Application of compound sophora flavescens extract in treatment or prevention of radioactive intestinal injury

Abstract

The invention relates to the field of medicines, and particularly provides a compound sophora flavescens extract and application thereof in preparation of medicines for preventing or treating radioactive intestinal injury. The compound sophora flavescens extract effectively improves radiation enteritis and symptoms such as diarrhea and hematochezia caused by radiation enteritis, and provides a medicine choice for treatment of radiation enteritis.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to application of a compound sophora flavescens extract in preparing a medicine for treating or preventing radiation-induced intestinal damage. Background Art

[0002] Radiation therapy often causes severe side effects for cancer patients. Adverse reactions caused by radiation damage not only limit the therapeutic dose of radiotherapy but also impact patients' quality of life. Gastrointestinal mucositis (GIM), or damage to the gastrointestinal mucosa, is a major side effect for patients with tumors in the abdominal or pelvic regions, and there is currently no effective treatment.

[0003] Although the molecular and cellular mechanisms underlying radiotherapy-induced mucosal damage are not fully understood, radiotherapy induces cell death in both tumors and healthy tissues. Gastrointestinal epithelial cells are highly proliferative and are therefore more susceptible to damage, leading to mucositis. The pathogenesis of mucositis begins with radiation-induced production of reactive oxygen species (ROS), which trigger mucositis by causing DNA damage and epithelial cell apoptosis. Simultaneously, several transcription factors (such as NF-κB) are activated and upregulated. A group of genes targeted by NF-κB are proinflammatory cytokines. Multiple studies have demonstrated that mRNA and / or protein levels of cytokines (such as IL-1b, IL-6, and tumor necrosis factor (TNF-α)) are upregulated in the gastrointestinal mucosa of irradiated animals. Once proinflammatory cytokines are upregulated, they provide positive feedback by activating NF-κB in other cells, leading to further activation of inflammatory pathways. Furthermore, other studies have also provided evidence that upregulation of these inflammatory cytokines can lead to the activation of innate proinflammatory leukocytes, which produce more tissue-damaging enzymes, leading to expanded tissue damage and ultimately, the progression of mucositis to the ulcerous stage. The final stage is healing, but the mucosal changes caused by mucositis are long-lasting and increase the risk of subsequent injury from radiation exposure. Although significant progress has been made in mucositis research over the past decade using novel animal models, which have provided some clues to the pathology of mucositis, effective treatments are limited, and there is an urgent need for therapeutics that can inhibit one or more of these pathological pathways.

[0004] Radiation enteritis is intestinal damage caused by radiation therapy for pelvic, abdominal, or retroperitoneal malignancies. It can affect the small intestine, colon, and rectum, hence the name "radiation rectum, colon, and enteritis." During treatment, while radioactive elements inhibit cancer, they also damage normal tissues. Damage to the intestinal mucosa can cause radiation enteritis. Symptoms include abdominal pain, diarrhea, bloody stools, and mucus, pus, and blood in the stool, severely impacting patients' quality of life.

[0005] Radiation enteritis currently lacks a standardized treatment strategy and process. While many treatments have demonstrated some effectiveness, these have mostly been based on small, single-center, non-double-blind studies, and few treatments are supported by high-quality research evidence. Current clinical treatment outcomes for chronic radiation enteritis are generally poor.

[0006] Compound Sophora flavescens extract is composed of Sophora flavescens and Poria cocos. Sophora flavescens, the main herb in the formula, is bitter in taste and cold in nature. It enters the liver, kidney, large intestine, and small intestine meridians, clearing heat, drying dampness, killing insects, and acting as a diuretic. Sophora flavescens was first mentioned in the Shennong Bencao Jing, which states that it primarily treats "heart and abdominal qi stagnation, accumulation of masses, and carbuncles." These "heart and abdominal qi stagnation" and "accumulation of masses" are similar to modern references to lung and chest tumors. The Yaoxing Lun (Treatise on the Properties of Materia Medica) explicitly states that Sophora flavescens can "treat heat-toxic wind and accumulation of heart and abdominal masses." Similar references are also found in the Diannan Materia Medica, which states that Sophora flavescens can "relieve heat-toxic wind and eliminate swelling-toxic and phlegm-toxic." Modern research has also shown that Sophora flavescens can not only significantly reduce the expression of TGF-β and TNF-α, but also increase T lymphocyte counts, stabilize the CD4+ / CD8+ ratio, enhance immunity, and effectively reduce ROS generated by radiation.

[0007] Compound Kushen Injection (CKI) was launched in 1995. Its primary functions include clearing heat and dampness, cooling blood and detoxifying, and dispersing nodules and relieving pain. It is used to treat cancer pain and bleeding. The formula consists of two Chinese herbs, Kushen and Bai Tuling. The main herb, Kushen, is bitter and cold in nature, entering the liver, kidney, large intestine, and small intestine meridians to clear heat and dissipate dampness. The auxiliary herb, Bai Tuling, is sweet and mild in taste and neutral in nature, entering the liver, spleen, and bladder meridians to invigorate the spleen, eliminate dampness, dispel wind, and detoxify. The combination of the two herbs has the effects of clearing heat and dampness, cooling blood and detoxifying, and dispersing nodules and relieving pain. Bai Tuling is neutral in nature and sweet and mild in taste, and has the function of detoxifying and removing dampness. Currently, there is no consensus in Traditional Chinese Medicine (TCM) on the classification of RP syndromes. Traditional Chinese Medicine (TCM) considers the lungs to be a delicate organ, and they also govern the skin and fur. Radiation-induced lung injury (RLI) is a pulmonary disease in TCM, and can be categorized as a cough, pulmonary distension, and pulmonary atrophy. Its pathogenesis is primarily related to pathogenic heat and fire toxicity, and its pathological features often demonstrate the presence of intense heat toxicity. Furthermore, intense heat toxicity can damage the lung meridian, leading to stasis in the lung meridian. Therefore, its pathogenesis is primarily characterized by intense heat toxicity, combined with stasis in the lung meridian. Ancient texts provide strong TCM theoretical support for the treatment of RP with a combination of Sophora flavescens and Poria cocos. Based on this analysis, the efficacy of Compound Sophora flavescens Injection, with its heat-clearing and dampness-removing, blood-cooling and detoxifying, and pain-relieving properties, should be the primary treatment for RP.

[0008] Modern medical research indicates that the development of RP is associated with the massive production of cytokines and reactive oxygen free radicals. The cytokines that have been extensively studied are primarily transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α). TGF-β can attract inflammatory cells and fibroblasts, thereby promoting the formation of fibrosis; TNF-α is a pro-inflammatory factor that participates in the development of inflammatory reactions along with other inflammatory factors. Reactive oxygen free radicals produced by lung tissue exposed to ionizing radiation can cause cell damage, alveolar wall edema, and increased vascular permeability. Proteins that infiltrate the alveolar cavity further reduce alveolar septum and vascular integrity, leading to apoptosis of type I alveolar cells and alveolar re-epithelialization driven by type II alveolar epithelial cells, reducing microvascular density and oxygen perfusion, ultimately leading to radiation-induced lung damage.

[0009] Studies have shown that compound sophora flavescens injection can not only significantly reduce the expression of TGF-β and TNF-α, but also increase the number of T lymphocytes, stabilize CD4+ / CD8+, and improve the body's immunity[5]. In addition, compound sophora flavescens injection can effectively reduce the ROS generated by radiation[6]; improve pulmonary fibrosis in mice with radiation-induced lung injury model, and its mechanism may be related to inhibiting the expression of TGF-β1, Smad3, GSK-3β and β-catenin mRNA and channel proteins related to epithelial-mesenchymal transition (EMT), thereby promoting E-cadheren expression, inhibiting Vimentin protein expression, and ultimately inhibiting the occurrence of EMT. In addition, compound sophora flavescens injection can significantly downregulate the lung coefficient and Notch1 and Jagged1 expression in rats with radiation-induced pneumonia, and the activation of the Notch / Jagged signaling pathway is closely related to the occurrence and development of radiation-induced pneumonia.

[0010] So far, there are no reports on the use of compound Sophora flavescens extract in the treatment or prevention of radiation-induced intestinal injury. Summary of the Invention

[0011] In view of the above technical status, the present invention provides an application of a compound Sophora flavescens extract in the preparation of a medicament for treating or preventing radiation-induced intestinal injury, wherein the compound Sophora flavescens extract comprises Sophora flavescens and Poria cocos.

[0012] In the present invention, as one of the implementation schemes, the mass ratio of Sophora flavescens to White Tuling is 7:3.

[0013] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0014] Weigh the prescribed amount of the medicinal materials Sophora flavescens and Poria cocos, add 8 to 10 times the volume of 50% to 90%, preferably 60% ethanol, respectively, and reflux and extract 1 to 3 times, preferably 2 times. The extracts are filtered, combined, and concentrated; the concentrate is transferred to a water bath and steamed to a thick extract, and the thick extract is placed in a vacuum drying oven and dried under reduced pressure at 60°C to 80°C, preferably 70°C.

[0015] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0016] Weigh the prescribed amount of medicinal materials Sophora flavescens and Poria cocos, add 10 times and 8 times the amount of 60% ethanol respectively, reflux and extract twice, filter the extracts, combine, and concentrate; transfer the concentrate to a water bath and steam it to a thick extract, place the thick extract in a vacuum drying oven, and dry it under reduced pressure at 70°C to obtain the product.

[0017] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0018] Weigh the prescribed amount of medicinal materials Sophora flavescens and Phellodendron chinense (7:3), add 10 times and 8 times the amount of 60% ethanol respectively, and reflux extract twice, each extraction for 1 hour. Filter the extracts, combine, and concentrate; transfer the concentrate to a water bath and steam it to a thick extract, place the thick extract in a vacuum drying oven, and dry it under reduced pressure at 70°C to obtain the product.

[0019] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0020] Weigh the prescribed amount of Sophora flavescens and Poria cocos (7:3), add 8-10 times the amount of water, reflux extract, filter, combine, and concentrate the extracts. Slowly add 50% to 90% (preferably 60%) ethanol to the concentrate, stirring until the alcohol content reaches 60%. Let it stand and refrigerate overnight. Filter the supernatant of the remaining solution, concentrate the filtrate, and evaporate it to dryness in a water bath to a thick extract. Place the thick extract in a vacuum drying oven at 70°C.

[0021] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0022] Take the prescribed amount of medicinal materials Sophora flavescens and Poria cocos (7:3), add 10 times and 8 times the amount of water respectively, reflux extraction, filter the extracts, combine, and concentrate; slowly add ethanol to the concentrate while stirring to make the alcohol content of the medicinal solution reach 60%, and let it stand and refrigerate overnight; take the supernatant of the medicinal solution after standing, filter, concentrate the filtrate, place it in a water bath and evaporate to a thick extract, place the thick extract in a vacuum drying oven, and dry it under reduced pressure at 70°C.

[0023] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0024] Take the prescribed amount of medicinal materials Sophora flavescens: Poria cocos (7:3), add 10 times and 8 times the amount of water respectively, reflux extraction twice, each extraction is 1 hour, the extracts are filtered, combined, and concentrated; slowly add ethanol to the concentrate while stirring to make the alcohol content of the medicinal solution reach 60%, and let it stand and refrigerate overnight; take the supernatant of the medicinal solution after standing, filter, concentrate the filtrate and place it in a water bath to evaporate to a thick extract, place the thick extract in a vacuum drying oven, and dry it under reduced pressure at 70℃.

[0025] Another aspect of the present invention provides a use of a preparation containing the compound Sophora flavescens extract in preparing a medicine for treating or preventing radiation-induced intestinal damage.

[0026] In the present invention, as one embodiment, the preparation is a tablet, capsule, oral preparation, liquid injection or injection.

[0027] In a further aspect, the present invention provides a compound Sophora flavescens extract for treating or preventing radiation-induced intestinal damage, wherein the compound Sophora flavescens extract consists of Sophora flavescens and Psoralea corylifolia.

[0028] In the present invention, as one of the embodiments, the mass ratio of Sophora flavescens to Psoralea corylifolia in the compound Sophora flavescens extract is 7:3.

[0029] In the present invention, as one of the embodiments, the compound Sophora flavescens extract is prepared by the following method:

[0030] Weigh the prescribed amount of Sophora flavescens and Poria cocos according to the ratio, add 10 times and 8 times the amount of 60% ethanol respectively, reflux and extract twice, each extraction for 1 hour, filter the extract, combine, and concentrate. Transfer the concentrate to a water bath and steam until it becomes a thick extract. Place the thick extract in a vacuum drying oven and dry it at 70℃ under reduced pressure to obtain; or

[0031] Weigh the prescribed amount of Sophora flavescens and Poria cocos according to the prescribed ratio. Add 10-fold and 8-fold amounts of water, respectively, and reflux extract twice, each time for 1 hour. Filter the extracts, combine, and concentrate. Slowly add ethanol to the concentrate while stirring until the alcohol content reaches 60%. Let it stand and refrigerate overnight. Filter the supernatant of the remaining solution. Concentrate the filtrate and evaporate it to dryness in a water bath to a thick extract. Place the thick extract in a vacuum drying oven at 70°C.

[0032] The compound Sophora flavescens extract of the present invention differs substantially from the compound Sophora flavescens injection in their composition. The compound Sophora flavescens extract of the present invention is prepared by alcohol extraction, while the compound Sophora flavescens injection is prepared by a series of treatments, including percolation with 0.1% acetic acid followed by water decoction and alcohol precipitation. The compound Sophora flavescens extract and the compound Sophora flavescens injection differ in their production processes and composition. The compound Sophora flavescens extract of the present invention contains a high concentration of flavonoids, while the compound Sophora flavescens injection contains virtually no flavonoids.

[0033] In the present invention, the mechanism of radiation-induced intestinal injury is essentially different from that of lung injury: First, the mechanism of radiation-induced intestinal injury: histological changes can occur within a few hours after radiation acts on the intestine. Early manifestations include epithelial cell apoptosis, lamina propria inflammation, and crypt abscesses. Later changes include vasculitis, small vessel ischemia, submucosal fibrosis, intestinal wall thickening, etc. The subsequent pathological development process follows two typical development paths: one is mucosal ulcers, perforations, fistulas, and intra-abdominal abscesses, and the other is intestinal wall fibrosis, stenosis, and intestinal obstruction. The mechanism of radiation-induced lung injury: the effect of radiation on body tissues is mainly manifested in direct damage to DNA and indirect damage through ionization of water molecules leading to DNA breakage. Cells in the G2 / M phase are more sensitive to damage. Radiation acts on lung tissue, manifesting as damage to alveolar epithelial cells and endothelial cells, causing destruction of the alveolar mucosal barrier, thereby generating a series of inflammatory cascade reactions. Within a few days or weeks, inflammatory cells aggregate, vascular permeability increases, inflammatory cytokines are released, macrophages are activated and aggregated, causing local hypoxia, leading to the generation and release of reactive oxygen and reactive nitrogen species, producing pro-inflammatory, pro-fibrotic, and pro-angiogenic effects, resulting in prolonged lung tissue damage and the formation of chronic radiation-induced lung injury.

[0034] Secondly, the clinical manifestations of radiation-induced intestinal injury encompass virtually all possible gastrointestinal symptoms and signs. The main clinical manifestations of chronic radiation-induced intestinal injury include intestinal obstruction, intestinal fistula, diarrhea, gastrointestinal bleeding, anemia, and malnutrition. The clinical symptoms of radiation pneumonitis are nonspecific, generally presenting with cough, shortness of breath, and fever. Cough is the most common, an irritating, dry cough; shortness of breath is second, with varying degrees of severity, ranging from mild cases only occurring after exertion to severe cases with significant dyspnea even at rest. Approximately 50% of patients experience fever, often preceding the onset of symptoms such as cough and shortness of breath, with temperatures ranging from 37.0 to 38.5°C. High fever may also occur with concurrent infection. Signs of radiation pneumonitis are nonspecific, but some patients may present with coarse, dry, or damp breath sounds, decreased breath sounds, and a pleural friction rub.

[0035] Finally, there are differences in the animal modeling. In radiation pneumonitis studies, irradiation is applied to the lungs, while in the present method, irradiation is applied to the intestines. Irradiation to the lungs primarily targets radiation-induced pneumonitis in chest tumors, while irradiation to the intestines primarily targets radiation-induced intestinal damage in patients undergoing radiotherapy for abdominal tumors.

[0036] The present invention provides a new drug option for the treatment of radiation enteritis, and effectively improves symptoms such as diarrhea and bloody stools caused by radiation enteritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1: In Experimental Example 1, the effect of alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The rat abdomen was irradiated 5 times, each time with 4Gy. The rats were orally administered with alcohol-extracted compound Sophora flavescens preparation (A-CKP) at three different doses from day 0 to day 6 after irradiation. (4A) Body weight changes of irradiated rats administered with alcohol-extracted compound Sophora flavescens oral preparation. (4B) Changes in food intake of irradiated rats administered with alcohol-extracted compound Sophora flavescens oral preparation. (4C) Diarrhea severity scores of irradiated rats administered with alcohol-extracted compound Sophora flavescens oral preparation. (4D) Average diarrhea severity scores from day 5 to day 11 of irradiation. Data are expressed as mean ± standard deviation (4A, 4C and 4D) and mean value per cage (4B).

[0038] Figure 2 : In Experimental Example 1, the effect of water-extracted compound Sophora flavescens oral preparation (W-CKP) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The abdomen of the rats was irradiated for a total of 5 times, each time with 4Gy. The rats were orally administered with water-extracted compound Sophora flavescens preparation (A-CKP) at three different doses from day 0 to day 6 after irradiation. (5A) Changes in body weight of irradiated rats administered with water-extracted compound Sophora flavescens oral preparation. (5B) Changes in food intake of irradiated rats administered with water-extracted compound Sophora flavescens oral preparation. (5C) Diarrhea severity scores of irradiated rats administered with water-extracted compound Sophora flavescens oral preparation. (5D) Average diarrhea severity scores from day 5 to day 11 of irradiation. Data are expressed as mean ± standard deviation (5A, 5C and 5D) and average value per cage (5B);

[0039] Figure 3 : In Experimental Example 2, the effect of a high dose of alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The rat abdomen was irradiated 5 times, each time with 4 Gy. The rats were orally administered with alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) at a dose of 140 mg / kg from day 0 to day 6 after irradiation. (8A) Body weight changes in irradiated rats administered with alcohol-extracted compound Sophora flavescens oral preparation. (8B) Changes in food intake in irradiated rats administered with alcohol-extracted compound Sophora flavescens oral preparation. (8C) Diarrhea severity scores in irradiated rats administered with alcohol-extracted compound Sophora flavescens oral preparation. (8D) Average diarrhea severity scores on days 5 to 7 of irradiation. (8E) Changes in FITC fluorescent-labeled dextran levels in irradiated rats treated with alcohol-extracted compound Sophora flavescens oral preparation throughout the course of the disease. Data are expressed as mean ± standard deviation.

[0040] Figure 4: In Experimental Example 2, the effect of compound sophora flavescens injection (CKI) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The abdomen of the rats was irradiated for a total of 5 times, each time with 4 Gy. From day 0 to day 6 after irradiation, the rats were treated with compound sophora flavescens injection at a dose of 2.5 ml / kg by intraperitoneal injection. (10A) Changes in body weight of irradiated rats given compound sophora flavescens injection. (10B) Changes in food intake of irradiated rats given compound sophora flavescens injection. (10C) Diarrhea severity scores of irradiated rats given compound sophora flavescens injection. (10D) Average diarrhea severity scores from day 5 to day 7 of irradiation. Data are expressed as mean ± standard deviation.

[0041] Figure 5 : Comparison of alkaloid components in Test Example 2, the curves from bottom to top are compound sophora flavescens injection, compound sophora flavescens ethanol extract, and compound sophora flavescens water extract.

[0042] Figure 6 : Comparison of flavonoid components in Test Example 2, the curves from bottom to top are compound sophora flavescens injection, compound sophora flavescens ethanol extract, and compound sophora flavescens water extract. DETAILED DESCRIPTION

[0043] The following examples are used to further illustrate the present invention, but are not intended to limit the effective scope of the present invention in any way.

[0044] Preparation of pharmaceutical compositions

[0045] Example 1

[0046] Compound Sophora flavescens alcohol extract (A-CKP): Weigh the prescribed amount of Sophora flavescens: Poria cocos (7:3) and add 10-fold and 8-fold amounts of 60% ethanol, respectively, for 1 hour each time. Filter, combine, and concentrate. Transfer the concentrate to a water bath and evaporate to a thick extract. Place the thick extract in a vacuum drying oven at 70°C.

[0047] Example 2

[0048] Compound Sophora flavescens Water Extraction and Alcohol Precipitation (W-CKP): Weigh the prescribed amount of Sophora flavescens: Poria cocos (7:3) and add 10-fold and 8-fold amounts of water, respectively, to the mixture. Reflux and extract twice, each time for 1 hour. Filter, combine, and concentrate the extracts. Slowly add ethanol to the concentrate while stirring until the alcohol content reaches 60%. Allow to stand and refrigerate overnight. Filter the supernatant of the remaining solution. Concentrate the filtrate and evaporate to dryness in a water bath to a thick extract. Place the thick extract in a vacuum drying oven at 70°C.

[0049] Preparation of formulations

[0050] Example 3 Capsule Preparation:

[0051]

Prescription 1

[0052]

[0053]

Preparation method

[0054] Dry mix: weigh the mixture prepared in Example 1

[0055] Compound Sophora flavescens dry powder 300g, dextrin 100g, high-speed stirring at 200r / min for 2min;

[0056] Making soft material: Use a peristaltic pump to spray 31.4g of 90% ethanol at 20r / min to make soft material. During the making process, there are many lumps, which are sticky and difficult to crush. The rest of the material forms a soft material that can be grasped and dispersed when squeezed.

[0057] Granulation: The above soft material was passed through an 18-mesh sieve and granulated by shaking. There were many lumps that could not be sieved into granules. The granules obtained had poor viscosity and contained more fine powder. They were dried at 60°C for 1 hour.

[0058] Whole grain: pass the above granules through a 14-mesh sieve to obtain whole grains (about 200 g).

[0059]

result

[0060] 1.60 g of magnesium stearate was added to the granules prepared above and mixed thoroughly. The properties of the granules were as follows:

[0061]

[0062] The above granules were encapsulated, packaged, and anhydrous calcium chloride was added as a desiccant. An accelerated stability test (40°C, 75%) was conducted. The results are as follows:

[0063]

[0064] Example 3 Tablet Preparation Method:

[0065] Method 1

[0066] [Prescription 1]

[0067]

[0068] [Particle preparation method]

[0069] Dry mixing: Weigh 600 g of the compound Sophora flavescens dry powder prepared in Example 2 and 200 g of dextrin, and stir at 500 rpm for 2 min;

[0070] Preparation of soft material: Use a peristaltic pump at 25 r / min to spray 100.10 g of 92% ethanol (containing 20% povidone K-30) into the soft material to make a soft material that can be grasped and dispersed when squeezed;

[0071] Granulation: Pass the above soft material through a 20-mesh sieve and granulate it by shaking. The granules should not stick to the sieve. The granules should have appropriate viscosity and be dried at 60℃ for 1h.

[0072] Granulation: The above granules were sieved through an 18-mesh sieve and finally sieved to obtain granules (718 g).

[0073] result:

[0074] 5.70 g of magnesium stearate was added to the granules prepared above and mixed thoroughly. The properties of the granules were as follows:

[0075]

[0076] [Tableting process]

[0077] The above granules were tableted. The control tablet weight was about 0.5g. No abnormal phenomenon occurred during the tableting process. The results of relevant inspection items are as follows:

[0078]

[0079] Coating process 1:

[0080] The plain tablets prepared according to the above-mentioned [Prescription 1] were coated. The coating process parameters and test results are as follows:

[0081]

[0082] Example 4 Tablet Preparation Method:

[0083] Method 2

[0084] [Prescription 2]

[0085]

[0086] [Particle preparation method]

[0087] Dry mixing: weigh 600 g of the compound Sophora flavescens dry powder prepared in Example 1 and 200 g of dextrin, and stir at 500 rpm for 2 min;

[0088] Preparation of soft material: Use a peristaltic pump at 25 r / min to spray 93.5 g of 92% ethanol (containing 20% povidone K-30) to prepare a soft material that can be grasped and dispersed when squeezed;

[0089] Granulation: The above soft material is passed through a 20-mesh sieve and granulated by shaking. The granules do not stick to the sieve. The granules obtained have appropriate viscosity and are dried at 60℃ for 1.5h.

[0090] Granulation: The above granules were sieved through an 18-mesh sieve and finally sieved to obtain granules (776 g).

[0091] result:

[0092] 2.9 g of magnesium stearate was added to the granules prepared above and mixed thoroughly. The properties of the granules were as follows:

[0093]

[0094] [Tableting process]

[0095] The above granules were tableted and relevant parameters were adjusted to control the tablet weight to about 0.5g. No abnormal phenomena occurred during the tableting process. The relevant inspection results are as follows:

[0096]

[0097] Coating process 2:

[0098] The plain tablets prepared according to the above-mentioned [Prescription 2] were coated. The coating process parameters and test results are as follows:

[0099]

[0100]

[0101] Experimental Example 1 Effect of Compound Sophora flavescens Extract on Radiation-induced Intestinal Damage

[0102] 1. Sample:

[0103] Compound Sophora flavescens alcohol extract (A-CKP): prepared according to Example 1.

[0104] Compound Sophora flavescens water extraction and alcohol precipitation (W-CKP): prepared according to Example 2.

[0105] 2. Experimental Methods

[0106] 2.1 Animals

[0107] Experimental animals: Sprague-Dawley rats were maintained at the Biological Resource Centre of the South Australian Health and Medical Research Institute (SAHMRI) in individually ventilated cages with ad libitum access to food and water.

[0108] 2.2 Instruments

[0109] RS2000 small animal irradiator (Rad Source Technologies, GA USA)

[0110] Victor X multifunctional microplate reader (PerkinElmer, MA USA)

[0111] NanoDrop 2000 micro-volume spectrophotometer (Thermo Fisher)

[0112] 2100 Bioanalyzer (Agilent Technologies)

[0113] DNBSEQ (MGI, Shenzhen, China; operated by BGI Genomics)

[0114] 2.3 Animal Model

[0115] Eight-week-old Sprague-Dawley rats were anesthetized with isoflurane inhalation according to standard SAHMRI anesthesia protocols. After anesthesia, the rats were placed facedown in a lead capsule, with their abdomens exposed to the radiation field through the capsule window while the rest of the body was shielded from radiation. Rats were irradiated for 5 consecutive days at 4 Gy / day for a total of 20 Gy to induce radiation enteritis. After each irradiation, the animals were returned to their original housing and allowed to recover according to standard SAHMRI anesthesia protocols.

[0116] 2.4 Administration

[0117] In the prevention and treatment model, rats were orally administered A-CKP, W-CKP, or water from day 0 to day 6 after irradiation. During abdominal irradiation (day 0 to day 4), administration was performed approximately 2 hours before irradiation.

[0118] 2.5 Grouping

[0119] positive control (water) (NT), 7 ml / kg body weight;

[0120] A-CKP, 11.75g crude drug / kg body weight

[0121] A-CKP, 8.40 g crude drug / kg body weight;

[0122] A-CKP, 5.88 g crude drug / kg body weight;

[0123] W-CKP, 16.06 g crude drug / kg body weight;

[0124] W-CKP, 11.47 g crude drug / kg body weight;

[0125] W-CKP, 8.03 g crude drug / kg body weight;

[0126] 2.6 Experimental observation indicators:

[0127] Animals were monitored daily for GIM symptoms, including weight change, food intake, and diarrhea severity. Diarrhea severity was scored as follows: 0 = normal stool, 0.5 = mucous stool, 1 = mild diarrhea with soft stools, 1.5 = mild diarrhea with loose stools and slight staining of the perianal fur, 2 = moderate diarrhea with loose stools and staining of the perianal fur, 2.5 = moderate diarrhea with loose stools and staining of the perianal fur, loose stools and extended staining of the perianal fur, and 3 = severe diarrhea with watery stools and staining of the hind legs.

[0128] 3. Experimental Results

[0129] Figure 1 Effects of an alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) on irradiation-induced gastrointestinal mucositis in SD rats when administered concurrently with irradiation. Rats were irradiated to the abdomen for a total of five times, each dosed at 4 Gy. Three different doses of alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) were administered orally to the rats from day 0 to day 6 after irradiation. (4A) Body weight changes in irradiated rats administered A-CKP. (4B) Food intake changes in irradiated rats administered A-CKP. (4C) Diarrhea severity scores in irradiated rats administered A-CKP. (4D) Average diarrhea severity scores from day 5 to day 11 of irradiation. Data are presented as mean ± SD (4A, 4C, and 4D) and as the mean per cage (B).

[0130] The data showed that rats given A-CKP (140 mg / kg) experienced faster weight loss and early recovery compared to the control group or other dose groups (4A). Food intake also recovered more quickly (4B). Diarrhea data showed that rats receiving the high dose of A-CKP (140 mg / kg) experienced significantly less diarrhea during the course of the disease compared to the control group (4C).

[0131] Figure 2 Effects of a water-extracted compound Sophora flavescens oral preparation (W-CKP) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The rats were irradiated to the abdomen for a total of five times, each time with 4 Gy. The rats were orally administered with a water-extracted compound Sophora flavescens preparation (A-CKP) at three different doses from day 0 to day 6 after irradiation. (5A) Body weight changes in irradiated rats administered with a water-extracted compound Sophora flavescens oral preparation. (5B) Food intake changes in irradiated rats administered with a water-extracted compound Sophora flavescens oral preparation. (5C) Diarrhea severity scores in irradiated rats administered with a water-extracted compound Sophora flavescens oral preparation. (5D) Average diarrhea severity scores from day 5 to day 11 of irradiation. Data are expressed as mean ± SD (5A, 5C, and 5D) and mean per cage (5B).

[0132] The data showed that the body weight of rats administered with W-CKP (140 mg / kg or 100 mg / kg) was significantly reduced compared with the control group ( Figure 5 A), and food intake also showed the same trend ( Figure 5 B). The diarrhea results showed that the W-CKP group had no significant effect on the severity of diarrhea ( Figure 5 C and D). Experimental Example 2: Comparison of Compound Sophora flavescens Extract and Compound Sophora flavescens Injection in Preventing and Treating Radiation-induced Mucosal Injury

[0133] 1. Sample

[0134] Compound Sophora flavescens alcohol extract (A-CKP): prepared according to Example 1;

[0135] Compound Sophora flavescens injection (also known as "CKI"), batch number: 20210519; provided by Shanxi Zhendong Pharmaceutical Co., Ltd.

[0136] 2. Experimental Methods

[0137] 2.1 Animals

[0138] Experimental animals: Sprague-Dawley rats were maintained at the Biological Resource Centre of the South Australian Health and Medical Research Institute (SAHMRI) in individually ventilated cages with ad libitum access to food and water.

[0139] 2.2 Instruments

[0140] RS2000 small animal irradiator (Rad Source Technologies, GA USA)

[0141] Victor X multifunctional microplate reader (PerkinElmer, MA, USA)

[0142] NanoDrop 2000 micro-volume spectrophotometer (Thermo Fisher)

[0143] 2100 Bioanalyzer (Agilent Technologies)

[0144] DNBSEQ (MGI, Shenzhen, China; operated by BGI Genomics)

[0145] 2.3 Animal Model

[0146] Eight-week-old Sprague-Dawley rats were anesthetized with isoflurane inhalation according to standard SAHMRI anesthesia protocols. After anesthesia, the rats were placed facedown in a lead capsule, with their abdomens exposed to the radiation field through the capsule window while the rest of the body was shielded from radiation. Rats were irradiated for 5 consecutive days at 4 Gy / day for a total of 20 Gy to induce radiation enteritis. After each irradiation, the animals were returned to their original housing and allowed to recover according to standard SAHMRI anesthesia protocols.

[0147] 2.4 Administration

[0148] In the prevention and treatment model, rats were orally administered A-CKP, CKI, vehicle (0.25% Tween-80 + 25 mM HEPES) (VC), or water from day 0 to day 6 after irradiation. During abdominal irradiation (days 0 to 4), administration was performed approximately 2 hours before irradiation.

[0149] 2.5 Grouping

[0150]

[0151]

[0152] 2.6 Experimental observation indicators:

[0153] Animals were monitored daily for GIM symptoms, including weight change, food intake, and diarrhea severity. Diarrhea severity was scored as follows: 0 = normal stool, 0.5 = mucous stool, 1 = mild diarrhea with soft stools, 1.5 = mild diarrhea with loose stools and slight staining of the perianal fur, 2 = moderate diarrhea with loose stools and staining of the perianal fur, 2.5 = moderate diarrhea with loose stools and staining of the perianal fur, loose stools and extended staining of the perianal fur, and 3 = severe diarrhea with watery stools and staining of the hind legs.

[0154] On days 4 and 7 after irradiation, rats were given FITC-dextran at a dose of 600 mg / kg by oral gavage to measure gastrointestinal permeability and barrier damage.

[0155] 3. Experimental Results

[0156] Figure 3 Effects of high doses of alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The rat abdomen was irradiated 5 times with 4 Gy each time. The rats were orally administered with alcohol-extracted compound Sophora flavescens oral preparation (A-CKP) at a dose of 140 mg / kg from day 0 to day 6 after irradiation. (8A) Body weight changes in irradiated rats given alcohol-extracted compound Sophora flavescens oral preparation. (8B) Changes in food intake in irradiated rats given alcohol-extracted compound Sophora flavescens oral preparation. (8C) Diarrhea severity scores in irradiated rats given alcohol-extracted compound Sophora flavescens oral preparation. (8D) Average diarrhea severity scores on days 5 to 7 of irradiation. (8E) Changes in FITC fluorescent-labeled dextran levels in irradiated rats treated with alcohol-extracted compound Sophora flavescens oral preparation throughout the course of the disease. Data are expressed as mean ± standard deviation.

[0157] The high-dose A-CKP-treated group lost significantly less weight compared to the control group (8A), while food intake did not differ significantly between the A-CKP and control groups (8B). The effect of A-CKP on the extent of diarrhea was significantly less severe compared to the control group (8C and 8D). However, there were no significant differences in FITC-dextran levels in the serum of A-CKP- and control-treated rats, indicating that A-CKP had no effect on the extent of intestinal radiation damage when measured in this manner (8E).

[0158] Note: NIR=(No irradiation); NT=(No treatment); VC=Vehicle control).

[0159] Figure 4 Effects of compound sophora flavescens injection (CKI) on irradiation-induced gastrointestinal mucositis in SD rats when administered simultaneously with irradiation. The abdomen of rats was irradiated 5 times with 4 Gy each time. Rats were treated with compound sophora flavescens injection at a dose of 2.5 ml / kg by intraperitoneal injection from day 0 to day 6 after irradiation. (10A) Body weight changes in irradiated rats given compound sophora flavescens injection. (10B) Food intake changes in irradiated rats given compound sophora flavescens injection. (10C) Diarrhea severity scores in irradiated rats given compound sophora flavescens injection. (10D) Average diarrhea severity scores from day 5 to day 7 of irradiation. Data are expressed as mean ± standard deviation.

[0160] Compared with the control group, CKI administration had no significant effect on body weight and food intake (10A and 10B). However, CKI treatment only resulted in a slight improvement in the severity of diarrhea (10C and 10D).

[0161] 1. Sample:

[0162] Compound Sophora flavescens injection (CKI), batch number 20210519, was provided by Shanxi Zhendong Pharmaceutical Co., Ltd.

[0163] Compound Sophora flavescens alcohol extract (A-CKP): prepared according to Example 1.

[0164] Compound Sophora flavescens water extraction and alcohol precipitation (W-CKP): prepared according to Example 2.

[0165] 2. Methods

[0166] Figure 5 Testing conditions:

[0167] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel as the packing (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm), methanol as mobile phase A, 0.2% potassium dihydrogen phosphate solution (pH adjusted to 3.0 with phosphoric acid) as mobile phase B, gradient elution as specified in the table below; column temperature 30°C; detection wavelength 211 nm; flow rate 0.6 ml / min. The number of theoretical plates calculated based on the oxymatrine peak should be no less than 5000.

[0168]

[0169] Figure 6 Testing conditions:

[0170] Chromatographic conditions and system suitability test

[0171] Chromatographic column: Thermo BDS Hypersil™ C18250*4.6mm, 5μm

[0172] Mobile phase: methanol as mobile phase A, water as mobile phase B, gradient elution program:

[0173] Time / min Mobile phase A (%) Mobile phase B (%) 0～15 40→50 60→50 15～20 50→60 50→40 20～25 60 40 25～35 60→70 40→30 35～45 70 30 55 70→90 30→10 60 90→40 10→60 65 40 60

[0174] Flow rate: 1.0ml / min

[0175] Injection volume: 10 μl

[0176] Column temperature: 35°C

[0177] Detection wavelength: 295nm

[0178] in conclusion

[0179] The compositions of samples CKI, A-CKP, and A-CKP are as follows: Figure 5 and Figure 6 As shown by Figure 5 and 6 It can be seen that the compound Sophora flavescens extract of the present invention and the compound Sophora flavescens injection are essentially different in their composition. The compound Sophora flavescens extract of the present invention is prepared by alcohol extraction, while the compound Sophora flavescens injection is prepared by a series of treatments including 0.1% acetic acid percolation and then water decoction and alcohol precipitation.

[0180] The compound sophora flavescens extract and the compound sophora flavescens injection of the present invention have different processes and ingredients. The compound sophora flavescens extract of the present invention is rich in flavonoids, while the compound sophora flavescens injection contains almost no flavonoids.

Claims

1. A use of a compound Sophora flavescens extract in the preparation of a medicament for treating or preventing radiation-induced intestinal damage, wherein the compound Sophora flavescens extract is composed of Sophora flavescens and Poria cocos.

2. The application according to claim 1, characterized in that The mass ratio of Sophora flavescens to Psoralea corylifolia is 7:

3.

3. The application according to claim 1, characterized in that The compound Sophora flavescens extract is prepared by the following method: Weigh the prescribed amount of medicinal materials Sophora flavescens and Poria cocos, add 8 to 10 times the volume of ethanol to a concentration of 50% to 90%, preferably 60%, and reflux and extract 1 to 3 times, preferably 2 times. The extracts are filtered, combined, and concentrated; and dried under reduced pressure at 60°C to 80°C, preferably 70°C to obtain the product.

4. The application according to claim 3, characterized in that The compound Sophora flavescens extract is prepared by the following method: Weigh the prescribed amount of medicinal materials Sophora flavescens and Poria cocos, add 10 times and 8 times the amount of 60% ethanol respectively, reflux and extract twice, filter the extracts, combine, concentrate, and dry under reduced pressure at 70°C to obtain the product.

5. The application according to claim 4, characterized in that: The compound Sophora flavescens extract is prepared by the following method: Weigh the prescribed amount of Sophora flavescens and Psoralea corylifolia (7:3), add 10 times and 8 times the amount of 60% ethanol respectively, reflux and extract twice, each time for 1 hour, filter the extracts, combine, concentrate, and dry under reduced pressure at 70°C to obtain the product.

6. The application according to claim 1, characterized in that: The compound Sophora flavescens extract is prepared by the following method: Weigh the prescribed amount of Sophora flavescens and Poria cocos (7:3) and add 8-10 times the amount of water to each, reflux extract. Filter, combine, and concentrate the extracts. Slowly add 50% to 90% (preferably 60%) ethanol to the concentrate, stirring until the alcohol content reaches 60%. Allow to stand and refrigerate overnight. Filter the supernatant of the remaining solution, concentrate, and dry under reduced pressure at 70°C.

7. The use according to claim 6, characterized in that The compound Sophora flavescens extract is prepared by the following method: Take the prescribed amount of the medicinal materials Sophora flavescens and Psoralea corylifolia (7:3), add 10 times and 8 times the amount of water respectively, reflux extraction, filter the extracts, combine, and concentrate; add ethanol to the concentrate, stir to make the alcohol content of the medicinal solution reach 60%, let it stand and refrigerate overnight; take the supernatant of the medicinal solution after standing, filter, and dry under reduced pressure at 70°C.

8. The application according to claim 7, characterized in that: The compound Sophora flavescens extract is prepared by the following method: Take the prescribed amount of medicinal materials Sophora flavescens: Phellodendron chinense (7:3), add 10 times and 8 times the amount of water respectively, reflux extraction twice, each extraction is 1 hour, the extracts are filtered, combined, and concentrated; ethanol is added to the concentrate, stirred to make the alcohol content of the medicinal solution reach 60%, and refrigerated overnight; take the supernatant of the medicinal solution after standing, filter, concentrate, and dry under reduced pressure at 70°C.

9. Use of a preparation containing the compound Sophora flavescens extract according to claims 1 to 8 in the preparation of a drug for treating or preventing radiation-induced intestinal injury, characterized in that: The preparation is a tablet, capsule, oral preparation, liquid injection or injection.

10. A compound Sophora flavescens extract for treating or preventing radiation-induced intestinal damage, characterized in that: The compound Sophora flavescens consists of Sophora flavescens and Poria cocos.

11. The compound Sophora flavescens extract according to claim 10, characterized in that The mass ratio of Sophora flavescens to Psoralea corylifolia is 7:

3.

12. The compound Sophora flavescens extract according to claim 11, which is prepared by the following method: Weigh the prescribed amount of Sophora flavescens and Poria cocos according to the ratio, add 10 times and 8 times the amount of 60% ethanol respectively, reflux and extract twice, each extraction for 1 hour, filter the extracts, combine, and concentrate; transfer the concentrate to a water bath and steam to a thick extract, place the thick extract in a vacuum drying oven, and dry it under reduced pressure at 70°C to obtain; or Weigh the prescribed amount of Sophora flavescens and Poria cocos according to the prescribed ratio. Add 10-fold and 8-fold amounts of water, respectively, and reflux extract twice, each time for 1 hour. Filter the extracts, combine, and concentrate. Slowly add ethanol to the concentrate while stirring until the alcohol content reaches 60%. Let it stand and refrigerate overnight. Filter the supernatant of the remaining solution. Concentrate the filtrate and evaporate it to dryness in a water bath to a thick extract. Place the thick extract in a vacuum drying oven at 70°C.

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