A composition for regulating overall metabolic balance, inhibiting cancer cell growth, and anti-tumor

Abstract

The application discloses a kind of composition of adjusting overall metabolic balance, inhibiting cancer cell growth, antitumor, raw material includes ganoderma lucidum, crocodile peptide, astragalus, polypodium pinnatum, ginseng, turmeric, clove, poria cocos, camellia sinensis, rich glutathione yeast, cordyceps militaris, gizzard gold, peach kernel, compound probiotics, brassica campestris l.var.rapa.The application fuses modern medical scientific concept and traditional Chinese medicine "blood stasis and stagnation and cause mass" theory, raw material synergistic effect, by improving blood oxygen content, anti-inflammatory, improve metabolic function, blood circulation and other ways, realize protection mitochondria from damage, anti-mutation;And through the way such as supplementing compound probiotics, regulate intestinal flora, enhance immunity, finally realize soft and hard knot, adjust overall metabolic balance, inhibit cancer cell growth, antitumor, relieve tumor treatment side effect, application prospect is wide.

Description

Technical Field

[0001] This invention belongs to the field of composition technology, specifically relating to a composition, formulation, preparation method, and application for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors. Background Technology

[0002] Cancer, as a malignant disease, seriously threatens human life, health, and quality of life. Modern medicine has developed a diverse range of treatment options for cancer, including surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. However, these methods generally suffer from significant side effects (such as nausea and vomiting, bone marrow suppression, and intestinal mucosal damage), are prone to drug resistance, and are difficult to effectively improve quality of life.

[0003] Traditional Chinese medicine attributes the pathogenesis of cancer to "blood stasis and qi stagnation, phlegm and blood stasis." Poor blood and qi circulation leads to obstruction of the meridians, and abnormal body fluid metabolism generates phlegm and dampness. Over time, phlegm and blood stasis accumulate to form "masses and lumps." Therefore, traditional Chinese medicine often uses "softening and dispersing masses" as its core principle. However, single Chinese herbs or traditional compound formulas have limitations such as slow onset of action, single target, and difficulty in simultaneously regulating metabolic balance and enhancing immune function.

[0004] With the deepening of modern medical research, studies have found that the occurrence and development of tumors are not only related to gene abnormalities, but also closely related to metabolic disorders. Mitochondria, as the cell's "energy factory," are a key inducing factor for abnormal tumor metabolism when their function is impaired. Mitochondrial damage leads to a decrease in oxidative phosphorylation capacity, forcing tumor cells to switch to inefficient glycolysis for energy, producing large amounts of lactic acid, acidifying the microenvironment, and increasing reactive oxygen species production, exacerbating genomic instability and gene mutations. Furthermore, metabolic diseases such as diabetes and obesity, as well as hypoxemia and chronic inflammation, can all induce gene mutations by damaging mitochondria, promoting cancer cell proliferation. In a hypoxic microenvironment, hypoxia-inducible factor-1α (HIF-1α) released by tumor cells further upregulates glycolysis and promotes angiogenesis, thereby enhancing the tumor's invasive and metastatic capabilities.

[0005] Currently, existing technologies lack compositions that can simultaneously target multiple aspects and multiple targets, including metabolic abnormalities, inflammatory responses, mitochondrial damage caused by low blood oxygen levels, immunosuppression, and blood stasis. Therefore, integrating modern medical theories of metabolic regulation with traditional Chinese medicine's methods of promoting blood circulation and removing blood stasis to develop a synergistic, safe, and effective combination of traditional Chinese and Western medicine is of great value in enhancing anti-tumor effects and reducing treatment-related side effects. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a composition, preparation method, and application for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors. This invention integrates modern medical science concepts with the traditional Chinese medicine theory of "blood stasis and qi stagnation leading to lumps," with scientifically formulated and synergistically combined ingredients. The overall composition is safe and highly effective, effectively softening and dispersing lumps, regulating overall metabolic balance, inhibiting cancer cell growth, fighting tumors, and alleviating the side effects of tumor treatment.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] In a first aspect, the present invention provides a composition for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors. The composition comprises the following raw materials: 10-30 parts of Ganoderma lucidum, 0.05-0.2 parts of crocodile peptide, 15-35 parts of Astragalus membranaceus, 5-20 parts of Anoectochilus roxburghii, 10-35 parts of ginseng, 1-5 parts of turmeric, 5-15 parts of clove, 10-25 parts of Poria cocos, 5-20 parts of Camellia chrysantha, 0.1-0.5 parts of glutathione-enriched yeast, 5-20 parts of Cordyceps militaris, 10-25 parts of chicken gizzard lining, 5-15 parts of peach kernel, 0.1-0.5 parts of compound probiotics, and 1-5 parts of Viola yedoensis and Citrus aurantium.

[0009] Further, the raw material composition of the composition is as follows: 15-25 parts of Ganoderma lucidum, 0.08-0.18 parts of crocodile peptide, 20-30 parts of Astragalus membranaceus, 10-18 parts of Anoectochilus roxburghii, 20-30 parts of ginseng, 2-5 parts of turmeric, 8-12 parts of clove, 15-22 parts of Poria cocos, 10-18 parts of Camellia chrysantha, 0.2-0.4 parts of glutathione-enriched yeast, 10-18 parts of Cordyceps militaris, 15-22 parts of chicken gizzard lining, 8-12 parts of peach kernel, 0.2-0.4 parts of compound probiotics, and 2-4 parts of Viola yedoensis.

[0010] Further, the raw material composition of the composition is as follows: 20-25 parts of Ganoderma lucidum, 0.1-0.15 parts of crocodile peptide, 25-30 parts of Astragalus membranaceus, 12-16 parts of Anoectochilus roxburghii, 25-30 parts of ginseng, 2-4 parts of turmeric, 10-12 parts of clove, 18-22 parts of Poria cocos, 12-16 parts of Camellia chrysantha, 0.3-0.4 parts of glutathione-enriched yeast, 12-16 parts of Cordyceps militaris, 18-22 parts of chicken gizzard lining, 10-12 parts of peach kernel, 0.3-0.4 parts of compound probiotics, and 3-4 parts of Viola yedoensis and Citrus aurantium.

[0011] The compound probiotics consist of the following: Pediococcus pentosaceus MPL5, Lactobacillus plantarum La10, Lactobacillus casei C7-2, Lactobacillus reuteri HBM11-69, and Lactobacillus rhamnosus HBM11-35 in a live bacteria ratio of (1-5):(1-5):(1-5):(1-5):(1-5).

[0012] The aforementioned Pediococcus pentosaceus MPL5 is classified and named as follows:Pediococcus pentosaceus Lactobacillus plantarum La10, deposited at the Guangdong Provincial Center for Microbial Culture Collection, accession number GDMCC No:63606, deposited on June 30, 2023, at the Institute of Microbiology, Guangdong Academy of Sciences, No. 59, 100 Xianlie Middle Road, Guangzhou. Lactiplantibacillus plantarum The specimen, Lactobacillus casei C7-2, is deposited at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No. 63605) on June 30, 2023. The address of the depository is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences, Institute of Microbiology. Lacticaseibacillus casei The sample, Lactobacillus reuteri HBM11-69, is deposited at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No. 63609) on June 30, 2023. The address of the depository is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences, Institute of Microbiology. Limosilactobacillus reuteri The specimen, Lactobacillus rhamnosus HBM11-35, is deposited at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No. 63608) on June 30, 2023. The address of the depository is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences, Institute of Microbiology. Lacticaseibacillus rhamnosus It is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No:63607, deposited on June 30, 2023. The address of the depository is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Academy of Sciences Institute of Microbiology.

[0013] Furthermore, the viable count ratios of Pediococcus pentosaceus MPL5, Lactobacillus plantarum La10, Lactobacillus casei C7-2, Lactobacillus reuteri HBM11-69, and Lactobacillus rhamnosus HBM11-35 are 1:1:1:1:1, or 1:2:1:1:1, or 1:1:2:1:1, or 1:1:1:2:1, or 1:1:1:1:1:2, or 1:2:2:1:1, or 1:2:1:2:1:2, or 1:2:1:1:2.

[0014] Furthermore, the compound probiotics may also include, but are not limited to, the addition or replacement of bacterial strains.

[0015] Furthermore, the compound probiotics, whether in live or inactivated form, are applicable to this invention.

[0016] Furthermore, the molecular weight of the crocodile peptide is preferably 500-1000 Da.

[0017] Furthermore, the sources of the Ganoderma lucidum include, but are not limited to, Ganoderma lucidum fruiting bodies, Ganoderma lucidum extracts, and Ganoderma lucidum fermentation broth.

[0018] Furthermore, the sources of Astragalus membranaceus, Anoectochilus roxburghii, ginseng, turmeric, clove, Poria cocos, Camellia chrysantha, Cordyceps militaris, chicken gizzard lining, peach kernel, and Viola yedoensis include, but are not limited to, their roots, stems, leaves, extracts, and fermentation products.

[0019] Secondly, the present invention provides a compound preparation for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors, characterized in that the raw materials include the aforementioned composition.

[0020] Furthermore, the compound formulation also includes a pharmaceutically acceptable carrier or excipient.

[0021] Furthermore, the carrier or excipient includes one or more of the following: diluent, wetting agent, lubricant, adhesive, disintegrant, encapsulating agent, flavoring agent, sustained-release agent, dispersant, plasticizer, and light-blocking agent.

[0022] Furthermore, the compound preparation is an oral preparation, and its dosage form includes powder, granule, pill, drop pill, tablet, capsule, gel, ointment, and oral liquid. The compound preparation can also be used as a topical preparation as needed.

[0023] Thirdly, the present invention provides a method for preparing the composition, comprising the following steps:

[0024] Step 1: Grind the following ingredients separately: Ganoderma lucidum, Astragalus membranaceus, Anoectochilus roxburghii, ginseng, turmeric, clove, Poria cocos, Camellia chrysantha, Cordyceps militaris, chicken gizzard lining, peach kernel, and Viola yedoensis. Then, sift them through an 80-100 mesh sieve to obtain the raw material powder.

[0025] Step 2: Add 5-15 times the weight of purified water to each raw material powder, soak for 0.5-1h, heat to boiling, maintain a gentle boil and extract 2-3 times, each extraction for 30-45min, combine the extracts of each raw material, filter to remove the residue, and obtain a mixed extract.

[0026] Step 3: The mixed extract is concentrated under reduced pressure at 60-70℃ and vacuum degree of -0.08 to -0.09MPa to obtain an extract with a relative density of 1.15-1.25. The extract is then spray-dried at an inlet air temperature of 180-200℃ and an outlet air temperature of 80-90℃ to obtain a fine powder.

[0027] Step 4: Add crocodile peptide, glutathione-enriched yeast, and compound probiotics to the fine powder of extract, and mix evenly in an environment of 30-35℃ and relative humidity ≤45% to obtain the composition.

[0028] Furthermore, in step 2, the first extraction is done with 10 times the weight of the raw material in purified water, and the second and third extractions are done with 8 times the weight of the raw material in purified water, respectively; in step 3, the inlet air temperature of the spray dryer is 190℃ and the outlet air temperature is 85℃.

[0029] Furthermore, after mixing evenly in step 4, 4-8 times the amount of water can be added, and fermentation can be carried out at 30-45℃ for 2-12 hours to obtain a compound fermentation preparation.

[0030] Fourthly, the present invention provides the use of the composition or the compound preparation described herein in the preparation of preparations that regulate overall metabolic balance, inhibit cancer cell growth, and have anti-tumor effects.

[0031] The formulation concept and the function of each raw material component of this invention are briefly explained below:

[0032] For decades, the pathogenesis of cancer has been primarily attributed to genetic abnormalities. However, with the advancement of clinical research, it has been discovered that tumorigenesis involves not only genetic alterations but is also closely related to metabolic disorders. Current research indicates that the genomic defects observed in most cancer cells are actually secondary changes caused by mitochondrial metabolic dysfunction; that is, mitochondrial damage is the cause, and genomic abnormalities are the effect.

[0033] Under normal physiological conditions, when oxygen supply is sufficient, mitochondria break down glucose to produce pyruvate, which is then converted into carbon dioxide and a large amount of ATP via oxidative phosphorylation. When mitochondrial function is impaired, its oxidative phosphorylation capacity is compromised, failing to meet the high energy demands of rapid tumor cell proliferation. This forces cells to rely on the less efficient glycolysis for energy. This process produces large amounts of lactic acid, leading to acidification of the tumor microenvironment and promoting tumor growth. Furthermore, mitochondrial damage increases the generation of reactive oxygen species (ROS), exacerbating the instability of the cell nucleus genome and thus increasing the probability of gene mutations.

[0034] Metabolic diseases (such as diabetes, hyperuricemia, and obesity) are caused by abnormal metabolism in the body and can lead to mitochondrial damage by altering the body's microenvironment. Hypoxia is also a significant factor affecting mitochondrial respiratory function. In a hypoxic microenvironment, tumor cells release hypoxia-inducible factor-1α (HIF-1α). This transcription factor not only induces the expression of glycolysis-related enzymes and inhibits mitochondrial oxidative metabolism, thereby enhancing aerobic glycolysis, but also promotes angiogenesis, thus increasing the invasive and metastatic capabilities of tumor cells. Chronic inflammation, by continuously upregulating the levels of inflammatory factors, causes long-term damage to mitochondrial function. In summary, metabolic abnormalities, hypoxia, and inflammation can all impair mitochondrial function, induce genomic mutations, and thus promote cancer cell formation.

[0035] The immune system, as an important defense mechanism of the body, relies on innate and adaptive immunity to recognize and eliminate abnormal cells such as cancer cells. In other words, even if mitochondrial damage is caused by factors such as metabolic abnormalities, hypoxia, or inflammation, a healthy immune system can still act as a terminal barrier to eliminate abnormally generated cancer cells.

[0036] Based on the above mechanisms, this formula innovatively integrates modern medical scientific perspectives with the traditional Chinese medicine theory of "blood stasis and qi stagnation leading to lumps," employing a multi-component synergistic effect: Crocodile peptides are rich in eight essential amino acids and can be derived from one or more of crocodile blood, meat, and bones; *Anoectochilus roxburghii*, crocodile peptides, and glutathione-rich yeast enhance the body's anti-inflammatory and free radical scavenging capabilities; *Corydalis yanhusuo* is a selenium-rich plant with a high organic selenium content, possessing regulatory and antioxidant effects on oxidative stress; *Astragalus membranaceus*, ginseng, and *Camellia sinensis* improve blood oxygen supply and saturation; turmeric, peach kernel, and chicken gizzard membrane work together to soften and disperse lumps, promote blood circulation, remove blood stasis, and relieve pain. The anti-tumor potential of curcumin, the active ingredient in turmeric, has been proven, with core mechanisms including inhibiting cancer cell proliferation, invasion, and metastasis, and blocking tumor angiogenesis. Its anti-inflammatory and antioxidant properties can also help reduce the side effects of tumor treatment. Building upon this foundation, the formula incorporates Ganoderma lucidum, Poria cocos, Cordyceps militaris, and cloves to enhance the body's immunity, strengthen the killing effect on tumor cells, inhibit their immune escape, suppress cancer cell growth, and thus block tumor proliferation. Furthermore, the introduction of compound probiotics, whether live or inactivated, strengthens the mucosal barrier, improves the intestinal microecology, alleviates cancer treatment-related side effects such as nausea, diarrhea, and loss of appetite, and improves patients' quality of life. Attached Figure Description

[0037] Figure 1 The graph shows the effect of AGS proliferation.

[0038] Figure 2 This is a bar chart showing the effect of [amount] on AGS proliferation. Note: Compared to 0 mg / mL, [the percentage of each component] is [percentage]. * P <0.05, ** P <0.01, *** P <0.001; compared to 8 mg / mL, # P <0.05, ## P <0.01, ### P <0.001; comparison at the same concentration & P <0.05, && P <0.01, &&& P <0.001.

[0039] Figure 3 The effect on weight before and after treatment. Note: Compared with the model group, * P <0.05, ** P <0.01, *** P <0.001; compared with the positive control group (cisplatin), # P <0.05, ## P <0.01, ### P <0.001.

[0040] Figure 4 This represents the impact on the general state score. Note: Compared to the model group, * P <0.05, ** P <0.01, *** P <0.001; compared with the positive control group (cisplatin), # P <0.05, ## P <0.01, ### P <0.001.

[0041] Figure 5 This represents the average tumor weight for each group. Note: Compared to the model group, * P <0.05, ** P <0.01, *** P <0.001; compared with the positive control group (cisplatin), # P <0.05, ## P <0.01, ### P <0.001.

[0042] Figure 6 The tumor inhibition rate is represented by the value of each group. Detailed Implementation

[0043] Example 1:

[0044] A composition for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors: 25 parts Ganoderma lucidum, 0.15 parts alligator peptide, 30 parts Astragalus membranaceus, 16 parts Anoectochilus roxburghii, 30 parts Ginseng, 4 parts Curcuma longa, 12 parts Clove, 22 parts Poria cocos, 16 parts Camellia chrysantha, 0.4 parts glutathione-rich yeast, 16 parts Cordyceps militaris, 22 parts chicken gizzard lining, 12 parts Prunus persica, 0.4 parts compound probiotics (Vibrio vulgaris MPL5, Lactobacillus plantarum La10, Lactobacillus casei C7-2, Lactobacillus reuteri HBM11-69, Lactobacillus rhamnosus HBM11-35 live bacteria count ratio 1:2:1:1:1), and 4 parts Viola yedoensis.

[0045] The method for preparing the composition includes the following steps:

[0046] Step 1: Grind the following ingredients separately: Ganoderma lucidum, Astragalus membranaceus, Anoectochilus roxburghii, ginseng, turmeric, clove, Poria cocos, Camellia chrysantha, Cordyceps militaris, chicken gizzard lining, peach kernel, and Viola yedoensis. Then, sift them through a 100-mesh sieve to obtain the raw material powder.

[0047] Step 2: Add 10 times the weight of purified water to each raw material powder, soak for 1 hour, boil, and extract by simmering for 40 minutes. Filter to obtain the first extract. Add 8 times the weight of purified water, boil, and extract by simmering for 30 minutes. Filter to obtain the second extract. Combine the two extracts and filter.

[0048] Step 3: The mixed extract was concentrated under reduced pressure at 65℃ and a vacuum of -0.085MPa to a paste with a relative density of 1.15-1.25. The paste was then spray-dried at an inlet air temperature of 190℃ and an outlet air temperature of 85℃ to obtain a fine powder.

[0049] Step 4: Add crocodile peptide, glutathione-enriched yeast, and compound probiotics to the fine powder of extract, and mix evenly at 35°C and 40% relative humidity to obtain the composition.

[0050] Example 2:

[0051] A composition for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors: 20 parts Ganoderma lucidum, 0.12 parts alligator peptide, 25 parts Astragalus membranaceus, 14 parts Anoectochilus roxburghii, 25 parts ginseng, 3 parts turmeric, 10 parts clove, 20 parts Poria cocos, 14 parts Camellia chrysanthemi, 0.35 parts glutathione-rich yeast, 14 parts Cordyceps militaris, 20 parts chicken gizzard lining, 10 parts peach kernel, 0.35 parts compound probiotics (Vibrio vulgaris MPL5, Lactobacillus plantarum La10, Lactobacillus casei C7-2, Lactobacillus reuteri HBM11-69, Lactobacillus rhamnosus HBM11-35 live bacteria count ratio 1:2:1:1:1), and 3 parts Viola yedoensis.

[0052] Preparation steps: Same as in Example 1, except that the inlet air temperature of spray drying in step 3 is adjusted to 185°C and the outlet air temperature is adjusted to 82°C.

[0053] Example 3:

[0054] A composition for regulating overall metabolic balance, inhibiting cancer cell growth, and fighting tumors: 15 parts Ganoderma lucidum, 0.08 parts alligator peptide, 20 parts Astragalus membranaceus, 10 parts Anoectochilus roxburghii, 20 parts Ginseng, 2 parts Curcuma longa, 8 parts Clove, 15 parts Poria cocos, 10 parts Camellia chrysantha, 0.2 parts glutathione-rich yeast, 10 parts Cordyceps militaris, 15 parts chicken gizzard lining, 8 parts Prunus persica, 0.2 parts compound probiotics (live count ratio of each strain of Pediococcus pentosaceus MPL5, Lactobacillus plantarum La10, Lactobacillus casei C7-2, Lactobacillus reuteri HBM11-69, and Lactobacillus rhamnosus HBM11-35 is 1:2:1:1:1), and 2 parts Viola yedoensis.

[0055] Preparation steps: Same as in Example 1, except that the extraction in step 1 is adjusted by using 8 times the amount of water for the first extraction and extracting for 40 minutes; and using 6 times the amount of water for the second extraction and extracting for 35 minutes.

[0056] Example 4:

[0057] Preparation of compound formulation (capsules): Take 100 parts of the composition prepared in Example 1, add 20 parts of lactose and 1 part of magnesium stearate, mix evenly and fill into empty capsules, each capsule containing 0.5g of the composition, thus obtaining the capsule formulation.

[0058] Example 5:

[0059] Preparation of compound preparation (oral liquid): Take 50 parts of the composition prepared in Example 1, add 500 parts of purified water, 0.5 parts of steviol glycoside, and 0.1 parts of ethylparaben. After stirring and dissolving, filter and fill into 10mL oral liquid bottles. Sterilize at 121℃ for 15min to obtain the oral liquid.

[0060] Comparative Example 1:

[0061] Compared to Example 1, the composition lacks glutathione-enriched yeast, and the amount of corydalis leaf fragments is adjusted to 4.4 parts.

[0062] Comparative Example 2:

[0063] Compared to Example 1, the composition lacks *Corydalis yanhusuo* and the amount of glutathione-enriched yeast is adjusted to 4.4 parts.

[0064] Experimental Example Composition Efficacy Verification

[0065] Experiment 1: Cell Experiment

[0066] 1. Experimental Materials and Methods

[0067] 1.1 Experimental Materials

[0068] Cell line: Human gastric cancer cell line AGS

[0069] Experimental sample: The composition of Example 1

[0070] Experimental reagents and consumables: CCK8 kit, FBS, PBS, RPMI 1640 medium, 0.25% trypsin

[0071] Main instruments: -80℃ freezer, ultrapure water system, magnetic stirrer, inverted microscope, low-speed centrifuge, electronic balance, carbon dioxide incubator, microplate reader, pipettes

[0072] Preparation of experimental samples: The composition of Example 1 was obtained according to the preparation method described above, and then prepared into a solution with a concentration of 1.2 g / mL for later use. Different concentrations can be prepared as needed for the experiment.

[0073] 1.2 Experimental Methods

[0074] CCK-8 test method

[0075] (1) A single-cell suspension was prepared by digesting the logarithmic growth phase human gastric cancer cell line AGS with trypsin, centrifuging, and then using complete culture medium.

[0076] (2) Cell counting, adjusting the cell suspension density to 1×10⁻⁶. 4 100 μl was inoculated into each well of a 96-well plate and incubated in a carbon dioxide incubator.

[0077] (3) Based on the preliminary experimental results, the sample concentration gradients were set at 0 mg / mL, 8 mg / mL, and 16 mg / mL. After 24 hours, the cell condition was observed. Once the adherent growth density reached 50%, a control group and an experimental group were set up, with 5 replicates in each group. The experimental group was given the gradient experimental samples and continued to be cultured in a carbon dioxide incubator for 24 hours, 48 ​​hours, and 72 hours, respectively.

[0078] (4) Three hours before the end, discard the supernatant, add culture medium containing 10% CCK-8 solution to each 96-well plate, continue culturing for 3 hours, and then terminate the culture.

[0079] (5) Discard the old culture medium, wash gently with PBS, add DMSO, and use an ELISA reader to detect the absorbance (OD value) of each well at a wavelength of 490 nm.

[0080] (6) Calculate the cell growth inhibition rate for each group: Cell proliferation inhibition rate = (OD 对照组 - OD 实验组 ) / OD 对照组 ×100%.

[0081] 2. Experimental Results

[0082] In the human gastric cancer cell line AGS, compared with the initial level, the inhibition rate of the sample on gastric cancer cell proliferation was positively correlated with both concentration and time, and the difference was statistically significant. The results indicate that the experimental sample has a significant inhibitory effect on the proliferation of gastric cancer AGS cells, and this effect is concentration- and time-dependent. (See [link to relevant documentation]). Figure 1 , Figure 2 .

[0083] Experiment 2 Animal Experiment

[0084] 1. Experimental Materials and Methods

[0085] 1.1 Experimental Materials

[0086] Cell line: Human gastric cancer cell line MKN45

[0087] Laboratory animals: SPF grade BALB / c nude mice, 4-6 weeks old, female, weighing 20±2g, housed separately in an environment with a temperature of 20-25℃ and a humidity of 50%±10%, with free access to water and standard feed.

[0088] Experimental sample: The composition of Example 1

[0089] Experimental reagents and consumables: FBS, PBS, RPMI 1640 medium, 0.25% trypsin

[0090] Main instruments: -80℃ freezer, ultrapure water system, magnetic stirrer, inverted microscope, low-speed centrifuge, electronic balance, carbon dioxide incubator, pipettes

[0091] Preparation of experimental samples: The composition of Example 1 was obtained according to the preparation method described above, and then prepared into a solution with a concentration of 1.2 g / mL for later use. Different concentrations can be prepared as needed for the experiment.

[0092] 1.2 Experimental Methods

[0093] 1.2.1 Animal Modeling

[0094] The gastric cancer cell line MKN45 was removed from the liquid nitrogen storage tank. After cell resuscitation, routine culture, and passage, the MKN45 gastric cancer cells were resuspended in PBS and viable cell counts were performed. The cells were then inoculated into the first batch of 10 BALB / c nude mice, 0.2 mL per mouse. The mice were allowed free access to food and water, and subcutaneous tumor growth was observed. Once a tumor developed and was confirmed, the mice were euthanized, the tumor was removed, and a small piece (approximately 5 mm) was cut. 3 The tumor was then transplanted into a second batch of mice, and mice that successfully developed the model were selected.

[0095] 1.2.2 Animal grouping and administration

[0096] Mice bearing gastric cancer were randomly divided into three groups: model group, cisplatin group, and drug administration group (Example 1 group, Example 2 group, Comparative Example 1 group, and Comparative Example 2 group).

[0097] The experiment lasted 14 days. The dosage of the composition was calculated using the conversion factor between humans and mice. The dosage for mice was nine times that of adults (70 kg). The daily dose for mice was set at 27 g / kg, administered via gavage daily. The model group received an equal volume of 0.9% saline via gavage daily. The cisplatin group received 10 mg / kg via intraperitoneal injection every other day, with an injection volume of 0.1 mL / 10 g.

[0098] 1.2.3 Recording and Evaluation

[0099] Observe the mice's mental state, activity level, fur, etc., record the observations, and evaluate and score them.

[0100] Table 1. General Condition Scoring Table for Mice

[0101]

[0102] 1.2.4. Collection of materials

[0103] Mice were fasted but allowed free access to water the day before sampling. On day 16, the mice were sacrificed, and samples were collected for testing. The dissected tumors were gently rinsed with physiological saline to remove excess blood, blotted dry on filter paper, and then weighed.

[0104] Tumor inhibition rate = [1 - (average tumor weight in the treatment group / average tumor weight in the model group)] × 100%

[0105] 1.2.5 Statistical Analysis

[0106] All data were statistically analyzed using SPSS 26.0 software. Results are expressed as mean ± standard deviation (x̄ ± s). For two groups of data that are normally distributed and have homogeneous variances, a t-test was used. For three or more groups of data, one-way ANOVA was used. P <0.05 indicates that the difference is statistically significant.

[0107] 2. Experimental Results

[0108] 2.1 Body weight and general condition of tumor-bearing mice

[0109] No mice died during the experiment, but some exhibited behaviors such as biting each other and tail injuries. Mice in all groups showed signs of deterioration, reduced activity, hair loss, and impaired feeding and drinking, with the model group showing the most pronounced effects.

[0110] Compared to the model group, the weight gain in each treatment group was relatively greater after treatment. Compared to the cisplatin positive control group, the weight gain in each treatment group was significantly higher, possibly due to the side effects of cisplatin. Examples 1 and 2 showed better weight gain than Comparative Examples 1 and 2. Compared to the model group, the scores in each treatment group were higher, suggesting that the treatment groups improved the general condition of tumor-bearing mice. The effects of treatment on mouse weight and general condition scores before and after treatment are shown in Table 2 and... Figure 3-4 .

[0111] Table 2. Assessment of mouse body weight and general condition (x̄±s, n=10)

[0112]

[0113] 2.2 Tumor Suppression Effect

[0114] Compared with the model group, the tumor growth rate of each group was relatively slowed down. The tumor weight of the treatment groups (Examples 1 & 2), Comparative Example 2, and the positive drug cisplatin group was reduced, showing statistically significant differences. Compared with the positive drug cisplatin group, there were statistically significant differences in the treatment groups (Examples 1 & 2) and Comparative Examples 1 & 2. It can be seen that all treatment groups were able to inhibit tumor growth, suggesting an in vivo antitumor effect. The average tumor weight and tumor inhibition rate of each group are shown in Table 3. Figure 5-6 .

[0115] Table 3. Tumor weight and tumor inhibition rate in each group

[0116]

[0117] 3. Discussion

[0118] This invention integrates modern medical science concepts with the traditional Chinese medicine theory of "blood stasis and qi stagnation leading to lumps". It targets several cancer-related factors, including metabolic abnormalities, low blood oxygen content, inflammation-induced mitochondrial damage, low immune function, and blood stasis, achieving a synergistic effect of "1+1>2" and providing more comprehensive improvement advantages than single-component or traditional compound formulas.

[0119] The formula of this invention includes crocodile peptides, which are rich in eight essential amino acids and can be derived from one or more of crocodile blood, meat, and bones. *Anoectochilus roxburghii*, crocodile peptides, and glutathione-rich yeast are used to enhance the body's anti-inflammatory and free radical scavenging abilities. *Corydalis yanhusuo* is a selenium-rich plant with a high organic selenium content, possessing regulatory and antioxidant effects on oxidative stress. *Astragalus membranaceus*, ginseng, and *Camellia sinensis* improve blood oxygen supply and saturation. *Curcuma longa*, peach kernel, and chicken gizzard lining are used together to soften and disperse nodules, promote blood circulation, remove blood stasis, and relieve pain. The anti-tumor potential of curcumin, the active ingredient in turmeric, has been proven. Its core mechanisms include inhibiting cancer cell proliferation, invasion, and metastasis, and blocking tumor angiogenesis. Its anti-inflammatory and antioxidant properties can also help reduce the side effects of tumor treatment. Based on this, *Ganoderma lucidum*, *Poria cocos*, *Cordyceps militaris*, and cloves are added to enhance the body's immunity, strengthen the killing effect on tumor cells, inhibit their immune escape, and thus block tumor proliferation. In addition, the introduction of compound probiotics, whether live or inactivated, can strengthen the mucosal barrier, improve the intestinal microecology, alleviate tumor treatment-related side effects, and improve patients' quality of life.

[0120] This invention aims to protect mitochondria from damage and prevent mutations by increasing blood oxygen levels, reducing inflammation, improving metabolic function, and promoting blood circulation. Furthermore, by supplementing with a variety of probiotics (either live or inactivated), it regulates the intestinal flora, enhances immunity, and ultimately achieves the effects of softening and dispersing nodules, regulating overall metabolic balance, inhibiting cancer cell growth, fighting tumors, and alleviating the side effects of tumor treatment. The raw materials are all medicinal and edible or natural active ingredients, ensuring high safety and no significant toxic side effects. They can be prepared into various dosage forms, such as oral liquids, to meet the needs of different patients (e.g., those with swallowing difficulties or those requiring long-term use), and have broad application prospects.

[0121] 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. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition for regulating overall metabolic balance, inhibiting the growth of gastric cancer cells, and having anti-tumor effects, characterized in that, The raw material composition of the composition is as follows: 20-25 parts of Ganoderma lucidum, 0.1-0.15 parts of crocodile peptide, 25-30 parts of Astragalus membranaceus, 12-16 parts of Anoectochilus roxburghii, 25-30 parts of ginseng, 2-4 parts of turmeric, 10-12 parts of clove, 18-22 parts of Poria cocos, 12-16 parts of Camellia chrysantha, 0.3-0.4 parts of glutathione-enriched yeast, 12-16 parts of Cordyceps militaris, 18-22 parts of chicken gizzard lining, 10-12 parts of peach kernel, 0.3-0.4 parts of compound probiotics, and 3-4 parts of Viola yedoensis and Citrus aurantium. The compound probiotics consist of the following probiotics: Pediococcus pentosaceus MPL5, Lactobacillus plantarum La10, Lactobacillus casei C7-2, Lactobacillus reuteri HBM11-69, and Lactobacillus rhamnosus HBM11-35, with a live bacteria ratio of 1:2:1:1:

1. The preservation number of *Pediococcus pentosaceus* MPL5 is GDMCC NO: 63606, the preservation number of *Lactobacillus plantarum* La10 is GDMCC NO: 63605, the preservation number of *Lactobacillus casei* C7-2 is GDMCC NO: 63609, the preservation number of *Lactobacillus reuteri* HBM11-69 is GDMCC NO: 63608, and the preservation number of *Lactobacillus rhamnosus* HBM11-35 is GDMCC NO: 63607.

2. A compound preparation that regulates overall metabolic balance, inhibits the growth of gastric cancer cells, and has anti-tumor effects, characterized in that, The raw material composition of the compound preparation is: the composition according to claim 1, a pharmaceutically acceptable carrier or excipient.

3. The compound formulation according to claim 2, characterized in that, The carrier or excipient includes one or more of the following: diluent, wetting agent, lubricant, adhesive, disintegrant, encapsulating agent, flavoring agent, sustained-release agent, dispersant, plasticizer, and opaque agent.

4. The compound formulation according to claim 2 or 3, characterized in that, The compound preparation is an oral preparation, and its dosage form is powder, granules, pills, tablets, capsules, ointments, or oral liquid.

5. A method for preparing the composition according to claim 1, characterized in that, Includes the following steps: Step 1: Grind the following ingredients separately: Ganoderma lucidum, Astragalus membranaceus, Anoectochilus roxburghii, ginseng, turmeric, clove, Poria cocos, Camellia chrysantha, Cordyceps militaris, chicken gizzard lining, peach kernel, and Viola yedoensis. Then, sift them through an 80-100 mesh sieve to obtain the raw material powder. Step 2: Add 5-15 times the weight of purified water to each raw material powder, soak for 0.5-1h, heat to boiling, maintain a gentle boil and extract 2-3 times, each extraction for 30-45min, combine the extracts of each raw material, filter to remove the residue, and obtain a mixed extract. Step 3: The mixed extract is concentrated under reduced pressure at 60-70℃ and vacuum degree of -0.08 to -0.09MPa to obtain an extract with a relative density of 1.15-1.

25. The extract is then spray-dried at an inlet air temperature of 180-200℃ and an outlet air temperature of 80-90℃ to obtain a fine powder. Step 4: Add crocodile peptide, glutathione-enriched yeast, and compound probiotics to the fine powder of extract, and mix evenly in an environment of 30-35℃ and relative humidity ≤45% to obtain the composition.

6. The preparation method according to claim 5, characterized in that, In step 2, the first extraction is done with 10 times the weight of the raw material in purified water, and the second and third extractions are done with 8 times the weight of the raw material in purified water, respectively. In step 3, the inlet air temperature of the spray dryer is 190℃ and the outlet air temperature is 85℃.

7. The use of a composition as described in claim 1 or a compound formulation as described in any one of claims 2-4 in the preparation of a formulation for treating gastric cancer.

Images