Preparation method of white jade powder soup

Abstract

The invention relates to the field of traditional Chinese medicine preparations and pharmaceutical science, and discloses a preparation method of white jade powder decoction, which comprises the following steps: (1) gradient directional extraction: a, mixing and extracting 15-30 parts of sculellaria barbata, 15-30 parts of oldenlandia diffusa, 6-9 parts of curcuma zedoary, 3-10 parts of ground beetle and 70-80% ethanol; b, mixing and decocting 9-30 parts of radix astragali, 6-12 parts of roasted rhizoma atractylodis macrocephalae, 5-10 parts of radix saposhnikoviae and a buffer solution; (2) mixing the two extracting solutions in the step (1) to construct a core-shell structure; (3) carrying out spray drying on the mixed solution to prepare a solid dispersion; and (4) performing enteric coating modification on the powder. Through scientific drug release kinetic analysis, the effect of optimizing the ratio and bioavailability of the effective components is achieved, and compared with the scheme of single-component drugs in the prior art, the defects that the absorption rate of the effective components is low and the drug effect is unstable are overcome. Therefore, the medicine can better exert the treatment effect in clinical application, and the medication experience of a patient is improved.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicine preparations and pharmaceutical science, and in particular to a method for preparing Baiyusan decoction. Background Art

[0002] In modern medicine, the use of traditional Chinese medicine has received increasing attention, especially for the treatment of chronic diseases and complex conditions. Chinese herbal medicines have become an important choice for disease treatment due to their complex ingredients and diverse mechanisms of action. With the development of science and technology, researchers have gradually realized the close relationship between the activity of active ingredients and the bioavailability of drugs, which provides new opportunities for the modernization of Chinese medicine and its clinical application.

[0003] Existing technologies mainly focus on improving the release and bioavailability of effective ingredients in traditional Chinese medicine. However, most studies have focused on single ingredients, which are often limited to traditional extraction and preparation processes. This has resulted in many traditional Chinese medicine compounds failing to achieve the expected effects in clinical applications. In addition, the lack of diversity in drug dosage forms has limited their role in the treatment process, affecting the overall treatment effect.

[0004] The biggest problem with the existing technology is the lack of research on drug release kinetics. The release behavior of many drugs in the body is difficult to accurately predict, especially in a complex physiological environment. This leads to uncertainty in the clinical application of drugs on the one hand, and makes it difficult to optimize treatment plans for individual differences in patients on the other hand. The problem raised by Quan Yi, namely optimizing the dosage form design and release rate of traditional Chinese medicine compound, is precisely to improve the absorption efficiency and overall efficacy of drugs in the body and achieve better clinical application effects. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides a method for preparing Baiyusan decoction, which solves the problems in the prior art of insufficient research on drug release kinetics, resulting in low drug bioavailability and unstable clinical application effect.

[0006] To achieve the above purpose, the present invention is implemented by the following technical scheme: a method for preparing Baiyusan soup, comprising the following steps:

[0007] (1) Gradient directional extraction:

[0008] a. The barbata 15-30 parts, Hedyotis diffusa 15-30 parts, Curcuma 6-9 parts, Eupolyphaga 3-10 parts and 70-80% ethanol mixed extract;

[0009] b. Mix 9-30 parts of Astragalus, 6-12 parts of fried Atractylodes macrocephala, 5-10 parts of Saposhnikovia divaricata and decoct with buffer;

[0010] (2) mixing the two extracts from step (1) to construct a core-shell structure;

[0011] (3) Spray-dry the mixed solution to prepare a solid dispersion;

[0012] (4) Modify the powder with enteric coating.

[0013] Preferably, in step (1)a, 3-10 parts of Gleditsia sinensis thorn, 3-10 parts of Curculigo orchioides, 6-10 parts of Epimedium brevicornu, and 6-12 parts of Polyporus umbellatus are also added.

[0014] Preferably, step (1)a is specifically:

[0015] Mix the medicinal materials with 70-80% ethanol at a material-liquid ratio of 1:8-12, extract at 40-60°C and an ultrasonic power of 250-350W for 30-90min, and simultaneously add hydroxypropyl-β-cyclodextrin, the addition amount of which is 5-10% of the ethanol volume, and the substitution degree of the hydroxypropyl-β-cyclodextrin is 0.6-0.9.

[0016] Preferably, step (1)b is specifically:

[0017] Mix the medicinal materials with a pH5.5-6.5 buffer solution at a material-liquid ratio of 1:10-15, decoct at 80-95°C for 1.5-2.5h, and simultaneously add polyoxyethylene (35) castor oil, the addition amount of which is 2-5% of the buffer solution volume, and the HLB value of the polyoxyethylene (35) castor oil is 12-14.

[0018] Preferably, step (2) specifically includes:

[0019] Mix the extraction solutions of a and b at a volume ratio of 1:1.5-2.5, add a freeze-drying protectant accounting for 5-10% of the total mass of the original medicinal materials, stir at 55-65°C and 200-400rpm for 1.0-2.0h to form a core-shell structure with a particle size of 80-120nm and a Zeta potential of -20 to -30mV. The freeze-drying protectant is mannitol, with a particle size <50μm and a water activity ≤0.30.

[0020] Preferably, the specific parameters of step (3) are:

[0021] Inlet air temperature 140-160°C, outlet air temperature 60-70°C, atomization disk rotation speed 18000-22000rpm, feed rate 5-10mL / min, and the obtained powder has a glass transition temperature of 80-85°C and a moisture content ≤3%.

[0022] Preferably, step (4) specifically includes:

[0023] Dissolve the methyl methacrylate-ethyl acrylate copolymer in 95% ethanol to prepare an 8-12% coating solution, add triethyl citrate, and the dosage is 15-25% of the weight of the coating material. Coating is carried out in a fluidized bed at 40-50 °C with an atomization pressure of 1.2-1.8 bar and a spraying rate of 2-5 g / min. The thickness of the coating film is 10-15 μm and the weight gain is 8-15%.

[0024] Preferably, the mass ratio of hydroxypropyl-β-cyclodextrin to polyoxyethylene (35) castor oil is 3:1-5:1.

[0025] Preferably, the mass ratio of the freeze-drying protectant to the enteric coating material is 1:0.5-1:1.5.

[0026] Preferably, in the powder after enteric coating, hydroxypropyl-β-cyclodextrin accounts for 10-20% of the total mass of the original medicinal materials, and polyoxyethylene (35) castor oil accounts for 3-8% of the total mass of the original medicinal materials.

[0027] The present invention provides a preparation method of Baiyu powder soup. It has the following beneficial effects:

[0028] 1. The present invention adopts the extraction of traditional Chinese medicine compound, achieving the effects of optimizing the ratio of active ingredients and bioavailability. Compared with the single-component drug scheme in the prior art, it solves the deficiencies of low absorption rate of active ingredients and unstable drug efficacy. This enables the drug to better exert its therapeutic effect in clinical applications and improves the patient's medication experience.

[0029] 2. The present invention introduces an improved dosage form design, achieving a faster dissolution rate and a sustained release effect of active ingredients. Compared with the traditional dosage form in the prior art, this scheme solves the deficiencies of uneven drug release and long treatment cycle. This innovative design improves the bioavailability of the drug in the body, enabling patients to feel the drug effect in a shorter time.

[0030] 3. The present invention adopts high-performance liquid chromatography as the analysis technique, achieving the technical effect of accurately determining the drug concentration. Compared with the other analysis method schemes in the prior art, it solves the deficiencies of low detection sensitivity and poor repeatability. Through this advanced technique, the reliability of bioavailability and stability tests is ensured, providing a solid scientific basis for the safety evaluation of the drug.

[0031] 4. The present invention combines the modern drug release kinetics model, achieving the effect of accurately describing the dynamic release behavior. Compared with the prior art schemes that ignore the pharmacokinetic characteristics, it solves the limitations in the research of drug action mechanisms. This innovation enables drug development to more precisely target patient needs, optimize treatment plans, and improve the clinical effect and competitiveness in the pharmaceutical market. Description of the Drawings

[0032] Figure 1 This is a schematic flow chart of the method of the present invention. Specific embodiments

[0033] Next, in combination with the accompanying drawings of the present invention specification, the technical solutions in the embodiments of the present invention will be clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0034] Please refer to the appendix Figure 1 , the embodiment of the present invention provides a method for preparing Baiyu powder soup, including:

[0035] Scutellaria barbata 15g - 30g, with pungent, bitter, and cold properties; belonging to the lung, liver, and kidney meridians. Its functions are to clear heat and detoxify, and promote blood circulation to remove stasis and induce diuresis. It is applicable to furuncles and carbuncles, sore throats, traumatic injuries, edema, jaundice, and snake and insect bites.

[0036] Oldenlandia diffusa 15g - 30g, slightly bitter, sweet, and cold. Belonging to the stomach, large intestine, and small intestine meridians. Efficacy: clearing heat and detoxifying, promoting diuresis and removing dampness. Indications: carbuncles and sores, sore throats, snake bites; stranguria with pain due to heat, etc.

[0037] Chemical composition: mainly containing hentriacontane, stigmasterol, ursolic acid, oleanolic acid, β - sitosterol, β - sitosterol - D - glucoside, p - coumaric acid, etc.

[0038] This product has an anti - tumor effect. Its antibacterial effect in vitro is not significant, and the high - concentration decoction has a weak inhibitory effect on Staphylococcus aureus and Shigella dysenteriae; in vivo, it can enhance the phagocytic ability of white blood cells and has an anti - inflammatory effect. It also has the effects of inhibiting spermatogenesis ability and protecting the liver and promoting bile secretion.

[0039] Gleditsia sinensis thorn 3g - 10g, with pungent and warm properties; belonging to the liver and stomach meridians. Functions: promoting pus discharge to resolve carbuncles, expelling pus, and killing parasites. Applicable to the initial stage of carbuncles or non - ulceration of abscesses, and externally treating scabies and leprosy.

[0040] Curcuma zedoaria 6g - 9g, pungent, bitter, and warm. Belonging to the liver and spleen meridians. Efficacy: promoting blood circulation to remove stasis, regulating qi, and relieving pain by reducing accumulation. Indications: mass in the abdomen, amenorrhea due to blood stasis, chest pain; abdominal distension and pain due to food stagnation and qi stagnation, etc.

[0041] Chemical composition: mainly containing volatile oils: germacrone, curdione, curcumol, curcolactone, wenyujinol, zingiberene, borneol, curzerenone, terpinene, eugenol, etc.; phenolic components: curcumin, etc. The Chinese Pharmacopoeia stipulates that this product should contain no less than 1.5% (mL / g) of volatile oil, and the cut crude drug should contain no less than 1.0% (mL / g).

[0042] Pharmacological effects: The volatile oil preparation of Curcuma zedoaria has anti-cancer effects. The volatile oil of Curcuma wenyujin can inhibit the growth of various pathogenic bacteria; Curcuma oil has anti-inflammatory, anti-gastric ulcer, liver-protecting and anti-early pregnancy effects. The water extract of Curcuma zedoaria can inhibit platelet aggregation, promote the restoration of microarterial blood flow, and promote the restoration of local microcirculation; the water extract and ethanol precipitation of Curcuma zedoaria have an inhibitory effect on in vivo thrombus formation. In addition, Curcuma zedoaria has a direct inactivating effect on respiratory syncytial virus.

[0043] Eupolyphaga seu Steleophaga: 3-10 g, salty, cold; slightly poisonous. It acts on the Liver Meridian. Efficacy: Promote blood circulation to remove blood stasis, and reconnect tendons and bones. Indications: traumatic injury, tendon and bone fracture; blood stasis amenorrhea, postpartum blood stasis abdominal pain, mass and abdominal mass. Modern research:

[0044] Chemical constituents: mainly contain fatty acid components: palmitoleic acid, oleic acid, palmitic acid, myristic acid, stearic acid and a small amount of linoleic acid. It also contains uracil, allantoin, alkaloids, amino acids, etc. 2. Pharmacological effects The water extract of Eupolyphaga seu Steleophaga has the effects of regulating lipid metabolism, antioxidant free radicals, and protecting vascular endothelial cells; the streptokinase of Eupolyphaga seu Steleophaga has anticoagulant and thrombolytic effects; the fibrinolytic active protein has the effect of inhibiting tumor cells in vitro, and can significantly inhibit the growth of various tumor cells such as melanoma, gastric cancer, and primary liver cancer. In addition, Eupolyphaga seu Steleophaga can promote the healing of bone injuries.

[0045] Adverse reactions: Eupolyphaga seu Steleophaga is likely to cause allergic reactions in patients with drug allergies, mainly manifested as general itching, bright red skin lesions or dense small papules on the skin, and even cause exfoliative dermatitis.

[0046] Curculigo orchioides: 3-10 g, pungent, hot; poisonous. It acts on the Kidney, Liver and Spleen Meridians. Efficacy: Tonify kidney yang, strengthen muscles and bones, dispel cold dampness. Indications: Kidney yang deficiency, decline of life gate fire, impotence and sperm coldness, frequent urination; soreness and cold pain in the waist and knees, weakness of muscles and bones; yang deficiency cold diarrhea.

[0047] Chemical constituents: mainly contain phenolic glycoside components: curculigoside; triterpenoid components: curculigosaponins A-M, curculigine A, B, C, etc.; alkaloid components: lycorine, etc.; sterol components: cycloartenol, stigmasterol, etc. The Chinese Pharmacopoeia stipulates that this product should contain no less than 0.080% of curculigoside (C22H26O11).

[0048] Pharmacological effects: Curculigo orchioides can prolong the average survival time of experimental animals. The alcohol extract of Curculigo orchioides can significantly increase the phagocytosis percentage and phagocytosis index of peritoneal macrophages in mice; the water decoction of Curculigo orchioides can significantly increase the weights of the anterior pituitary, ovary and uterus of rats, and significantly increase the specific binding capacity of ovarian hCG / LH receptors; the alcohol extract of Curculigo orchioides can significantly prolong the sleeping time of mice, and antagonize the convulsions induced by picrotoxin in mice, with sedative and anti-convulsant effects.

[0049] Adverse Reactions: This product has an inhibitory effect on the central nervous system. Overdose can cause cardiac depression, arrhythmia and paralysis. The main manifestations of poisoning are profuse sweating all over the body, cold limbs, numbness, tongue swelling protruding out of the mouth, restlessness, and then coma. The main reasons for Curculiginis Rhizoma poisoning are, on the one hand, long-term large-dose administration leading to toxic reactions, and on the other hand, its pungent and hot characteristics. To ensure the safety of medication, on the one hand, it must be used strictly in accordance with the specified dosage and usage, and on the other hand, it must be used dialectically and cannot be misused.

[0050] Epimedii Folium 6g - 10g, pungent, sweet, warm. It belongs to the liver and kidney meridians. Tonifies kidney yang, strengthens tendons and bones, dispels wind-dampness. It is mainly used for treating kidney yang deficiency, impotence and spermatorrhea, flaccidity of tendons and bones; wind-cold-damp arthralgia, numbness and contracture.

[0051] Chemical Constituents: It mainly contains flavonoid components: icariin, baohuoside I, II, icariside I, II, epimedin A, rhamnosyl icariside II, sagittatoside A, B, C, hyperoside, etc.; it also contains polysaccharides, etc. The Chinese Pharmacopoeia stipulates that the total flavonoids contained in this product, calculated as icariin, shall not be less than 5.0%, the content of icariin calculated as dry product shall not be less than 0.50%, the content of icariin in the cut crude drug shall not be less than 0.40%, and the total content of icariin and baohuoside I in the stir-fried Epimedii Folium shall not be less than 0.60%.

[0052] Pharmacological Effects: Epimedii Folium has androgen-like and phytoestrogen-like activities and can enhance the sexual function of animals; subcutaneous injection of Epimedii Folium polysaccharide to female mice can cause thymus shrinkage while stimulating the function of peripheral T cells. The total flavonoids of Epimedii Folium have a regulatory effect on the abnormally increased immune function in androgen-deficient model mice; icariin can improve the serum SOD activity and androgen level in subacute aging model rats, reduce germ cell apoptosis, improve the degenerative changes of testicular tissue and inhibit the expression of the germ cell aging gene P16 protein, thereby delaying gonadal aging. In addition, Epimedii Folium also has effects on the cardiovascular system, bone marrow and hematopoietic system functions, anti-osteoporosis, improving learning and memory, anti-radiation, anti-tumor, etc.

[0053] Polyporus 6g - 12g, sweet, light, flat. It belongs to the kidney and bladder meridians. Promotes diuresis and percolates dampness. It is mainly used for treating edema, dysuria, diarrhea, stranguria, leukorrhea.

[0054] Chemical Constituents: It mainly contains polysaccharides: polyporus glucan I, polyporus polysaccharide, etc.; sterol components: ergosterol, etc. It also contains organic acids, proteins, etc. The Chinese Pharmacopoeia stipulates that this product contains ergosterol (C28H44O) not less than 0.070%, and the cut crude drug not less than 0.050%.

[0055] Pharmacological effects: This product has a diuretic effect, and its diuretic mechanism is caused by inhibiting the reabsorption of water and electrolytes in the renal tubules. Polyporus polysaccharide has anti-tumor and hepatitis prevention and treatment effects. The water and alcohol extracts of Polyporus have effects such as promoting immunity, preventing stone formation, anti-mutagenesis, and antibacterial

[0056] Astragalus membranaceus 9g - 30g, sweet, slightly warm. It acts on the spleen and lung meridians. Tonifies qi and raises yang, benefits the defensive qi and consolidates the exterior, promotes diuresis and reduces swelling, generates body fluid and nourishes blood, promotes qi movement to relieve pain, expels pus by supporting healthy qi, and astringes sores and promotes granulation. It is mainly used for qi deficiency and weakness, poor appetite and loose stools, edema with scanty urine, qi sinking, chronic diarrhea and rectal prolapse, hematochezia and metrorrhagia. This product is sweet and warm, enters the spleen meridian, and is an important herb for tonifying the spleen qi. It treats weakness of the lung qi, cough and shortness of breath; spontaneous sweating due to exterior deficiency; internal heat and thirst; sallow complexion due to blood deficiency, deficiency of both qi and blood; qi deficiency and blood stasis, hemiplegia, pain and numbness in the bi syndrome; qi and blood deficiency, difficult ulceration of carbuncles and sores, and non-healing of long-term ulcers

[0057] Chemical constituents: It mainly contains triterpenoid saponin components: astragaloside I, II, III, IV (astragaloside A), astragalocystin I, II, etc.; flavonoid components: formononetin, calycosin - 7 - O - β - D - glucoside, etc.; it also contains polysaccharides, amino acids, etc. The Chinese Pharmacopoeia stipulates that this product should contain no less than 0.020% of calycosin - 7 - O - β - D - glucoside, and the cut crude drug should contain no less than 0.020%; it should contain no less than 0.040% of astragaloside A, and the cut crude drug should contain no less than 0.040%. Prepared Astragalus membranaceus should contain no less than 0.030% of astragaloside A and no less than 0.020% of calycosin - 7 - O - β - D - glucoside

[0058] Pharmacological effects: Astragalus polysaccharide can promote the synthesis of RNA and proteins, make cells grow vigorously and have an extended lifespan, and can also resist fatigue, tolerate low temperatures, and resist influenza virus

[0059] The water decoction, polysaccharide, and saponin of Astragalus membranaceus have a protective and promoting effect on hematopoietic function. Total astragalus saponin has a positive inotropic effect, and total astragalus flavonoids and total saponin can protect the myocardium from ischemia and hypoxia. The water decoction of Astragalus membranaceus has a protective effect on the kidneys, eliminates proteinuria and has a diuretic effect, and has a dual - regulating effect on blood pressure. In addition, Astragalus membranaceus has effects such as anti - aging, anti - radiation, anti - inflammation, reducing blood lipid, reducing blood sugar, enhancing immunity, anti - tumor, and protecting the liver

[0060] Atractylodes macrocephala (parched) 6g - 12g, sweet, bitter, warm. It acts on the spleen and stomach meridians. Efficacy: Tonifies qi and strengthens the spleen, dries dampness and promotes diuresis, stops sweating, and prevents miscarriage. Indications: Spleen qi weakness, poor appetite and lassitude, abdominal distension and diarrhea, phlegm - fluid dizziness and palpitations, edema, leukorrhea; spontaneous sweating due to qi deficiency; restlessness of the fetus due to spleen deficiency

[0061] Chemical constituents: It mainly contains volatile oils such as atractylon, atractylol, atractylodether, cadinol, atractylenolide, etc., lactone compounds such as atractylenolide I - IV, bisatractylenolide, etc., and also contains fructose, inulin, atractylodes polysaccharide, various amino acids, atractylotriol, vitamin A and other components

[0062] Pharmacological effects: The water decoction of Atractylodes macrocephala can promote gastric emptying and small intestine propulsion function in mice, and can prevent and treat experimental gastric ulcers. Atractylenolide I has the effects of enhancing the activity of salivary amylase, promoting the absorption of nutrients, and regulating gastrointestinal function. The water decoction and fluid extract of Atractylodes macrocephala both have obvious and lasting diuretic effects. Atractylodes macrocephala polysaccharide and volatile oil can enhance cellular immune function. The water decoction of Atractylodes macrocephala has anti-aging effects. The alcohol extract and petroleum ether extract of Atractylodes macrocephala can inhibit the contraction of uterine smooth muscle in experimental animals. In addition, Atractylodes macrocephala has effects such as protecting the liver, promoting bile secretion, lowering blood sugar, antibacterial, anti-tumor, sedative, antitussive, and expectorant.

[0063] Saposhnikovia divaricata 5g - 10g, pungent, sweet, slightly warm. It belongs to the bladder, liver, and spleen meridians. Efficacy: Dispel wind and relieve exterior syndrome, relieve pain due to dampness, stop convulsions. Indications: Cold, headache; Rheumatic arthralgia; Rubella itching; Tetanus.

[0064] Chemical constituents: It mainly contains chromone components: saposhnikoviadin, 5-O-methylvisammioside, cimifugin, cimifugin glycoside; coumarin components: bergapten. It also contains acidic polysaccharides, volatile oils, etc. The Chinese Pharmacopoeia stipulates that the total amount of cimifugin glycoside (C22H28O11) and 5-O-methylvisammioside (C22H28O10) contained in this product shall not be less than 0.24%.

[0065] This product has effects such as antipyretic, anti-inflammatory, sedative, analgesic, anti-convulsive, and anti-allergic. The fresh juice of Saposhnikovia divaricata has certain antibacterial effects against Pseudomonas aeruginosa and Staphylococcus aureus, and the decoction has varying degrees of inhibitory effects on Shigella dysenteriae, hemolytic streptococcus, etc. It also has the effect of enhancing the phagocytic function of peritoneal macrophages in mice.

[0066] This formula follows the theory of "sovereign, ministerial, adjuvant, and guiding herbs". Scutellaria barbata and Hedyotis diffusa are the sovereign herbs (clearing heat and promoting blood stasis removal), Curcuma zedoaria and Eupolyphaga sinensis are the ministerial herbs (activating blood circulation and dissipating stasis), and Astragalus membranaceus and Atractylodes macrocephala are the adjuvant and guiding herbs (strengthening healthy qi and consolidating the root). The proportions of each herb are optimized through orthogonal experiments to ensure the synergistic effect of heat-clearing and tonifying components.

[0067] Functional excipients:

[0068] Hydroxypropyl-β-cyclodextrin (HP-β-CD): Degree of substitution 0.8, dosage is 15% of the total mass of the herbs;

[0069] Polyoxyethylene (35) castor oil (Cremophor EL): HLB value 13.5, dosage is 5% of the total mass of the herbs;

[0070] Eudragit L100-55 (enteric coating material): Dosage is 10% of the total mass of the final powder;

[0071] Mannitol (lyoprotectant): Particle size <50μm, dosage is 8% of the total mass of the herbs.

[0072] HP-β-CD incorporates lipophilic components such as curcumol through its hydrophobic cavity (inclusion constant Kf = 2.3×10 3 L / mol), forming a molecular dispersion to inhibit the precipitation of crystals;

[0073] Cremophor EL forms micelles in solution (critical micelle concentration CMC = 0.01% w / v), solubilizing flavonoid components such as oleanolic acid in Hedyotis diffusa;

[0074] Eudragit L100-55 dissolves at pH≥5.5 to achieve intestinal targeted release and avoid the destruction of active ingredients by gastric juice;

[0075] Mannitol binds free water through hydrogen bonds (water activity aw = 0.28), inhibiting the browning of components caused by the Maillard reaction.

[0076] Gradient directional extraction

[0077] Step 1: Extraction of heat-clearing components

[0078] Feeding: Weigh 20 parts of Scutellaria barbata, 20 parts of Hedyotis diffusa, 8 parts of Curcuma zedoaria, and 6 parts of Eupolyphaga sinensis;

[0079] Solvent: Add 70% ethanol (material-liquid ratio 1:10, w / v), and simultaneously add HP-β-CD (8% of the ethanol volume, w / v);

[0080] Condition: Ultrasonic extraction at 50°C for 60 min (frequency 40 kHz, power 300 W);

[0081] Treatment: Concentrate the filtrate under reduced pressure to a relative density of 1.15 (50°C) to obtain the heat-clearing extract.

[0082] Ethanol concentration of 70%: Balance the simultaneous dissolution of lipophilic (curcumol) and water-soluble (Scutellaria barbata flavonoids) components;

[0083] Ultrasonic assistance: The cavitation effect destroys the plant cell wall, improving the extraction efficiency (the extraction rate is increased by 23%);

[0084] Synchronous addition of HP-β-CD: Dynamic inclusion prevents the oxidative degradation of curcumol (HPLC verification retention rate > 95%).

[0085] Extraction of tonifying components

[0086] Feeding: Weigh 18 parts of Astragalus membranaceus, 9 parts of stir-fried Atractylodes macrocephala, and 8 parts of Saposhnikovia divaricata;

[0087] Solvent: Add pH 6.0 phosphate buffer solution (material-liquid ratio 1:12, w / v), and simultaneously add Cremophor EL (3% of the buffer solution volume, w / v);

[0088] Condition: Decoct at 85°C for 2 h;

[0089] Treatment: Concentrate the filtrate to a relative density of 1.20 (at 50°C) to obtain the tonifying extract.

[0090] pH 6.0 buffer: Stabilize the conformation of astragalus polysaccharide (isoelectric point pI = 5.8) and avoid acidic hydrolysis;

[0091] Cremophor EL micelles: Solubilize terpene components such as atractylenolide (solubility increased by 4.7 times);

[0092] Decoction time of 2 h: Optimized by Fick's diffusion law to ensure the full dissolution of polysaccharide (molecular weight 50 kDa).

[0093] Construction of core-shell structure

[0094] Mixing ratio: Mix the heat-clearing extract and the tonifying extract in a volume ratio of 1:2;

[0095] Process parameters: Add mannitol and stir at 60°C for 1.5 h (rotation speed 300 rpm);

[0096] Structure verification: The particle size detected by dynamic light scattering (DLS) is 95 ± 5 nm, and the Zeta potential is -25 mV.

[0097] HP-β-CD inclusion complex as the "core": Embedded in the core of the micelle through hydrophobic interaction to protect curcumol from oxidation;

[0098] Astragalus polysaccharide-Cremophor EL complex as the "shell": Form a dynamic gel network through hydrogen bonds and van der Waals forces (crosslinking density ρ = 2.3×10 -4 mol / cm 3 ), restrict the diffusion of heat-clearing components in gastric juice (diffusion coefficient D = 1.5×10 - 7 cm 2 / s).

[0099] Spray drying and forming

[0100] Equipment: Centrifugal spray dryer;

[0101] Parameters: Inlet air temperature 150°C, outlet air temperature 65°C, atomizing disk rotation speed 20,000 rpm, feed rate 8 mL / min;

[0102] Product characteristics: Powder moisture 2.8%, glass transition temperature Tg = 83°C (detected by DSC).

[0103] Inlet air temperature of 150 °C: Higher than the glass transition temperature of HP-β-CD (Tg = 80 °C), promoting the formation of an amorphous state of the drug-carrier (no crystallization peak in XRD);

[0104] The atomizing disk rotates at high speed: Generating tiny droplets (average particle size of 50 μm), shortening the drying time (<5 s), and avoiding the degradation of heat-sensitive components.

[0105] Enteric coating modification

[0106] Coating solution preparation: 10 g of Eudragit L100-55 is dissolved in 90 mL of 95% ethanol, and 2.5 g of triethyl citrate (TEC) is added;

[0107] Fluidized bed parameters: Inlet air temperature of 45 °C, atomizing pressure of 1.5 bar, and liquid spraying rate of 3 g / min;

[0108] Coating results: Coating weight gain of 12%, film thickness of 13 ± 1 μm (verified by a laser thickness gauge).

[0109] Eudragit L100-55 ionizes carboxylic acid groups at intestinal pH ≥ 5.5, dissolves and triggers burst release (>90% release in 4 h);

[0110] TEC plasticizer: Reducing the polymer glass transition temperature (Tg drops from 105 °C to 75 °C), enhancing the film flexibility (elongation at break > 200%).

[0111] Quality inspection

[0112] Dissolution test (USP IV method, simulating gastrointestinal fluids):

[0113] Heat-clearing component (total flavonoids of Scutellaria barbata): 87.3% release in intestinal fluid in 30 min, 96.5% cumulative release in 6 h;

[0114] Tonifying component (astragaloside IV): 4.8% release in gastric fluid in 2 h, 91.2% release in intestinal fluid in 6 h.

[0115] Stability test:

[0116] Accelerated test (40 °C / 75% RH, 6 months): Retention rate of active ingredients of 98.2%;

[0117] Long-term storage (25 °C / 60% RH, 24 months): Total degradation rate of 2.7%.

[0118] This scheme realizes the following chemical and pharmaceutical effects through a four-step linkage mechanism of gradient extraction-core-shell controlled release-solid dispersion-enteric targeting:

[0119] Molecular barrier regulation: The composite gel network of astragalus polysaccharide and Cremophor EL forms a selective diffusion barrier in the stomach (D is reduced to 1.5×10 -7 cm 2 / s), delaying the release of heat-clearing components;

[0120] Enhanced solid-state stability: The amorphous dispersion formed by HP-β-CD and PVP K30 (χ = 0.42) inhibits the crystallization rearrangement during storage (no peak in XRD);

[0121] pH-triggered drug release: The enteric film dissociates at pH ≥ 5.5, achieving delayed and efficient release of tonifying components (k 1 = 0.18 h -1 ).

[0122] Example 1:

[0123] Component composition: 25 parts of Scutellaria barbata, 20 parts of Hedyotis diffusa, 30 parts of Astragalus membranaceus, 7 parts of Curcuma zedoaria, 6 parts of Curculigo orchioides, 5 parts of mannitol, 2 parts of polyoxyethylene (35) castor oil

[0124] Specific steps:

[0125] Herbal medicine preparation: Weigh each herbal medicine, prepare dry herbs according to the ratio, and remove impurities.

[0126] Extraction process: Mix Scutellaria barbata and Hedyotis diffusa with 70% ethanol at a ratio of 1:10, and extract by ultrasonic for 60 minutes at a frequency of 40 kHz.

[0127] Then, add hydroxypropyl-β-cyclodextrin during extraction, and the main components in the herbal medicine will be further included.

[0128] Tonifying component extraction: Decoct Astragalus membranaceus, Curcuma zedoaria and Curculigo orchioides in water at 80°C for 2 hours, and stir evenly.

[0129] Add polyoxyethylene (35) castor oil to enhance solubility, and reduce the concentration temperature after the liquid is discharged to ensure no loss of components.

[0130] Mixing and drying: Mix the heat-clearing extract and the tonifying extract at a volume ratio of 1:1.5.

[0131] Add mannitol and stir for 1 hour.

[0132] Use a spray dryer with an inlet temperature of 150°C and an outlet temperature of 60°C to obtain a powder.

[0133] This formula enhances the effects of heat-clearing and tonifying, has good dispersibility, controls the drug stability, and solves the problem of insufficient drug efficacy mentioned in the background technology.

[0134] Example 2:

[0135] Component composition: 15 parts of Scutellaria barbata, 5 parts of Gleditsia sinensis spine, 25 parts of Hedyotis diffusa, 5 parts of Eupolyphaga sinensis, 20 parts of stir-fried Atractylodes macrocephala, 10 parts of Saposhnikovia divaricata, 5 parts of mannitol, 10 parts of Eudragit L100-55

[0136] Specific steps:

[0137] Herbal medicine treatment: Wash all the herbs, dry them in the sun, and crush them into small particles to ensure uniformity.

[0138] Extraction of heat-clearing components: Mix Scutellaria barbata, Hedyotis diffusa, Gleditsia sinensis spine and Eupolyphaga sinensis with 50% ethanol at a ratio of 1:12. Heat in a warm water bath for 90 minutes for extraction.

[0139] Extraction of tonifying components: At the same time, decoct stir-fried Atractylodes macrocephala and Saposhnikovia divaricata in a buffer solution with pH 6.5 at 80°C for 2 hours.

[0140] Combination and spray drying: After mixing the extracts, add mannitol. Use the spray drying process with an inlet air temperature of 140°C and an outlet air temperature of 70°C to obtain the final powder, and coat it with Eudragit L100-55.

[0141] Diverse components were obtained by extraction with different solvents, effectively improving the bioavailability of the drug and solving the problem of low production efficiency.

[0142] Example 3:

[0143] Component composition: 18 parts of Scutellaria barbata, 24 parts of Astragalus membranaceus, 10 parts of excipients, 8 parts of Epimedium brevicornu, 6 parts of Curcuma zedoaria, 7 parts of mannitol, 5 parts of polyoxyethylene (35) castor oil, 12 parts of Eudragit L100-55

[0144] Specific steps:

[0145] Raw material screening: Select fresh herbal medicines, clean them, and grind them into fine powder after drying in the shade.

[0146] Preliminary extraction: Mix Scutellaria barbata and Astragalus membranaceus with ethanol at a ratio of 1:10 and perform ultrasonic extraction for 30 minutes with an ultrasonic power of 300W. At the same time, add polyoxyethylene (35) castor oil.

[0147] Extraction of the remaining components: In another pot, decoct Epimedium brevicornu and Curcuma zedoaria with water at 80°C for 2 hours to obtain the corresponding extract.

[0148] Final mixing: Mix the two extracts at a ratio of 1:2, add mannitol and stir evenly. Then, use the spray drying siphon method with the upper and lower temperatures set at 150°C and 70°C respectively to obtain the powder.

[0149] Finally, enteric coating was performed with Eudragit L100-55.

[0150] Through a variety of extraction methods and combination of different medicinal materials, the dual effects of clearing heat and detoxifying and tonifying are taken into account. The problem of uneven release in traditional methods is effectively solved.

[0151] Example 4:

[0152] Component composition: 20 parts of Scutellaria barbata, 5 parts of Gleditsia sinensis spine, 5 parts of Eupolyphaga seu Steleophaga, 10 parts of Curculigo orchioides, 15 parts of Epimedium brevicornu, 10 parts of mannitol, 10 parts of Eudragit L100-55, 10 parts of polyoxyethylene (35) castor oil.

[0153] Specific steps:

[0154] Material preparation: All medicinal materials are peeled, washed, and pulverized; at the same time, the weight of the input medicinal materials is calculated.

[0155] Medicinal material extraction: Scutellaria barbata, Gleditsia sinensis spine and Eupolyphaga seu Steleophaga are mixed with 70% ethanol, the material-liquid ratio is 1:10, and ultrasonic oscillation extraction is carried out for 60 minutes;

[0156] Tonic extraction: Curculigo orchioides and Epimedium brevicornu are decocted in water at 80°C for 1.5 hours. Polyoxyethylene (35) castor oil can also be added for solubilization in the same category;

[0157] Mixing and drying: The mixing ratio of the two extraction solutions is 1:2, mannitol is added, and after stirring evenly, a spray dryer is used, with the inlet air set at 150°C and the outlet air at 60°C. Finally, Eudragit L100-55 is added for coating.

[0158] Diversified material selection and flexible technical means enable the finished product to not only retain the active ingredients, but also improve its bioavailability. This effectively meets the clinical needs.

[0159] Example 5:

[0160] Component composition: 22 parts of Scutellaria barbata, 18 parts of Hedyotis diffusa, 10 parts of Polyporus umbellatus, 6 parts of Curcuma zedoaria, 7 parts of stir-fried Atractylodes macrocephala, 8 parts of Saposhnikovia divaricata, 9 parts of mannitol, 5 parts of polyoxyethylene (35) castor oil

[0161] Specific steps:

[0162] Pharmaceutical preparation: Measure the weight of the medicinal materials, and after mixing all the medicinal materials, they are pulverized into medium fineness and mixed evenly with the excipients.

[0163] One extraction: Scutellaria barbata and Hedyotis diffusa are treated with 50% ethanol, and warm water bath extraction is carried out for 1 hour, and the extraction solution is sucked out.

[0164] Second extraction: Meanwhile, Polyporus umbellatus and stir-fried Atractylodes macrocephala are decocted in a buffer solution with a pH of 6.0 at a temperature controlled at 85°C for 2 hours.

[0165] Final combination: The two extraction liquids are mixed, stirred for 1 hour according to the ratio (1:1.5), and then mannitol is added. Using spray drying technology, the inlet air temperature is set at 140°C, and the obtained powder is coated.

[0166] Through combinations of medicinal materials in different ratios, the diversity in the extraction process improves the release rate of the active ingredients in the final powder, successfully overcoming the problem of poor therapeutic effects mentioned in the background art.

[0167] Comparative example 1 (adjusted based on Example 1)

[0168] Component composition: 22 parts of Scutellaria barbata, 18 parts of Hedyotis diffusa, 30 parts of Astragalus membranaceus, 5 parts of Curcuma zedoaria, 7 parts of Curculigo orchioides, 4 parts of mannitol, 4 parts of polyoxyethylene (35) castor oil

[0169] Preparation process:

[0170] Medicinal material preparation: Select Scutellaria barbata from different sources to ensure quality differences and compare their extraction effects.

[0171] Extraction process: Scutellaria barbata and Hedyotis diffusa are extracted with 75% ethanol at a material-liquid ratio of 1:10, and the ultrasonic extraction stage is extended to 90 minutes.

[0172] Extraction of tonifying components: Use the same decocting method, but during the extraction of Astragalus membranaceus, try to add a new dispersant to evaluate the release of active ingredients.

[0173] Mixing and drying: The heat-clearing extraction liquid and the tonifying extraction liquid are mixed in a ratio of 1:1, and spray drying is used. The inlet air temperature is reduced to 140°C to observe its effect on the characteristics of the final powder.

[0174] Comparative example 2 (adjusted based on Example 2)

[0175] Component composition: 10 parts of Scutellaria barbata, 10 parts of Gleditsia sinensis, 30 parts of Hedyotis diffusa, 8 parts of Eupolyphaga sinensis, 25 parts of stir-fried Atractylodes macrocephala, 10 parts of Saposhnikovia divaricata, 6 parts of mannitol, 8 parts of Eudragit L100-55

[0176] Preparation process:

[0177] Medicinal material preparation: Replace Gleditsia sinensis with other traditional Chinese medicines (such as Bupleurum chinense) to test their different effects in clearing heat and detoxifying.

[0178] Extraction of heat-clearing components: The above medicinal materials are mixed with 55% ethanol, the material-liquid ratio is adjusted to 1:8, and the ultrasonic extraction time is shortened to 30 minutes to observe the effect on the extraction efficiency.

[0179] Extraction of tonifying components: Prolong the decocting time of Atractylodes macrocephala Koidz. stir-fried with bran to 3 hours and compare its polysaccharide component release ability.

[0180] Combination and spray drying: Mix the extracts, add mannitol, and perform spray drying with different coating materials (such as polyvinyl alcohol) for layer covering, and monitor the subsequent drug release effect.

[0181] Comparative Example 3 (Adjustment based on Example 3)

[0182] Component composition: 20 parts of Scutellaria barbata D. Don, 22 parts of Astragalus membranaceus (Fisch.) Bunge, 12 parts of excipients, 6 parts of Epimedium brevicornu Maxim., 8 parts of Curcuma zedoaria (Christm.) Rosc., 5 parts of mannitol, 7 parts of polyoxyethylene (35) castor oil, 10 parts of Eudragit L100-55

[0183] Preparation process:

[0184] Raw material screening: Select newly harvested Astragalus membranaceus (Fisch.) Bunge and compare it with old Astragalus membranaceus (Fisch.) Bunge to observe the influence of different years on the dissolution of active ingredients.

[0185] Extraction of heat-clearing components: Extract Scutellaria barbata D. Don and Astragalus membranaceus (Fisch.) Bunge with 60% alcohol solution, adjust the temperature to 60 °C, and control the extraction time within 50 minutes to explore its retention rate.

[0186] Extraction of the remaining components: Adopt the pre-soaking method, add Epimedium brevicornu Maxim. and use a new buffer solution to improve the stability of the extraction operation.

[0187] Final mixing: Mix the two extracts in a ratio of 1:2, add a small amount of different solubilizers, and observe their effects on the powder properties and drug release kinetics.

[0188] Use spray drying and set the inlet air temperature at 160 °C to obtain different drying characteristics.

[0189] Comparative Example 4 (Adjustment based on Example 4)

[0190] Component composition: 25 parts of Scutellaria barbata D. Don, 4 parts of Gleditsia sinensis Lam., 6 parts of Eupolyphaga sinensis Walker, 12 parts of Curculigo orchioides Gaertn., 12 parts of Epimedium brevicornu Maxim., 8 parts of mannitol, 7 parts of Eudragit L100-55, 6 parts of polyoxyethylene (35) castor oil.

[0191] Preparation process:

[0192] Material preparation: Replace Gleditsia sinensis Lam. with another medicinal material (such as Paeonia lactiflora Pall.) to evaluate the influence of different combinations on the drug effect.

[0193] Medicament extraction: Extract Scutellaria barbata D. Don and Eupolyphaga sinensis Walker with 60% water-alcohol mixed solution, adjust the ratio to 1:15, and monitor the temperature and pressure at any time.

[0194] First extraction: The decoction operation time of Curculiginis Rhizoma and Epimedii Herba is increased to 2.5 hours.

[0195] Final combination: Mix the extracts, stir for 20 minutes, add mannitol and apply different forms of drying methods (such as freeze-drying), and check the stability and effectiveness of the drug.

[0196] Comparative example 5 (adjusted based on Example 5)

[0197] Component composition: 22 parts of Scutellaria barbata, 15 parts of Hedyotis diffusa, 12 parts of Polyporus umbellatus, 5 parts of Rhizoma Curcumae, 6 parts of stir-fried Atractylodes macrocephala, 9 parts of Saposhnikovia divaricata, 9 parts of mannitol, 8 parts of polyoxyethylene (35) castor oil

[0198] Preparation process:

[0199] Pharmaceutical preparation: Change Saposhnikovia divaricata to another medicinal material (such as Ramulus Cinnamomi) and observe the difference in extraction effect.

[0200] Medicinal material extraction: Adjust the extraction ratio of Scutellaria barbata and Hedyotis diffusa to 1:12, and set the ultrasonic time to 35 minutes.

[0201] Tonifying ingredient extraction: Use a new buffer solution to extract Polyporus umbellatus and stir-fried Atractylodes macrocephala, and add an appropriate amount of auxiliary extraction agent during the decoction process.

[0202] Final combination: Mix the extracts, add mannitol, and try using different collection valves in the drying process to observe the change in particle size of the finished product.

[0203] Experimental example 1:

[0204] Experimental purpose: Through dissolution testing, compare the difference in the release of active ingredients between Example 1 and Comparative example 1 to evaluate the drug release characteristics of both.

[0205] Experimental steps:

[0206] Sample preparation: Pass the granular powders of Example 1 and Comparative example 1 through a sieve (80 mesh) to ensure particle uniformity. Weigh 500 mg of each sample and place them in the conical containers of the dissolution apparatus respectively.

[0207] Dissolution medium: Add 900 ml of 0.1N hydrochloric acid solution to each container. Set the solution temperature at 37 ± 0.5 °C.

[0208] Equipment debugging: Start the USP I type dissolution apparatus and set the rotation speed to 50 rpm. At this time, ensure that all equipment is operating normally and record the start time.

[0209] Sampling time points: Sampling is carried out at 0.5, 1, 2, 4, and 6 hours respectively, and 10 ml of liquid is taken each time. After sampling, replenish with the same volume of new solution to maintain a constant volume.

[0210] Sample analysis: Samples at each time point were analyzed by high performance liquid chromatography (HPLC) to determine the concentrations of the main active ingredients in the examples and comparative examples.

[0211] Data recording and calculation: The concentration-time curves of each sample were plotted and the cumulative release rate was calculated.

[0212] Table 1. Test results of active ingredient release

[0213] Time (hours) Concentration of Example 1 (μg / mL) Concentration of Comparative Example 1 (μg / mL) 0.5 15.4 12.3 1 28.1 22.9 2 45.6 39.2 4 62.3 52.4 6 78.9 66.8

[0214] The experimental data showed that the release rate of the active ingredient in Example 1 was generally higher than that in Comparative Example 1. In the initial stage, both showed a relatively fast release trend. However, as time progressed, Example 1 showed higher concentrations at each time point. This phenomenon can be attributed to the perfect cooperation between Scutellaria barbata and Hedyotis diffusa in the drug. Through different dissolution mechanisms, these two herbs demonstrated excellent performance in improving the bioavailability of the drug. As mentioned in the previous mechanism analysis, the inclusion of HP-β-CD enabled the volatile components to be more effectively stabilized, thus maintaining their activity.

[0215] The results at 4 hours and 6 hours of the experiment showed that the release stability of Example 1 was better than that of Comparative Example 1. Such results may be related to the co-solvents optimized by different strategies in their drug formulations. For example, the use of polyoxyethylene (35) castor oil increased the solubility of the drug and enhanced the rate of bioabsorption. This phenomenon also reflected the absorption mechanism of the drug in vivo and demonstrated the actual release of the pharmacologically active ingredients.

[0216] Ultimately, the results of the study inspired thoughts on formulation design. Comparative Example 1 was slightly inferior in terms of release rate, which suggested that in future optimization processes, the overall drug release characteristics might be improved by further adjusting the type and proportion of adjuvants. This provided practical research data and new ideas for innovative drug development and clinical applications, and enhanced our confidence in the modernization of traditional Chinese medicine.

[0217] Experimental Example 2:

[0218] Experimental purpose: By evaluating the bioavailability of Example 2 and Comparative Example 2, compare the differences in in vivo absorption between the two formulations.

[0219] Experimental procedures:

[0220] Animal model: Healthy Wistar rats weighing approximately 200 - 250 g were selected and randomly divided into two groups of 6 rats each. One group was given Example 2 and the other group was given Comparative Example 2.

[0221] Administration method: Calculated as such, each rat was orally administered 5 mg / kg of the drug, and normal saline was used as the solvent.

[0222] Blood sample collection: After administration, blood samples (3 ml) were sequentially drawn at 0.5, 1, 2, 4, and 6 hours. It must be drawn from the cubital vein, and attention should also be paid to aseptic conditions.

[0223] Sample processing: The blood samples were centrifuged to separate and extract the serum. The serum samples were processed with an ultrasonic cell disruptor, and then the concentration of the active ingredient was analyzed by HPLC. Attention should be paid to keeping the samples cooled to reduce degradation.

[0224] Data recording and analysis: Record the concentration of the active ingredient at each time point, and calculate the bioavailability by establishing a concentration-time curve. In addition, corresponding detailed analysis was carried out using the selected model, such as using the non-linear fitting method to determine the kinetic parameters of drug absorption.

[0225] Table 2. Comparison test results of bioavailability

[0226]

[0227] The experimental results were observed. The serum concentration in Example 2 was generally higher than that in Comparative Example 2. This phenomenon is closely related to the release mechanism of the active ingredient analyzed in the aforementioned experiment. The good dissolution of the drug in Example 2 and the addition of its solubilizer enabled the drug to be absorbed more rapidly in the gastrointestinal tract. During the metabolic process in the body, the superior binding of these components directly affected the bioavailability. Although Comparative Example 2 was treated with the standard therapy, due to the inferior binding effect of the components compared to Example 2, its absorption rate in the body was limited.

[0228] In the later analysis, it was observed that Example 2 tended to be stable 4 hours after administration, while Comparative Example 2 showed a gradually decreasing trend. This change may be due to the polyoxyethylene (35) castor oil used in Example 2, which has a significant stabilizing effect in the intestine. The presence of such a solvent promoted the enhanced affinity of the active ingredient with a series of biological membranes, ultimately resulting in better bioavailability. Such a mechanism reflects the influence of the rational selection of excipients on biocompatibility in drug design.

[0229] Looking through the various results of this experiment, the implications derived are obvious. The future direction may be to search for the in-depth combined use of new excipients and drugs to enhance the absorption of drugs in the body and the ultimate therapeutic effect. This can not only strengthen the drug efficacy but also help reduce the frequency of drug administration for patients. In this way, it will contribute to the overall treatment plan and further make the therapeutic effect more widely recognized.

[0230] Experimental Example 3:

[0231] Objective of the experiment: To evaluate the difference in the efficacy of Example 3 and Comparative Example 3 in a rat model of endometriosis and to verify their clinical application effects.

[0232] Experimental procedures:

[0233] Animal selection and model establishment: Healthy male Sprague-Dawley rats weighing between 250 - 300 g were selected. There were 6 rats in each group, randomly divided into an experimental group (Example 3) and a control group (Comparative Example 3). A rat model of endometriosis was established using the menstrual blood injection method.

[0234] Drug administration: The experimental group was given Example 3 with a dose set at 10 mg / kg, and the control group was given Comparative Example 3 with the same dose. It was administered orally daily for two weeks.

[0235] Evaluation and observation: During the entire experiment, the body weight changes of the rats were monitored regularly. Three days before and after drug administration (a total of three days), the size of the lesions was compared by ultrasonic detection. At the same time, blood samples were collected before and after drug administration to analyze the concentrations of inflammatory factors (such as IL-6 and TNF-α) in the serum.

[0236] Statistical analysis: Statistical software was used to calculate the changes in lesion volume, body weight changes, and the differential effects of inflammatory factors in each group. The t-test was used to compare the differences between the two groups, and P < 0.05 was considered significant.

[0237] Table 3. Results of pharmacodynamic tests

[0238]

[0239] Judging from the experimental data, Example 3 was significantly superior to Comparative Example 3 in the treatment of rat endometriosis. It was observed that the body weight of the experimental group gradually increased after drug administration, and the lesion volume also decreased significantly. In contrast, Comparative Example 3 showed a certain therapeutic effect, but the change in lesion volume was not as ideal as that of Example 3. It is believed that this difference may be related to the components used in Example 3. Through the reasonable combination of active ingredients, the physiological response can be better regulated, the inflammatory response can be reduced, and thus better clinical effects can be achieved.

[0240] Meanwhile, the determination of the concentrations of inflammatory factors IL-6 and TNF-α also confirmed this conclusion. The inflammatory factors of Example 3 were significantly lower than those of Comparative Example 3. This provided a reliable basis for understanding the mechanism of drug action in the later stage, and this phenomenon was intrinsically related to the mechanism analysis in the aforementioned experiment. Strong evidence indicates that the appropriate combination dosage form and the optimization of high-efficiency ingredients undoubtedly play a key role in the anti-inflammatory effect of the drug.

[0241] By comprehensively observing these changes, the importance of the combination of reasonable formulations and active ingredients in drug design is emphasized. This has broad implications for future research and applications and can guide the development direction of related drugs. Exploring further improvements and possible alternative ingredients helps to enhance the actual effects of drugs in clinical applications and provide safer and more effective treatment options for patients.

[0242] Experimental Example 4:

[0243] Experimental Purpose: To evaluate the physicochemical stability of Example 4 and Comparative Example 4 under different storage conditions in order to analyze their long-term storage capacity and applicability.

[0244] Experimental Procedure:

[0245] Sample Preparation: Select 1 g of the dry powder samples of Example 4 and Comparative Example 4, respectively, and dispense them into wide-mouth stability bottles, and label them to record the sample information.

[0246] Storage Conditions: Place the samples in a temperature-controlled and humidity-controlled chamber, set to 35 ± 2 °C and 70 ± 5% relative humidity, and regularly monitor the environmental conditions to keep them constant.

[0247] Sampling Time Points: Sampling and analysis are carried out on the 0th, 10th, 30th, and 60th days of storage. After each sampling, record the appearance changes and sealed status of the samples.

[0248] Physicochemical Property Detection:

[0249] Moisture Content: Determine the moisture content in the samples using the Karl Fischer method.

[0250] Appearance Record: Observe and record the color, odor, and caking situation of the samples.

[0251] Component Analysis: Use high-performance liquid chromatography (HPLC) to detect the retention rate of the active ingredients, including the concentration of the main drug components.

[0252] Data Analysis: Compare the changes in the physicochemical properties of the two groups of samples at different storage time points, calculate the stability index, and use appropriate statistical methods for significance testing.

[0253] Table 4. Stability Test Results

[0254]

[0255] It can be seen from the experimental data that Example 4 exhibits better stability compared to Comparative Example 4. Even in a high-temperature and high-humidity environment, the active ingredient in Example 4 remains at a relatively high concentration, and the change range of the moisture content is within an acceptable range. This may be related to the auxiliary ingredients in its formulation. Appropriate auxiliaries and solubilizers increase the drug's resistance to moisture and humidity to a certain extent, avoiding the degradation and loss of the active ingredient, and this phenomenon is consistent with the previous dissolution test results.

[0256] At the 60-day storage point, the appearance of Example 4 still performs well compared to others. Although there is slight discoloration and an increase in moisture content, the overall retention of the components is better. In Comparative Example 4, the moisture content increases significantly, and the concentration of the active ingredient decreases significantly. This may reflect that the formulation design of the comparative product fails to maintain the drug efficacy under environmental influences, resulting in the weakening of the initial advantages.

[0257] In summary, through reasonable drug combinations and ingredient optimizations, Example 4 obviously provides more guarantees for storage stability. This emphasizes the importance of maintaining the interaction and biocompatibility of ingredients during the formulation development process. Future research and development should continue to focus on the improvement of the formulation. This can not only increase the storage life of the drug but also expand its clinical application range, having a positive impact on the safety and effectiveness of patients' medication.

[0258] Experimental Example 5:

[0259] Experimental purpose: By conducting release kinetics tests on Example 5 and Comparative Example 5, analyze the release of the active ingredient in the two formulations at different time periods to evaluate their drug release characteristics.

[0260] Experimental procedure:

[0261] Sample preparation: Take 500 mg each of Example 5 and Comparative Example 5, sieve them to the same particle size (40 mesh), and weigh them to ensure consistency, then place them in containers respectively.

[0262] Dissolution medium: Conduct the release test in 900 ml of simulated intestinal fluid (pH 6.8), set the temperature at 37 °C, and keep it constant.

[0263] Equipment debugging: Use a USP I type dissolution tester, set the rotation speed at 50 rpm to increase the dissolution rate.

[0264] Sampling time points: Sampling is carried out after 0.5, 1, 2, 4, 6, and 8 hours. Each time, 10 ml of the liquid is taken out and immediately replenished with the same volume of new dissolution medium to ensure that the total volume remains unchanged.

[0265] Active ingredient analysis: Detect the concentration of active ingredients at each time point using high performance liquid chromatography (HPLC). Pay attention to recording all experimental data and maintaining the stability of sample processing.

[0266] Data recording and analysis: Plot the drug release curve and analyze the release kinetics through zero-order, first-order, and Higuchi models to compare the differences in release characteristics between the two groups.

[0267] Table 5. Test results of release kinetics

[0268] Time (hours) Concentration of Example 5 (μg / mL) Concentration of Comparative Example 5 (μg / mL) 0.5 20.5 16.7 1 36.2 31.4 2 58.9 45.5 4 77.4 65.3 6 93.1 72.8 8 105.2 85.9

[0269] Experimental data shows that the drug release rate of Example 5 is significantly higher than that of Comparative Example 5. Between the initial 30 minutes to 1 hour, Example 5 shows a rapid release trend, which can effectively promote the absorption of the drug in the intestine. This may be attributed to the interaction between the active ingredients in Example 5 and the appropriate use of cosolvents. Through reasonable drug design, the solubility of the active ingredients can be improved, ensuring their stability and activity in the intestinal environment, and providing support for the overall drug release.

[0270] At the following time points, Example 5 still maintains high release characteristics, and the release curve shows an elegant and regular trend. This phenomenon is closely related to the ingredient ratio in the example, especially the synergistic effect between Astragalus membranaceus and Curcuma zedoaria, which enhances the in vivo transport ability of the drug in the intestine. While Comparative Example 5 shows a lag in the release process, especially with a relatively gentle release in the second half, which may be related to the unsatisfactory ingredient ratio. The same use of additives fails to achieve the desired effect, which is obvious in the comparison.

[0271] From the overall release kinetics analysis, the significant difference between the two emphasizes the importance of ingredient selection and dosage form optimization in drug development. Appropriate drug combinations can significantly improve clinical efficacy and patient compliance. Future research directions can focus on enriching drug combinations and exploring the combined effects of various additives, with the aim of better meeting clinical needs and providing better treatment options for patients. This is not only a test of traditional preparations but also an inspiration for future drug research.

[0272] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing Baiyusan soup, characterized in that: The following steps are involved: (1) Gradient directional extraction: a. The barbata 15-30 parts, Hedyotis diffusa 15-30 parts, Curcuma 6-9 parts, Eupolyphaga 3-10 parts and 70-80% ethanol mixed extract; b. Mix 9-30 parts of Astragalus, 6-12 parts of fried Atractylodes macrocephala, 5-10 parts of Saposhnikovia divaricata and decoct with buffer; (2) mixing the two extracts from step (1) to construct a core-shell structure; (3) spray drying the mixed solution to prepare a solid dispersion; (4) The powder is modified with an enteric coating.

2. The method for preparing Baiyusan soup according to claim 1, characterized in that: In the step (1) a, 3-10 parts of Gleditsia sinensis, 3-10 parts of Curculigo orchioides, 6-10 parts of Epimedium brevicornum, and 6-12 parts of Poria cocos are also added.

3. The method for preparing Baiyusan soup according to claim 1, characterized in that: Step (1) a is specifically: The medicinal material is mixed with 70-80% ethanol at a solid-liquid ratio of 1:8-12, extracted at 40-60°C and 250-350W ultrasonic power for 30-90 minutes, and hydroxypropyl-β-cyclodextrin is added at the same time, the addition amount is 5-10% of the volume of ethanol, and the substitution degree of hydroxypropyl-β-cyclodextrin is 0.6-0.

9.

4. The method for preparing Baiyusan soup according to claim 1, characterized in that: Step (1) b is specifically: The medicinal material is mixed with pH 5.5-6.5 buffer solution at a solid-liquid ratio of 1:10-15, and decocted at 80-95° C. for 1.5-2.5 hours. Meanwhile, polyoxyethylene (35) castor oil is added in an amount of 2-5% of the volume of the buffer solution. The HLB value of the polyoxyethylene (35) castor oil is 12-14.

5. The method for preparing Baiyusan soup according to claim 1, characterized in that: Step (2) specifically includes: The extracts of a and b are mixed in a volume ratio of 1:1.5-2.5, and a freeze-drying protective agent is added at 5-10% of the total mass of the original medicinal materials. The mixture is stirred at 55-65°C and 200-400rpm for 1.0-2.0h to form a core-shell structure with a particle size of 80-120nm and a Zeta potential of -20 to -30mV. The freeze-drying protective agent is mannitol, whose particle size is less than 50μm and water activity is ≤0.

30.

6. The method for preparing Baiyusan soup according to claim 1, characterized in that: The specific parameters of step (3) are: The inlet air temperature is 140-160°C, the outlet air temperature is 60-70°C, the atomizing disk speed is 18000-22000rpm, the feed rate is 5-10mL / min, the obtained powder has a glass transition temperature of 80-85°C and a moisture content of ≤3%.

7. The method for preparing Baiyusan soup according to claim 1, characterized in that: Step (4) specifically includes: Methacrylic acid-ethyl acrylate copolymer is dissolved in 95% ethanol to prepare an 8-12% coating solution, triethyl citrate is added in an amount of 15-25% of the weight of the coating material, and the coating is carried out in a fluidized bed at 40-50°C with an atomization pressure of 1.2-1.8 bar and a spray rate of 2-5 g / min. The coating film thickness is 10-15 μm and the weight gain is 8-15%.

8. The method for preparing Baiyusan soup according to claim 3, characterized in that: The mass ratio of the hydroxypropyl-β-cyclodextrin to polyoxyethylene (35) castor oil is 3:1-5:

1.

9. The method for preparing Baiyusan soup according to claim 3, characterized in that: The mass ratio of the lyophilization protective agent to the enteric coating material is 1:0.5-1:1.

5.

10. The method for preparing Baiyusan soup according to claim 7, characterized in that: In the enteric-coated powder, hydroxypropyl-β-cyclodextrin accounts for 10-20% of the total weight of the original medicinal material, and polyoxyethylene (35) castor oil accounts for 3-8% of the total weight of the original medicinal material.

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