A traditional Chinese medicine composition for treating colorectal cancer and its preparation method and application

Abstract

The present invention discloses a Chinese medicine composition called Fushao Diqin Fang for treating colorectal cancer (CRC) and its preparation method and application. It is made up of the following Chinese medicinal materials in parts by weight: 6-15 parts of white peony root, 2-10 parts of liquorice, 3-15 parts of aconite root, 3-10 parts of scutellaria root, 4.5-9 parts of sophora flavescens, 10-15 parts of raw rehmannia root, 10-15 parts of spinach seed, 3-10 parts of dried ginger, 9-30 parts of oyster, 3-6 parts of amomum, 9-15 parts of poria, 5-10 parts of earthworm. This Chinese medicine composition has the effect of treating CRC, and it is both strengthening the body and dispelling pathogenic factors. It is suitable for CRC patients whose TCM syndrome differentiation is "spleen and kidney yang deficiency, yin and blood deficiency, damp-heat and blood stasis" syndrome, and both deficiency of the body and excess of pathogenic factors, and has a definite therapeutic effect. The Chinese medicinal materials used in the present invention are all from natural plants, and raw materials are easy to obtain, with high safety, and are easily prepared into medicines such as tablets, pills, capsules and granules, suitable for clinical application and promotion.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a traditional Chinese medicine composition for treating colorectal cancer prepared from traditional Chinese medicine as raw materials, as well as a preparation method and application thereof. Background Art

[0002] Colorectal cancer (CRC) is the third most common cancer worldwide. Common treatments for CRC include surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy, which have prolonged patient survival to a certain extent. However, drug resistance often develops and is accompanied by serious side effects and adverse reactions, such as gastrointestinal reactions and bone marrow suppression, which restrict CRC treatment. Therefore, reducing the morbidity and mortality of CRC, prolonging the survival of CRC patients, and improving their quality of life have become major challenges that need to be addressed urgently.

[0003] In recent years, through extensive clinical and basic research, Traditional Chinese Medicine (TCM) has been shown to play an important role in intervening in CRC precancerous lesions, reducing toxicity and increasing efficacy, preventing recurrence and metastasis, improving patient quality of life, prolonging survival, and relieving toxic side effects and adverse reactions during treatment. TCM also offers advantages such as low cost, multiple targets, multiple components, low drug resistance, and minimal toxicity and adverse reactions. Therefore, TCM is an effective clinical treatment for CRC, with significant efficacy and a high safety profile. Summary of the Invention

[0004] The present invention provides a traditional Chinese medicine composition for treating colorectal cancer and a preparation method thereof. The composition is suitable for CRC patients whose TCM syndromes are "spleen and kidney yang deficiency, yin and blood deficiency, damp-heat, blood stasis and toxins" and who are in the stage of both deficiency of the body's vital energy and excess of pathogenic factors. The composition can exert the therapeutic effects of both strengthening the body's vital energy and eliminating pathogenic factors, and solve the technical difficulties of conventional colorectal cancer treatment in modern medicine, such as large toxic side effects and adverse reactions, high cost, easy drug resistance of tumors, high recurrence and metastasis rates, and low quality of life for patients.

[0005] The present invention adopts the following technical scheme: A traditional Chinese medicine composition for treating colorectal cancer is prepared by extracting the following traditional Chinese medicines in parts by weight: 6-15 parts of white peony root, 2-10 parts of liquorice, 3-15 parts of aconite root, 3-10 parts of scutellaria baicalensis, 4.5-9 parts of sophora flavescens, 10-15 parts of raw rehmannia root, 10-15 parts of spinach seed, 3-10 parts of dried ginger, 9-30 parts of oyster, 3-6 parts of amomum villosum, 9-15 parts of poria, and 5-10 parts of earthworm.

[0006] The traditional Chinese medicine composition for treating colorectal cancer is prepared by extracting the following traditional Chinese medicines in parts by weight: 10 parts of white peony root, 6 parts of liquorice, 9 parts of aconite root, 7 parts of scutellaria baicalensis, 7 parts of sophora flavescens, 12 parts of raw rehmannia root, 15 parts of spinach seed, 9 parts of dried ginger, 30 parts of oyster, 6 parts of amomum villosum, 15 parts of poria, and 9 parts of earthworm.

[0007] In the traditional Chinese medicine composition for treating colorectal cancer, the licorice is raw licorice, the aconite root is Heishunpian, the spinach seeds are stir-fried spinach seeds, and the oyster is raw oyster.

[0008] The preparation method of the traditional Chinese medicine composition for treating colorectal cancer comprises the following steps: weighing the above medicinal materials according to the dosage, adding 10-20 times of the weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 2-3 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.2-1.3 at 25°C, cooling, adding 85-90% ethanol to adjust the ethanol concentration to 65-75%, precipitating with alcohol, fully mixing, leaving overnight, taking the supernatant, recovering the ethanol under reduced pressure to obtain an extract, spray drying, sieving, and adding appropriate amounts of auxiliary materials to prepare a preparation.

[0009] The traditional Chinese medicine composition for treating colorectal cancer is in the form of granules, tablets, capsules, powders, pills, oral liquids or pastes.

[0010] The Chinese medicine composition for treating colorectal cancer belongs to the syndrome of "spleen and kidney yang deficiency, yin and blood deficiency, dampness, heat, blood stasis and toxins" in traditional Chinese medicine differentiation.

[0011] Formula basis and beneficial effects

[0012] Colorectal cancer is also known as large intestinal cancer, which falls into the category of diseases such as "visceral poison", "intestinal accumulation", "intestinal wind" and "locked anus hemorrhoids" in traditional Chinese medicine. Professor Zhou Zhongying, a master of traditional Chinese medicine, proposed the "cancer poison" pathogenesis theory based on the pathogenicity, intractability and stubbornness of malignant tumors. He believes that factors such as the six exogenous evils, improper diet or emotional imbalance lead to excessive evil or accumulate into cancer poison over time, or endogenous cancer poison due to dysfunction of internal organs and deficiency of vital energy. Cancer poison is mutually bonded with wind, fire (heat), phlegm, blood stasis, dampness and cold and other pathological factors. It is the key to the occurrence, development, metastasis and recurrence of malignant tumors, and the core pathogenesis of colorectal cancer is the mutual bond of dampness, heat and blood stasis. The applicant has been treating colorectal cancer with the cancer poison pathogenesis theory for many years, and believes that cancer poison is the key to the onset of colorectal cancer. Its location is in the large intestine and is related to internal organs such as the spleen, stomach, liver and kidneys. Spleen deficiency generates dampness or infection, which then transforms into fire or accumulates into heat over time, leading to earth deficiency and wood stagnation, liver qi stagnation, qi stagnation, and blood stasis. These evil qi, dampness, heat, and blood stasis, accumulate and become cancerous toxins, manifesting as dampness, heat, and blood stasis. These evil qi, intermingled with common pathogens like dampness, heat, and blood stasis, flow into the large intestine, causing cancer. "The Origin of Miscellaneous Diseases: Xizhu: The Origin of Accumulation Syndromes, Tumors, and Abnormalities" states: "A healthy individual will not have accumulations. This is because they are deficient in vital energy, allowing the retention of evil qi." "Accumulation" in "Medical Essentials: Accumulation" states: "Accumulations are caused by insufficient vital energy, which then allows the accumulation of evil qi." Furthermore, according to the theory of vital energy deficiency, "deficiency of vital energy leads to the formation of rocks." Cancerous toxins form on the basis of vital energy deficiency, which then depletes vital energy and blood for self-sustaining, further depleting vital energy. This deficiency of vital energy must be explained using the theory of yin and yang, which manifests as a deficiency of yin (essence, blood, and body fluids) and yang (qi) in the body's internal organs. According to Huang Yuanyu's theory of Zhongqi, "the space between clear and turbid is called Zhongqi. Zhongqi, also known as earth, is the pivot of the rise and fall of yin and yang, the foundation of the transformation of yin and yang, the five elements, and the life and growth of all things." Zhongqi in the body is the qi of the spleen and stomach, transporting essential substances to nourish the internal organs, meridians, and bones. It is the foundation of the acquired organs and controls the rise and fall of qi throughout the body. The leftward rise of the liver and the rightward descent of the lungs depend entirely on the smooth flow of qi from the spleen and stomach, circulating the middle earth and unifying qi. Malignant tumors deplete vital energy, primarily affecting the body's Zhongqi, leading to a deficiency of spleen and stomach yang qi. Yin and yang are interdependent, and without yang, yin cannot exist. This damages the yin fluid of the spleen and stomach, which gradually affects the liver, kidneys, and other organs, resulting in a deficiency of both yin and yang. The applicant combined the classic theories of cancer poison theory, vital energy deficiency theory, yin and yang theory, and central qi theory to diagnose and treat colorectal cancer. At the same time, in many years of clinical practice in tumors, they found that many colorectal cancer patients had symptoms of damaged yang qi, such as cold body, fear of cold, cold stomach, poor appetite, frequent and long urination at night, and symptoms of insufficient yin blood, such as weight loss, dry stool, and dry eyes. At the same time, they had symptoms of dampness, heat, blood stasis, and toxins, such as bitter taste in the mouth, yellow urine, sticky stool, and purple tongue. This confirmed the core pathogenesis of colorectal cancer patients, "deficiency of vital energy and excess of evil, damaged yang qi, insufficient yin blood, and coexistence of dampness, heat, blood stasis and toxins." Clinical treatment should strengthen the vital energy, eliminate evil, replenish yin and yang, and eliminate cancer and detoxify."Essential Readings in the Medical Ancestral Book: Accumulation" points out: "In the initial stage, when the pathogenic factors first appear, the vital energy is still strong and the pathogenic factors are still shallow, then the Ren Qi will be attacked; in the middle stage, when the disease lasts for a long time, the pathogenic factors are deeper and the vital energy is weaker, then the Ren Qi will be attacked and supplemented; in the final stage, when the disease lasts for a long time, the pathogenic factors invade and the vital energy is exhausted, then the Ren Qi will be supplemented." The applicant pointed out that the clinical diagnosis and treatment of colorectal cancer should combine the core pathogenesis with the stage of the disease. Although the cancer toxins are all manifested as the combination of dampness, heat, blood stasis and toxins, the degree of vital energy deficiency, pathogenic factors prevalence, and internal organs yin and yang deficiency varies in different stages of the disease. In the early stage of colorectal cancer, dampness, heat, blood stasis and toxins are intertwined, blocking and damaging the vital energy, causing deficiency of spleen and stomach yang, and later leading to insufficient yin and fluid. At this time, the deficiency of the vital energy is not serious, and the cancer toxins are just beginning. In the middle stage of colorectal cancer, the deficiency of the vital energy and the prevalence of evil are equally serious. The accumulation of cancer toxins leads to further deficiency of the vital energy, deficiency of both yin and yang of the spleen and stomach, and inability of the acquired to nourish the innate. Dampness, heat, blood stasis and toxins further deplete the innate foundation, leading to loss of kidney yang and insufficient kidney yin. The kidney's transpiration, gasification and moisturizing functions are weakened, and water cannot contain wood. The mother's disease affects the child, leading to insufficient liver yin and blood. In the late stage of colorectal cancer, the vital energy becomes further deficient, and the yin and yang of the spleen, stomach, liver and kidney are all deficient, while the cancer toxins are rampant. At this time, the vital energy is exhausted and the evil energy is strong.

[0013] The mid-stage of colorectal cancer presents a stage of both deficiency of vital energy and prevalence of pathogenic factors. Based on the aforementioned core pathogenesis, the TCM syndrome is summarized as "spleen and kidney yang deficiency, yin and blood deficiency, and the accumulation of dampness, heat, stasis, and toxins." Clinical treatment should warm and tonify the spleen and kidney yang, nourish yin and blood, and dissolve the accumulation of dampness, heat, stasis, and toxins, thereby achieving the effects of strengthening vital energy, dispelling pathogenic factors, eliminating cancer, strengthening the middle, replenishing essence, harmonizing yin and yang, and promoting the circulation of qi. To warm and tonify the spleen and kidney yang, a warming approach is used, consisting of warming herbs that warm yang, replenish qi, promote qi circulation, and regulate qi. To nourish yin and blood, a nourishing approach is used, consisting of moistening herbs that nourish yin, nourish blood, promote fluid production, and replenish essence. To dissolve the accumulation of dampness, heat, stasis, and toxins, a detoxifying approach is used, consisting of dampness-removing, heat-clearing, blood-activating, stasis-resolving, and knot-dispersing herbs that dispel dampness and detoxify, clear heat and detoxify, resolve stasis and detoxify, and dissolve cancer and knots. The Chinese medicine composition of the present invention, Fushao Diqin prescription, is a prescription that emphasizes both attack and tonification, and is formulated under the guidance of the cancer-toxicity theory, the vital energy deficiency theory, the yin-yang theory, and the central qi theory, based on the traditional Chinese medicine symptoms of "spleen and kidney yang deficiency, yin-blood deficiency, and damp-heat, blood stasis and toxins" in patients with colorectal cancer. It is targeted at the stage of the disease where "the evil qi is deep and the vital energy is weak" where both vital energy deficiency and pathogenic factors are prevalent, so as to exert the therapeutic effects of both strengthening the vital energy and eliminating pathogenic factors, warm and tonify the yang of the spleen and kidney, nourish the yin and blood, and eliminate the accumulation of dampness, heat, blood stasis and toxins. The Fushao Diqin prescription is adapted from many classic small prescriptions such as Xuanwu Decoction (Zhenwu Decoction), Sini Decoction, Shaoyao Gancao Fuzi Decoction and Sanwu Huangqin Decoction. In the prescription, the aconite greatly replenishes the kidney's yang energy and restores yang to rescue the adverse condition, while the white peony root and the sour jujube seed nourish the yin and blood of the liver and spleen. The three medicines are used together. The aconite promotes the qi and blood of the internal organs and helps the white peony root and the sour jujube seed nourish yin and blood. The sourness of the white peony root and the sour jujube seed astringes yin and alleviates the dry and strong nature of the aconite. It combines movement and stillness, and is appropriate for gathering and dispersing. It uses both hardness and softness, and adopts both warming and nourishing methods to restore yang and benefit yin. They are all the main medicines. Dried ginger warms the middle and strengthens the spleen, restoring yang and unblocking the meridians. It assists aconite in warming and tonifying kidney yang and also replenishing the yang energy of the spleen and stomach. Raw rehmannia nourishes the yin and blood of the liver and kidneys, assisting white peony root and Chinese jujube seed in nourishing yin and blood. The combined effects of dried ginger and raw rehmannia are harmonious, enhancing the warming and nourishing effects of the main herb. Together, they achieve the functions of warming and tonifying the yang of the spleen and kidneys and nourishing yin and blood, providing a strong strengthening effect. Furthermore, raw rehmannia clears heat and cools the blood to resolve heat and toxicity. Combined with scutellaria baicalensis and sophora flavescens, they clear heat and dampness, purge fire and detoxify, eliminating heat and dampness toxins and significantly dispelling pathogenic factors. These herbs, acting simultaneously as both strengthening and dispelling pathogenic factors, serve as assistant herbs. Oyster, with its salty and cold properties, enters the liver and kidneys. Combined with aconite, it warms and subdues yin and yang, while also dispelling cancer and dispersing nodules. Earthworm unblocks the meridians, activates blood circulation, and resolves stasis. Poria cocos calms the mind and invigorates the spleen and promotes warmth. All three serve as adjuvant herbs. Amomum villosum is pungent and warm, warming the middle and promoting qi, strengthening the spleen and eliminating dampness, supporting spleen yang and removing dampness and stasis. Raw licorice root, combined with white peony root, spinach seed, and aconite root, transforms yin with sour and sweet, transforms yang with pungent and sweet, clearing away heat and detoxifying, tonifying qi and strengthening the spleen while also regulating the spleen and stomach, and harmonizing the other herbs. These two herbs serve as guiding herbs. The combined use of warming, nourishing, and detoxifying methods in the formula can achieve the effects of warming and tonifying spleen and kidney yang qi, nourishing yin and blood, and eliminating dampness, heat, stasis, and toxicity. It emphasizes both strengthening the body and removing pathogenic factors, with potent and specific medicinal properties, and is safe and feasible when combined.This prescription differs from traditional Chinese medicine prescriptions that primarily invigorate the spleen and replenish qi, such as Astragalus, Atractylodes macrocephala, and Codonopsis pilosula, which have mild properties and are used to strengthen the body's health. It also differs from traditional Chinese medicine prescriptions that primarily invigorate the spleen and replenish qi, such as Hedyotis diffusa and Paris polyphylla, which have been shown in modern pharmacological studies to have anti-cancer and anti-toxic effects. This prescription focuses on the mid-stage colorectal cancer, where both deficiency of the body's health and excess of pathogenic factors are prevalent. It firmly grasps the core pathogenesis of this stage, characterized by "spleen and kidney yang deficiency, yin and blood deficiency, and the interplay of damp-heat, blood stasis, and toxins." It prioritizes both strengthening the body's health and dispelling pathogenic factors, warming and tonifying the spleen and kidney's yang, nourishing yin and blood, and dissolving the interplay of damp-heat, blood stasis, and toxins. Its potent properties, combined with appropriate combinations, and potent and specific efficacy directly target the affected area, resulting in rapid, safe, and feasible results. The Fushao Diqin prescription (hereinafter referred to as FSDQ) of this invention has excellent clinical efficacy, minimal toxic and adverse reactions, and is highly safe and suitable for long-term use, effectively improving the clinical efficacy of traditional Chinese medicine in preventing and treating CRC. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 HE staining of mouse tumor tissue (A: model group; B: oxaliplatin group; C: Fushao Diqin prescription (FSDQ) low-dose group; D: FSDQ medium-dose group; E: FSDQ high-dose group);

[0015] Figure 2 It is the migration inhibitory effect of FSDQ on colorectal cancer cell CT-26;

[0016] Figure 3 It is the migration inhibitory effect of FSDQ on colorectal cancer cell RKO;

[0017] Figure 4 The result is that FSDQ promotes apoptosis of colorectal cancer cells CT-26. DETAILED DESCRIPTION

[0018] The present invention will be further described below with reference to the embodiments, but the present invention is not limited to these specific examples.

[0019] Example 1

[0020] The invention discloses a preparation method for preparing 100g of white peony root, 60g of liquorice root, 90g of aconite root, 70g of scutellaria root, 70g of sophora flavescens, 120g of raw rehmannia root, 150g of spinach seed, 90g of dried ginger, 300g of oyster, 60g of amomum villosum, 150g of poria and 90g of earthworm. The preparation method comprises the following steps: weighing the above medicinal materials according to the amount, adding 15 times the weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 2 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.25 at 25°C, cooling, adding 90% ethanol to adjust the ethanol concentration to 65%, precipitating with alcohol, fully mixing, standing overnight, taking the supernatant, recovering the ethanol under reduced pressure to obtain an extract, spray drying, sieving, adding dextrin and preparing granules.

[0021] Example 2

[0022] The invention discloses a preparation method for preparing 60g of white peony root, 100g of liquorice root, 30g of aconite root, 100g of scutellaria root, 4.50g of sophora flavescens, 150g of raw rehmannia root, 100g of spinach seed, 120g of dried ginger, 90g of oyster, 90g of amomum villosum, 90g of poria and 150g of earthworm. The preparation method comprises the following steps: weighing the above medicinal materials according to the amount, adding 20 times of their weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 2 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.3 at 25°C, cooling, adding 85% ethanol to adjust the concentration to 75%, precipitating with alcohol, fully mixing, standing overnight, taking the supernatant, recovering the ethanol under reduced pressure to obtain an extract, spray drying, sieving, adding an appropriate amount of starch and preparing tablets.

[0023] Example 3

[0024] Take 150g of white peony root, 20g of liquorice, 150g of aconite root, 30g of scutellaria baicalensis, 90g of sophora flavescens, 100g of raw rehmannia root, 150g of spinach seed, 30g of dried ginger, 400g of oyster, 30g of amomum villosum, 250g of poria, and 50g of earthworm. The preparation method comprises the following steps: weighing the above medicinal materials according to the amount, adding 10 times the weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 3 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.2 at 25°C, cooling, adding 90% ethanol to adjust the ethanol concentration to 65%, precipitating with alcohol, fully mixing, standing overnight, taking the supernatant, reducing the pressure to recover ethanol to obtain an extract, spray drying, sieving, adding microcrystalline cellulose, and preparing capsules.

[0025] Example 4

[0026] The invention discloses a preparation method for 100g of white peony root, 60g of liquorice root, 90g of aconite root, 70g of scutellaria root, 70g of sophora flavescens, 120g of raw rehmannia root, 150g of spinach seed, 90g of dried ginger, 300g of oyster, 60g of amomum villosum, 150g of poria and 90g of earthworm. The preparation method comprises the following steps: weighing the above medicinal materials according to the amount, adding 10-20 times of their weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 2-3 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.2-1.3 at 25°C, cooling, adding 85-90% ethanol to adjust the ethanol concentration to 65-75%, precipitating with alcohol, fully mixing, standing overnight, collecting the supernatant, recovering the ethanol under reduced pressure to obtain an extract, and adding a flavoring agent to prepare an oral solution.

[0027] Example 5

[0028] The invention discloses a preparation method for preparing 100g of white peony root, 60g of liquorice root, 90g of aconite root, 70g of scutellaria root, 70g of sophora flavescens, 120g of raw rehmannia root, 150g of spinach seed, 90g of dried ginger, 300g of oyster, 60g of amomum villosum, 150g of poria and 90g of earthworm. The preparation method comprises the following steps: weighing the above medicinal materials according to the amount, adding 10-20 times of their weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 2-3 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.2-1.3 at 25°C, cooling, adding 85-90% ethanol to adjust the ethanol concentration to 65-75%, precipitating with alcohol, fully mixing, standing overnight, taking the supernatant, recovering the ethanol under reduced pressure to obtain an extract, spray drying, passing through a 100-mesh sieve, adding appropriate amounts of auxiliary materials and preparing pills.

[0029] Example 6: In vivo anti-CRC effect of the present invention

[0030] To investigate the in vivo anti-CRC effect of the traditional Chinese medicine composition of the present invention [name: Fu Shao Di Qin Fang (abbreviation: FSDQ)], a CT-26 cell CRC xenograft mouse model was constructed. The experiment was divided into blank control group, model group, oxaliplatin group, and FSDQ low-, medium-, and high-dose groups.

[0031] 1. Experimental Procedure

[0032] 1.1 Establishment, grouping, and drug administration of subcutaneous colorectal cancer transplants

[0033] 0.1 ml of CT-26 single cell suspension in the logarithmic growth phase was extracted and inoculated subcutaneously on the right side of the mouse back. The cell density of the single cell suspension was 1×10 7 When the volume of mouse subcutaneous transplanted tumor is 100mm 3 -300mm 3 When the model is successfully established, the drug can be started.

[0034] A blank control group (Control) was set up, and the mice with successful modeling were randomly divided into the model group, oxaliplatin group, and Example 1 Fu Shao Di Qin Fang low-dose group, medium-dose group and high-dose group. Fu Shao Di Qin Fang low-dose, medium-dose and high-dose groups were 4.49, 8.97 and 17.94 g of crude drug kg -1 ·d -1 The positive control group was given oxaliplatin 1.5 mg kg -1 ·d -1 The blank control group and the model group were intraperitoneally injected with an equal amount of normal saline, once a day for 21 consecutive days, with a dosing volume of 0.2 ml / 10 g body weight.

[0035] 1.2 Monitoring of mouse body weight and tumor volume

[0036] Mouse weight and tumor volume were measured every three days after dosing. The major and minor diameters of the transplanted tumors were measured using a vernier caliper, and tumor volume and tumor inhibition rate were calculated. Relative tumor volume (RTV) = Vt / V0 (V0 is the tumor volume at the start of dosing, and Vt is the tumor volume on day t of dosing). Tumor inhibition rate (TGI) = (1 - RTV experimental group / RTV model group) × 100%.

[0037] 1.3 HE staining

[0038] After the tumor tissue was peeled off, it was fixed with 4% paraformaldehyde solution, dehydrated, embedded, sectioned, and then routinely stained with HE and observed under a microscope.

[0039] 2. Experimental Results

[0040] Table 1 Changes in body weight and tumor volume of mice in each group ( n=6)

[0041]

[0042] Note: Compared with the model group, *P<0.05, **P<0.01

[0043] By observing the changes in tumor volume (Table 1), it was found that the tumor in the model group grew rapidly. Compared with the model group, the tumor volume of the low, medium and high dose groups of FSDQ and the oxaliplatin group of the present invention was reduced to varying degrees. The tumor inhibition rates of mice in each group were calculated by relative tumor volume, and were 33.30%, 42.02%, 54.86% and 43.36%, respectively. The results showed that FSDQ can inhibit tumor growth. The effect of the medium dose of FSDQ was roughly the same as that of the oxaliplatin group, and the high dose of FSDQ had the strongest tumor inhibition effect. The morphology of tumor tissue cells in each group was further observed by HE staining to evaluate the in vivo anti-CRC effect of FSDQ ( Figure 1 ) revealed that tumor tissue cells in each group were irregularly arranged and morphologically diverse, exhibiting characteristics of tumor cells, with visible areas of tumor necrosis. In the model group, tumor cells were intact, tightly packed, with abundant cytoplasm and numerous mitotic figures. Compared with the model group, all treatment groups showed varying degrees of nuclear pyknosis, a decreased nuclear-cytoplasmic ratio, and enlarged necrotic areas. Tumor tissue was loosely arranged and its density decreased in a dose-dependent manner. These results confirm the potent anti-CRC activity of FSDQ in vivo.

[0044] Example 7: In vitro anti-CRC effect of the present invention

[0045] The in vitro anti-CRC effect of the Fushaodiqin prescription (FSDQ) of the present invention was studied using two CRC cell lines, CT-26 and RKO.

[0046] 1. Experimental Procedure

[0047] 1.1 Recovery and culture of colorectal cancer CT-26 cells

[0048] Mouse colorectal cancer CT-26 cells cryopreserved in liquid nitrogen were quickly rewarmed in a 37°C water bath and added to RPMI-1640 medium containing 10% FBS and 1% penicillin-streptomycin. The cells were incubated at 37°C in a 5% CO2 incubator. Cell experiments were performed when the cells reached the logarithmic growth phase.

[0049] 1.2MTT IC test 50 , detecting cell proliferation

[0050] CT-26 cells and RKO cells in the logarithmic growth phase were cultured at 5×10 3 and 8×10 3 After the cells adhered to the wall, different concentrations (0, 0.4, 0.8, 1.2, 1.6, 2.0, 2.4, 2.8 mg mL) were added to each well. -1 ) Radix Paeoniae Rubra and Radix Rehmanniae lyophilized powder (preparation method: combine the two medicinal solutions according to the steps of Example 1, filter and concentrate to obtain the Radix Paeoniae Rubra and Radix Rehmanniae lyophilized powder, freeze it at -80°C for 48 hours, quickly transfer it to a freeze dryer, and vacuum dry it to obtain the Radix Paeoniae Rubra and Radix Rehmanniae lyophilized powder) culture medium, set up 3 replicate wells for each group, culture for 48 hours, add MTT solution, continue incubation for 4 hours, discard the culture medium, add 150 μL DMSO solution to each well, and detect the absorbance value of each group at 490 nm with a microplate reader.

[0051] 1.3 Transwell assay for CT-26 cell migration

[0052] The cells were divided into blank control group (Control), positive control group (oxaliplatin group), low-dose, medium-dose and high-dose groups of Fushao Diqin prescription. CT-26 cells in the logarithmic growth phase were cultured in RPMI-1640 to prepare 1×10 6 mL -1 For cell suspension, 200 μL of cell suspension was added to the upper chamber, and 800 μL of 1640 containing 10% FBS was added to the lower chamber. After 24 h, the cells were fixed with paraformaldehyde for 10 min and stained with 1% crystal violet for 20 min. The migration of cells in the lower chamber was recorded under a microscope.

[0053] 1.4 RKO cell scratch assay

[0054] The cells were divided into blank control group, positive control group, low-, medium- and high-dose groups of Fushao Diqin prescription. RKO cells in the logarithmic growth phase were seeded in 6-well plates. Scratches were made when the cells grew to 80% density. After 24 hours of drug intervention, photos were taken to record the cell migration of each group.

[0055] 1.5 Flow cytometry detection of CT-26 cell apoptosis

[0056] The cells were divided into blank control group, Fushao Diqin prescription 1mg / ml, 2mg / ml, 4mg / ml groups, and flow cytometry was used to detect the apoptosis-promoting effect of FSDQ on colorectal cancer cell CT-26.

[0057] 2. Experimental Results

[0058] Table 2 Inhibitory effect of FSDQ on proliferation of colorectal cancer cells CT-26 and RKO ( n=3)

[0059]

[0060] Note: Compared with the Control group, *P<0.05, **P<0.01

[0061] MTT assay results showed that compared with the Control group, FSDQ could inhibit the proliferation of CT-26 and RKO cells in a concentration-dependent manner (Table 2). Transwell and cell scratch assay results showed that compared with the Control group, FSDQ could reduce the migration ability of CT-26 and RKO cells in a concentration-dependent manner, and the effect of the medium-dose FSDQ group was comparable to that of the oxaliplatin group ( Figure 2-3 Flow cytometry was used to determine the apoptosis rate of CT-26 cells induced by FSDQ. The results showed that compared with the 5.95% apoptosis rate in the Control group, FSDQ induced apoptosis of CT-26 cells in a dose-dependent manner, with apoptosis rates reaching 12.24%, 25.16% and 46.70% respectively ( Figure 4 The above results indicate that FSDQ has a good anti-CRC effect in vitro.

[0062] Example 8: Anti-CRC effect of the present invention in clinical trials

[0063] 1. Research Methods

[0064] 1.1 Case Source

[0065] This study enrolled 46 patients with stage III postoperative CRC diagnosed with "spleen and kidney yang deficiency, yin and blood deficiency, and damp-heat, blood stasis, and toxins" at the Jiangsu Provincial Hospital of Traditional Chinese Medicine's outpatient clinic or ward between February 2021 and March 2024. Clinical symptoms included symptoms of yang deficiency (fear of cold, a cold stomach, poor appetite, frequent and long nocturnal urination) combined with yin and blood deficiency (emaciation, weight loss, hard stools, and dry eyes). These symptoms were accompanied by a bitter taste in the mouth, dark urine, sticky stools, and a dark purple tongue, reflecting a complex mix of dampness, heat, blood stasis, and toxins.

[0066] 1.2 Treatment options

[0067] All patients underwent conventional anti-tumor treatment for CRC, with 23 cases in the observation group and the control group. The control group underwent FOLFOX chemotherapy, with oxaliplatin, folinate calcium combined with fluorouracil as the commonly used drugs. The observation group took the granules of the present invention (prepared by the method of Experimental Example 1, with a daily prescription of: 10g of white peony root, 6g of liquorice, 9g of aconite root, 7g of scutellaria root, 7g of sophora flavescens, 12g of raw rehmannia root, 15g of spinach seed, 9g of dried ginger, 30g of oyster, 6g of amomum villosum, 15g of poria, and 9g of earthworm) on the basis of the control group, 1 dose per day, taken half an hour after breakfast, lunch, and dinner.

[0068] 1.3 Statistical methods

[0069] The data were imported into the Excel database and statistically analyzed using SPSS 23.0. If the data are in accordance with the normal distribution, the t test is used; if they are not in accordance with the normal distribution, the rank sum test is used and expressed as percentiles. The count data are expressed as percentages and the chi-square (χ 2 ) test. P < 0.05 indicated statistically significant differences.

[0070] 2. Research Results

[0071] 2.1 Comparison of Karnofsky (KPS) functional status scores between the two groups of CRC patients

[0072] To assess patients' physical condition, the KPS scores of the two groups of CRC patients were compared (see Table 3). The KPS scores of the observation group before and after treatment showed a statistically significant difference (P < 0.05); the KPS scores of the control group before and after treatment showed no statistically significant difference (P > 0.05). The KPS scores of the two groups after treatment showed a statistically significant difference (P < 0.05). This suggests that conventional CRC treatment alone does not significantly improve patients' physical condition, while combined treatment with the Fushao Diqin formula can significantly improve the physical condition of CRC patients.

[0073] Table 3 KPS score rank sum test results of two groups of CRC patients (percentile, points)

[0074]

[0075] 2.2 Comparison of body weight between the two groups of CRC patients

[0076] Body weight was compared between the two groups of CRC patients before and after treatment. There was no statistically significant difference in body weight between the observation group and the control group (P>0.05). There was also no statistically significant difference in body weight after treatment between the two groups (P>0.05), as shown in Table 4. Although Fushao Diqin prescription did not significantly improve body weight in CRC patients after treatment, the majority of patients (17 / 23) experienced stable or increased body weight, confirming to some extent its ability to stabilize body weight.

[0077] Table 4 T-test results of body weight of two groups of CRC patients ( kg)

[0078]

[0079] 2.3 Comparison of tumor markers between the two groups of CRC patients

[0080] To evaluate the objective therapeutic effect, carcinoembryonic antigen (CEA) and carbohydrate antigen 199 (CA199), tumor markers closely associated with CRC, were measured (see Tables 5 and 6). There were statistically significant differences in CEA and CA199 before and after treatment in the observation group (P < 0.05); there were no statistically significant differences in CEA and CA199 before and after treatment in the control group (P > 0.05). There was a statistically significant difference in CEA after treatment between the two groups (P < 0.05), but no statistically significant difference in CA199 after treatment between the two groups (P > 0.05). This suggests that combined treatment with Fushao Diqin decoction can significantly reduce CEA and CA199, tumor markers in CRC patients, and its CEA-lowering effect is significantly superior to conventional treatment alone.

[0081] Table 5 CEA rank sum test results for two groups of CRC patients (percentile, ng / ml)

[0082]

[0083] Table 6 Rank sum test results of CA199 in two groups of CRC patients (percentile, u / ml)

[0084]

[0085] 2.4 Comparison of TCM symptom scores between the two groups of CRC patients

[0086] To assess patients' quality of life, a custom-made Traditional Chinese Medicine (TCM) symptom scoring scale for CRC syndromes characterized by "Yin and Yang Deficiency, Internal Cancer Toxins" was used to score key symptoms, including changes in stool consistency, abdominal pain, diarrhea, and hematochezia (see Tables 7-10). There were statistically significant differences in the TCM symptom scores between the observation group and the control group before and after treatment (P < 0.05). There were also statistically significant differences in stool consistency, hematochezia, and diarrhea after treatment between the two groups (P < 0.05). There was no statistically significant difference in abdominal pain after treatment between the two groups (P > 0.05). This suggests that combined treatment with the Fushao Diqin formula significantly alleviates key clinical symptoms of CRC patients, including changes in stool consistency, hematochezia, diarrhea, and abdominal pain, and is significantly more effective than conventional treatment alone in alleviating these symptoms.

[0087] Table 7 Rank sum test results of stool characteristics change scores of two groups of CRC patients (percentile, points)

[0088]

[0089] Table 8 Rank sum test results of abdominal pain scores in two groups of CRC patients (percentile, points)

[0090]

[0091] Table 9 Rank sum test results of diarrhea scores in two groups of CRC patients (percentile, points)

[0092]

[0093] Table 10 Rank sum test results of blood in stool scores in two groups of CRC patients (percentile, points)

[0094]

[0095] 3. Conclusion

[0096] Clinical trials have demonstrated that the Fushao Diqin formula of the present invention can enhance the physical condition of CRC patients, stabilize their weight to a certain extent, reduce objective efficacy indicators such as tumor markers CEA and CA199, improve major clinical symptoms such as diarrhea, bloody stools, and abdominal pain, and enhance their quality of life. Therefore, the Fushao Diqin formula of the present invention has a clear anti-CRC effect.

Claims

1. A Chinese medicine composition for treating colorectal cancer, characterized in that: The medicine is extracted from the following Chinese medicinal materials in parts by weight: 6-15 parts of white peony root, 2-10 parts of liquorice, 3-15 parts of aconite root, 3-10 parts of scutellaria baicalensis, 4.5-9 parts of sophora flavescens, 10-15 parts of raw rehmannia root, 10-15 parts of jujube seed, 3-12 parts of dried ginger, 9-40 parts of oyster, 3-9 parts of amomum villosum, 9-25 parts of poria, and 5-15 parts of earthworm.

2. The Chinese medicine composition for treating colorectal cancer according to claim 1, wherein The medicine is extracted from the following Chinese medicinal materials in parts by weight: 10 parts of white peony root, 6 parts of liquorice, 9 parts of aconite root, 7 parts of scutellaria baicalensis, 7 parts of sophora flavescens, 12 parts of raw rehmannia root, 15 parts of spinach seed, 9 parts of dried ginger, 30 parts of oyster, 6 parts of amomum villosum, 15 parts of poria, and 9 parts of earthworm.

3. The Chinese medicine composition for treating colorectal cancer according to claim 1, wherein The licorice is raw licorice, the aconite is Heishunpian, the jujube kernel is fried jujube kernel, and the oyster is raw oyster.

4. The Chinese medicine composition for treating colorectal cancer according to claim 1, wherein The preparation method comprises the following steps: weighing the above medicinal materials according to the amount, adding 10-20 times the weight of water, soaking for 2 hours, heating to boiling, decocting twice, each time for 2-3 hours, combining the decoctions, filtering, evaporating and concentrating to a relative density of 1.2-1.3 at 25°C, cooling, adding 85-90% ethanol to adjust the ethanol concentration to 65-75%, precipitating with alcohol, fully mixing, leaving overnight, taking the supernatant, recovering ethanol under reduced pressure to obtain an extract, spray drying, sieving, adding appropriate amounts of auxiliary materials and preparing the preparation.

5. The Chinese medicine composition for treating colorectal cancer according to claim 4, characterized in that: The dosage form of the preparation is one of granules, tablets, capsules, powders, pills or oral liquid.

6. The Chinese medicine composition for treating colorectal cancer according to claim 1, wherein: Colorectal cancer belongs to the syndrome of "spleen and kidney yang deficiency, yin and blood deficiency, dampness, heat, blood stasis and toxins" in traditional Chinese medicine.

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