Pharmaceutical composition for treating multiple myeloma and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a pharmaceutical composition for treating multiple myeloma and application thereof, the pharmaceutical composition is prepared from the following raw materials: turmeric, turtle shell, leech, curcuma zedoary, caulis perllae, rhizoma sparganii, ginseng, agilawood, wolfberry fruit, liquorice, centella asiatica and bungarus parvus. The pharmaceutical composition disclosed by the invention can directly reach the focus of infection, and realizes the treatment of multiple myeloma in a mode of breaking first and standing second by taking the main effect of removing excess symptoms and the auxiliary effects of clearing heat and promoting diuresis, detoxifying and diminishing swelling, tonifying spleen and kidney and tonifying kidney and absorbing qi, so that the characteristic of treating both symptoms and root causes is achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a pharmaceutical composition for treating multiple myeloma and application thereof. Background Art

[0002] Traditional Chinese medicine believes that cancer is caused by insufficient positive energy, which is invaded and gathered by evil energy. The above two factors cause imbalance of yin and yang, thus forming cancer. Specifically, multiple myeloma is a type of bone malignant tumor disease, which generally refers to multiple myeloma, bone tumors, and bone tumor metastasis. It is a tumor that occurs in bones or their attached tissues. The tumor develops rapidly, with a poor prognosis and a high mortality rate. The external pain is mild and intermittent at the beginning of the disease. As the disease progresses, the pain may gradually increase and develop into a continuous pain. Most patients experience increased pain at night, which affects their sleep. The pain may radiate to distant places. Existing Western medical treatments are mainly chemotherapy and radiotherapy. Multiple myeloma has the characteristic of metastasis in the early stage, which leads to the problem of poor cure rate and prognosis. Existing Chinese medicine treatment plans are mainly based on clearing away heat and detoxification, invigorating qi and nourishing blood, or nourishing the liver and kidneys, that is, by replenishing the positive energy to resist external evil to achieve the purpose of curing multiple myeloma. The above methods are in line with the traditional concept of Chinese medicine. However, due to the rapid development of multiple myeloma and the gradual progress of traditional Chinese medicine, the above treatment principles cannot achieve satisfactory results, so patients and their families always have doubts about the diagnosis and treatment plans of traditional Chinese medicine. Summary of the invention

[0003] The purpose of the present invention is to provide a pharmaceutical composition for treating multiple myeloma and its application, so as to overcome the defects in the prior art.

[0004] In order to achieve the above object, the technical solution adopted by the present invention is: A pharmaceutical composition for treating multiple myeloma, the pharmaceutical composition is prepared from the following raw materials in parts by weight: 10-20 parts of turmeric, 25-45 parts of turtle shell, 25-45 parts of leech, 10-30 parts of zedoaria, 10-30 parts of scutellaria baicalensis, 10-28 parts of triangularis lanceolata, 20-46 parts of ginseng, 5-19 parts of agarwood, 20-50 parts of wolfberry, 10-30 parts of liquorice, 20-50 parts of centella asiatica; and also comprises 1 golden snake.

[0005] Preferably, the pharmaceutical composition is prepared from the following raw materials in parts by weight: 10-16 parts of turmeric, 25-35 parts of turtle shell, 25-35 parts of leech, 10-20 parts of curcuma, 10-15 parts of sedge stem, 15-28 parts of triangularis, 20-40 parts of ginseng, 5-10 parts of agarwood, 20-30 parts of wolfberry, 10-18 parts of licorice, and 20-40 parts of Centella asiatica; and also includes 1 golden snake.

[0006] Preferably, the pharmaceutical composition is prepared from the following raw materials in parts by weight: 12-16 parts of turmeric, 28-35 parts of turtle shell, 30-35 parts of leech, 18-20 parts of curcuma, 10-12 parts of sedge stem, 17-20 parts of triangularis, 25-30 parts of ginseng, 9-10 parts of agarwood, 16-30 parts of wolfberry, 10-13 parts of licorice, and 25-30 parts of Centella asiatica; and also includes 1 golden snake.

[0007] Specifically, the pharmaceutical composition is prepared from the following raw materials in parts by weight: 15 parts of turmeric, 30 parts of turtle shell, 30 parts of leech, 18 parts of curcuma, 12 parts of sophora japonica stem, 18 parts of triangularis, 30 parts of ginseng, 9 parts of agarwood, 30 parts of wolfberry, 12 parts of licorice, and 30 parts of Centella asiatica; and also includes 1 golden snake.

[0008] Preferably, the drug is prepared into a pharmaceutically commonly used preparation by using the medicinal powder of the raw material drug corresponding to the pharmaceutical composition or the water or / and organic solvent extract of the raw material drug as the active ingredient, and pharmaceutically acceptable excipients, including diluents, lubricants, adhesives, disintegrants, sweeteners, stabilizers, preservatives, etc., and can be specifically prepared into a decoction, or crushed and sieved, mixed, made into a powder or granule, or the above-mentioned powder or granule is loaded into a capsule or compressed into a tablet, capsule, or soft capsule. When used specifically, taking the decoction for the treatment of multiple myeloma as an example, one dose per day, decocted in water three times to take about 840ml of juice, and divided into three times in the morning, noon and evening. One month is a course of treatment, and a 4-day break is taken between two courses of treatment, and then the next course of treatment is carried out. Generally, 3 courses of treatment are taken continuously; until the clinical symptoms and signs disappear, three courses of treatment are consolidated; in particular, special populations such as pregnant women need to follow the doctor's advice when taking, and no special adverse reactions have been found. The treatment described in the present invention means treatment after onset, not intervention before onset.

[0009] A use of a pharmaceutical composition in preparing a pharmaceutical composition for treating multiple myeloma.

[0010] Pharmacology: The pharmaceutical composition of the present invention is used to treat multiple myeloma, specifically multiple myeloma, bone tumors and bone tumor metastasis. In the present invention, turmeric and turtle shell are used as the main drugs to achieve the effects of breaking blood and promoting qi, softening and dispersing nodules, and removing tumors. At the same time, leeches, zedoaria, scutellaria, trillium, and centella are selected as the assistant drugs. The combination of leeches, zedoaria and trillium can strengthen the function of promoting blood circulation and removing blood stasis, so as to achieve the purpose of removing blood stasis and removing symptoms and relieving pain in cooperation with the main drugs. At the same time, scutellaria and centella are used. To achieve the effects of regulating qi and relieving rush, clearing away heat and dampness, and detoxifying and reducing swelling; ginseng, agarwood, wolfberry, and licorice are used as adjuvants to achieve the effects of tonifying the spleen and kidney and tonifying the kidney and absorbing qi, and at the same time can make up for the organ deficiency caused by the above-mentioned main and auxiliary drugs of breaking blood, promoting qi, removing blood stasis and eliminating symptoms; the present invention specially selects white flower snake as the guiding drug, which can enable the drug to quickly reach the lesion while also having the effect of quickly dispersing nodules and eliminating symptoms. In addition, by breaking blood, promoting qi, dispersing nodules and eliminating symptoms, it can accelerate the dredging of qi and blood meridians and the efficacy of the drug. From the above content, it can be seen that the present invention overcomes the technical prejudice in traditional Chinese medicine, adopts the combination of monarch medicine and assistant medicine to directly eliminate the real symptoms as the main purpose, and on this basis, the assistant medicine and adjuvant medicine are matched to enhance the efficacy of the monarch medicine and reduce the side effects of the drugs of breaking blood, promoting qi, removing blood stasis and eliminating symptoms on the patient's organs, so as to achieve the purpose of promoting drug absorption and quickly stabilizing the condition of the disease. Compared with the prior art, the present invention adopts the method of enabling the drugs to reach the lesions directly, mainly eliminating the real symptoms, supplemented by clearing away heat and dampness, detoxifying and reducing swelling, tonifying the spleen and kidney, and tonifying the kidney and absorbing qi, and achieves the treatment of multiple myeloma in the form of breaking first and then establishing, thereby achieving the characteristics of treating both the symptoms and the root causes.

[0011] The present invention is used to treat multiple myeloma, and comprises 12 Chinese medicines, which can be directly obtained from the market, and their pharmacological properties are as follows: Turmeric: pungent, bitter, warm; enters the spleen and liver meridians; breaks blood, promotes qi, promotes menstruation, and relieves pain. It is used to treat fullness and pain in the heart and abdomen, arm pain, lumps, amenorrhea due to blood stasis in women, abdominal pain due to postpartum blood stasis, injuries from falls, and carbuncles; it is used for chest and abdominal pain, dysmenorrhea, and limb pain caused by qi stagnation and blood stasis. The present invention uses turmeric as the main drug, mainly using its effects of breaking blood, promoting qi, promoting menstruation, and relieving pain. It can not only eliminate solid symptoms, but also has an analgesic effect.

[0012] Turtle shell: salty, cold. Enter the liver and kidney meridians. Efficacy: nourish the kidney and suppress yang, soften and dissipate stagnation; remove blood and qi, break up knots, bad blood, abort, eliminate sores and swellings, and eliminate blood loss, malaria, intestinal carbuncle. Indications: bone steaming, fever, malaria, hardness under the ribs, low back pain, amenorrhea, and lumps. The present invention uses turtle shell and turmeric as the main medicine to achieve the effects of breaking blood and promoting qi, softening and dissipating stagnation, and removing lumps; at the same time, turtle shell is a liver-reducing agent, and turmeric used for tonifying the liver is used in the main medicine to avoid damaging the patient's liver.

[0013] Leech: salty and bitter, neutral, poisonous; enters the liver and bladder meridians. Effects: amenorrhea, abdominal pain caused by abdominal masses, blood stasis, pain caused by blood stasis due to injury, carbuncle, erysipelas, etc.

[0014] Curcuma: pungent, bitter, warm. Enters the liver and spleen meridians. Efficacy: breaks blood and promotes qi, eliminates stagnation and relieves pain. Used for abdominal pain caused by blood stasis, hepatosplenomegaly, abdominal distension and pain, accumulation, amenorrhea caused by blood stasis in women, pain caused by injuries and food stagnation. Promotes qi and relieves pain, breaks blood and eliminates stagnation: used for amenorrhea caused by qi stagnation and blood stasis, chest and flank pain, abdominal pain, and lumps and masses.

[0015] Sparganium: bitter, flat. Enter the liver and spleen meridians. Efficacy: breaking blood and promoting qi, eliminating stagnation and relieving pain. Indications: used for lumps and masses, dysmenorrhea, amenorrhea due to blood stasis, chest pain, and distension and pain due to food stagnation. The present invention uses leech, Curcuma and Sparganium as ministerial drugs to achieve the purpose of removing stagnation and relieving symptoms and eliminating stagnation and relieving pain in coordination with the monarch drug.

[0016] Su Geng: pungent, warm. Enters the lung and spleen meridians. Efficacy: regulating qi, relieving fullness, relieving pain, and stabilizing pregnancy. Indications: used for chest and diaphragm fullness, stomach pain, belching and vomiting, and restless fetus. Pungent, warm and dispersing, the leaves are as light as the lungs, which can dissipate wind and cold, clear the lungs and relieve cough; the stems are like the spleen and stomach, which are good at promoting qi and harmonizing the middle, regulating qi and stabilizing pregnancy. Therefore, it is often used for wind-cold cough and spleen and stomach qi stagnation. In addition, it can also detoxify fish and crabs.

[0017] Ginseng: neutral in nature, sweet, slightly bitter, slightly warm. Enters the spleen and lung meridians. Efficacy: greatly replenishes vital energy, restores pulse and consolidates weakness, replenishes spleen and lungs, promotes body fluid and quenches thirst, calms the mind and improves intelligence. Indications: fatigue due to overwork, poor appetite, fatigue, nausea and vomiting, loose stools, weak cough and shortness of breath, spontaneous sweating and sudden loss of body fluids, palpitations, forgetfulness, dizziness and headache, impotence, frequent urination, thirst, metrorrhagia in women, slow shock in children and long-term weakness, all symptoms of insufficient qi, blood and body fluids.

[0018] Agarwood: pungent, bitter, warm. Enters the spleen, stomach, kidney, and lung meridians. Efficacy: Lowers qi and warms the middle, warms the kidney and absorbs qi. Indications: Treats qi reversal and wheezing, vomiting and hiccups, abdominal distension and pain, cold waist and knees, constipation of the large intestine, stranguria with urination, and cold semen in men.

[0019] Goji berries: neutral in nature, sweet in taste, and enter the liver, kidney, and lung meridians. Efficacy: nourishes the liver, kidneys, and lungs; Indications: Suitable for symptoms such as dizziness, vertigo, soreness of the waist and knees, spermatorrhea, premature ejaculation, and excessive leucorrhea caused by liver and kidney yin deficiency and insufficient essence and blood.

[0020] Licorice: neutral in nature, sweet in taste, belongs to the twelve meridians. Efficacy: has pharmacological effects such as detoxification, expectoration, analgesia, antispasmodic and even anti-cancer. Indications: tonifying the spleen and replenishing qi, clearing away heat and detoxification, expectoration and cough relief, relieving pain, and harmonizing various medicines. It is used for spleen and stomach weakness, fatigue, palpitations and shortness of breath, cough with sputum, abdominal and limb cramps and pain, carbuncle, sore and ulcer, and relieve drug toxicity and potency.

[0021] Centella asiatica: cold in nature, bitter and pungent in taste. Enters the liver, spleen, and kidney meridians. Efficacy: clears away heat and detoxifies, promotes dampness and reduces swelling, and enhances memory; Indications: damp-heat jaundice, heatstroke diarrhea, sand stranguria and blood stranguria, carbuncle, ulcer, fall injury, restlessness, forgetfulness, calming the mind, promoting blood circulation, insomnia, dreaminess, and palpitations.

[0022] Goldenrod: sweet, salty, warm; poisonous. Enters the liver meridian. Functions and indications: dispel wind, dredge meridians, and stop spasms. Used for rheumatism, numbness, cramps, stroke, hemiplegia, convulsions, tetanus, leprosy, scabies, scrofula, and malignant ulcers.

[0023] The present invention is used for treating multiple myeloma, and is significantly different from the treatment method of replenishing vital energy to resist external evils adopted in traditional Chinese medicine. According to the characteristics of rapid development of multiple myeloma and early metastasis, the main drug and the assistant drug are matched to achieve the main purpose of removing the entity and eliminating the real symptoms. On this basis, the assistant drug and the adjuvant drug are matched to achieve the characteristics of enhancing the efficacy of the main drug and reducing the side effects. Furthermore, the main drug used in the present invention can promote the effects of breaking blood and promoting qi, softening and dispersing nodules, and removing lumps. At the same time, the liver-tonifying effect of turmeric is utilized to eliminate the liver-scaling agent of turtle shell, so as to achieve rapid stabilization of the disease, thereby achieving the characteristics of treating both the symptoms and the root causes. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a graph showing the toxicity test results of the Chinese herbal decoction of the present invention on human multiple myeloma MM.1S cells.

[0025] Figure 2 This is a morphological diagram of MM.1S cells after being treated with the Chinese medicinal decoction of the present invention.

[0026] Figure 3 This is a diagram showing the results of measuring the activity of caspases in MM.1S cells after the Chinese medicine decoction of the present invention was used to treat the cells for 24 hours.

[0027] Figure 4 This is a graph showing the western blot detection results of MM.1S cells after the Chinese herbal decoction of the present invention was treated for 24 hours.

[0028] Figure 5 This is a graph showing the results of detecting changes in mitochondrial membrane potential (MMP) in MM.1S cells after the Chinese medicine decoction of the present invention was used to treat the cells for 24 hours.

[0029] Figure 6 This is a photo of the establishment results of the MM.1S cell nude mouse model in the present invention.

[0030] Figure 7 This is a graph showing the detection results of tumor volume and weight indicators of mice with multiple myeloma treated with Chinese medicine by intragastric administration in the present invention.

[0031] Figure 8This is a graph showing the results of detecting changes in caspase-3 expression in vivo in mice with multiple myeloma after intragastric administration of traditional Chinese medicine in the present invention.

[0032] Fig. 9 This is a diagram for detecting and analyzing the changes in tissue structure of mice with multiple myeloma after intragastric administration of traditional Chinese medicine for treatment in the present invention. DETAILED DESCRIPTION

[0033] Example 1 A pharmaceutical composition for treating multiple myeloma, the pharmaceutical composition comprises the following raw materials in parts by weight: 15 parts of turmeric, 30 parts of turtle shell, 30 parts of leech, 18 parts of zedoaria, 12 parts of scutellaria baicalensis, 18 parts of triangularis lancea, 30 parts of ginseng, 9 parts of agarwood, 30 parts of wolfberry, 12 parts of liquorice and 30 parts of centella asiatica; and also comprises 1 golden snake.

[0034] The drug is made of the medicinal material powder of the raw material drug corresponding to the pharmaceutical composition or the water or / and organic solvent extract of the raw material drug as the active ingredient, and is prepared into a pharmaceutically commonly used preparation with pharmaceutically acceptable excipients, and the pharmaceutically acceptable excipients include diluents, lubricants, adhesives, disintegrants, sweeteners, stabilizers, preservatives, etc., and can be specifically prepared into a decoction, or crushed and sieved, mixed, made into a powder or granule, or the above-mentioned powder or granule is loaded into a capsule or compressed into a tablet, capsule, or soft capsule. When used specifically, taking the decoction for the treatment of multiple myeloma as an example, one dose per day, decocted in water three times to take about 840ml of juice, and divided into three times in the morning, noon and evening. One month is a course of treatment, and a 4-day break is taken between two courses of treatment, and then the next course of treatment is carried out. Generally, 3 courses of treatment are taken continuously; until the clinical symptoms and signs disappear, three courses of treatment are consolidated; in particular, special populations such as pregnant women need to follow the doctor's advice when taking, and no special adverse reactions have been found so far. The treatment described in the present invention means treatment after onset, not intervention before onset.

[0035] Example 2 The difference between Example 2 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 10 parts of turmeric, 25 parts of turtle shell, 25 parts of leech, 10 parts of Curcuma, 10 parts of Sophora flavescens, 10 parts of Trigonum multiflorum, 20 parts of ginseng, 5 parts of agarwood, 20 parts of wolfberry, 10 parts of licorice and 20 parts of Centella asiatica; and also includes 1 Golden Snake.

[0036] Example 3 The difference between Example 3 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 20 parts of turmeric, 45 parts of turtle shell, 45 parts of leech, 30 parts of Curcuma, 30 parts of Sophora flavescens, 28 parts of Trigonum multiflorum, 46 parts of ginseng, 19 parts of agarwood, 50 parts of wolfberry, 30 parts of licorice and 50 parts of Centella asiatica; and also includes 1 Golden Snake.

[0037] Example 4 The difference between Example 4 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 15 parts of turmeric, 35 parts of turtle shell, 35 parts of leech, 20 parts of Curcuma, 20 parts of Sophora flavescens, 17 parts of Trigonum multiflorum, 33 parts of ginseng, 12 parts of agarwood, 35 parts of wolfberry, 20 parts of licorice and 35 parts of Centella asiatica; and also includes 1 Golden Snake.

[0038] Example 5 The difference between Example 5 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 10 parts of turmeric, 35 parts of turtle shell, 25 parts of leech, 10 parts of Curcuma, 10 parts of Sophora flavescens, 15 parts of Trigonum multiflorum, 20 parts of ginseng, 5 parts of agarwood, 20 parts of wolfberry, 10 parts of licorice and 20 parts of Centella asiatica; and also includes 1 Golden Snake.

[0039] Example 6 The difference between Example 6 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 16 parts of turmeric, 25 parts of turtle shell, 35 parts of leech, 20 parts of Curcuma, 15 parts of Sophora flavescens, 28 parts of Trigonum multiflorum, 40 parts of ginseng, 10 parts of agarwood, 30 parts of wolfberry, 18 parts of licorice and 40 parts of Centella asiatica; and also includes 1 Golden Snake.

[0040] Example 7 The difference between Example 7 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 13 parts of turmeric, 30 parts of turtle shell, 30 parts of leech, 15 parts of Curcuma, 13 parts of Sophora flavescens, 22 parts of Trigonum multiflorum, 30 parts of ginseng, 8 parts of agarwood, 25 parts of wolfberry, 14 parts of licorice and 30 parts of Centella asiatica; and also includes 1 Golden Snake.

[0041] Example 8 The difference between Example 8 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 12 parts of turmeric, 28 parts of turtle shell, 35 parts of leech, 20 parts of Curcuma, 10 parts of Sophora flavescens, 18.5 parts of Trigonum multiflorum, 27 parts of ginseng, 9 parts of agarwood, 16 parts of wolfberry, 10 parts of licorice and 25 parts of Centella asiatica; and also includes 1 Golden Snake.

[0042] Example 9 The difference between Example 9 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 16 parts of turmeric, 35 parts of turtle shell, 30 parts of leech, 18 parts of Curcuma, 12 parts of Sophora flavescens, 20 parts of Trigonum multiflorum, 30 parts of ginseng, 10 parts of agarwood, 30 parts of wolfberry, 13 parts of licorice and 30 parts of Centella asiatica; and also includes 1 Golden Snake.

[0043] Example 10 The difference between Example 10 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 14 parts of turmeric, 31.5 parts of turtle shell, 32.5 parts of leech, 19 parts of Curcuma, 11 parts of Sophora flavescens, 17 parts of Trigonum multiflorum, 25 parts of ginseng, 9.5 parts of agarwood, 23 parts of wolfberry, 11.5 parts of licorice and 27 parts of Centella asiatica; and also includes 1 Golden Snake.

[0044] Embodiment 11 The difference between Example 11 and Example 1 is that the pharmaceutical composition is prepared from the following raw materials in parts by weight: 25 parts of turmeric, 50 parts of turtle shell, 24 parts of leech, 9 parts of Curcuma, 25 parts of Sophora flavescens, 9 parts of Trigonum multiflorum, 50 parts of ginseng, 4 parts of agarwood, 55 parts of wolfberry, 21 parts of licorice and 19 parts of Centella asiatica; and also includes 1 Golden Snake.

[0045] Application Examples Case 1: Name: Chen, gender: female, age: 60 years old, from Huangjiang Town, Dongguan City, Guangdong Province. The patient sought medical treatment for low back and leg pain, especially severe low back pain in 2017. She was treated for lumbar disc herniation, but the effect was not significant after three months of treatment. The pain worsened and she was unable to walk. She came to our clinic for treatment in 2021 and was diagnosed with multiple myeloma through bone marrow puncture. She took the medicine (decoction) of Example 2 for treatment and was able to walk after one course of treatment. The symptoms disappeared after two courses of treatment and she was cured after examination in the hospital. She insisted on taking the medicine for three courses of treatment and has not relapsed so far.

[0046] Case 2: Name: Lei, gender: male, age: 50 years old, from Jinji Town, Tianchang City, Anhui Province. The patient was diagnosed with multiple myeloma locally in 2017. After four months of treatment, the condition recurred and the condition was never controlled. In 2018, the patient came to our hospital for treatment and was diagnosed with late-stage multiple myeloma. The symptoms were pale complexion, anemia, fatigue, swelling of the limbs, hepatosplenomegaly, and pain in both lower limbs. The pharmaceutical composition (decoction) of Example 1 was used for treatment. After two courses of treatment, the symptoms were significantly improved, and the lower limbs were no longer painful. The symptoms disappeared after the eighth course of treatment and were cured by hospital examination. After continuing to take the medicine for three courses, there has been no recurrence so far.

[0047] Case 3: Name: Liu, Gender: Male, Age: 32 years old, from Tanghe County, Nanyang City, Henan Province. The patient was diagnosed with POEMS syndrome (IgG Lambda type, multiple myeloma comprehensive systemic disease) in a large hospital in Henan Province in 2021; in November 2021, he was admitted to a large hospital in Henan Province with intermittent edema of both eyes and lower limbs for more than two months, dizziness, hepatomegaly, splenomegaly, polyserosal effusion, acute upper respiratory tract infection, chronic nephritis syndrome, lumbosacral pain, etc. After more than three months of treatment, the effect was not significant, and he was transferred to our hospital for treatment. The pharmaceutical composition (decoction) of Example 6 was used for treatment. After two courses of treatment, the lumbosacral pain was relieved and the appetite increased; after four courses of treatment, all symptoms disappeared, and after six courses of treatment, the patient was cured by hospital examination. After three courses of medication, there has been no recurrence so far.

[0048] Pharmacological performance test of the composition (1) Decoction treatment: The decoction sample in Example 1 was subjected to ultracentrifugation in a laboratory centrifuge, and the obtained supernatant was further filtered (treated with a 0.22 μm membrane filter), aliquoted and frozen for later use.

[0049] (2) Toxicity test The purchased human multiple myeloma cells MM.1S were expanded and cultured (RPMI-1640+FBS, 10%, 5% CO 2 MM.1S cells were inoculated into 96-well plates, and then the above-mentioned Chinese medicine decoction was diluted in a series of concentrations to prepare multiple gradients of original solution, 1 / 2 of the original solution, 1 / 4 of the original solution, 1 / 8 of the original solution, and 1 / 16 of the original solution for research. At the same time, a blank control group (i.e., the control group, i.e., the group without any drug treatment and allowing tumor cells to grow freely) was set up, and cells were added to the 96-well plate for continued culture. After 24 hours, the MTT experiment was performed. The results showed that the Chinese medicine decoction ( Figure 1 ) has a significant inhibitory effect on human multiple myeloma MM.1S cells in a dose-dependent manner. Specifically, Figure 1 Each column represents the effect of Chinese herbal decoction on the viability of myeloma MM.1S cells. The higher the column, the greater the cell viability. As can be seen from the figure, the greater the dilution multiple of the Chinese herbal sample and the lower the concentration, the greater the viability of the cells after treatment. In other words, the higher the concentration of the Chinese herbal sample, the smaller the viability of the cells after treatment, which means that the high-concentration Chinese herbal sample has a greater killing effect on MM.1S cells, that is, it is dose-dependent.

[0050] (3) Morphological observation After MM.1S cells were inoculated into a 6-well plate, different dilution ratios of Chinese herbal decoction (concentrations were original solution, 1 / 2 original solution, 1 / 4 original solution, 1 / 8 original solution, and 1 / 16 original solution) were used to treat MM.1S for different periods of time, and then the changes in cell morphology were observed using an inverted microscope. The results showed that as time went on, the number of cells in the normal control group (not treated with Chinese herbal decoction) gradually increased and grew normally; while the cells in the group treated with the original Chinese herbal decoction were severely fragmented, fell off, and the number of cells was scarce. Although the number of cells in the group treated with the 1 / 16 diluted original solution increased, it was far less than that of the control group. The results showed that the inhibitory effect of Chinese herbal decoction treatment on the growth of MM.1S cells was concentration- and time-dependent ( Figure 2 ),Depend on Figure 2 It can be seen that the MM.1S cells in the control group (blank control) grew normally, and the number of cells increased from 0 h to 48 h (that is, there were more and more bright spots in the figure), indicating that their proliferation rate was very fast. After treatment with Chinese herbal decoctions in each group, the trend of cell increase from 0 h to 48 h was significantly reduced compared with the control group, and the smaller the dilution multiple (that is, the higher the concentration), the stronger the inhibitory effect on the trend of cell increase, especially for the undiluted stock solution samples. The number of cells after treatment was significantly less at 48 h, indicating that the high-concentration samples have a stronger tumor inhibitory effect. It also shows that the inhibitory effect of Chinese herbal decoction treatment on MM.1S cell growth is concentration-dependent, and Figure 1 The results are consistent.

[0051] (4) Cell signaling pathway analysis I After MM.1S cells were inoculated into culture dishes, they were treated with the original solution of Chinese herbal medicine and 1 / 2 dilution of the original solution for 24 hours, and then samples were collected. The cell caspases activity was determined according to the operating procedures of the caspases activity detection kit (Beyotime, China). The results showed (eg Figure 3 ), the activities of cellular caspase-3 and caspase-9 were significantly enhanced, while the intracellular caspase-8 was enhanced but not significantly after treatment with the original solution.

[0052] Combined with the cell apoptosis pathway, caspase-3 and caspase-9 will be activated in the cell mitochondrial apoptosis pathway, while caspase-8 is an effector molecule of the death receptor pathway. In this experiment, the Chinese medicine treatment of MM.1S cells stimulated the activity of these two enzymes, indicating that Chinese medicine can inhibit tumor cell proliferation by inducing the cell mitochondrial apoptosis pathway. Other core factors of the mitochondrial apoptosis pathway can be verified in the future to further confirm it.

[0053] (5) Cell signaling pathway analysis II After MM.1S cells were inoculated into culture dishes, they were treated with the original solution of Chinese herbal medicine and 1 / 2 dilution of the original solution for 24 h, and then samples were collected for protein extraction and subsequent western blotting analysis. Expression changes of apoptosis-related proteins such as PARP and Bax were detected (e.g. Figure 4 ), and the changes of other important factors in the related apoptosis pathway can be further detected as needed.

[0054] (6) Cell signaling pathway analysis III After inoculating MM.1S cells into culture dishes, the MM.1S cells were treated with the original solution of Chinese herbal decoction and 1 / 2 dilution of the original solution for 24 hours, and then the samples were collected. After JC-1 staining, the mitochondrial membrane potential (MMP) of the cells was detected (CCCP diluted 1:1000 as a positive control). When the cellular MMP content is high, JC-1 aggregates in the mitochondrial matrix to form polymers and can produce red fluorescence. In contrast, JC-1 cannot aggregate in the matrix of mitochondria to form monomers that can produce green fluorescence. The current results show that the green fluorescence of MM.1S cells after Chinese medicine treatment was significantly enhanced compared with the control group (such as Figure 5 ). These data indicate that Chinese medicine treatment leads to a decrease in mitochondrial membrane potential in cells, upregulation of ROS levels, and ultimately cell apoptosis.

[0055] Through the above experimental analysis data, it can be found that after treating human multiple myeloma cells MM.1S, the Chinese herbal decoction can induce apoptosis by inducing the mitochondrial (endogenous) pathway of the cells, thereby inhibiting the proliferation of tumor cells. This can be further confirmed through in vivo experiments.

[0056] (7) Establishment of human multiple myeloma model in NOD / SCID mice SPF-grade female NOD / SCID mice (Beijing Weitong Lihua Company, n=6, 4-6 weeks old, weighing 20-25g) were used and housed in an SPF-grade sterile laminar flow animal breeding system. The mice were given free access to water and food and fed at room temperature of 25±2℃. The feed and drinking water were sterilized, and the bedding and cages were sterilized at high temperature. The density of the human multiple myeloma cell line MM.1S was adjusted to 7×10 7 Each mouse was inoculated with 150 μL of the drug subcutaneously on the back of the left hind limb, and then tumors were observed. Tumors were observed on the back of the mice on the 10th day after feeding. The tumor volume gradually increased from 11 to 14 days, and the model was successfully established ( Figure 6 ), in preparation for subsequent anti-tumor in vivo effect studies.

[0057] (8) Grouping, treatment and index observation of myeloma model mice The mice with successful modeling were randomly divided into two groups - control group and traditional Chinese medicine group. The traditional Chinese medicine group was gavaged with traditional Chinese medicine, and the control group was intravenously injected with the same dose of normal saline, once every two days from 1 to 10 days, with a dose of 50 μL, and once a day from 11 to 15 days, with a dose of 300 μL. All mice were observed for tumors on the 1st day, 8th day, and 15th day of treatment, and the tumors of the mice were weighed after dissection on the 16th day, and the tumor inhibition rate was calculated (tumor inhibition rate = (average tumor weight of the control group - average tumor weight of the experimental group) / average tumor weight of the control group × 100%). The current results show that there is no significant difference in the body weight of the two groups of mice after treatment. Although the tumor volume of the traditional Chinese medicine group increased, it was smaller than that of the model group, and the tumor inhibition rate was 16.67% (the average tumor weight of the control group was 2.7523 g, and the average tumor weight of the traditional Chinese medicine group was 2.2321 g, Figure 7 ).

[0058] (9) Changes in caspase-3 levels in myeloma model mice treated with traditional Chinese medicine Tumor tissues from mice treated with traditional Chinese medicine were collected and frozen ultrathin sections (3-5 μm thick) were made. Immunofluorescence analysis was performed on tumor tissue sections, and DAPI cell nuclei were stained after incubation with fluorescently labeled caspase-3 antibodies. The results showed that compared with the control group, the positive signals of caspase-3 protein in tissue cells in the traditional Chinese medicine treatment group increased significantly, and the cell nuclei appeared condensed and irregular ( Figure 8 ), showing apoptosis characteristics, and in vivo experiments once again confirmed that this Chinese herbal decoction has the effect of stimulating cell apoptosis, laying a certain theoretical foundation for future clinical testing.

[0059] (10) HE staining of myeloma model mouse tissues treated with traditional Chinese medicine The heart, liver, spleen, lung, kidney and other tissues of mice treated with traditional Chinese medicine were sliced ​​ultrathinly, and then stained with hematoxylin-eosin (HE). The results showed that the tissue structure of normal organs in the traditional Chinese medicine treatment group was relatively intact and the holes were relatively reduced; in terms of tumor tissue, the tissue layers of the control group were relatively intact and the cells were densely connected, while the tumor tissue structure of the traditional Chinese medicine decoction gavage group had gaps and holes in the cell layers ( Fig. 9 ). These results indicate that after 15 days of intragastric administration of Chinese herbal decoction, tumor proliferation can be effectively inhibited while protecting normal tissues, and tumor cell death can be initially induced.

[0060] In summary, the in vitro and in vivo experiments after treating human multiple myeloma cells MM.1S and NOD / SCID mouse multiple myeloma models with the above-mentioned Chinese herbal decoction confirmed that the Chinese herbal decoction can effectively inhibit the proliferation of myeloma cells and the proliferation of tumor tissue in vivo, which are all related to the Chinese herbal decoction's ability to effectively induce the apoptosis pathway of tumor cells.

[0061] Based on the above clinical practice examples, the pharmaceutical compositions of Example 1, Example 2, and Example 6 (all decoctions) were taken by multiple multiple myeloma patients for several courses of treatment, and none of them relapsed. Therefore, the pharmaceutical composition of the present invention is simple to prepare, easy to use, has clear efficacy, can be efficiently used for the treatment of multiple myeloma, and its anti-tumor mechanism has been basically clarified, and has very good prospects for large-scale development and application.

Claims

1. A pharmaceutical composition for treating multiple myeloma, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 10-20 parts of turmeric, 25-45 parts of turtle shell, 25-45 parts of leech, 10-30 parts of zedoaria, 10-30 parts of sedge stem, 10-28 parts of triangularis, 20-46 parts of ginseng, 5-19 parts of agarwood, 20-50 parts of wolfberry, 10-30 parts of liquorice, and 20-50 parts of centella asiatica; and also includes 1 golden snake.

2. The pharmaceutical composition for treating multiple myeloma according to claim 1, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 10-16 parts of turmeric, 25-35 parts of turtle shell, 25-35 parts of leech, 10-20 parts of zedoaria, 10-15 parts of sedge stem, 15-28 parts of triangularis, 20-40 parts of ginseng, 5-10 parts of agarwood, 20-30 parts of wolfberry, 10-18 parts of liquorice, and 20-40 parts of Centella asiatica; it also includes one golden snake.

3. The pharmaceutical composition for treating multiple myeloma according to claim 2, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 12-16 parts of turmeric, 28-35 parts of turtle shell, 30-35 parts of leech, 18-20 parts of zedoaria, 10-12 parts of sedge stem, 17-20 parts of triangularis, 25-30 parts of ginseng, 9-10 parts of agarwood, 16-30 parts of wolfberry, 10-13 parts of liquorice, and 25-30 parts of Centella asiatica; and also includes 1 golden snake.

4. The pharmaceutical composition for treating multiple myeloma according to claim 3, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 15 parts of turmeric, 30 parts of turtle shell, 30 parts of leech, 18 parts of zedoaria, 12 parts of scutellaria, 18 parts of trigonosciadium japonicum, 30 parts of ginseng, 9 parts of agarwood, 30 parts of wolfberry, 12 parts of licorice, and 30 parts of Centella asiatica; and also includes 1 golden snake.

5. The pharmaceutical composition for treating multiple myeloma according to any one of claims 1 to 4, characterized in that: It is prepared from the medicinal material powder of the raw material medicine corresponding to the pharmaceutical composition or the water or / and organic solvent extract of the raw material medicine as active ingredients, and pharmaceutically acceptable excipients to form a commonly used pharmaceutical preparation.

6. The pharmaceutical composition for treating multiple myeloma according to claim 5, characterized in that: The preparation is decoction, powder, tablet, granule, capsule or soft capsule.

7. Use of the pharmaceutical composition according to any one of claims 1 to 4 in the preparation of a pharmaceutical composition for treating multiple myeloma.

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