Preparation and application of hedyotis diffusa, sculellaria barbata, lucid ganoderma, houttuynia cordata and astaxanthin compound preparation for adjuvant therapy of tumors

Abstract

The invention belongs to the field of biological medicines, and particularly discloses preparation and application of a hedyotis diffusa, sculellaria barbata, lucid ganoderma, houttuynia cordata and astaxanthin compound preparation for adjuvant therapy of tumors. The compound preparation is prepared from the following raw materials in parts by weight: 5-30 parts of oldenlandia diffusa, 3-10 parts of sculellaria barbata, 3-6 parts of houttuynia cordata, 2-4 parts of lucid ganoderma, 0.5-1 part of astaxanthin and the like through gradient decoction, nano emulsification and component synergistic compatibility. The bioavailability of thermosensitive components (such as astaxanthin) is remarkably improved by combining gradient temperature control decoction with a nano emulsification technology (the particle size is 50-200 nm), and various dosage forms (tablets, capsules and pills) are designed to meet different clinical requirements. In-vitro experiments show that the IC50 on HepG2 liver cancer cells is as low as 12.5 mu g / mL, and animal models confirm that the compound can enhance the curative effect of cytarabine and cis-platinum and reduce chemotherapy toxicity (ALT / AST is recovered to a normal level) at the same time. According to the invention, multi-target anti-tumor, immunoregulation and dosage form stability are integrated, and an innovative solution is provided for tumor adjuvant therapy.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically discloses the preparation and application of a compound preparation of Hedyotis diffusa, Scutellaria barbata, 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, the main clinical treatments are surgery, radiotherapy, chemotherapy, etc., but such therapies are often accompanied by significant side effects, such as decreased immunity, organ damage, etc., seriously affecting the quality of life of patients. In this context, the adjuvant treatment of traditional Chinese medicine has gradually attracted attention. Due to the characteristics of the synergistic action of multiple components, traditional Chinese medicine compound preparations 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 traditional Chinese medicines such as Hedyotis diffusa and Scutellaria barbata have been recognized as having anti-tumor activity; components such as Ganoderma lucidum polysaccharide 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 the traditional decoction process is difficult to effectively retain thermosensitive components, resulting in limited curative effects.

[0003] Furthermore, the following technical bottlenecks exist in the current traditional Chinese medicine compound preparations for adjuvant tumor treatment: First, the ingredient compatibility lacks innovation, and most compound preparations only simply stack known anti-tumor medicinal materials without introducing unconventional components with synergistic effects; second, the preparation process is extensive, and the research on nano-delivery technology for fat-soluble components (such as astaxanthin) is insufficient, affecting bioavailability; third, the dosage form design is single, making it difficult to meet the medication needs of different patients. For example, traditional decoctions are inconvenient to carry, while modern solid preparations may lose some volatile active substances. In addition, the existing compound preparations have less targeted research on hematological tumors and lack a broad-spectrum plan that takes into account both solid tumors and hematological tumors. In view of the above problems, there is an urgent need to develop a traditional Chinese medicine compound preparation with innovative components, advanced technology, and diverse dosage forms to improve the adjuvant tumor treatment effect through multi-target synergistic action, while reducing side effects and improving patient compliance. Summary of the Invention

[0004] In view of the above problems, the present invention discloses the preparation and application of a compound preparation of Hedyotis diffusa, Scutellaria barbata, Ganoderma lucidum, Houttuynia cordata, and astaxanthin for adjuvant treatment of tumors.

[0005] The object of the present invention is achieved by the following technical solutions.

[0006] A compound preparation of Hedyotis diffusa, Scutellaria barbata, Ganoderma lucidum, Houttuynia cordata, and astaxanthin for adjuvant treatment of tumors is prepared from the following raw materials in parts by weight:

[0007] Hedyotis diffusa 5 - 30 parts

[0008] Scutellaria barbata 3 - 10 parts

[0009] 3 - 6 parts of Houttuynia cordata

[0010] 2 - 4 parts of Ganoderma lucidum

[0011] 0.5 - 1 part of astaxanthin

[0012] 0.2 - 0.8 part of sea buckthorn oil

[0013] 0.1 - 0.5 part of Gynostemma pentaphyllum extract

[0014] 0.1 - 0.3 part of Ranunculus ternatus Thunb. extract

[0015] 0.1 - 0.2 part of oridonin

[0016] 0.05 - 0.2 part of andrographolide.

[0017] The present invention also discloses a preparation method of the above compound preparation, comprising the following steps:

[0018] 1) By weight, grind Hedyotis diffusa, Scutellaria barbata, Houttuynia cordata, and Ganoderma lucidum in the form of traditional Chinese medicine beverages into powders respectively. After passing through a 50 - mesh sieve, mix them, decoct with water for 2 - 3 times, filter with 5 - 10 layers of gauze, and combine the decoction and concentrate it to a clear paste with a relative density of 1.10 - 1.25;

[0019] 2) Mix astaxanthin and sea buckthorn oil evenly and carry out nano - emulsification treatment under nitrogen protection;

[0020] 3) Mix the Gynostemma pentaphyllum extract, Ranunculus ternatus Thunb. extract, oridonin, andrographolide and the emulsion obtained in step 2) evenly, and add them to the clear paste in step 1) to obtain the compound preparation.

[0021] Further, in the above - mentioned preparation method, the decoction in step 1) adopts gradient temperature control: the first decoction is maintained at 100 °C for 30 minutes and then cooled to 80 °C and continued to decoct for 1 hour, and the second decoction is continued at 90 °C for 1.5 hours.

[0022] Further, in the above - mentioned preparation method, the nano - emulsion particle size in step 2) is controlled within 50 - 200 nm, and high - pressure homogenization combined with ultrasonic treatment is adopted.

[0023] Further, the above - mentioned preparation method further includes step 4) adding pharmaceutically acceptable excipients for formulation.

[0024] Further, the above - mentioned preparation method is made into tablets, which contain the following excipients: 10 - 30% of microcrystalline cellulose, 2 - 5% of croscarmellose sodium, and 0.5 - 1.5% of magnesium stearate.

[0025] Furthermore, the above preparation method is made into a capsule, which contains an enteric coating material selected from hydroxypropyl methylcellulose phthalate or hydroxypropyl methylcellulose acetate succinate.

[0026] Furthermore, the above preparation method is made into a pill, which contains honey refining agent, the pill diameter is controlled at 3 - 8 mm, and the moisture content ≤ 12%.

[0027] Furthermore, the above preparation method is made into a dan agent, which includes the following process: prepared by the ascending and descending dan method, the calcination temperature is controlled at 200 - 300 °C, and the time is 2 - 4 hours.

[0028] The present invention also discloses the application of the above compound preparation in the preparation of tumor adjuvant therapy drugs, which is characterized in that the tumors include solid tumors and hematological system tumors.

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

[0030] The present invention forms a basic anti - tumor matrix through Hedyotis diffusa (clearing heat and detoxifying), Scutellaria barbata (anti - inflammation and anti - proliferation) and Ganoderma lucidum (immune regulation), and constructs a multi - target synergistic network by combining astaxanthin (high - efficiency antioxidant) and oridonin (apoptosis induction), breaking through the limitation of the single - action mode of traditional compound preparations; the gradient temperature - controlled decoction process (high temperature quickly extracts lipophilic components → low temperature retains thermosensitive active substances) is combined with the nano - emulsification technology (particle size < 200 nm) to solve the technical problem of low bioavailability of lipophilic components in traditional Chinese medicine compound preparations; in terms of dosage form design, enteric capsules achieve intestinal - targeted drug release through pH - sensitive coating, and dan agents improve stability by calcination eutectification, taking into account both traditional medication habits and modern precision medical needs. When combined with chemotherapy drugs, the gynostemma pentaphyllum extract in the compound preparation inhibits the expression of tumor cell resistance proteins, and andrographolide reduces chemotherapy - induced liver toxicity, and the synergy index (CI) > 1.3, significantly reducing the chemotherapy dose requirement. This scheme is applicable to the adjuvant treatment of solid tumors and hematological tumors, and can be flexibly adapted to various administration methods such as oral and enteric, providing an innovative preparation with standardization and high compatibility for the integrated traditional Chinese and Western medicine tumor treatment. Description of the Drawings

[0031] Figure 1 Comparison of serum IL - 2 (pg / mL) concentration in the evaluation of immune regulation effect;

[0032] Figure 2 Comparison of data of spleen lymphocyte proliferation (OD450) in the evaluation of immune regulation effect;

[0033] Figure 3 Comparison of data of thymus index (mg / g) in the evaluation of immune regulation effect;

[0034] Figure 4Comparison of average survival time (days) in the test of synergistic effect on hematological tumors;

[0035] Figure 5 Comparison of tumor volume (mm³, on the 21st day) in the test of synergistic effect on solid tumors;

[0036] Figure 6 Comparison of apoptosis rate (%) in the test of synergistic effect on solid tumors. Detailed implementation mode

[0037] To make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below. However, it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the scope of the present invention. In addition, in the following description, the descriptions of well-known structures and technologies are omitted to avoid unnecessarily confusing the concepts 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, without conflict, the embodiments in the present application and the features in the embodiments 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 embodiments 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] Oldenlandia diffusa 5 parts, Scutellaria barbata 3 parts, Houttuynia cordata 3 parts, Ganoderma lucidum 2 parts, astaxanthin 0.5 part, seabuckthorn oil 0.2 part, Gynostemma pentaphyllum extract 0.1 part, cat's claw grass extract 0.1 part, oridonin 0.1 part, andrographolide 0.05 part.

[0045] Preparation method:

[0046] 1) Weigh the four traditional Chinese medicines such as Oldenlandia diffusa according to the ratio, grind them and pass through a 50-mesh sieve. After mixing, decoct them with water twice: the first time at 100 °C for 30 minutes, then cool down to 80 °C and continue for 1 hour; the second time at 90 °C for 1.5 hours; combine the decoction and concentrate it to a clear paste with a relative density of 1.10;

[0047] 2) Mix astaxanthin and seabuckthorn oil, and under nitrogen protection, perform high-pressure homogenization (100 MPa) combined with ultrasonic treatment (20 kHz) to prepare a nanoemulsion with an average particle size of 50 nm;

[0048] 3) Mix the remaining components such as the extract of Gynostemma pentaphyllum with the nanoemulsion, and add it to the clear extract 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 part of sea buckthorn oil, 0.5 part of Gynostemma pentaphyllum extract, 0.3 part of cat's claw herb extract, 0.2 part of oridonin, 0.2 part of andrographolide.

[0051] The preparation method is basically the same as that of Example 1, the difference is that after decocting, it 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 part of astaxanthin, 0.5 part of sea buckthorn oil, 0.3 part of Gynostemma pentaphyllum extract, 0.2 part of cat's claw herb extract, 0.15 part of oridonin, 0.12 part of andrographolide.

[0054] The preparation method is basically the same as that of Example 1, with an average nanoemulsion particle size of 120 nm, and made into tablets: adding 20% microcrystalline cellulose, 3% cross-linked carboxymethylcellulose sodium, and 1% magnesium stearate.

[0055] Example 4

[0056] The raw material ratio is the same as that of Example 3. During preparation, in step 4), hydroxypropyl methylcellulose phthalate is added for enteric coating to make a capsule.

[0057] Example 5

[0058] The raw material ratio is the same as that of Example 2. In step 4), it is calcined by the method of Shengjiang Dan (250 °C, 3 hours) to make a Dan preparation with a moisture content of 10%.

[0059] Comparative Example 1

[0060] Astaxanthin and sea buckthorn oil are missing, and the remaining components and processes are the same as those of Example 3.

[0061] Comparative Example 2

[0062] Gradient decoction is not carried out: decoct at 100 °C for 3 hours throughout the process, and the rest is the same as that of Example 1.

[0063] Comparative Example 3

[0064] The nanoemulsion particle size is 400 nm (only high-pressure homogenization treatment, without ultrasonic treatment), and the rest is the same as that of Example 2.

[0065] Comparative Example 4

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

[0067] Comparative Example 5

[0068] Conventional tablet excipients (50% starch) were used, and cross-linked carboxymethyl cellulose sodium was not added. The rest was the same as in Example 3.

[0069] Test Example 1

[0070] In vitro anti-tumor activity test

[0071] 1. Test method:

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

[0073] Drug administration treatment: Treated with the compound preparation of Example 3 (0 - 100 μg / mL) for 48 hours, and the cell viability was measured by the MTT method.

[0074] 2. Grouping: Blank control group, Example 3 group (gradient concentration), Comparative Example 1 group (without astaxanthin), Comparative Example 4 group (lacking oridonin).

[0075] The results are shown in Table 3.

[0076]

[0077] Analysis of the tabular results:

[0078] 1. Dose-dependent effect:

[0079] The compound preparation showed significant concentration-dependent inhibition in the range of 10 - 100 μg / mL, with an IC50 of 12.5 μg / mL, which was much lower than that of traditional Chinese medicine compound preparations (usually IC50 > 50 μg / mL), indicating its high efficiency.

[0080] 2. Component synergistic mechanism:

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

[0082] Oridonin: 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. Influence of the missing components:

[0084] The IC50 of Comparative Example 1 (without astaxanthin) was missing, indicating that astaxanthin is the key to overcoming the antioxidant defense of tumor cells; the IC50 of Comparative Example 4 (without oridonin) increased by 2.8 times, confirming the irreplaceability of this component in the apoptosis pathway.

[0085] 4. Horizontal comparison:

[0086] Compared with the literature-reported single use of Hedyotis diffusa extract (IC50 ≈ 45 μg / mL), the activity of this compound preparation increased by 3.6 times, reflecting the advantage of multi-component integration.

[0087] Test Example 2

[0088] Evaluation of immunomodulatory effect

[0089] 1. Test purpose

[0090] To verify the immune recovery effect of the compound preparation on immunosuppressed mice after chemotherapy, and to focus on evaluating the IL-2 secretion level and the proliferative ability of splenic lymphocytes.

[0091] 2. Test method

[0092] Animal model: ICR mice (male, 8 weeks old, n = 40), an immunosuppression model was established by intraperitoneal injection of cyclophosphamide (80 mg / kg, for 3 consecutive days).

[0093] 3. Grouping and administration (10 mice in each group):

[0094] Normal control group: Not modeled, gavaged with normal saline

[0095] Model control group: Gavaged with normal saline after modeling

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

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

[0098] 4. Detection indexes:

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

[0100] Proliferation of splenic lymphocytes: OD450 value measured by CCK-8 method after stimulation with ConA

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

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

[0103]

[0104] Result Analysis

[0105] 1. Immunological Reconstitution Effect:

[0106] The high-dose compound restored IL-2 to the normal level of 85.2 pg / mL and the thymus index reached 3.5 mg / g, approaching the healthy state, indicating its ability to repair the thymic microenvironment, possibly achieved by upregulating the expression of FoxN1 in thymic epithelial cells.

[0107] 2. Analysis of the Role of Core Components:

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

[0109] Astaxanthin: Inhibits the expansion of Treg cells caused by chemotherapy and reverses immune tolerance.

[0110] 3. Dose-Effect Relationship:

[0111] The low-dose group only partially restored immune function (IL-2 was 68.4 pg / mL), indicating that 200 mg / kg / d is the optimized dose, which has the safety advantage of natural components compared with commonly used clinical immunomodulators (such as thymosin).

[0112] 4. Clinical Transformation Significance:

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

[0114] Test Example 3

[0115] Bioavailability Evaluation

[0116] 1. Test Purpose

[0117] To compare the bioavailability differences between the astaxanthin nanoemulsion and the traditional preparation in the compound and verify the advantages of the nanoemulsification process.

[0118] 2. Test Method

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

[0120] Traditional Preparation Group: Suspension of astaxanthin without nanoemulsification (0.5 mg / kg)

[0121] Nanoemulsion Group: Compound of Example 1 (containing 0.5 mg / kg astaxanthin)

[0122] Control Group 3: Coarse emulsion of astaxanthin (particle size 400 nm, 0.5 mg / kg)

[0123] Administration and Sampling: Blood samples were collected at 0.5, 1, 2, 4, 8, 12, and 24 hours after single intragastric administration, and the plasma astaxanthin concentration was determined by HPLC (Agilent 1260, C18 column, detection wavelength 476 nm).

[0124] Data Analysis: Calculate AUC 0-24h and C max .

[0125] The results are shown in Table 5

[0126]

[0127] Analysis of Tabular Results

[0128] 1. Pharmacokinetic Advantages of Nanoemulsion:

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

[0130] Increased Exposure: AUC increased by 2.3 times, related to the promotion of bile secretion by sea buckthorn oil in nanoemulsion and the increase in the solubility of lipophilic 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 a particle size <200 nm is the key threshold for breaking through the intestinal mucus layer (referring to the consensus in the field of nano-drug delivery).

[0133] 2. Breakthrough of Technical Barriers:

[0134] The bioavailability of lipophilic components (such as andrographolide) in traditional Chinese medicine compound is usually <10%, while this process increased it to 28.7 μg·h / mL, reaching the level of chemical drugs.

[0135] Test Example 4

[0136] Release Test of Enteric Capsules

[0137] 1. Test Purpose

[0138] Verify the protective effect of enteric coating on thermosensitive components (such as gypenosides) in the compound preparation and ensure targeted release in the intestine.

[0139] 2. Test Method

[0140] Sample Preparation: Capsules of Example 4 (containing hydroxypropyl methylcellulose phthalate coating) and uncoated control group.

[0141] Release Conditions:

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

[0143] Simulated intestinal fluid: phosphate buffer (pH 6.8), for subsequent 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] Analysis of tabular results

[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), and avoiding the hydrolysis and destruction of the glycosidic bond of gypenosides by gastric acid.

[0150] 2. Release kinetics analysis:

[0151] The enteric-coated capsules showed zero-order release characteristics (R² = 0.992) in intestinal fluid, conforming to the Higuchi model, indicating that the release was controlled by diffusion, which was beneficial for maintaining a steady-state blood drug concentration.

[0152] 3. Comparison of the defects of conventional preparations:

[0153] The uncoated capsules released 85% in 1 hour in gastric juice, 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 coating design.

[0154] Test Example 5

[0155] Accelerated long-term stability test

[0156] 1. Test purpose

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

[0158] 2. Test method

[0159] Samples and conditions:

[0160] Pills of Example 5 (prepared by the method of Shengjiang Pills) and tablets of Comparative Example 5 (starch excipients)

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

[0162] Detection indicators:

[0163] Retention rate of oridonin: HPLC method (mobile phase: acetonitrile - water, detection wavelength: 242 nm)

[0164] Retention rate of andrographolide: same as above, detection wavelength: 225 nm.

[0165] The results are shown in Table 7

[0166]

[0167] Analysis of the tabular results:

[0168] 1. Physical and chemical basis for the stability of Danji:

[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: Moisture in Danji ≤ 12% inhibits hydrolysis reactions, while the moisture in tablets > 15% after starch absorbs moisture, accelerating the degradation of glycoside components.

[0171] 2. Inspiration from the scientific compatibility of excipients:

[0172] Using starch in Comparative Example 5 resulted in a retention rate < 65%, suggesting that in future development, hydrophobic excipients (such as microcrystalline cellulose) or anti - caking agents such as silica should be used.

[0173] 3. Shelf - life calculation:

[0174] According to the ICH Q1A guideline, the predicted shelf - life of Danji at 25 °C / 60% RH is > 36 months, while that of tablets is only 12 months, significantly enhancing the commercial value of the product

[0175] Test Example 6

[0176] Synergistic enhancement test for the combined treatment of leukemia (hematological tumor) with cytarabine

[0177] 1. Test purpose

[0178] Verify the synergistic therapeutic effect of the compound preparation combined with the chemotherapeutic drug cytarabine on the L1210 leukemia mouse model, and evaluate the effect of extended survival period and reduced tumor burden.

[0179] 2. Test method

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

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

[0182] Control group: intragastric administration of normal saline

[0183] Compound single-drug group: Compound in Example 3 (200 mg / kg / d, intragastric administration)

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

[0185] Combined group: Compound (200 mg / kg / d) + Cytarabine (10 mg / kg, half dose)

[0186] Detection indexes:

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

[0188] Peripheral blood leukemia cell count (proportion of CD45+ cells detected by flow cytometry)

[0189] Spleen weight (tumor infiltration index)

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

[0191]

[0192] Result analysis

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

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

[0195] Pharmacokinetic synergy: Sea buckthorn oil increases the liposolubility 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 combined treatment with cisplatin on Lewis lung cancer (solid tumor)

[0200] 1. Test purpose

[0201] To evaluate the tumor inhibition rate and the remission effect of chemotherapy toxicity of the compound combined with cisplatin in mice with Lewis lung cancer.

[0202] 2. Test method

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

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

[0205] Control group: gavaged with normal saline

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

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

[0208] Combined group: Compound (200 mg / kg / d) + cisplatin (1.5 mg / kg, half dose)

[0209] Detection indexes:

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

[0211] Serum ALT / AST (liver function injury index)

[0212] Apoptosis rate of tumor tissue (TUNEL staining)

[0213] The results are shown in Table 9 and Figures 5-6 as follows.

[0214]

[0215] Result analysis

[0216] The tumor volume of the combined group (410 mm³) was significantly smaller than that of the cisplatin single-drug group (620 mm³), the apoptosis rate increased to 43.8% (p < 0.01), and the ALT / AST level was close to the normal value, indicating that the compound enhanced cisplatin-induced apoptosis of tumor cells through oridonin, while andrographolide reduced liver toxicity.

[0217] Summarize the key points of the above test examples as shown in Table 10 below

[0218]

[0219] It can be seen from the above test examples that the solution of the present invention has the following outstanding progressiveness:

[0220] 1. Multi-component synergistic effect: Through the scientific compatibility of Hedyotis diffusa (heat-clearing and detoxifying), Ganoderma lucidum polysaccharide (immune activation), astaxanthin (antioxidant), and oridonin (apoptosis induction), the multi-pathway synergy of "immune regulation - oxidative stress inhibition - apoptosis induction" is achieved. The in vitro anti-tumor activity (IC50 = 12.5 μg / mL) is 3.6 times higher than that of single components, and it is effective against both solid tumors and hematological tumors.

[0221] 2. Process innovation: Gradient decoction (100°C → 80°C → 90°C) is used to retain the activity of heat-sensitive components. Combining high-pressure homogenization (100 MPa) and ultrasound (20 kHz) to prepare nanoemulsion (particle size 50 - 200 nm), which increases the bioavailability of astaxanthin (AUC = 28.7 μg·h / mL) by 2.3 times compared with traditional preparations, and shortens Tmax to 2 hours.

[0222] 3. Dosage form diversification and stability: Design enteric-coated capsules (coating material HPMCP) to achieve intestinal targeted release (gastric juice release rate < 7%). The Dan preparation forms a stable eutectic structure through the method of raising and lowering Dan (calcination at 250°C). After 6 months of accelerated testing, the retention rate of active ingredients > 90%, which is significantly better than that of conventional tablets (retention rate < 65%).

[0223] 4. Chemotherapy synergy and toxicity reduction: Combining with cytarabine prolongs the survival period of leukemia mice by 35.8 days (CI = 1.32) and reverses drug resistance by inhibiting P-gp. Combining with cisplatin increases the apoptosis rate of Lewis lung cancer to 43.8% (single drug 27.5%), and the liver toxicity markers ALT / AST return to normal levels (42 U / L vs 68 U / L).

[0224] 5. Wide clinical applicability: Covers various dosage forms such as tablets (microcrystalline cellulose excipient), capsules (enteric coating), and Dan preparations (moisture ≤ 12%), meeting the compliance needs of different patients, and having both 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 rather than to limit the protection scope of the present invention. Therefore, based on the innovative concept of the present invention, any changes and modifications made to the embodiments described herein, or equivalent structural or equivalent process transformations made using the content of the specification of the present invention, directly or indirectly applying the above technical solutions to other related technical fields, are all included in the protection scope of the present invention patent.

Claims

1. A compound preparation of Hedyotis diffusa, Scutellaria barbata, Ganoderma lucidum, Houttuynia cordata and astaxanthin for adjuvant treatment of tumors, characterized in that, It is made from the following raw materials in parts by weight: Oldenlandia diffusa 5 - 30 parts Scutellaria barbata 3 - 10 parts Houttuynia cordata 3 - 6 parts Ganoderma lucidum 2 - 4 parts Astaxanthin 0.5 - 1 part Sea buckthorn oil 0.2 - 0.8 part Gynostemma pentaphyllum extract 0.1 - 0.5 part Ranunculus ternatus Thunb. extract 0.1 - 0.3 part Oridonin 0.1 - 0.2 part Andrographolide 0.05 - 0.2 part.

2. The preparation method of the compound agent according to claim 1, characterized in that, It includes the following steps: 1) By weight, grind Oldenlandia diffusa, Scutellaria barbata, Houttuynia cordata, and Ganoderma lucidum in the form of traditional Chinese medicine beverages into powders respectively. After passing through a 50 - mesh sieve, mix them, decoct with water 2 - 3 times, filter with 5 - 10 layers of gauze, and combine the decoctions and concentrate them into a clear paste with a relative density of 1.10 - 1.25; 2) Mix astaxanthin and sea buckthorn oil evenly and conduct nano - emulsification treatment under nitrogen protection; 3) Mix the Gynostemma pentaphyllum extract, Ranunculus ternatus Thunb. extract, oridonin, andrographolide and the emulsion obtained in step 2) evenly, and add them to the clear paste in step 1) to obtain the compound preparation.

3. The preparation method according to claim 2, characterized in that, In step 1), the decoction is controlled by gradient temperature: for the first decoction, maintain at 100 °C for 30 minutes and then reduce the temperature to 80 °C and continue decocting for 1 hour. For the second decoction, continue at 90 °C for 1.5 hours.

4. The preparation method according to claim 2, characterized in that, In step 2), the nano - emulsion particle size is controlled within 50 - 200 nm, and high - pressure homogenization combined with ultrasonic treatment is adopted.

5. The preparation method according to claim 2, characterized in that, It also includes step 4) adding pharmaceutically acceptable excipients for formulation.

6. The preparation method according to claim 5, characterized in that, When made into tablets, it contains the following excipients: microcrystalline cellulose 10 - 30%, croscarmellose sodium 2 - 5%, magnesium stearate 0.5 - 1.5%.

7. The preparation method according to claim 5, characterized in that When made into capsules, it contains an enteric - coating material, and the coating material is selected from hydroxypropyl methylcellulose phthalate or hydroxypropyl methylcellulose acetate succinate.

8. The preparation method according to claim 5, characterized in that, When made into pills, it contains honey refining agent, the pill diameter is controlled within 3 - 8 mm, and the moisture content ≤ 12%.

9. The preparation method according to claim 5, characterized in that, When made into pellets, it includes the following process: prepared by the rising and falling pellet method, control the calcination temperature at 200 - 300 °C for 2 - 4 hours.

10. Use of the compound preparation according to claim 1 in the preparation of a drug for adjuvant cancer treatment, characterized in that, The tumors include solid tumors and hematological system tumors.

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