in multiple organs such as the lungs, intestines, and hematopoietic system through nano-technology, can efficiently remove radiation-induced reactive oxygen species (ROS), inhibit excessive inflammatory reactions, and achieve the prevention and treatment of multi-organ radiation damage (such as radiation pneumonia, enteritis, and bone marrow suppression). Experiments have confirmed that the efficacy of this nano-preparation is significantly improved compared to traditional preparations, and the dosage is greatly reduced, with no obvious toxic side effects, providing a safe and effective new solution for radiation protection.

[0007] Table 1

[0008] Pk 1Mean Int Pk 2Mean Int Pk 3Mean Int d.nm d.nm d.nm 3936 606.2 11.23 2389 531.6 22.85 3256 746.3 9.424

[0009] (1) Combination of active ingredients in American cockroach extract

[0010] Peptides: Contains a certain amount of peptides and their acetylated derivatives, which have anti-inflammatory and epithelial repair promoting functions.

[0011] Acidic polysaccharide complex: contains a certain amount of sulfated polysaccharides, which can inhibit fibrosis factors such as TGF-β1.

[0012] Nucleoside active substances: contain a certain amount of uracil, hypoxanthine and inosine, which play an antioxidant and microcirculation improving role.

[0013] (2) Nanoparticle formulation (weight ratio)

[0014] PAE: 10-50%;

[0015] Nano drug delivery systems (such as polymer nanocarriers, liposomes, nanoemulsions, etc.): 5-30%;

[0016] Antioxidant synergistic ingredients (such as sulfur-containing amino acids and polyphenols): 3-15%;

[0017] Mucosal protective agents (such as natural polysaccharide copolymers, etc.): 5-25%;

[0018] Pharmaceutical excipients (such as lyoprotectants, buffers, etc.): balance.

[0019] (III) Innovative preparation technology (including nano-steps, such as Figure 1 shown)

[0020] Ultra-high pressure and low temperature extraction: Blattidae insect bodies are taken and ultra-low temperature crushed to 100-300 mesh at -80°C. They are extracted 1-3 times (5-15 minutes each time) with 50-70% ethanol at 2-50 MPa pressure to retain the active ingredients and purify to obtain American cockroach extract (PAE). The PAE extract contains antioxidant synergistic components.

[0021] Preparation of nano-drug delivery system: PAE is combined with nano-carrier materials through emulsification-solvent evaporation method or ultrasonic encapsulation method. Among them, the preparation of polymer nanoparticles is as follows: PAE and natural or synthetic polymers (such as PLGA, PEG and their copolymers, etc.) are dissolved in dichloromethane, and polyvinyl alcohol aqueous solution is added dropwise. Ultrasonic emulsification is performed at 50W-300W for 5-30 minutes. After evaporation of the solvent, centrifugation is performed to collect the particles and control the particle size to 10-3000nm. Liposomes are prepared by dissolving PAE, phospholipids and cholesterol in chloroform in a ratio of 5:4:1. After film formation under reduced pressure, a hydration medium is added, and ultrasonic treatment at 5-30kHz is performed to form liposomes with a particle size of 10-3000nm. The resulting nano-drug delivery system can exert mucosal protective function.

[0022] Dosage form: nano atomizer (mist droplets 0.5-10μm account for ≥60%), oral nano microspheres, nano liposome injection or external nano gel preparation.

[0023] Dosage form preparation:

[0024] (1) Preparation of nano-atomizer

[0025] PAE is mixed with polymer nanoparticles (e.g., polylactic acid-co-glycolic acid copolymer PLGA-polyethylene glycol PEG copolymer nanoparticles) in a weight ratio of 2-5:1, and selenocysteine ​​(5%-10%, based on the weight of the extract) and a buffer (e.g., phosphate buffer, citric acid buffer) are added;

[0026] Ultrasonic emulsification (power 100-600W, time 5-15 minutes) forms a stable dispersion, and the particle size distribution (100-3000nm) is monitored by dynamic light scattering. The qualified nano-dispersion is filled into a special container for the nebulizer, and the droplet size is regulated by a laser particle size analyzer (0.5-10μm accounts for ≥60%).

[0027] (2) Preparation of oral nanoparticles

[0028] PAE and PLGA (molecular weight 5000-100000) are dissolved in an organic solvent (such as dichloromethane, ethyl acetate), and then dropped into an emulsifier aqueous solution (such as polyvinyl alcohol, povidone aqueous solution) for emulsification. After evaporation of the solvent, the microspheres are collected by centrifugation with a particle size of 3-50 μm.

[0029] The surface of the microspheres is coated with a natural polysaccharide nanolayer (such as chitosan, sodium alginate nanolayer, particle size 10-200nm) to enhance mucosal adhesion. They are made into oral capsules or granules and administered 1-2 times a day.

[0030] (III) Preparation of nanoliposome injection

[0031] PAE extraction: Take an appropriate amount of American cockroach bodies, add water and ultrasonically extract, precipitate with alcohol, purify by centrifugation and freeze-dry to obtain PAE extract;

[0032] Liposome preparation: Dissolve phospholipid:cholesterol (molar ratio) = 3-6:1 (can contain auxiliary lipids such as soybean lecithin, hydrogenated phospholipids) in an organic solvent (such as chloroform, ether), add PAE, evaporate under reduced pressure to form a film, add a hydrating medium (such as water for injection, normal saline), hydrate, and then sonicate (15-30kHz) to form nanoliposomes. The particle size is controlled to be 10-3000nm.

[0033] An osmotic pressure regulator (such as sodium chloride or glucose) was added to adjust the osmotic pressure to 280-300 mOsm / kg, and the solution was sterilized with a 0.22 μm filter membrane to prepare a PAE nanoliposome injection.

[0034] (IV) Nanogel preparations for external use

[0035] PAE is mixed with a nanostructured matrix material, which includes: nano-scale natural polymers (such as hyaluronic acid, sodium carboxymethyl cellulose, chitosan, with a particle size of 50-500nm), synthetic nano-hydrogel materials (such as polyethylene glycol (PEG)-polylactic acid (PLA) copolymers, polyethylene oxide nanogels); nano-carriers (such as nano-liposomes, nano-emulsions) can be further added and dispersed therein to enhance skin penetration efficiency;

[0036] Through cross-linking, swelling and other processes, it is prepared into local preparations such as nano hydrogels, nano gel patches or nano ointments, which are suitable for application or application on the body surface.

[0037] Application of preparations

[0038] (1) Scope of radiation damage treatment

[0039] Including but not limited to acute radiation sickness, local radiation damage, delayed radiation damage, and radiation-induced hematopoietic system damage.

[0040] The advantages of the present invention are as follows:

[0041] Nano-enhanced: Nanocarriers (such as polymeric nanoparticles, liposomes, nanoemulsions, etc.) can improve the enrichment efficiency of PAE in multiple organ tissues (such as lung, intestine, bone marrow, etc.), reduce systemic exposure, and increase the drug efficacy by 1.2-3 times compared with non-nano formulations.

[0042] Multi-path targeting: Pulmonary aerosolized nano-formulations can penetrate deep into the alveoli, oral nano-formulations are resistant to digestion fluid degradation, and injection nano-formulations can enhance tissue penetration, adapting to the protection needs of different injury sites.

[0043] Stability enhancement: Nano-embedding technology makes the activity loss of PAE less than 8% after 6-12 months of storage at room temperature, effectively solving the problem of easy oxidation and inactivation of natural extracts.

[0044] Low toxicity and safety: Nano-formulations can reduce the dosage (such as reducing the dosage by 30%-60% compared with traditional oral administration), have no obvious hemolytic reaction and abnormal liver and kidney function, and have good safety. BRIEF DESCRIPTION OF DRAWINGS

[0045] Figure 1 The preparation method flowchart of the present application.

[0046] Figure 2 The scavenging effect diagram of PAE of the present application on 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radicals.

[0047] Figure 3 The scavenging effect diagram of PAE of the present application on superoxide free radicals (O2 - ).

[0048] Figure 4 The scavenging effect diagram of PAE of the present application on TA (OH - ) free radicals under irradiation.

[0049] Figure 5 The scavenging effect diagram of PAE of the present application on DCF (ROS) free radicals under irradiation.

[0050] Figure 6 The HE staining of PAE of the present application in the lung tissues of mice at 7, 14, 28, and 56 days.

[0051] Figure 7 The inflammatory indicators of PAE of the present application in the lavage fluid and serum of mice at 7, 14, 28, and 56 days;

[0052] (a) TNF-α in serum, (b) IL-1β in serum, (c) IL-6 in serum, (d) TNF-α in lung lavage fluid, (e) IL-1β in lung lavage fluid, and (f) IL-6 in lung lavage fluid.

[0053] Figure 8 This is a picture of TGF and Masson staining pathological analysis of the lungs of mice with PAE on day 56 of the present invention;

[0054] (a) TGF-β staining section of the lung on day 56, (b) Masson staining section of the lung on day 56, (c) statistical graph of TGF-β staining, (d) statistical graph of Masson staining. DETAILED DESCRIPTION

[0055] The present invention will be described in further detail below with reference to the accompanying drawings. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0056] The present invention proposes a novel use of an American cockroach extract as a free radical scavenger and radiation protectant. To further clarify the advantages and technical solutions of the present invention, the present invention is described in detail below with reference to specific examples. These examples are intended only to facilitate understanding of the present invention and should not be construed as limiting the present invention.

[0057] 1. The raw materials required for the present invention can be purchased through commercial channels.

[0058] Example 1: Preparation of PAE nano-atomizer

[0059] Raw materials: PAE 5g, polymer nanocarrier (such as PLGA-PEG copolymer) 3g, selenocysteine ​​0.3-0.5g,

[0060] Add an appropriate amount of buffer (such as phosphate buffer at pH 6.5-7.0) and water for injection to 50 mL;

[0061] step:

[0062] Dissolve PAE and polymer nanocarriers in an organic solvent (such as dichloromethane), add 3%-7% polyvinyl alcohol aqueous solution, perform ultrasonic emulsification at 200-400W for 3-8 minutes, and collect 100-500nm nanoparticles by centrifugation;

[0063] Add selenocysteine ​​and buffer solution, and sonicate at 400-600W for 8-15 minutes to form a dispersion;

[0064] Freeze-dry (-30 to -50°C, 5-15Pa) to obtain freeze-dried powder, and after reconstitution, the proportion of droplets 0.5-10μm is ≥80%.

[0065] The nanoparticles obtained in the steps were characterized by transmission electron microscopy and Fourier transform infrared spectroscopy. The results showed that the average particle size of the nanoparticles prepared in this example was 50-300 nm; the infrared spectrum showed that they contained a large number of groups with free radical scavenging effects.

[0066] Example 2: Preparation of PAE Oral Sustained-Release Microspheres

[0067] Preparation of microspheres: PAE and PLGA (molecular weight 5000-100000, refer to CN 112791071 B) are dissolved in an organic solvent (such as dichloromethane), dropped into an emulsifier aqueous solution (such as polyvinyl alcohol aqueous solution) for emulsification, and after evaporation of the solvent, the microspheres are collected by centrifugation. The particle size is 3-20 μm.

[0068] Preparation molding: The microspheres are mixed with a freeze-drying protectant (such as mannitol) and made into oral capsules or granules for administration 1-2 times a day.

[0069] Example 3: PAE topical nanogel preparation for the treatment of radiation-induced skin damage

[0070] Preparation of nanogel: PAE is mixed with a matrix material (such as hyaluronic acid, carbomer, polyethylene glycol hydrogel), and nanocarriers (such as nanoliposomes, nanoemulsions, particle size 50-500nm) are added and dispersed evenly; the pH is adjusted to 5.0-7.0 with triethanolamine, and ultrasonic treatment is performed at 20-30kHz for 10-20 minutes to form a gel with a nano-network structure; a preservative (such as benzalkonium chloride, potassium sorbate) is added and stirred until uniform; filtration is performed through a 0.45μm filter membrane and the mixture is dispensed into sterile containers to obtain a PAE topical nanogel preparation.

[0071] Example 4: Preparation of PAE nanoliposome injection

[0072] PAE extraction: Take an appropriate amount of American cockroach body, add water and ultrasonic extraction, precipitate with alcohol, purify by centrifugation and then freeze-dry to obtain PAE extract.

[0073] Preparation of liposomes: Dissolve phospholipids and cholesterol in an organic solvent (such as chloroform) at a ratio of 3-6:1 (can contain auxiliary lipids such as soybean lecithin), add PAE, evaporate under reduced pressure to form a film, add a hydrating medium (such as water for injection) and hydrate, then sonicate to form nanoliposomes, with a particle size controlled at 50-500 nm.

[0074] Preparation molding: Add an osmotic pressure regulator (such as sodium chloride) to adjust the osmotic pressure to 280-300mOsm / kg, sterilize through a 0.22μm filter membrane, and prepare PAE nanoliposome injection.

[0075] It is worth noting that the radiation used below is all based on soft X-ray models, but the radiation protection targeted by the present invention is not limited to the soft X-ray model. Other radiation models that can cause free radical damage to the human body, such as β rays, and even ionizing radiation with shorter wavelengths, such as ultraviolet rays, all fall within the scope of protection and disclosure of the present invention.

[0076] 2. The following specific experiments are used to illustrate the application effect of PAE in free radical scavenging.

[0077] (1) Study on the scavenging effect of PAE on DPPH free radicals

[0078] DPPH is a classic in vitro free radical model with a characteristic absorption peak at 517nm. After reacting with free radical scavengers, the absorption value of the characteristic absorption peak will decrease, which is used to evaluate the scavenging ability of PAE on DPPH free radicals. Figure 2 , Figure 2 The figure shows the scavenging effect of PAE on DPPH free radicals, where the concentration of DPPH free radicals is 100 μM. The figure shows that PAE has a good scavenging effect on DPPH free radicals, and this effect is concentration-dependent. The higher the PAE concentration, the stronger the scavenging ability of DPPH free radicals.

[0079] (2) PAE to O2 - Study on the effect of clearance

[0080] O2 - It is another important free radical that causes cell damage. Similarly, the evaluation material's resistance to O2 - The scavenging ability of O2 is of great significance for protecting the body. - , the generated O2 - It can react with nitro blue tetrazolium chloride (NBT) to form a product with characteristic absorption at 560nm. - If the scavenger is added, the degree of oxidation will be reduced and the generated products will be reduced, which is reflected in the lower absorbance at 560nm. Figure 3 , Figure 3 O2 produced by PAE to non-enzymatic system - Clear effect, by Figure 3 It can be seen that when there is no PAE in the reaction system, the absorption value is the largest, indicating that O2 - The maximum amount of O2 was generated. When 25 μg / mL PAE was present in the reaction system, the absorption value decreased, indicating that O2 - The amount of O2 generated was reduced. When 200 μg / mL of PAE was present in the reaction system, the absorption value was even lower, indicating that O2 - The amount of O2 generated is even lower, proving that PAE can remove O2 - , and the effect is concentration-dependent. The higher the material concentration, the better the effect on O2 - The stronger the cleaning ability.

[0081] (3) Study on the scavenging effect of PAE on ·OH produced by enzyme system

[0082] OH -Free radicals are an important type of free radical that causes DNA damage in cells, so the evaluation of the material's OH - The scavenging ability of PAE is of great significance for resisting ROS-induced damage to the body. The present invention uses terephthalic acid (TA) method to detect the OH - The scavenging effect. - Produced by radiation irradiation of water, the generated OH - It will generate fluorescent products with TA. The generated fluorescent substance has a fluorescence absorption peak at 435nm. If OH is added to the reaction system - When the scavenger is added, the fluorescence intensity decreases, which is reflected by the decrease of the absorbance value at 435nm.

[0083] See also Figure 4 , Figure 4 PAE to OH - The scavenging effect is shown in the figure. When there is no PAE in the reaction system, the absorption curve of the obtained product has the largest value at 435nm. When there is 2μg / mLPAE in the reaction system, the absorption curve of the obtained product at 435nm decreases. When there is 10μg / mLPAE in the reaction system, the absorption curve of the obtained product at 435nm decreases more, which proves that PAE can scavenge OH - , and the effect is concentration-dependent. The higher the material concentration, the greater the effect on OH - The stronger the cleaning ability.

[0084] (4) PAE free radical scavenging test

[0085] In this example, the free radical scavenging activity of PAE was determined using the following method: a DCFH-DA (2',7'-dichlorofluorescein diacetate) probe assay was used. The specific experimental steps were as follows: 0.01 mol / L NaOH (aq) and phosphate buffer solution (pH = 7.2, prepared using Na2HPO4·12H2O and NaH2PO4·2H2O) were prepared. 12.5 μL of DCFH-DA (10 mM) was dissolved in 0.5 mL of DMSO, followed by the addition of 2 mL of NaOH (0.01 mol / L). The reaction was incubated in the dark for 30 minutes, and then 10 mL of phosphate buffer solution was added to terminate the reaction. This resulted in a DCFH working solution, which was stored on ice in the dark. The test was then divided into four groups: DCFH + ultrapure water + X-ray group (0.1 mL DCFH working solution + 2.9 mL ultrapure water, X-ray irradiation for 10 minutes), DCFH + PAE + X-ray group (0.1 mL DCFH working solution + PAE (2.9 mL, 1 mg / mL), X-ray irradiation for 10 minutes, and fluorescence detection was performed on a fluorescence spectrometer with an excitation wavelength of 488 nm. (X-ray generating instrument: X-ray tube (50 kV, 75 μA), manufacturer: AMPTEK Inc.; fluorescence spectrometer manufacturer: Horiba FluoroLog-3).

[0086] Test principle: Water is a good absorber of soft X-rays. When soft X-rays interact with water molecules, they cause radiolysis of the water molecules, thereby generating a large number of free radicals. These free radicals can oxidize non-fluorescent DCFH (2',7'-dichlorofluorescein diacetate) to generate fluorescent DCF (2',7'-dichlorofluorescein). By detecting the fluorescence intensity of the generated DCF, the generation of free radicals can be determined. Finally, the free radical scavenging test results of the PAE prepared in Example 1 are obtained, as shown in the figure. Figure 5 shown. Figure 5 This reflects the scavenging effect of PAE on free radicals based on the DCFH fluorescence probe method. The fluorescence signal generated by the DCFH+ultrapure water+X-ray group is the strongest, while when PAE is present (i.e., the DCFH+PAE+X-ray group), the fluorescence signal is significantly reduced, indicating that PAE has good scavenging activity on free radicals.

[0087] The above-mentioned free radical scavenging experimental results show that the PAE described in the present invention can effectively scavenge free radicals and therefore has the potential to be used as a radiation protection agent.

[0088] 3. The present invention also provides the application of the above-mentioned PAE in radiation protection.

[0089] Because high-energy radiation in radiotherapy can produce a large amount of reactive oxygen species (ROS, such as OH -、H2O2、O2 - These ROS can attack DNA, proteins, and cell membranes, causing DNA breakage, protein denaturation, and lipid peroxidation, ultimately leading to cell apoptosis or necrosis and radiation damage to multiple organs. The aforementioned experiments have demonstrated the ability of PAEs (especially nanoformulations) to efficiently scavenge ROS. Based on this, the present invention further applies them to radiation protection. Using nanotechnology, the present invention achieves a balance between targeting and low toxicity, addressing the poor targeting and significant side effects of existing protective agents (such as amifostine).

[0090] This invention is the first to systematically apply PAE (particularly nanoformulations) to multi-organ radiation damage protection, expanding the biomedical application of PAE and providing an innovative approach of "natural ingredients + nanotechnology" for the development of radiation protectants. The following specific experiments further validate its effectiveness:

[0091] Research on PAE's radiation protection of the lungs:

[0092] To evaluate the radiation protective effect of PAE on the lungs, first, 64 C57 female mice were purchased from Spectrum Biotechnology Co., Ltd. (Beijing) and randomly divided into four groups (16 mice in each group): (i) PBS buffer group (control group); (ii) PAE group; (iii) X-ray irradiation alone group (X-ray group); (iv) PAE and X-ray irradiation group (PAE+X-ray group). Mice in groups (i) and (ii) were treated with nebulized PBS or PAE, while mice in groups (iii) and (iv) were irradiated with X-ray (16 Gy). After 7, 14, 28, and 56 days, the mice were killed and their lung tissues were removed for analysis.

[0093] Figure 6 The following is an H&E staining image showing the protective effect of PAE on the lung tissue of irradiated mice. H&E staining analysis was performed on the lung tissues of mice in four groups: the control group, the PAE group, the X-ray irradiation group alone, and the PAE and X-ray irradiation group. As can be seen from the figure, for the control and PAE groups, the lung structure was generally normal, the glands were regularly arranged and morphologically regular, the cells had no obvious atypia, and a few scattered neutrophil infiltrations. For the X-ray irradiation group alone, the lung structure was disordered, with inflammatory cell infiltration, structural damage, and partial swelling and degeneration of cells. For the PAE and X-ray irradiation group, the lung structure was relatively normal, with no inflammatory cell infiltration, and the cell morphology was generally normal. This shows that compared with the control group, the X-ray group had the most severe lesions, and some lung tissues in the PAE+X-ray group had shown significant repair and were close to normal. This shows that PAE can protect against radiation-induced lung damage.

[0094] Figure 7This is an ELISA of PAE for inflammatory indicators in the lung lavage fluid and serum of irradiated mice. Lavage fluid and serum were collected from mice in the control group, PAE group, X-ray irradiation group, and PAE and X-ray irradiation group on days 7, 14, 28, and 56, respectively. As shown in the figure, there was no significant difference in inflammatory indicators in the lung lavage fluid and serum between the control and PAE groups. In the X-ray irradiation group, inflammatory indicators in the lung lavage fluid and serum were significantly increased. In the PAE and X-ray irradiation group, inflammatory indicators in the lung lavage fluid and serum were significantly decreased compared to the X-ray irradiation group. This demonstrates that PAE can alleviate the inflammatory response caused by subsequent radiation damage.

[0095] Figure 8 The images show the protective effect of PAE on the lung tissue of irradiated mice using TGF-β and Masson staining. TGF-β and Masson staining were performed on the lung tissues of mice in the control, PAE, X-ray irradiation alone, and PAE and X-ray irradiation groups. As can be seen from the figures, there was no obvious collagen deposition in the lung tissues of the control and PAE groups. In the X-ray irradiation alone group, TGF-β staining in the lung tissue was diffuse and severe. Masson staining was significantly enlarged and severely deposited. In the PAE and X-ray irradiation group, TGF-β staining in the lungs was mild, with no obvious TGF-β markers. Masson staining was mild, with no obvious collagen deposition.

[0096] The above radiation protection studies indicate that the PAE of the present invention can effectively protect lung damage caused by radiation, and therefore is expected to be used as a new type of radiation protection agent.

[0097] The present invention provides a nanoformulation system based on American cockroach extract (PAE). Its preparation process combines high-pressure and low-temperature extraction with nanocarrier technology. It is not only simple to operate and cost-controlled, but also uses aqueous extraction and biocompatible carriers (such as PLGA and chitosan) throughout the process, which conforms to the concept of green environmental protection. This system achieves the effect of OH by the precise combination of active ingredients (peptides, polysaccharides, nucleosides) and the synergy of nano-delivery technology. - 、H2O2、O2 - It can efficiently remove various free radicals, providing an innovative paradigm of "natural ingredients + nano-enhancement" for the development of new free radical scavengers.

[0098] Meanwhile, the present application breaks through the protection limitation of a single organ (such as skin) in the prior art, and through multi-dosage form designs such as lung-targeted nano-aerosol preparations, intestinal-targeted nano-oral preparations, and systemic delivery nano-liposomes, can effectively resist radiation-induced lung damage (radiation pneumonitis, pulmonary fibrosis), intestinal barrier damage, hematopoietic system inhibition, and other multi-organ damage, especially in reducing the incidence of radiation pneumonitis, promoting intestinal mucosa repair, and improving bone marrow hematopoietic function, and provides a new solution covering the "acute-subacute-chronic" whole cycle and "local-systemic" multiple scenarios for the development of radiation protection agents.

[0099] The applicant declares that the present application realizes multi-organ targeted radiation protection of PAE through a nano delivery system, and the core innovation point lies in the synergistic design of "PAE active ingredients + nano carrier". Routine adjustment of the type of nano carrier, targeting ligand, and dosage form parameters all fall within the protection scope of the present application.

[0100] The core value of the present application lies in combining the natural radiation tolerance characteristics of Periplaneta americana with the targeted delivery advantages of nanotechnology, and providing a new strategy with effectiveness, safety and practicality for the prevention and treatment of radiation damage, and the clinical conversion potential of which is not limited to tumor radiotherapy patients, but can also be extended to nuclear emergency, radiation occupational protection and other fields, and has wide social and economic value.

[0101] Although specific embodiments of the present application are disclosed for the purpose of illustrating the present application, the purpose is to help understand the content of the present application and to implement the same, and those skilled in the art can understand that various substitutions, changes and modifications are possible without departing from the spirit and scope of the present application and the appended claims. Therefore, the present application should not be limited to the disclosed content of the best mode, and the scope of the present application claimed is defined by the scope of the claims.

Claims

1. An American cockroach extract preparation, characterized in that: The invention comprises an American cockroach extract, a nano drug delivery system, an antioxidant synergistic component, a mucosal protective agent and pharmaceutical excipients; wherein the weight ratio of the American cockroach extract is 10-50%, the weight ratio of the nano drug delivery system is 5-30%, the weight ratio of the antioxidant synergistic component is 3-15%, the weight ratio of the mucosal protective agent is 5-25%, and the balance is pharmaceutical excipients.

2. The American cockroach extract preparation according to claim 1, characterized in that The nano drug delivery system is a polymer nano carrier, liposome or nanoemulsion.

3. The American cockroach extract preparation according to claim 2, characterized in that The substrate of the polymer nanocarrier is a natural or synthetic polymer, and the particle size is 10-3000 nm.

4. The American cockroach extract preparation according to claim 1, 2 or 3, characterized in that: The antioxidant synergistic component is a compound of amino acids and polyphenols; the mucosal protective agent is a natural polysaccharide copolymer; and the pharmaceutical excipient is a freeze-drying protective agent or a buffer.

5. The American cockroach extract preparation according to claim 1, characterized in that The American cockroach extract preparation is a nano atomizer, an oral nano microsphere, a nano liposome injection or an external nano gel preparation.

6. The American cockroach extract preparation according to claim 1, characterized in that The active ingredients of the American cockroach extract include polypeptides and their acetylated derivatives, sulfated polysaccharides, uracil, hypoxanthine and inosine.

7. A method for preparing the American cockroach extract preparation according to claim 1, comprising the steps of: 1) Ultra-high pressure and low temperature extraction of American cockroach extract; 2) combining the American cockroach extract with a nanocarrier material through an emulsification-solvent evaporation method or an ultrasonic encapsulation method to obtain a nano drug delivery system; 3) Forming the dosage form to obtain the American cockroach extract preparation.

8. The method according to claim 7, characterized in that The American cockroach extract preparation is a nano atomizer, an oral nano microsphere, a nano liposome injection or an external nano gel preparation; wherein, The preparation method of the nano atomizer comprises: mixing the American cockroach extract and polymer nanoparticles in a weight ratio of 2-5:1, adding selenocysteine ​​and a buffer; then ultrasonically emulsifying to form a stable dispersion, filling the dispersion into a dedicated atomizer container, and controlling the droplet size of 0.5-10 μm to account for ≥60% by a laser particle size analyzer to obtain the nano atomizer; The preparation method of the oral nano-microspheres is as follows: dissolving the American cockroach extract and PLGA in an organic solvent, dripping into an emulsifier aqueous solution for emulsification, volatilizing the solvent, and then centrifuging to collect the microspheres, preparing oral capsules or granules to obtain oral nano-microspheres; The preparation method of the nanoliposome injection comprises: dissolving phospholipid and cholesterol in an organic solvent at a molar ratio of 3-6:1, adding the American cockroach extract, and then evaporating under reduced pressure to form a film, adding a hydrating medium for hydration, and then ultrasonically forming nanoliposomes, with the particle size controlled at 10-500 nm; and then adding an osmotic pressure regulator to adjust the osmotic pressure to 280-300 mOsm / kg to prepare the nanoliposome injection. The preparation method of the external nanogel preparation comprises the following steps: mixing the American cockroach extract with a nanometer matrix material, and then adding a nanocarrier; and then preparing the nanohydrogel, nanogel patch or nanopaint through a crosslinking or swelling process to obtain the external nanogel preparation.

9. Use of the American cockroach extract preparation according to claim 1 in preventing and treating radiation damage.

10. The use according to claim 9, characterized in that The radiation damage includes but is not limited to acute radiation sickness, local radiation damage, delayed radiation damage, and radiation-induced hematopoietic system damage.

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