A traditional Chinese medicine prescription for treating bone marrow suppression after chemotherapy, and its preparation method and application

Abstract

The present invention discloses a traditional Chinese medicine formula for treating post-chemotherapy bone marrow suppression, as well as a preparation method and application thereof, belonging to the technical field of traditional Chinese medicine. The traditional Chinese medicine formula of the present invention is prepared from ten traditional Chinese medicines: Millettia reticulata, Clove, Astragalus, Cimicifuga heracleifolia, Sanguisorba officinalis, Agrimoniae scabra, Rhodiola rosea, Codonopsis pilosula, Cinnamon bark, and Angelica sinensis, and has a significant effect on improving post-chemotherapy bone marrow suppression. The present invention also has good therapeutic effects on four types of post-chemotherapy bone marrow suppression, including leukopenia, neutropenia, thrombocytopenia, and low hemoglobin. The formula has good stability, high safety, and significant efficacy.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine preparations, and in particular to a traditional Chinese medicine prescription for treating post-chemotherapy bone marrow suppression, and a preparation method and application thereof. Background Art

[0002] Chemotherapy-induced myelosuppression (CIM) is a common, dose-limiting toxic side effect of cancer chemotherapy. Its mechanism of action is primarily due to the non-selective damage of chemotherapy drugs to bone marrow hematopoietic stem cells. While most chemotherapy drugs inhibit tumor growth by interfering with the cell division cycle, they also have toxic effects on actively proliferating bone marrow hematopoietic cells, leading to impaired red blood cell, white blood cell, and platelet production.

[0003] Post-chemotherapy bone marrow suppression is the most common and most serious toxic side effect after chemotherapy, with a detection rate as high as 50-98.6%. It is an important factor that forces chemotherapy drug dosage reduction and treatment discontinuation.

[0004] Currently, CIM treatment remains limited to the use of hematopoietic growth factors to stimulate bone marrow production, and is associated with serious drug-related adverse reactions. Currently, clinical approaches primarily focus on drug intervention and alternative therapies. These include: cytokine therapy, including granulocyte colony-stimulating factor (G-CSF) and thrombopoietin (TPO), which can temporarily boost peripheral blood cell counts but carries the risk of bone marrow overdraft; component transfusions, for severe anemia or thrombocytopenia, which can rapidly correct indicators but may trigger an immune response and have a short-lived effect; and comprehensive Traditional Chinese Medicine (TCM) treatment, which utilizes a combination of Qi-tonifying and blood-nourishing Chinese herbs (such as Astragalus and Angelica) combined with acupuncture. However, these treatments suffer from slow onset and lack of standardization.

[0005] Many traditional Chinese medicine formulas used in comprehensive TCM treatments, such as Guilu Erxian Jiao, Danggui Buxue Tang, and Buxue Sijun Tang, have demonstrated unique advantages in the treatment of CIM by combining toxicity reduction with beneficial effects. However, due to the complex mechanisms of action of traditional Chinese medicine, their overall efficacy in improving CIM is relatively weak, and the stability of their efficacy remains controversial. Therefore, research into novel traditional Chinese medicine formulas for the treatment of post-chemotherapy myelosuppression is urgently needed. Summary of the Invention

[0006] One of the purposes of the present invention is to provide a traditional Chinese medicine prescription for treating bone marrow suppression after chemotherapy, which has the effects of strengthening the spleen and kidney, nourishing blood and producing marrow.

[0007] The second object of the present invention is to provide a method for preparing the Chinese medicine prescription.

[0008] The third object of the present invention is to provide the use of the traditional Chinese medicine formula in preparing medicine for treating bone marrow suppression after chemotherapy.

[0009] In order to achieve the above object, the technical solution adopted by the present invention is:

[0010] A traditional Chinese medicine prescription for treating post-chemotherapy bone marrow suppression comprises the following raw materials in parts by weight: 1-10 parts of Millettia reticulata, 5-15 parts of Codonopsis pilosula, 1-8 parts of Astragalus membranaceus, 1-8 parts of Angelica sinensis, 5-15 parts of Cimicifuga heracleifolia, 1-8 parts of Rhodiola rosea, 1-8 parts of Agrimoniae herb, 1-8 parts of Sanguisorba officinalis, 5-15 parts of cinnamon bark, and 1-8 parts of cloves.

[0011] Furthermore, the Chinese medicine prescription is composed of the following raw materials in parts by weight: 5 parts of Millettia reticulata, 10 parts of Codonopsis pilosula, 4 parts of Astragalus membranaceus, 4 parts of Angelica sinensis, 10 parts of Cimicifuga heracleifolia, 4 parts of Rhodiola rosea, 4 parts of Agrimoniae herb, 4 parts of Sanguisorba officinalis, 10 parts of cinnamon bark, and 4 parts of cloves.

[0012] Preferably, the Millettia reticulata is produced in Hunan Honghua, and the purchase batch number is 240201.

[0013] Preferably, the origin of the Codonopsis pilosula is Hunan Honghua, and the purchase batch number is 240509.

[0014] Preferably, the astragalus is produced in Hunan Honghua, and the purchase batch number is 240613.

[0015] Preferably, the angelica is produced in Hunan Honghua, and the purchase batch number is 240309.

[0016] Preferably, the Cimicifuga heracleifolia is wine Cimicifuga heracleifolia, produced in Hunan Honghua, with the purchase batch number being 240601.

[0017] Preferably, the Rhodiola rosea is produced in Hunan Honghua, and the purchase batch number is 240201.

[0018] Preferably, the agrimony is produced in Hunan Honghua, and the purchase batch number is 240702.

[0019] Preferably, the origin of the Sanguisorba officinalis is Hunan Honghua, and the purchase batch number is 240401.

[0020] Preferably, the cinnamon is produced in Hunan Honghua, and the purchase batch number is 240802.

[0021] Preferably, the cloves are produced in Hunan Honghua, and the purchase batch number is 240501.

[0022] The Chinese medicine prescriptions include Garcinia Shengbai San powder, Garcinia Shengbai San water extract, and Garcinia Shengbai San prescription hot paste.

[0023] Furthermore, other auxiliary materials can be added to the traditional Chinese medicine prescription to prepare corresponding dosage forms.

[0024] Furthermore, the dosage form is powder, oral preparation, granule, tablet or capsule.

[0025] The present invention also provides a method for preparing the Chinese medicine prescription, which is specifically:

[0026] (1) Garcinia cambogia and whitening powder: weigh the raw materials according to the weight of the formula, dry, crush and sieve the raw materials of Millettia spatholobi, Codonopsis pilosula, Astragalus membranaceus, Angelica sinensis, Cimicifuga heracleifolia, Rhodiola rosea, Agrimoniae herb, Sanguisorba officinalis, Cinnamomum cassia and Clove to obtain the Garcinia cambogia and whitening powder;

[0027] (2) Garcinia Shengbai Powder Water Extract: Garcinia Shengbai Powder powder is added to water to obtain a mixed solution, which is extracted three times using a water reflux method, and then boiled and cooled with water to obtain a water extract; the water extract is added with water, boiled, and extracted several times, and all extracts are combined, filtered, and vacuum concentrated to obtain the Garcinia Shengbai Powder Water Extract;

[0028] (3) Hot ointment of the prescription of Garcinia Shengbai Powder: Add the powder of Garcinia Shengbai Powder into water at a preset temperature to obtain the hot ointment of the prescription of Garcinia Shengbai Powder;

[0029] (4) Acupuncture plaster of the formula of Garcinia Shengbai Powder: Mix the Garcinia Shengbai Powder with rice wine, honey and ginger juice in a ratio of 1:1:1 to obtain the acupuncture plaster of the formula of Garcinia Shengbai Powder. Prepare and use immediately, and heat to 45℃~60℃ before use.

[0030] Furthermore, in step (1), the pulverization is performed using an ultra-fine continuous pulverizer at a speed of at least 5200 r / min; and the screening is performed through a sieve of at least 180 mesh.

[0031] Furthermore, step (2) is specifically as follows: adding the garcinia shengbai powder to water, wherein the weight volume ratio of the garcinia shengbai powder to water is 1:82, to obtain a mixed solution; adding the mixed solution to an extractor, extracting it three times using a water reflux method, and then adding 10 times the amount of water based on the mass of the mixed solution at this time, boiling it for 2 hours, and cooling it to obtain an aqueous extract; adding 10 times the amount of water based on the mass of the obtained aqueous extract, boiling it for 1 hour, extracting it, repeating this several times, and combining all the extracts, filtering them, and concentrating them in a vacuum at 60°C to obtain the garcinia shengbai powder aqueous extract.

[0032] The pharmacological properties of the raw materials used in the present invention are analyzed as follows:

[0033] Millettia reticulata: [Nature and Flavor and Meridians] Sweet, warm, enters the Liver and Kidney meridians. [Functions and Indications] Removes blood stasis, promotes new blood production, and promotes smooth meridians. It is used to treat numbness of the hands and feet, anemia, and irregular menstruation.

[0034] Codonopsis: [Nature and Flavor] Warms the middle and dispels cold, regulates Qi and relieves pain, and enters the spleen and lung meridians. [Functions and Indications] It is mainly used for mild spleen and lung deficiency, and also promotes fluid production and blood nourishment, treating Qi and blood deficiency and Qi and fluid deficiency. It mainly treats loss of appetite, Qi and blood deficiency, palpitations and shortness of breath, asthma and cough due to lung deficiency, and physical weakness due to blood deficiency.

[0035] Astragalus: [Nature and Flavor Meridians] Astragalus is slightly warm in nature, sweet in flavor, and enters the Lung and Spleen Meridians. [Functions and Indications] It tonifies Qi, has the effects of tonifying Qi and raising Yang, promoting fluid production and nourishing blood, and expelling toxins and pus. It is mainly used to treat fatigue caused by Qi deficiency and sallow complexion caused by blood deficiency.

[0036] Angelica: [Nature and Flavor] Warm in nature, sweet and pungent in flavor, enters the Liver, Heart, and Spleen meridians. [Functions and Indications] Danggui Buxue Tang (Danggui Buxue Tang) replenishes Qi and blood, regulates menstruation and relieves pain. It is mainly used to treat sallow complexion due to blood deficiency, rheumatic pain, and constipation caused by dry intestines.

[0037] Cimicifuga: [Nature and Flavor] Slightly cold, pungent, slightly sweet. Enters the Lung, Spleen, Stomach, and Large Intestine meridians. [Functions and Indications] Detoxifies, promotes urination and clears away heat and toxins, and raises Yang Qi. It is primarily used to treat gastric mucosal damage, spleen and stomach collapse, wind-heat headaches, and flare-ups caused by Yang poison.

[0038] Rhodiola Rosea: [Nature and Flavor and Meridians] Cold in nature, sweet in flavor, enters the spleen and lung meridians. [Functions and Indications] Used for qi deficiency, blood stasis, fatigue and shortness of breath. It has the effects of replenishing qi and nourishing the heart, and replenishing qi and blood. It is mainly used to treat physical weakness, low immunity, and shortness of breath and fatigue after serious illness.

[0039] Agrimony: [Nature and Flavor Meridians] Bitter and astringent, enters the Heart, Lung, Spleen, and Liver meridians. [Functions and Indications] Hemostasis and tonification, astringent, hemostasis, tonification, mainly used to treat hemoptysis, carbuncle, sore, anti-cancer, lowering blood sugar, and anti-inflammatory.

[0040] Sanguisorba officinalis: [Nature and Flavor] Slightly cold, bitter and sour, enters the Liver, Lung, Kidney, and Large Intestine meridians. [Functions and Indications] Cools blood and stops bleeding, detoxifies and heals sores. Indications: Bloody dysentery, carbuncles, sores, and burns.

[0041] Cinnamon: [Nature and Flavor and Meridians] Warm in nature, pungent in taste, enters the kidney, spleen, and bladder meridians. [Functions and Indications] The Shennong Herbal Classic calls it "the first of all herbs to open up the meridians," warming the meridians and unblocking the blood vessels, guiding fire back to the source, and relieving vomiting and diarrhea due to deficiency of cold. It is mainly used to treat kidney yang deficiency, cold pain in the heart and abdomen, vomiting and diarrhea due to deficiency of cold, and cold pain in the waist and knees.

[0042] Clove: [Nature and Flavor and Meridians] Slightly pungent and warm, it enters the spleen, stomach, and kidney meridians. [Functions and Indications] Warms the kidneys and supports yang, promotes qi circulation and relieves pain. It is mainly used to treat kidney deficiency, cold uterus, and nausea and vomiting.

[0043] The TCM theoretical basis of the Chinese medicine prescription of the present invention is as follows:

[0044] This prescription is based on the TCM theory of "tonifying the spleen and kidney, nourishing blood and producing marrow". It is an innovative TCM drug developed in accordance with the "Qian Jin Fang", "Nei Jing", "Modern Pharmacological Research of Chinese Materia Medica" and "Guiding Principles for Clinical Research of New Chinese Medicines". It combines ten authentic Hunan medicinal materials, including Millettia reticulata, Astragalus membranaceus, and Cimicifuga heracleifolia. CIM is classified as "consumption" and "blood deficiency". The pathogenesis is one of kidney essence damage, kidney deficiency and marrow deficiency, and lack of blood production; the other is spleen and stomach imbalance, and disorder of qi and blood biochemistry. Therefore, the prescription of Garcinia Shengbai San can exactly address the root cause of CIM pathogenesis.

[0045] Furthermore, the formula "Dang Huang Sheng Bai San," based on the TCM principle of "all ministers assisting and guiding" in the "Inner Canon of Medicine," utilizes TCM therapies to tonify the spleen and strengthen the kidneys, effectively alleviating CIM symptoms. Millettia repens, Codonopsis pilosula, Astragalus membranaceus, and Angelica sinensis serve as "king herbs" that tonify the spleen and replenish qi, promoting blood production and kidney nourishment, promoting meridian circulation, and strengthening the spleen and replenishing qi. Cimicifuga heracleifolia and Rhodiola rosea serve as "assistant herbs," combining Cimicifuga heracleifolia's "heat-clearing, detoxifying, and yang-raising" effects with Rhodiola rosea's "qi-tonifying, yin-nourishing, and blood circulation-activating" properties to achieve a synergistic therapeutic effect. The "adjuvant herbs" Agrimony Root and Sanguisorba officinalis complement the other herbs by regulating blood flow, stopping bleeding, and promoting tissue regeneration. Cinnamon bark and Clove serve as "guiding herbs," enhancing the overall efficacy of the formula by harmonizing medicinal properties, warming the spleen and kidneys, and guiding the herbs to the meridians, thereby coordinating drug interactions.

[0046] Beneficial effects of the present invention:

[0047] The present invention provides a traditional Chinese medicine prescription for treating bone marrow suppression after chemotherapy, as well as a preparation method and application. This prescription has solid traditional Chinese medicine theory, scientific formulation, significant efficacy, and solid preliminary research. The successful application of the present invention will help to achieve a breakthrough in the bottleneck of creative transformation and innovative development of traditional Chinese medicine prescriptions, and has important clinical significance and academic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 This is the technical roadmap for Example 2;

[0049] Figure 2 This is the technical roadmap for Example 3;

[0050] Figure 3 This is a graph showing the estimated mean changes in the WBC impact profiles of different groups of group intervention in Example 3;

[0051] Figure 4 This is a graph showing the estimated mean changes in NEUT impact profiles for different groups of the group intervention in Example 3;

[0052] Figure 5 This is a graph showing the estimated mean changes in the PLT impact profiles of different groups of group intervention in Example 3;

[0053] Figure 6This is a graph showing the estimated mean changes in the HGB impact profiles of different groups of group intervention in Example 3;

[0054] Figure 7 The WBC effect and NEUT effect box plot of the intervention method in Example 3 to improve CIM; Figure 7 A is the box plot of WBC effects of different intervention groups. Figure 7 B is a box plot of the NEUT effects of different groups of group intervention;

[0055] Figure 8 This is a box plot of the effect of the intervention method in Example 3 on improving the PLT and HGB effects of CIM; Figure 8 A is the box plot of the PLT effects of different groups of group intervention, Figure 8 B is a box plot of the HGB effects of different groups of group intervention;

[0056] Figure 9 This is a graph showing the nucleated cell count in the bone marrow of mice after the intervention method in Example 3 improved CIM;

[0057] Figure 10 This is the HE staining result of mouse spleen before and after the intervention method improves CIM in Example 3; Figure 10 A is the HE staining result of the spleen of blank mice without modeling. Figure 10 B is the HE staining result of the spleen of blank model mice without intervention. Figure 10 C is the HE staining result of spleen in G-CSF intervention model mice. Figure 10 D is the HE staining result of spleen in mice treated with placebo by gavage. Figure 10 E is the HE staining result of spleen in mice treated with low-concentration intervention model. Figure 10 F is the HE staining result of spleen in mice treated with medium concentration intervention by oral gavage. Figure 10 G is the HE staining result of spleen in mice treated with high-concentration intervention model. Figure 10 H is the HE staining result of spleen in mice treated with acupoint application placebo. Figure 10 I is the HE staining result of spleen in mice modeled by low-concentration intervention with acupoint application. Figure 10 J is the HE staining result of spleen in mice modeled with medium concentration intervention of acupoint application. Figure 10 K is the HE staining result of spleen in mice modeled with high-concentration intervention after acupoint application;

[0058] Figure 11 The HE staining results of mouse liver before and after the intervention method in Example 3 improved CIM; Figure 11 A is the HE staining result of the liver of blank mice without modeling. Figure 11 B is the HE staining result of the liver of blank model mice without intervention. Figure 11C is the HE staining result of the liver of G-CSF-intervention model mice. Figure 11 D is the HE staining result of the liver of mice treated with placebo by gavage. Figure 11 E is the HE staining result of the liver of mice treated with low-concentration intervention by oral gavage. Figure 11 F is the HE staining result of the liver of mice treated with medium concentration by oral gavage. Figure 11 G is the HE staining result of the liver of mice treated with high-concentration intervention by gavage. Figure 11 H is the HE staining result of the liver of mice treated with acupoint application placebo. Figure 11 I is the HE staining result of the liver of mice modeled with low-concentration intervention at acupoint application. Figure 11 J is the HE staining result of the liver of mice modeled with intervention of medium concentration in acupoint application. Figure 11 K is the HE staining result of the liver of mice modeled with high-concentration intervention after acupoint application;

[0059] Figure 12 The ion current chromatogram of the base peak of quercetin, an active ingredient extracted from a traditional Chinese medicine prescription, includes Figure 12 A. Figure 12 B and Figure 12 C, Figure 12 A is the base peak ion current chromatogram of the quercetin reference substance. Figure 12 B is the base peak ion current chromatogram of quercetin alcohol extraction. Figure 12 C is the base peak ion current chromatogram of quercetin water extraction;

[0060] Figure 13 The ion flow chromatogram of the kaempferol-based peak of the active ingredient extracted from the traditional Chinese medicine prescription includes Figure 13 A. Figure 13 B and Figure 13 C, Figure 13 A is the base peak ion current chromatogram of kaempferol alcohol extraction. Figure 13 B is the base peak ion current chromatogram of the kaempferol reference substance. Figure 13 C is the base peak ion current chromatogram of kaempferol water extraction;

[0061] Figure 14 The ion flow chromatogram of the active ingredient daidzein extracted from traditional Chinese medicine prescriptions, including Figure 14 A. Figure 14 B and Figure 14 C, Figure 14 A is the base peak ion current chromatogram of the daidzein reference substance. Figure 14 B is the base peak ion current chromatogram of daidzein water extraction. Figure 14 C is the base peak ion current chromatogram of daidzein alcohol extraction;

[0062] Figure 15The ion flow chromatogram of ursolic acid base peak, an active ingredient extracted from a traditional Chinese medicine prescription, includes Figure 15 A. Figure 15 B and Figure 15 C, Figure 15 A is the base peak ion current chromatogram of ursolic acid water extraction. Figure 15 B is the base peak ion current chromatogram of ursolic acid alcohol extraction. Figure 15 C is the base peak ion current chromatogram of the ursolic acid reference substance;

[0063] Figure 16 The ion flow chromatogram of the oleanolic acid base peak of the active ingredient extracted from traditional Chinese medicine prescriptions includes Figure 16 A. Figure 16 B and Figure 16 C, Figure 16 A is the base peak ion current chromatogram of oleanolic acid alcohol extraction. Figure 16 B is the base peak ion current chromatogram of oleanolic acid water extraction. Figure 16 C is the base peak ion current chromatogram of the oleanolic acid reference substance;

[0064] Figure 17 The base peak ion chromatogram of luteolin, an active ingredient extracted from a traditional Chinese medicine prescription, includes Figure 17 A. Figure 17 B and Figure 17 C, Figure 17 A is the base peak ion current chromatogram of luteolin water extraction. Figure 17 B is the base peak ion current chromatogram of luteolin alcohol extraction. Figure 17 C is the base peak ion current chromatogram of the luteolin reference substance. DETAILED DESCRIPTION

[0065] The present invention is further described in detail by the following examples, but the scope of protection of the present invention is not limited by any specific examples. Based on the examples in the embodiments and in combination with experimental design, other examples obtained by those skilled in the art without making creative work are all within the scope of protection of the present invention.

[0066] Example 1: This example provides a traditional Chinese medicine prescription for treating bone marrow suppression after chemotherapy, which is composed of the following raw materials in parts by weight: 5 parts of Millettia reticulata, 10 parts of Codonopsis pilosula, 4 parts of Astragalus membranaceus, 4 parts of Angelica sinensis, 10 parts of Cimicifuga heracleifolia, 4 parts of Rhodiola rosea, 4 parts of Agrimoniae herb, 4 parts of Sanguisorba officinalis, 10 parts of cinnamon bark, and 4 parts of cloves.

[0067] This embodiment also provides a preparation method of the above-mentioned Chinese medicine prescription, which is as follows:

[0068] Weigh the raw materials according to the above weight parts, mix them evenly, and put them into the DLF-50S type classification ultrafine continuous grinder in the preparation room, rotate at least 5200r / min, and pass through at least 180 mesh to prepare Garcinia cambogia white powder;

[0069] Add 45g of Garcinia Shengbai Powder to 410ml of water to create a Garcinia Shengbai Powder mixture. Place the mixture in an extractor and extract three times with reflux. Add 10 times the weight of the mixed solution to the mixture and boil for 2 hours. After cooling, obtain an aqueous extract. Add 10 times the weight of the resulting aqueous extract to the mixture, boil for 1 hour, and extract twice. Combine all extracts, filter, and vacuum concentrate at 60°C to obtain the Garcinia Shengbai Powder aqueous extract. Store in a refrigerator at -20°C.

[0070] Take 45g of Garcinia Whitening Powder and pour it into 90ml of pure water at 75℃. Mix and use immediately. Finally, you can get Garcinia Whitening and Heat-Removing Paste, which should be stored at a constant temperature.

[0071] The garcinia shengbai powder is mixed with rice wine, honey and ginger juice in a ratio of 1:1:1 to obtain the garcinia shengbai powder acupoint application paste, which is prepared and used immediately and heated to 45°C to 60°C before use.

[0072] Example 2: A group intervention experiment for patients with bone marrow suppression after chemotherapy. This example is based on the electronic medical record database of a tertiary hospital in Hunan Province. The real-world retrospective electronic medical records of bone marrow suppression after chemotherapy in the Department of Traditional Chinese Medicine and Oncology in the past 5 years were retrieved to analyze the level of bone marrow suppression after chemotherapy, revealing the clinical efficacy of the prescription of Garcinia Shengbai San in treating bone marrow suppression after chemotherapy in patients with malignant tumors.

[0073] This example uses the medical records of patients with myelosuppression after chemotherapy for non-myeloid malignancies admitted to the Department of Traditional Chinese Medicine and the Department of Oncology of a tertiary hospital in Hunan Province from January 2019 to December 2024 as the research subjects.

[0074] Specifically, referring to the Expert Consensus on the Diagnosis and Treatment of Antitumor Drug-Induced Myelosuppression with Integrated Traditional Chinese and Western Medicine (2021) and the WHO Grading Standards for Acute and Subacute Toxicity of Anticancer Drugs (see Table 1), antitumor drug-related myelosuppression can be diagnosed if the following conditions are met: ① A history of malignant tumors and recent chemotherapy. ② Adult peripheral blood leukocyte count <4.0×10 9 / L indicates leukopenia; the absolute neutrophil count is <2.0×10 9 / L is neutropenia; a decrease in hemoglobin concentration below normal level is anemia; a platelet count <100×10 9 / L is thrombocytopenia.

[0075] The specific exclusion criteria are as follows: ① Patients with bone marrow suppression before chemotherapy should be excluded; ② Patients with incomplete medication records and unclear chemotherapy regimens or treatment status should be excluded; ③ Patients who are too young (such as children <12 years old) or too old (such as over 80 years old) should be excluded to ensure the consistency of the study population and avoid these factors from confounding the study results.

[0076] This study has been reviewed by the Ethics Committee of Xiangya School of Nursing, Central South University (Project No. E2024197).

[0077] The intervention was carried out according to the group intervention scheme in Table 1 below:

[0078] Table 1 Intervention plan

[0079]

[0080] Comparison of the therapeutic effects between the intervention group of the Tanghuang Shengbai San prescription and the granulocyte colony-stimulating factor treatment group (standard control group).

[0081] (1) The group treated with acupoint application of the formula of Garcinia Shengbai Powder: patients who received acupoint application of Garcinia Shengbai Powder but did not receive granulocyte colony-stimulating factor treatment were selected.

[0082] The acupoint application method of the Garcinia Shengbai Powder formula is as follows: make the Garcinia Shengbai Powder water extract into a medicine cake with a length and width of about 0.5cm×0.5cm and a thickness of about 0.1cm, place it on a 3cm×3cm sterile dressing, and select the acupoints according to syndrome differentiation.

[0083] (2) The group treated with the Garcinia Shengbai powder and heat-relieving pack: patients who received the Garcinia Shengbai powder and heat-relieving pack but did not receive granulocyte colony-stimulating factor treatment were selected.

[0084] The method of hot packing of the Garcinia Shengbai Powder is as follows: make the Garcinia Shengbai Powder into a medicine cake with a length and width of about 0.5cm×0.5cm and a thickness of about 0.1cm, place it on a 3cm×3cm sterile dressing, and select acupoints according to syndrome differentiation.

[0085] (3) Patients in the combined treatment group of Garcinia Shengbai San prescription: patients who received Garcinia Shengbai San acupoint application therapy combined with Garcinia Shengbai Heat-relieving Pack therapy, but did not receive granulocyte colony-stimulating factor treatment.

[0086] (4) Granulocyte colony-stimulating factor treatment group: patients who were treated with granulocyte colony-stimulating factor but did not receive acupoint plasters, hot packs, or combined treatments with the prescription of Garcinia Shengbai San were selected.

[0087] The treatment method of granulocyte colony-stimulating factor is: adjust the dosage range according to the patient's bone marrow suppression after chemotherapy, and inject subcutaneously.

[0088] The experimental results are shown in Tables 2~5.

[0089] Table 2 Comparison of the efficacy of granulocyte colony-stimulating factor and huanghuangshengbaisan in increasing white blood cell count

[0090]

[0091] Table 3 Comparison of the efficacy of granulocyte colony-stimulating factor and Tanghuang Shengbai San prescription in increasing granulocyte count

[0092]

[0093] Table 4 Comparison of the efficacy of granulocyte colony-stimulating factor and Tanghuang Shengbai San in increasing hemoglobin

[0094]

[0095] ( Indicates the significance level of the mean difference P is 0.05, The significance level of the difference between the means is <0.001)

[0096] Table 5 Comparison of the efficacy of granulocyte colony-stimulating factor and the prescription of Garcinia Shengbai San in increasing granulocyte count

[0097]

[0098] ( Indicates the significance level of the mean difference P is 0.05, The significance level of the difference between the means is <0.001)

[0099] From the results in Tables 2 to 5, it can be seen that there is no statistical difference in the four routes of administration in increasing white blood cell count, among which the acupoint application of the formula of Garcinia Shengbai San is relatively the best; there is no significant difference in the four routes of administration in increasing granulocyte count, among which the acupoint application of the formula of Garcinia Shengbai San is relatively the best; in increasing hemoglobin, the various Chinese medicine therapies of the formula of Garcinia Shengbai San are significantly better than granulocyte colony-stimulating factor, and the acupoint application of the formula of Garcinia Shengbai San is the best ( F =8.713, P =0.026), but there was no statistical difference among the three Chinese medicine treatments; in terms of increasing platelet count, the effects of the hot pack of the prescription of Garcinia Shengbai San and the combination group were significantly better than those of the granulocyte colony-stimulating factor, among which the hot pack of the prescription of Garcinia Shengbai San was the best ( F =46.972, P <0.001).

[0100] Therefore, the hot pack or acupoint application of the prescription of Garcinia Shengbai San is significantly better than granulocyte colony-stimulating factor in increasing hemoglobin and platelets. The prescription of Garcinia Shengbai San is as effective as granulocyte colony-stimulating factor in increasing white blood cell count and granulocyte count, and the administration effects of the three traditional Chinese medicines are comparable.

[0101] Example 3: Mouse group intervention experiment. This example uses mice as an example. Specifically, the prescription of Gammonium Shengbai Powder was applied to cyclophosphamide-induced mice to further confirm the effect of the prescription of Gammonium Shengbai Powder on improving bone marrow suppression after chemotherapy.

[0102] This example used 90 SPF-grade BALB / c mice (7 weeks old, half male and half female). All mice were handled according to standard guidelines approved by the Animal Ethics Committee of Central South University and maintained at room temperature (20 ± 5°C) and a humidity level of 50-70%. They were housed under a 24-hour daylight cycle (6:30 AM–6:30 PM) with free access to food and water.

[0103] This study has passed the ethical review of the Department of Laboratory Animal Science of Central South University (ethics number: APU-2025-0033).

[0104] A total of 30 mice were divided into 11 groups. The mice were divided into the internal use group and the external use group according to the random number table method.

[0105] (1) Model preparation and model evaluation

[0106] ① Model preparation

[0107] The mice were gavaged with normal saline (10 ml / kg·d) for 3 consecutive days. On the 4th day, the model was established and cyclophosphamide (CTX) 80 mg / kg·d was injected intraperitoneally for 3 consecutive days (Ying M, Yu Q, Zheng B, Wang H, Wang J, Chen S, Nie S, XieM. Cultured Cordyceps sinensis polysaccharides modulate intestinal mucosal immunity and gut microbiota in cyclophosphamide-treated mice. CarbohydrPolym. 2020 May 1;235:115957. doi: 10.1016 / j.carbpol.2020.115957.).

[0108] ②Model evaluation

[0109] Evaluation of model disease characteristics: If the WHO "Grading Standards for Acute and Subacute Toxicity of Anticancer Drugs" is met, a: white blood cell count (×109) ≤ 4.0 and / or b: granulocyte count (×109) ≤ 2.0 and / or c: hemoglobin (g / l) ≤ 110 and / or d: platelet ≤ 100, the model is successful.

[0110] (2) Animal handling

[0111] After the last administration, the rats in each group were killed by rapid dislocation of the head and neck, and blood and tissue samples were collected as required for relevant index detection, and primary cells were extracted.

[0112] (3) The specific group intervention plan is as follows:

[0113] 1) Blank control group (n=2): no treatment;

[0114] 2) Normal saline gavage group (n=2): Normal saline (10 mL / kg·d) was administered orally for 8 consecutive days, followed by intraperitoneal injection of cyclophosphamide (CTX) 30 mg / kg on the 4th day for 3 consecutive days;

[0115] 3) High-concentration Garcinia Shengbai San extract gavage (n=3): Normal saline (10 mL / kg·d) was administered gavage for 3 consecutive days. The model was established on the 4th day. On the 8th day, a high-dose (25 mg / kg) Garcinia Shengbai San aqueous extract was prepared by mixing powder with purified water in a 1:1 ratio. The mixture was then administered by gavage for 2 consecutive weeks.

[0116] 4) Medium-concentration Garcinia Shengbaisan extract administered orally (n=3): Compared with (3), the dose was changed to medium-dose (15 mg / kg) Garcinia Shengbaisan extract administered orally;

[0117] 5) Low-concentration Garcinia Shengbai San extract administered orally (n=3): Compared with (3), the dose was changed to low-dose (10 mg / kg) Garcinia Shengbai San administered orally;

[0118] 6) Placebo group (n=2): Normal saline was administered as a placebo by gavage at a dose of 15 mg / kg.

[0119] 7) High-frequency acupoint application of Garcinia shengbaisan (n=3): Normal saline (10 mL / kg·d) was administered orally for 8 consecutive days. The model was established on the 4th day. On the 8th day, the mice were fixed in a supine position. After hair removal and disinfection, the umbilicus of the mice was fully exposed. Then, the Garcinia shengbaisan acupoint application paste was placed in the center of a non-woven fabric patch (6×7 cm) and fixed on the acupoint for 4-6 hours. It was then wrapped with breathable anti-allergic tape and fixed three times a day for 2 consecutive weeks.

[0120] 8) Medium-frequency acupoint application of Garcinia Shengbai San (n=3): Compared with 7), the frequency was changed to twice a day;

[0121] 9) Low-frequency acupoint application of Garcinia Shengbai San (n=3): Compared with 7), the frequency was changed to once a day;

[0122] 10) Placebo group for the acupoint application of Garcinia Shengbai San (n=2): dextrin was used as the placebo. Compared with 7), the frequency was changed to twice a day.

[0123] 11) G-CSF intervention group (n=2): subcutaneous injection, dosage of 40 mg / kg, once a day for 5 consecutive days.

[0124] This study used animal experiments to construct a CIM mouse model. Low, medium and high concentrations of Garcinia Shengbai San were used for external treatment (acupoint application) and internal treatment (oral gavage) to compare granulocyte colony-stimulating factor, and the effects of the three therapies on mouse body weight, CIM status, bone marrow nucleated cell count, and liver and spleen tissue were observed.

[0125] The changes in the body weight of the mice are shown in Table 6.

[0126] Table 6 Body weight changes of mice in each group

[0127]

[0128] After group intervention, the changes in bone marrow suppression after chemotherapy in mice included the effects on WBC, NEUT, PLT, and HGB. The results are shown in Tables 7 to 10.

[0129] Table 7 Effects of the group intervention method in Example 3 on WBC in mice

[0130]

[0131] Table 8 Effects of the group intervention method in Example 3 on NEUT in mice

[0132]

[0133] Table 9 Effects of the group intervention method in Example 3 on PLT in mice

[0134]

[0135] Table 10 Effects of the group intervention method in Example 3 on HGB in mice

[0136]

[0137] Example 3 Group intervention method to improve the outline of CIM Figures 3 to 6 As shown, the box plot is Figures 7 and 8 The nucleated cell counts of mouse bone marrow are shown in Figure 9 The HE staining results of mouse spleen are shown in Figure 10 Mouse liver staining results are shown in Figure 11 shown.

[0138] From the results in Tables 7 to 10, we can see that in terms of increasing WBC and NEUT, the high-concentration prescription gavage group, the low-concentration acupoint application group and the G-CSF group had the best effects ( P <0.001), and different intervention methods of traditional Chinese medicine can effectively increase WBC; in terms of increasing PLT and HGB, there is no significant statistical difference among the groups. However, in terms of increasing HGB, the low-concentration acupoint application group has the best effect ( P =0.01).

[0139] In addition, by Figure 10 、 Figure 11 It can be seen that the prescription of the present invention can improve the liver fatty degeneration and increased number of spleen megakaryocytes caused by CIM.

[0140] In summary, the prescription of Tanghuang Shengbai San can significantly improve bone marrow suppression after chemotherapy, and its clinical efficacy has good promotion potential.

[0141] Example 4: Analysis of the core active ingredients of the prescription of Cambogia Shengbai Powder based on LC-MS / MS technology. Specifically, this example analyzes the core active ingredients of the water extract and alcohol extract of the THSBS prescription based on LC-MS / MS technology, and collects the base peak ion current mass spectrum and chromatogram.

[0142] In this example, the garcinia whitening powder prepared in Example 1 was used as a sample to extract the active ingredient. The specific extraction method is as follows:

[0143] Water Extraction Method: Pour 45g of Garcinia Shengbai Powder into 410ml of water to create a Garcinia Shengbai Powder mixture. Place the mixture in an extractor and extract three times with water reflux. Add 10 times the volume of water, boil for 2 hours, and cool to obtain an aqueous extract. Add another 10 times the volume of water, boil for 1 hour, and extract twice. Combine all extracts, filter, and vacuum concentrate at 60°C to obtain the Garcinia Shengbai Powder aqueous extract. Add 45ml of purified water to prepare a 1g / ml Garcinia Shengbai Powder aqueous extract test sample. Prepare 100μL of the test sample solution and add 900μL of methanol solution. After high-speed centrifugation, extract 200μL for injection.

[0144] Alcohol extraction method: Pour 3g of Garcinia Shengbai Powder into 40ml of methanol, add 1μL of DMSO solution, and sonicate for 30 minutes. Place 2mL of the extract in a centrifuge tube and centrifuge at 12,000 rpm for 10 minutes. Filter through a 0.22μm syringe filter. Take the filtrate and add methanol to the volume to 18ml to prepare the 0.17g / ml Garcinia Shengbai Powder alcohol extract test sample. Prepare 100μL of the test sample stock solution and add 900μL of the methanol solution (concentration of 0.17g / ml). After high-speed centrifugation, extract 200μL for injection.

[0145] This example uses the following method to prepare the reference substance for comparison:

[0146] To the precisely weighed quercetin reference substance, 10 mL of methanol was added in a brown bottle to prepare a quercetin stock solution with a concentration of 1.023 mg / mL, named "Quercetin", and stored in the bottom layer of a pharmacological refrigerator at -20°C.

[0147] To the precisely weighed kaempferol reference substance, add 10 mL of methanol in a brown bottle to prepare a 1.025 mg / mL kaempferol stock solution, named "kaempferol" and stored in the same location as above;

[0148] To the precisely weighed daidzein reference substance, add 10 mL of methanol in a brown bottle to prepare a 1.016 mg / mL daidzein stock solution, named "daidzein" and stored in the same location as above;

[0149] To the precisely weighed ursolic acid reference substance, add 10 mL of methanol in a brown medium to prepare a 1.024 mg / mL ursolic acid stock solution named "ursolic acid" and store it in the same location as above;

[0150] To the precisely weighed oleanolic acid reference substance, add 10 ml of methanol in a brown bottle to prepare an oleanolic acid stock solution with a concentration of 0.984 mg / mL, named "oleanolic acid" and stored in the same location as above.

[0151] Prepare mixed reference solution by taking the above stock solution separately, dilute it to 1 μg / mL with methanol, take 200 μL of mixed reference solution in a centrifuge tube and centrifuge at 12000 r / min for 10 min, filter it through a 0.22 μm syringe filter, and take 200 μL of the filtrate for injection.

[0152] The chromatographic conditions used in this example were: pressure 5.4 MPa, mobile phase: A (water extraction) 0.1% FA in water, B (alcohol extraction) 0.1% FA in MEOH. Column temperature 30°C; flow rate 0.2 mL min -1 ; Injection volume 2 μL; Water (containing 0.1% formic acid by volume, A) and acetonitrile (containing 0.1% formic acid by volume, B) were used as the mobile phases, and elution was performed isocratically.

[0153] The mass spectrometry conditions used in this example are as follows: an electrospray ion source was selected, and the TOF MS-IDA-PI mode was used to collect data in positive and negative ion modes respectively. The parameters were set as follows: spray voltage +5500V / -4500V, ion source temperature 500°C / 450°C, spray gas 344.74 kPa, auxiliary heating gas 344.74 kPa, curtain gas 275.79 kPa, and declustering voltage 100V; TOF MS scan range m / z 100-1000; dynamic background subtraction was set, and the 10 most responsive ions were selected for MS / MS scanning; PI scan range 50-1000 m / z, collision energy ±(40±20) eV.

[0154] Ion mode is as follows:

[0155] Quercetin: ESI-, 301.09→151.03, cone(v)=30, Collision=25;

[0156] Kaempferol: ESI-, 285.19→226.94, cone(v)=30, Collision=40;

[0157] Ursolic acid: ESI-, 455.35→455.35, cone(v)=20, Collision=20;

[0158] Oleanolic acid: ESI-, 455.23→455.23, cone(v)=20, Collision=20;

[0159] Luteolin: ESI+, 287.08→152.96, cone(v)=30, Collision=35;

[0160] Daidzein: ESI+, 417.26→255.01, cone(v)=20, Collision=10.

[0161] The analysis results of the active ingredients of the Garcinia Shengbai Powder prescription by water extraction are shown in Table 11, and the analysis results of the active ingredients of the Garcinia Shengbai Powder prescription by alcohol extraction are shown in Table 12.

[0162] Table 11 Analysis results of active ingredients in the water extraction method of the prescription of Dahuang Shengbai San

[0163]

[0164] Table 12 Analysis results of active ingredients in the alcohol extraction method of the prescription of Dahuang Shengbai San

[0165]

[0166] The obtained chromatogram is as Figures 12 to 17 shown.

[0167] As can be seen from the table, the mass fraction of active ingredients in the alcohol extraction method is significantly better than that in the water extraction method, and luteolin has the highest content in both water and alcohol extraction methods.

Claims

1. A Chinese medicine prescription for treating bone marrow suppression after chemotherapy, characterized in that: The traditional Chinese medicine prescription is composed of the following raw materials in parts by weight: 1-10 parts of Millettia reticulata, 5-15 parts of Codonopsis pilosula, 1-8 parts of Astragalus membranaceus, 1-8 parts of Angelica sinensis, 5-15 parts of Cimicifuga heracleifolia, 1-8 parts of Rhodiola rosea, 1-8 parts of Agrimoniae herb, 1-8 parts of Sanguisorba officinalis, 5-15 parts of cinnamon bark, and 1-8 parts of cloves.

2. The Chinese medicinal formula for treating myelosuppression after chemotherapy according to claim 1, characterized in that: The traditional Chinese medicine prescription is composed of the following raw materials in parts by weight: 5 parts of Millettia reticulata, 10 parts of Codonopsis pilosula, 4 parts of Astragalus membranaceus, 4 parts of Angelica sinensis, 10 parts of Cimicifuga heracleifolia, 4 parts of Rhodiola rosea, 4 parts of Agrimoniae herb, 4 parts of Sanguisorba officinalis, 10 parts of cinnamon bark, and 4 parts of cloves.

3. The Chinese medicinal formula for treating myelosuppression after chemotherapy according to claim 1, characterized in that: The Chinese medicine prescription is Garcinia Shengbai San powder, Garcinia Shengbai San water extract, and Garcinia Shengbai San prescription hot paste made from the raw materials.

4. The method for preparing the traditional Chinese medicine prescription for treating myelosuppression after chemotherapy according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) Garcinia cambogia and whitening powder: weigh the raw materials according to the weight of the formula, dry, crush and sieve the raw materials of Millettia spatholobi, Codonopsis pilosula, Astragalus membranaceus, Angelica sinensis, Cimicifuga heracleifolia, Rhodiola rosea, Agrimoniae herb, Sanguisorba officinalis, Cinnamomum cassia and Clove to obtain the Garcinia cambogia and whitening powder; (2) Garcinia Shengbai Powder Water Extract: Garcinia Shengbai Powder powder is added to water to obtain a mixed solution, which is extracted three times using a water reflux method, and then boiled and cooled with water to obtain a water extract; the water extract is added with water, boiled, and extracted several times, and all extracts are combined, filtered, and vacuum concentrated to obtain the Garcinia Shengbai Powder Water Extract; (3) Hot ointment of the prescription of Garcinia Shengbai Powder: Add the powder of Garcinia Shengbai Powder into water at a preset temperature to obtain the hot ointment of the prescription of Garcinia Shengbai Powder; (4) Acupuncture plaster of the formula of Garcinia Shengbai Powder: Mix the Garcinia Shengbai Powder with rice wine, honey and ginger juice in a ratio of 1:1:1 to obtain the acupuncture plaster of the formula of Garcinia Shengbai Powder. Prepare and use immediately, and heat to 45℃~60℃ before use.

5. The preparation method according to claim 4, characterized in that In step (1), the pulverization is performed by using an ultra-fine continuous pulverizer at a rotation speed of at least 5200 r / min; and the screening is performed by passing through a sieve of at least 180 mesh.

6. The preparation method according to claim 4, characterized in that Step (2) is specifically as follows: adding the garcinia shengbai powder to water, wherein the weight volume ratio of the garcinia shengbai powder to water is 1:82, to obtain a mixed solution; adding the mixed solution to an extractor, extracting it three times using a water reflux method, then adding 10 times the amount of water based on the mass of the mixed solution at this time, boiling it for 2 hours, and cooling it to obtain an aqueous extract; adding 10 times the amount of water based on the mass of the obtained aqueous extract, boiling it for 1 hour, extracting it, repeating this several times, and combining all the extracts, filtering them, and concentrating them in a vacuum at 60°C to obtain the garcinia shengbai powder aqueous extract.

7. The preparation method according to claim 4, characterized in that Add auxiliary materials to the traditional Chinese medicine prescription to prepare corresponding dosage forms.

8. The preparation method according to claim 7, characterized in that The dosage form is powder, oral preparation, granule, tablet or capsule.

9. Use of the traditional Chinese medicine formula according to any one of claims 1 to 3 or the traditional Chinese medicine formula prepared by the preparation method according to any one of claims 4 to 8 in preparing a drug for treating post-chemotherapy bone marrow suppression.

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