Preparation and application of a compound preparation of Herba Lycopodii, Scutellariae Scutellariae, Ganoderma Lucidum, Houttuynia Cordata and Astaxanthin for adjuvant treatment of tumors

Abstract

This invention belongs to the field of biopharmaceuticals and specifically discloses the preparation and application of a compound formulation of Hedyotis diffusa (5-30 parts), Scutellaria barbata (3-10 parts), Houttuynia cordata (3-6 parts), Ganoderma lucidum (2-4 parts), and astaxanthin for adjuvant tumor treatment. This compound is prepared by gradient decoction, nanoemulsification, and synergistic combination of ingredients, including Hedyotis diffusa (5-30 parts), Scutellaria barbata (3-10 parts), Houttuynia cordata (3-6 parts), Ganoderma lucidum (2-4 parts), and astaxanthin (0.5-1 parts). The combination of gradient temperature-controlled decoction and nanoemulsification (particle size 50-200 nm) significantly improves the bioavailability of heat-sensitive components (such as astaxanthin), allowing for the design of various dosage forms (tablets, capsules, and pills) to meet diverse clinical needs. Animal models have demonstrated that this compound enhances the efficacy of cytarabine and cisplatin while reducing chemotherapy toxicity (resumption of ALT / AST levels to normal). This invention combines multi-target anti-tumor and immunomodulatory properties with dosage form stability, providing an innovative solution for adjuvant tumor treatment.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and specifically discloses the preparation and application of a compound preparation of Herba Epimedii, Scutellariae Barbatae, Ganoderma Lucidum, Houttuynia Cordata and Astaxanthin for adjuvant treatment of tumors. Background Art

[0002] Malignant tumors have become one of the major diseases threatening human health. Currently, clinical treatments mainly rely on surgery, radiotherapy, and chemotherapy, but these therapies are often accompanied by significant side effects, such as decreased immunity and organ damage, which seriously affect the quality of life of patients. Against this background, traditional Chinese medicine adjuvant therapy has gradually gained attention. Traditional Chinese medicine compound prescriptions, due to the synergistic effects of multiple ingredients, have shown unique advantages in enhancing the body's immune function, reducing the toxicity of radiotherapy and chemotherapy, and inhibiting tumor metastasis. For example, heat-clearing and detoxifying Chinese medicines such as Hedyotis diffusa and Scutellaria barbata have been recognized to have anti-tumor activity; ingredients such as Ganoderma lucidum polysaccharides and astaxanthin can regulate the immune microenvironment. However, existing compound preparations are mostly limited to conventional medicinal material combinations, with a single active ingredient, and traditional decoction processes make it difficult to effectively retain heat-sensitive ingredients, resulting in limited efficacy.

[0003] Furthermore, current traditional Chinese medicine (TCM) compound formulations for adjuvant cancer therapy face the following technical bottlenecks: First, the compositional combinations lack innovation; most compound formulations simply combine known anti-tumor herbs without incorporating unconventional ingredients for synergistic effects. Second, the preparation process is crude, and research into nanoscale delivery technologies for fat-soluble components (such as astaxanthin) is insufficient, impacting bioavailability. Third, the dosage form design is limited, making it difficult to meet the needs of diverse patients. For example, traditional decoctions are inconvenient to carry, while modern solid preparations may lose some volatile active substances. Furthermore, existing compound formulations have limited specific research on hematologic malignancies, and there is a lack of broad-spectrum formulations that address both solid and hematologic malignancies. To address these challenges, there is an urgent need to develop TCM compound formulations with innovative components, advanced processes, and diverse dosage forms that could enhance the efficacy of adjuvant cancer therapy through multi-target synergy, while minimizing toxicity and side effects and improving patient compliance. Summary of the Invention

[0004] In order to solve the above problems, the present invention discloses the preparation and application of a compound preparation of Herba Epimedii, Scutellariae Barbatae, Ganoderma Lucidum, Houttuynia Cordata and Astaxanthin for adjuvant treatment of tumors.

[0005] The purpose of the present invention is achieved through the following technical solutions.

[0006] A compound preparation of Herba Lycopodii, Scutellariae Scutellariae, Ganoderma Lucidum, Houttuynia Cordata and Astaxanthin for adjuvant treatment of tumors, which is prepared from the following raw materials in parts by weight:

[0007] 5-30 portions of Hedyotis diffusa

[0008] 3-10 servings of Scutellaria barbata

[0009] 3-6 servings of Houttuynia cordata

[0010] 2-4 servings of Ganoderma lucidum

[0011] Astaxanthin 0.5-1 part

[0012] 0.2-0.8 parts sea buckthorn oil

[0013] Gynostemma pentaphyllum extract 0.1-0.5 parts

[0014] 0.1-0.3 parts of cat's claw extract

[0015] Rubescensine A 0.1-0.2 parts

[0016] Andrographolide 0.05-0.2 parts.

[0017] The present invention also discloses a method for preparing the compound, comprising the following steps:

[0018] 1) Grind Hedyotis diffusa, Scutellaria barbata, Houttuynia cordata, and Ganoderma lucidum into powders in the form of a Chinese medicinal beverage by weight, pass through a 50-mesh sieve, mix, add water and decoct 2-3 times, filter through 5-10 layers of gauze, combine the decoctions, and concentrate to a clear paste with a relative density of 1.10-1.25;

[0019] 2) mixing astaxanthin and sea buckthorn oil uniformly and then performing nanoemulsification treatment under nitrogen protection;

[0020] 3) Evenly mixing the Gynostemma pentaphyllum extract, Radix Cat's Claw extract, Rubescensine A, andrographolide and the emulsion obtained in step 2), and adding the mixture to the clear paste in step 1) to obtain the compound.

[0021] Furthermore, in the above preparation method, step 1) decoction is controlled by gradient temperature: the first decoction is maintained at 100°C for 30 minutes, then the temperature is lowered to 80°C and decoction is continued for 1 hour, and the second decoction is continued at 90°C for 1.5 hours.

[0022] Furthermore, in the above preparation method, in step 2), the nanoemulsified particle size is controlled to be 50-200 nm, and high-pressure homogenization combined with ultrasonic treatment is used.

[0023] Furthermore, the above preparation method further comprises step 4) adding pharmaceutically acceptable excipients to prepare the preparation.

[0024] Furthermore, the above preparation method is used to make tablets containing the following excipients: 10-30% microcrystalline cellulose, 2-5% cross-linked sodium carboxymethyl cellulose, and 0.5-1.5% magnesium stearate.

[0025] Furthermore, the above preparation method is used to prepare capsules, which contain enteric coating materials, and the coating materials are selected from hypromellose phthalate or hypromellose acetate succinate.

[0026] Furthermore, the above preparation method is used to make pills, which contain a honey blending agent, and the pill diameter is controlled at 3-8 mm and the moisture content is ≤12%.

[0027] Furthermore, the above preparation method, for making the pill, comprises the following process: preparing by the lifting pill method, controlling the calcination temperature at 200-300°C, and the time for 2-4 hours.

[0028] The present invention also discloses the use of the compound in preparing a tumor adjuvant therapeutic drug, which is characterized in that the tumor includes solid tumors and blood system tumors.

[0029] Compared with the existing technology, the present invention has the following advantages and beneficial effects:

[0030] This invention utilizes Hedyotis diffusa (heat-clearing and detoxifying), Scutellaria barbata (anti-inflammatory and anti-proliferative), and Ganoderma lucidum (immunomodulatory) to form a foundational anti-tumor matrix. Combined with astaxanthin (a highly effective antioxidant) and Rubescensine A (apoptosis-inducing), this formulation establishes a multi-target synergistic network, overcoming the limitations of traditional compound formulas with their single mode of action. A gradient temperature-controlled decoction process (high temperature for rapid precipitation of fat-soluble components, low temperature for retention of heat-sensitive active substances) combined with nanoemulsification technology (particle size <200 nm) addresses the technical challenge of low bioavailability of fat-soluble components in traditional Chinese medicine compounds. In terms of dosage form design, enteric-coated capsules utilize a pH-sensitive coating to achieve intestinal-targeted drug release, while the pill utilizes calcination co-crystallization to enhance stability, balancing traditional medication practices with the demands of modern precision medicine. When used in combination with chemotherapy drugs, the Gynostemma pentaphyllum extract in the compound inhibits the expression of drug-resistance proteins in tumor cells, while andrographolide mitigates chemotherapy hepatotoxicity. The synergy index (CI) exceeds 1.3, significantly reducing chemotherapy dosage requirements. This program is suitable for the adjuvant treatment of solid tumors and blood tumors, and can be flexibly adapted to various administration methods such as oral and enteric-coated, providing standardized, highly compatible innovative preparations for the combined treatment of tumors with Chinese and Western medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Comparison of serum IL-2 (pg / mL) concentrations in the evaluation of immunomodulatory effects;

[0032] Figure 2 Comparison of spleen lymphocyte proliferation (OD450) data in the evaluation of immunomodulatory effects;

[0033] Figure 3 Comparison of data on thymus index (mg / g) in the evaluation of immunomodulatory effects;

[0034] Figure 4Comparison of median survival (days) in hematologic malignancy synergy testing;

[0035] Figure 5 Comparison of tumor volume (mm³, day 21) in solid tumor synergy testing;

[0036] Figure 6 Comparison of apoptosis rate (%) in solid tumor synergy assay. DETAILED DESCRIPTION

[0037] In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is further described in detail below. However, it should be understood that the specific embodiments described herein are merely for the purpose of explaining the present invention and are not intended to limit the scope of the invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention. All raw materials in the embodiments of the present invention can be obtained through commercial channels.

[0038] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present invention will be described in detail below with reference to the embodiments.

[0039] The main reagents in the examples of the present invention are shown in Table 1, and the main instruments are shown in Table 2.

[0040]

[0041]

[0042] Example 1

[0043] Raw material ratio (parts by weight):

[0044] 5 parts of Hedyotis diffusa, 3 parts of Scutellaria barbata, 3 parts of Houttuynia cordata, 2 parts of Ganoderma lucidum, 0.5 parts of astaxanthin, 0.2 parts of sea buckthorn oil, 0.1 parts of Gynostemma pentaphyllum extract, 0.1 parts of Cat's Claw extract, 0.1 parts of Rubescensine A, and 0.05 parts of Andrographolide.

[0045] Preparation method:

[0046] 1) Weigh four Chinese herbs, including Hedyotis diffusa, according to the appropriate ratio, grind them, pass through a 50-mesh sieve, mix them, add water, and decoct them twice: first at 100°C for 30 minutes, then cool to 80°C for 1 hour, and then decoct them at 90°C for 1.5 hours. The combined decoctions are concentrated to a paste with a relative density of 1.10.

[0047] 2) Astaxanthin was mixed with sea buckthorn oil and treated with high-pressure homogenization (100 MPa) and ultrasound (20 kHz) under nitrogen protection to produce a nanoemulsion with an average particle size of 50 nm.

[0048] 3) Mixing the remaining ingredients such as the Gynostemma pentaphyllum extract with the nanoemulsion and adding the mixture to the clear paste to prepare a compound preparation.

[0049] Example 2

[0050] Raw material ratio: 30 parts of Hedyotis diffusa, 10 parts of Scutellaria barbata, 6 parts of Houttuynia cordata, 4 parts of Ganoderma lucidum, 1 part of astaxanthin, 0.8 parts of sea buckthorn oil, 0.5 parts of Gynostemma pentaphyllum extract, 0.3 parts of Cat's Claw extract, 0.2 parts of Rubescensine A, and 0.2 parts of Andrographolide.

[0051] The preparation method is basically the same as that of Example 1, except that the decoction is concentrated to a relative density of 1.25 and nanoemulsified to an average particle size of 200 nm.

[0052] Example 3

[0053] Raw material ratio: 15 parts of Hedyotis diffusa, 6.5 parts of Scutellaria barbata, 4.5 parts of Houttuynia cordata, 3 parts of Ganoderma lucidum, 0.75 parts of astaxanthin, 0.5 parts of sea buckthorn oil, 0.3 parts of Gynostemma pentaphyllum extract, 0.2 parts of Cat's Claw extract, 0.15 parts of Rubescensine A, and 0.12 parts of Andrographolide.

[0054] The preparation method is basically the same as that of Example 1, wherein the average nanoemulsified particle size is 120 nm, and tablets are prepared by adding 20% ​​microcrystalline cellulose, 3% croscarmellose sodium, and 1% magnesium stearate.

[0055] Example 4

[0056] The raw material ratio is the same as that in Example 3. In step 4), enteric coating is performed by adding hypromellose phthalate to prepare capsules.

[0057] Example 5

[0058] The raw material ratio is the same as that in Example 2. In step 4), the pellets are calcined using the lifting and lowering pellet method (250° C., 3 hours) to prepare pellets with a moisture content of 10%.

[0059] Comparative Example 1

[0060] Astaxanthin and sea buckthorn oil were omitted, and the remaining ingredients and process were the same as those in Example 3.

[0061] Comparative Example 2

[0062] No gradient decoction was performed: decoction was performed at 100° C. for 3 hours, and the rest was the same as in Example 1.

[0063] Comparative Example 3

[0064] The nanoemulsified particle size was 400 nm (only high-pressure homogenization treatment was performed, without ultrasound), and the rest was the same as in Example 2.

[0065] Comparative Example 4

[0066] Rubescensine A and andrographolide were not added, and the rest were the same as in Example 3.

[0067] Comparative Example 5

[0068] Conventional tablet excipients (50% starch) were used, and no cross-linked sodium carboxymethyl cellulose was added. Other modifications were the same as in Example 3.

[0069] Test Example 1

[0070] In vitro antitumor activity test

[0071] 1. Test method:

[0072] Cell line: HepG2 liver cancer cells, cultured in DMEM medium containing 10% FBS.

[0073] Treatment: The compound of Example 3 (0-100 μg / mL) was used for treatment for 48 hours, and the cell survival rate was measured by MTT assay.

[0074] 2. Grouping: blank control group, Example 3 group (gradient concentration), Comparative Example 1 group (without astaxanthin), and Comparative Example 4 group (lacking Rubescensine A).

[0075] The results are shown in Table 3.

[0076]

[0077] Table result analysis:

[0078] 1. Dose-dependent effect:

[0079] The compound showed significant concentration-dependent inhibition in the range of 10-100 μg / mL, which was much lower than that of traditional Chinese medicine compounds (usually IC50>50 μg / mL), indicating its high efficacy.

[0080] 2. Synergistic mechanism of ingredients:

[0081] Astaxanthin: As a potent antioxidant, it inhibits the NF-κB pathway in HepG2 cells by scavenging ROS and reduces the expression of the anti-apoptotic protein Bcl-2 (Western blot verification, data not shown);

[0082] Rubescensine A: Directly activates the Caspase-3 / 9 pathway to induce apoptosis, forming a dual-target synergy of "oxidative stress regulation-apoptosis execution" with astaxanthin.

[0083] 3. Impact of missing components:

[0084] The IC50 of comparative example 1 (without astaxanthin) was missing, suggesting that astaxanthin is the key to overcoming the antioxidant defense of tumor cells; the IC50 of comparative example 4 (without ordovicianine) increased by 2.8 times, confirming the irreplaceable role of this ingredient in the apoptosis pathway.

[0085] 4. Horizontal comparison:

[0086] Compared with the single Houttuynia cordata extract (IC50≈45 μg / mL) reported in the literature, the activity of this compound is increased by 3.6 times, reflecting the advantages of multi-component integration.

[0087] Test Example 2

[0088] Evaluation of immunomodulatory effects

[0089] 1. Test Purpose

[0090] To verify the immune recovery effect of the compound on immunosuppressed mice after chemotherapy, focusing on evaluating the IL-2 secretion level and spleen lymphocyte proliferation capacity.

[0091] 2. Test methods

[0092] Animal model: ICR mice (male, 8 weeks old, n=40) were injected intraperitoneally with cyclophosphamide (80 mg / kg, for 3 consecutive days) to establish an immunosuppressive model.

[0093] 3. Grouping and drug administration (10 mice per group):

[0094] Normal control group: no model, oral administration of normal saline

[0095] Model control group: after modeling, normal saline was gavaged

[0096] Low-dose compound group: Compound of Example 3 (100 mg / kg / d, gavage)

[0097] High-dose compound group: Compound of Example 3 (200 mg / kg / d, gavage)

[0098] 4. Testing indicators:

[0099] Serum IL-2: ELISA kit (R&D Systems)

[0100] Splenic lymphocyte proliferation: OD450 value was measured by CCK-8 method after ConA stimulation

[0101] Thymus index: thymus weight (mg) / body weight (g)

[0102] The results are shown in Table 4 and Figure 1-3

[0103]

[0104] Result Analysis

[0105] 1. Immune reconstruction effect:

[0106] The high-dose compound restored IL-2 to a normal level of 85.2 pg / mL and the thymic index to 3.5 mg / g, close to a healthy state, suggesting that its ability to repair the thymic microenvironment may be achieved by upregulating FoxN1 expression in thymic epithelial cells.

[0107] 2. Analysis of the functions of core components:

[0108] Ganoderma lucidum polysaccharide: activates the TLR4 receptor of dendritic cells, promotes IL-12 secretion, and enhances Th1 immune response;

[0109] Astaxanthin: Inhibits chemotherapy-induced Treg cell expansion and reverses immune tolerance.

[0110] 3. Dose-effect relationship:

[0111] The low-dose group only partially recovered immune function (IL-2 was 68.4 pg / mL), indicating that 200 mg / kg / d is the optimal dose. Compared with commonly used immunomodulators in clinical practice (such as thymosin), it has the safety advantage of natural ingredients.

[0112] 4. Clinical translational significance:

[0113] This compound can reduce the need for granulocyte colony-stimulating factor (G-CSF) use after chemotherapy and lower treatment costs.

[0114] Test Example 3

[0115] Bioavailability evaluation

[0116] 1. Test Purpose

[0117] The bioavailability of astaxanthin nanoemulsion in the compound preparation was compared with that of traditional preparation to verify the advantages of nanoemulsification technology.

[0118] 2. Test methods

[0119] Animal model: SD rats (male, 250-300 g, n=24) were randomly divided into 3 groups:

[0120] Traditional preparation group: non-nanoemulsified astaxanthin suspension (0.5 mg / kg)

[0121] Nanoemulsion group: the compound of Example 1 (containing 0.5 mg / kg astaxanthin)

[0122] Comparative Example 3: Astaxanthin crude emulsion (particle size 400 nm, 0.5 mg / kg)

[0123] Dosing and sampling: Blood was collected at 0.5, 1, 2, 4, 8, 12, and 24 hours after a single oral gavage, and the plasma astaxanthin concentration was determined by HPLC (Agilent 1260, C18 column, detection wavelength 476 nm).

[0124] Data analysis: calculating AUC 0-24h and C max .

[0125] The results are shown in Table 5

[0126]

[0127] Table result analysis

[0128] 1. Pharmacokinetic advantages of nanoemulsification:

[0129] Absorption acceleration: T max The reduction from 4 hours to 2 hours was attributed to the lymphatic absorption pathway of the nanoemulsion through the small intestinal M cells, avoiding the first-pass effect;

[0130] Increased exposure: AUC increased by 2.3 times, which was related to the sea buckthorn oil in the nanoemulsion promoting bile secretion and increasing the solubility of fat-soluble components (verified by the bile duct cannulation model).

[0131] Effect of particle size on bioavailability:

[0132] The AUC of comparative example 3 (400 nm) was only 64.8% of that of the nanoemulsion group, confirming that the particle size <200 nm is the key threshold for breaking through the intestinal mucus layer (refer to the consensus in the field of nanodrug delivery).

[0133] 2. Breakthrough of technical barriers:

[0134] The bioavailability of fat-soluble components (such as andrographolide) in traditional Chinese medicine compounds is usually <10%, but this process increases it to 28.7 μg·h / mL, reaching the level of chemical drugs.

[0135] Test Example 4

[0136] Enteric-coated capsule release test

[0137] 1. Test Purpose

[0138] Verify the protective effect of enteric coating on heat-sensitive ingredients (such as gypenosides) in compound dosage forms to ensure targeted release in the intestine.

[0139] 2. Test methods

[0140] Sample preparation: capsules of Example 4 (containing a hypromellose phthalate coating) and an uncoated control group.

[0141] Release conditions:

[0142] Simulated gastric fluid: 0.1 M HCl (pH 1.2), 37°C, 100 rpm, 2 hours

[0143] Simulated intestinal fluid: phosphate buffer (pH 6.8), followed by 4 hours

[0144] Detection method: The cumulative release rate of gypenosides was determined by ultraviolet spectrophotometry (UV 320 nm).

[0145] The results are shown in Table 6

[0146]

[0147] Table result analysis

[0148] 1. Scientific selection of coating materials:

[0149] Hydroxypropyl methylcellulose phthalate (HPMCP) dissolves at pH ≥ 5.0, perfectly matching the pH environment of the duodenum (5.0-6.5), preventing gastric acid from hydrolyzing the glycosidic bonds of gypenosides.

[0150] 2. Release kinetics analysis:

[0151] Enteric-coated capsules exhibited zero-order release characteristics in intestinal fluid (R²=0.992), which was consistent with the Higuchi model, indicating that the release was controlled by diffusion, which was beneficial to maintaining steady-state blood drug concentration.

[0152] 3. Comparison with conventional preparation defects:

[0153] Uncoated capsules released 85% of their drug in gastric fluid within 1 hour, resulting in a 40% decrease in the absolute bioavailability of gypenosides (confirmed by in vivo gastric perfusion experiments in rats), highlighting the necessity of enteric-coated design.

[0154] Test Example 5

[0155] Long-term stability accelerated test

[0156] 1. Test Purpose

[0157] Evaluate the stability of active ingredients in different dosage forms (pills and tablets) under high temperature and high humidity conditions and verify the superiority of the pill preparation process.

[0158] 2. Test methods

[0159] Samples and conditions:

[0160] Example 5 pill (prepared by the Shengjiang pill method) and comparative example 5 tablet (starch excipient)

[0161] Accelerated conditions: 40°C / 75% RH, for 6 months

[0162] Detection indicators:

[0163] Oridonin retention rate: HPLC method (mobile phase acetonitrile-water, detection wavelength 242 nm)

[0164] Andrographolide retention rate: same as above, detection wavelength 225 nm.

[0165] The results are shown in Table 7

[0166]

[0167] Table result analysis:

[0168] 1. The physical and chemical basis of the stability of pills:

[0169] Calcination process: According to the literature, calcination at 200-300°C can form a eutectic structure between oridonin and andrographolide, reducing molecular mobility;

[0170] Moisture control: The moisture content of the pill is ≤12%, which inhibits the hydrolysis reaction, while the moisture content of the starch in the tablet is >15% after absorbing moisture, which accelerates the degradation of glycoside components.

[0171] 2. Enlightenment of scientific compatibility of excipients:

[0172] The use of starch in Comparative Example 5 resulted in a retention rate of <65%, suggesting that future development requires the use of hydrophobic excipients (such as microcrystalline cellulose) or the addition of anti-caking agents such as silicon dioxide.

[0173] 3. Calculation of shelf life:

[0174] According to the ICH Q1A guidelines, the predicted shelf life of the pill at 25°C / 60% RH is >36 months, while that of the tablet is only 12 months, significantly improving the commercial value of the product.

[0175] Test Example 6

[0176] Testing synergistic efficacy in combination with cytarabine for the treatment of leukemia (blood cancer)

[0177] 1. Test Purpose

[0178] To verify the synergistic therapeutic effect of the combination of the compound and the chemotherapy drug cytarabine on the L1210 leukemia mouse model, and to evaluate the effects of prolonging survival and reducing tumor burden.

[0179] 2. Test methods

[0180] Animal model: BALB / c mice (female, 6 weeks old, n=50) were injected with L1210 leukemia cells (1×10^6 cells / mouse) via the tail vein.

[0181] Grouping and dosing (10 mice per group):

[0182] Control group: Oral administration of normal saline

[0183] Compound single-drug group: Compound of Example 3 (200 mg / kg / d, gavage)

[0184] Cytarabine single-drug group: cytarabine (20 mg / kg, intraperitoneal injection, once every 3 days)

[0185] Combination group: compound drug (200 mg / kg / d) + cytarabine (10 mg / kg, half dose)

[0186] Detection indicators:

[0187] Survival period (time from vaccination to death)

[0188] Peripheral blood leukemia cell count (CD45+ cell percentage by flow cytometry)

[0189] Spleen weight (an indicator of tumor infiltration)

[0190] The results are shown in Table 8 and Figure 4 shown.

[0191]

[0192] Result Analysis

[0193] 1. Multi-dimensional analysis of the synergy mechanism:

[0194] Pharmacodynamic synergy: The cat's claw extract in the compound can reverse cytarabine resistance;

[0195] Pharmacokinetic synergy: Sea buckthorn oil increases the lipid solubility of cytarabine, promotes transmembrane transport, and increases bioavailability.

[0196] 2. Clinical significance

[0197] This regimen can reduce the clinical dose of cytarabine from 100 mg / m² to 50 mg / m² and is expected to reduce the incidence of myelosuppression from 70% to <30%.

[0198] Test Example 7

[0199] Synergistic effect of combination with cisplatin in the treatment of Lewis lung cancer (solid tumors)

[0200] 1. Test Purpose

[0201] To evaluate the tumor inhibition rate and chemotherapy toxicity alleviation effects of the compound combined with cisplatin on Lewis lung cancer mice.

[0202] 2. Test methods

[0203] Animal model: C57BL / 6 mice (male, 8 weeks old, n=40) were subcutaneously inoculated with Lewis lung carcinoma cells (5×10^5 cells / mouse) in the right axilla.

[0204] Grouping and dosing (10 mice per group):

[0205] Control group: Oral administration of normal saline

[0206] Compound single-drug group: Compound of Example 3 (200 mg / kg / d, gavage)

[0207] Cisplatin monotherapy group: cisplatin (3 mg / kg, intraperitoneal injection, twice a week)

[0208] Combination group: compound drug (200 mg / kg / d) + cisplatin (1.5 mg / kg, half dose)

[0209] Detection indicators:

[0210] Tumor volume (measured with a vernier caliper, formula: V = 0.5 × length × width²)

[0211] Serum ALT / AST (indicator of liver damage)

[0212] Tumor tissue apoptosis rate (TUNEL staining)

[0213] The results are shown in Table 9 and Figure 5-6 shown.

[0214]

[0215] Result Analysis

[0216] The tumor volume in the combination group (410 mm³) was significantly smaller than that in the cisplatin monotherapy group (620 mm³), the apoptosis rate increased to 43.8% (p<0.01), and the ALT / AST levels were close to normal values, indicating that the combination enhanced cisplatin-induced tumor cell apoptosis through oridonin, while andrographolide reduced liver toxicity.

[0217] The key points of the above test cases are summarized in Table 10.

[0218]

[0219] From the above test examples, it can be seen that the solution of the present invention has the following outstanding progress:

[0220] 1. Multi-component synergistic effect: Through the scientific combination of Hedyotis diffusa (heat-clearing and detoxifying), Ganoderma lucidum polysaccharide (immune activation), astaxanthin (antioxidant), and Rubescensine A (apoptosis-promoting), it achieves a multi-pathway synergy of "immunomodulation-oxidative stress inhibition-apoptosis induction", with an in vitro anti-tumor activity IC50 of 25-50 μg / mL and is effective against both solid tumors and hematological tumors;

[0221] 2. Process Innovation: A gradient decoction (100°C → 80°C → 90°C) is used to preserve the activity of heat-sensitive ingredients. High-pressure homogenization (100 MPa) and ultrasound (20 kHz) are combined to prepare nanoemulsions (particle size 50-200 nm). This results in a 2.3-fold increase in astaxanthin bioavailability (AUC = 28.7 μg·h / mL) compared to traditional formulations, with a Tmax shortened to 2 hours.

[0222] 3. Dosage form diversification and stability: Enteric-coated capsules (coating material HPMCP) are designed to achieve intestinal targeted release (gastric fluid release rate <7%). The pills form a stable eutectic structure through the ascending-decanting method (calcination at 250°C). After 6 months of accelerated testing, the active ingredient retention rate is >90%, significantly better than conventional tablets (retention rate <65%).

[0223] 4. Chemosynergy and toxicity reduction: Combination with cytarabine prolonged the survival of leukemia mice by 35.8 days (CI=1.32) and reversed drug resistance by inhibiting P-gp. Combination with cisplatin increased the apoptosis rate of Lewis lung cancer to 43.8% (27.5% for single-agent therapy) and restored the hepatotoxicity markers ALT / AST to normal levels (42 U / L vs 68 U / L).

[0224] 5. Wide clinical applicability: It covers a variety of dosage forms, including tablets (microcrystalline cellulose excipient), capsules (enteric coating), and pills (moisture ≤ 12%), meeting the compliance needs of different patients and combining high efficiency and safety.

[0225] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Therefore, based on the innovative concept of the present invention, changes and modifications to the embodiments described herein, or equivalent structural or equivalent process transformations made using the contents of the present invention specification, directly or indirectly applying the above technical solutions to other related technical fields are all included in the scope of protection of the patent of the present invention.

Claims

1. Use of a compound in the preparation of a drug for treating leukemia in combination with cytarabine, characterized in that: The compound is made from the following raw materials in parts by weight: 15 parts of Hedyotis diffusa, 6.5 parts of Scutellaria barbata, 4.5 parts of Houttuynia cordata, 3 parts of Ganoderma lucidum, 0.75 parts of astaxanthin, 0.5 parts of sea buckthorn oil, 0.3 parts of Gynostemma pentaphyllum extract, 0.2 parts of Cat's Claw extract, 0.15 parts of Rubescensine A, and 0.12 parts of andrographolide; The preparation method of the above-mentioned compound comprises the following steps: 1) Grinding Hedyotis diffusa, Scutellaria barbata, Houttuynia cordata, and Ganoderma lucidum into powders in the form of Chinese herbal medicine slices by weight, filtering through a 50-mesh sieve, mixing, decocting with water 2-3 times, filtering through 5-10 layers of gauze, combining the decoctions, and concentrating to a clear paste with a relative density of 1.10-1.25; 2) mixing astaxanthin and sea buckthorn oil uniformly and then performing nanoemulsification treatment under nitrogen protection; 3) uniformly mixing the Gynostemma pentaphyllum extract, Cat's Claw extract, Rubescensine A, andrographolide and the emulsion obtained in step 2), and adding the mixture to the clear paste in step 1) to obtain the compound; The decoction in step 1) is controlled by gradient temperature: the first decoction is maintained at 100°C for 30 minutes, then the temperature is lowered to 80°C and continued to be decocted for 1 hour, and the second decoction is continued at 90°C for 1.5 hours; In the step 2), the nanoemulsified particle size is controlled at 50-200 nm, and high-pressure homogenization combined with ultrasonic treatment is used.

2. The use according to claim 1, characterized in that The method further comprises step 4) adding pharmaceutically acceptable excipients to prepare the formulation.

3. The use according to claim 2, characterized in that The tablets are made into tablets containing the following excipients: microcrystalline cellulose 10-30%, cross-linked carboxymethyl cellulose sodium 2-5%, and magnesium stearate 0.5-1.5%.

Images