Composition for preventing and treating leucopenia as well as preparation method and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for preventing and treating leucopenia as well as a preparation method and application thereof. The composition is prepared from fructus momordicae, fructus ligustri lucidi, ginseng leaves and roses as raw materials through extraction. Experiments show that the composition can significantly increase the number and vitality of leukocytes so as to play multiple roles in increasing leukocytes, improving pathological injury of spleen tissues and enhancing immunity, and the effects are significantly superior to those of a formula-lacking control or a formula-replacing control. Therefore, all the components in the composition cooperate with one another and take effect synergistically, and are indispensable.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a composition for preventing and treating leukopenia, its preparation method, and its application. Background Technology

[0002] Leukopenia is a condition in which the number of white blood cells or neutrophils in a blood test remains consistently below the normal range. White blood cells are a crucial component of the human immune system, reflecting not only the body's immune status but also forming the first line of defense. As the primary form of immune cells, they maintain bodily health through various mechanisms, including phagocytosis of pathogens, promotion of tissue repair, and defense against invading microorganisms. In recent years, with increased environmental pollution, widespread use of radiotherapy and chemotherapy, and the inappropriate use of antibiotics and various chemical drugs, the incidence of leukopenia has shown a significant upward trend. When the number of white blood cells decreases, the body's immune surveillance function declines significantly, leading to severely impaired resistance to infection. In this situation, patients are highly susceptible to recurrent infections, ranging from mild local inflammation to severe systemic sepsis or multiple organ dysfunction, even threatening life. Therefore, maintaining normal white blood cell levels is crucial for ensuring the body's immune function.

[0003] Western medicine treatment primarily focuses on symptomatic relief, vitamin B4 supplementation, white blood cell-boosting drugs, and anti-infective medications. While these can rapidly increase white blood cell counts, they have limitations such as high cost, side effects, and poor efficacy in some patients. Traditional Chinese medicine (TCM), on the other hand, has advantages such as fewer adverse reactions and reliable efficacy, making it superior in treating leukopenia. Therefore, there is an urgent need to provide a TCM composition that can improve leukopenia and enhance the body's immune function. Summary of the Invention

[0004] In view of this, the present invention provides a composition for preventing and treating leukopenia, a method for preparing the composition, and its application.

[0005] The composition comprises the following parts by weight of raw materials:

[0006] 1.5–90 parts monk fruit, 1–72 parts privet fruit, 0.5–54 parts ginseng leaf and 0.5–36 parts rose.

[0007] Through long-term research and systematic screening and optimization, this invention has yielded a composition with significant therapeutic effects on leukopenia. This composition consists of four medicinal ingredients. Luo Han Guo (monk fruit) clears heat and nourishes Yin, serving as the principal ingredient; Nu Li Zi (privet fruit) nourishes the liver and kidneys, serving as the assistant ingredient; and Ren Shen Ye (ginseng leaf) and Mei Gui Hua (rose flower) are used together to tonify deficiency and promote Qi circulation, serving as adjuvant ingredients. Compared with other traditional Chinese medicine compositions, this composition has a rational selection of medicinal ingredients and appropriate dosage, thus producing a significant synergistic effect. Experiments show that this composition can effectively increase the number and activity of white blood cells, thereby exerting multiple effects such as increasing white blood cell count, improving pathological damage to spleen tissue, and enhancing immunity, thus effectively preventing and treating leukopenia.

[0008] In some embodiments, the composition comprises the following raw materials in parts by weight:

[0009] 3–60 parts monk fruit, 2–48 parts privet fruit, 1–36 parts ginseng leaves, and 1–12 parts rose petals.

[0010] In some specific embodiments, the composition is made from the following raw materials in parts by weight: 9 parts monk fruit, 6 parts privet fruit, 3 parts ginseng leaf and 3 parts rose.

[0011] The present invention also provides a method for preparing the composition, comprising:

[0012] Monk fruit, privet fruit, ginseng leaf and rose petals were mixed and extracted with water. The extract was then filtered and concentrated to obtain the composition.

[0013] In some implementations, the extraction method is water decoction. The decoction is performed 2 to 3 times, specifically 2 or 3 times. Each decoction lasts 0.5 to 1 hour, specifically 0.5 hours or 1 hour. In some specific embodiments, the decoction is performed twice: the first decoction involves adding water, heating over high heat to boiling, and then simmering over low heat for 0.5 hours. The second decoction involves taking the filtered residue, adding water, and decocting for 0.5 hours.

[0014] In some implementations, during the decoction, the mass ratio of the total mass of monk fruit, privet fruit, ginseng leaves, and rose petals to water is 1:(8-10), meaning the amount of water used is 8-10 times the total mass of the raw materials. In some specific embodiments, the mass ratio may be 1:8 or 1:10.

[0015] The preparation method provided by this invention further includes a step of soaking monk fruit, privet fruit, ginseng leaves, and rose petals in water before extraction. The soaking time is 0.5 to 1 hour, specifically 0.5 hours or 1 hour.

[0016] The present invention also provides the use of the composition or the composition prepared by the method described herein in the preparation of a medicament for the prevention and treatment of leukopenia.

[0017] The prevention and treatment include: increasing the number of immune cells, increasing immune organ indices, and / or improving spleen tissue damage. The immune cells include leukocytes or neutrophils.

[0018] The present invention also provides a medicament for the prevention and treatment of leukopenia, comprising the composition described in the present invention or the composition prepared by the preparation method described in the present invention, and excipients.

[0019] The pharmaceutically acceptable excipients include at least one of the following: fillers, lubricants, suspending agents, emulsifiers, and diluents.

[0020] The dosage forms of the drug include, but are not limited to, capsules, tablets, granules, powders, oral liquids, or pills. For specific dosage forms, the present invention can be prepared using conventional excipients and processes conventional in the art.

[0021] The drug described in this invention also includes other drugs that can improve and prevent leukopenia, such as vitamin B4, leukogen, and shark liver oil.

[0022] This invention also provides a method for preventing and treating leukopenia, comprising: administering the composition or drug described in this invention. In this invention, the target of prevention and treatment is a human or non-human mammal; there are no specific limitations on the specific species of non-human mammals, and any species commonly found in the art are acceptable.

[0023] The composition of this invention is extracted from monk fruit, privet fruit, ginseng leaf, and rose petals. The components work synergistically. Animal functional experiments have demonstrated that the composition of this invention can significantly increase the spleen index, white blood cell and neutrophil counts, and improve pathological damage to spleen tissue, thereby enhancing immunity. The absence of one or two of the medicinal ingredients significantly reduces the effect, indicating that the components of this invention work synergistically and are indispensable. Attached Figure Description

[0024] Figure 1 The results of HE staining of mouse spleen tissue (×10) are shown below. A is a pathological section of spleen tissue from the normal group of mice in Example 4; B is a pathological section of spleen tissue from the model group of mice in Example 4; C is a pathological section of spleen tissue from the positive drug group of mice in Example 4; D is a pathological section of spleen tissue from the low-dose group of mice in Example 4; E is a pathological section of spleen tissue from the medium-dose group of mice in Example 4; and F is a pathological section of spleen tissue from the high-dose group of mice in Example 4. Red arrow: spleen corpuscle. Detailed Implementation

[0025] This invention provides a composition for preventing and treating leukopenia, its preparation method, and its application. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments, and those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0026] In this article, expressions such as “including,” “having,” and “containing,” as well as their grammatical synonyms, should be understood as open-ended expressions that are inclusive rather than exclusive. The option of “composed of” is also provided.

[0027] In addition to the publicly disclosed point values, the numerical ranges discussed in this article also cover all intermediate values ​​between any two endpoint values, including any numerical value between any two point values ​​and a small range within any two numerical ranges.

[0028] This invention provides a traditional Chinese medicine composition for improving leukopenia. The raw materials include, by weight, 1.5-90 parts of monk fruit, 1-72 parts of privet fruit, 0.5-54 parts of ginseng leaf, and 0.5-36 parts of rose.

[0029] Preferably made from the following raw materials in the following weight ratio: monk fruit: privet fruit: ginseng leaf: rose flower = 9:6:3:3.

[0030] Monk fruit, in particular, is sweet and cool in nature, and enters the lung and large intestine meridians. It can clear heat and moisten the lungs, soothe the throat and improve voice, and lubricate the intestines to relieve constipation. Studies have shown that monk fruit polysaccharides can significantly enhance the immune capacity of immunosuppressed mice.

[0031] Privet fruit has a sweet and bitter taste, is cool in nature, and enters the liver and kidney meridians. It can nourish the liver and kidneys, improve eyesight, and darken hair. Studies have shown that privet fruit can reduce white blood cell damage during chemotherapy and radiotherapy and increase the number of peripheral white blood cells.

[0032] Ginseng leaves have a bitter and sweet taste, are cold in nature, and enter the lung and stomach meridians. Ginseng leaves can replenish qi, benefit the lungs, relieve summer heat, and promote the production of body fluids. Studies have shown that ginseng leaf products have been used to treat leukopenia.

[0033] Rose petals have a sweet and slightly bitter taste, and are warm in nature. They enter the liver and spleen meridians, and can regulate qi, relieve depression, harmonize blood, and relieve pain. Studies have shown that rose petals have effects such as enhancing immunity and fighting viruses.

[0034] Furthermore, this invention also provides a method for preparing the above-mentioned traditional Chinese medicine composition for improving leukopenia, comprising:

[0035] S1. Mix monk fruit, privet fruit, ginseng leaves, and rose petals to obtain the extraction raw material;

[0036] S2. Extract the raw material with water.

[0037] The extraction method is water decoction.

[0038] Specifically, the water is boiled twice.

[0039] The first decoction is performed as follows: After adding water to all the ingredients, heat over high heat until boiling, then simmer over low heat for 0.5 hours. The mass ratio of the extracted raw materials to water is 1:10.

[0040] After the first decoction, filter while hot. Add water to the filtered residue and decoct for 0.5 hours, then filter again. The mass ratio of the filtered residue to water is 1:8.

[0041] The process includes a soaking step before decoction. The soaking time is 0.5 hours.

[0042] The test materials used in this invention are all common commercial products and can be purchased on the market.

[0043] The present invention will be further illustrated below with reference to the embodiments:

[0044] Example 1

[0045] To quantitatively assess the potential role of traditional Chinese medicine (TCM) in enhancing immune function, a component-level weighted probability model was used to systematically model the efficacy. Deep learning technology was employed to obtain the potential efficacy and efficacy probability values ​​of TCM components. For each efficacy of a TCM, all associated TCM components were statistically analyzed, and each component was assigned a corresponding weight value based on its efficacy probability value. The weighted probability value of the efficacy at the component level was calculated and summed to obtain the efficacy intensity characterization value for each TCM. Based on this, to achieve focused analysis of immune enhancement, the relative weight of each efficacy was assigned upwards to the TCM level; that is, the product of the efficacy weight corresponding to immune enhancement and the efficacy intensity value of the TCM was used as the contribution index of TCM in enhancing immunity. Finally, the above indicators were summarized to construct a TCM immune enhancement scoring system for subsequent functional formulation design and screening. As shown in Table 1, the contribution of the four-herb group to enhancing immune function was higher than that of the non-herb group and the replacement herb group; therefore, the four-herb group was selected for subsequent formulation validation.

[0046] Table 1. Contribution of Traditional Chinese Medicine and Combinations to Enhancing Immunity

[0047] Traditional Chinese medicine (composition) Contribution Monk fruit + Privet fruit + Ginseng leaf + Rose 0.2308 Monk fruit + Privet fruit + Ginseng leaf + Cimicifuga 0.2304 Monk fruit + Privet fruit + Ginseng leaf + White peony root 0.2295 American ginseng + privet fruit + ginseng leaves + rose petals 0.2113 Monk fruit + ginseng leaves + rose petals 0.1735 Monk fruit + Privet fruit + Ginseng leaf 0.1730 Monk fruit + Cimicifuga rhizome + White peony root 0.1730 Monk fruit + rose 0.1169 Monk fruit + Privet fruit 0.1164 Monk fruit + ginseng leaf 0.1157 Privet + Rose 0.1152 Ginseng leaves + rose petals 0.1144 Privet fruit + Ginseng leaf 0.1139

[0048] Example 2

[0049] This embodiment provides a traditional Chinese medicine composition for improving leukopenia, comprising 9 parts of monk fruit, 6 parts of privet fruit, 3 parts of ginseng leaf, and 3 parts of rose. The raw materials are weighed as described above, and 10 times the amount of distilled water is added. The mixture is soaked for 30 minutes, heated to boiling over high heat, then simmered over low heat for 30 minutes. The mixture is filtered while hot. The residue is then added to 8 times the amount of distilled water and simmered for 30 minutes, followed by filtration. The two filtrates are combined and concentrated under reduced pressure to 1.4 g / mL.

[0050] Example 3

[0051] 1) Experimental materials:

[0052] Cells: Human promyelocytic leukemia cells HL-60 were purchased from Beina Chuanglian Biotechnology Co., Ltd.

[0053] Test drug: The traditional Chinese medicine composition of Example 1 of the present invention was diluted with culture medium to a final concentration of 1 mg / mL. The drug-deficient compositions 1 to 2 were prepared in corresponding proportions and diluted with the same culture medium to final concentrations of 0.57 mg / mL and 0.43 mg / mL, respectively.

[0054] Group 1 of the drug-deficient composition: 9 parts of monk fruit and 3 parts of ginseng leaf.

[0055] Group 2 of the drug-deficient composition: 6 parts of privet fruit and 3 parts of ginseng leaf.

[0056] Instruments and materials: Cell incubator, ELISA reader.

[0057] 1) Experimental methods

[0058] HL-60 cells were cultured in IMDM complete medium containing 20% ​​fetal bovine serum and incubated at 37°C in a 5% CO2 cell culture incubator. HL-60 cells in logarithmic growth phase were seeded at 10,000 cells / well in 96-well cell culture plates, 100 μL per well. After 24 h of culture, the cells in the 96-well plates were divided into control, model, and drug treatment groups. Cyclophosphamide (8.8 μM) was added to the model and drug treatment groups to establish an immunosuppression model. The drug treatment groups were each given 100 μL of the three tested drugs, while the control and model groups were each given 100 μL of culture medium. After 24 h of culture, cell viability was assessed using a CCK-8 assay. The absorbance (A) of each well was measured using a microplate reader at a wavelength of 450 nm. Cell viability was calculated from the measured absorbance, reflecting the effect of each drug on cell survival. Viability = (A / well) * ... 药物组 -A 空白组 ) / (A 对照组 -A 空白组 )×100%.

[0059] Table 2. Effects of the herbal composition and drug-deficient compositions 1-2 in Example 1 on HL-60 cell viability.

[0060] Experimental Groups Cell vitality control group 1.00±0.14 Model group 0.89±0.04 Example 1: Traditional Chinese Medicine Composition 1.18±0.10 Group 1 of drug-deficient composition <![CDATA[0.85±0.07 ** ]]> Group 2 of drug-deficient composition <![CDATA[0.99±0.07 * ]]>

[0061] Note: Compared with the herbal composition group in Example 1. * P<0.05, ** P<0.01.

[0062] As shown in Table 2, compared with the herbal composition group in Example 1, the cell viability of the drug-deficient composition groups 1 and 2 was significantly reduced. This indicates that the four medicinal ingredients in the herbal composition of the present invention exerted a synergistic effect.

[0063] Example 4

[0064] 1) Experimental materials:

[0065] Animals: 42 male mice, BALB / c, 18-22g. They were housed separately in independently ventilated cage systems at the Animal Experiment Center of Beijing University of Chinese Medicine.

[0066] Test drug: The traditional Chinese medicine composition of Example 1 of the present invention was prepared into a gavage volume containing 6 times, 8 times, and 10 times the minimum human dose of raw drug, which were 16.38 g / kg, 21.84 g / kg, and 27.3 g / kg, respectively.

[0067] Instruments and materials: analytical balance, blood cell analyzer.

[0068] 2) Experimental methods

[0069] Forty-two mice were randomly divided into six groups of seven each: a control group, a model group, a positive control group, and low-, medium-, and high-dose groups. The control groups received daily doses of drug solution equal to 6, 8, and 10 times the lowest human dose, respectively, while the positive control group received 200 mg / kg. -1 Astragalus polysaccharide was administered to both the control and model groups with equal volumes of physiological saline for a total of 12 days. On days 8 and 10 of administration, both the model and treatment groups received an intraperitoneal injection of 50 mg / kg. -1 Cyclophosphamide was administered. On day 13, mice were weighed using an analytical balance. Whole blood was collected by enucleating the mice, and 1-2 drops of blood were added to an EDTA-containing anticoagulant tube. White blood cell and neutrophil counts were analyzed using a blood cell analyzer; the results are shown in Table 4. Mice were euthanized, and the spleen was harvested and weighed. The organ index was calculated using the formula: Organ Index = Organ Mass (mg) / Mouse Body Mass (g); the results are shown in Table 3. The mouse spleen tissue was preserved at room temperature in a general-purpose tissue fixative for subsequent pathological analysis; the results are shown in Table 3. Figure 1 .

[0070] Table 3. Effects of traditional Chinese medicine composition on spleen / body weight ratio in mice.

[0071]

[0072] Note: Compared to the model group ** P<0.01, *** P<0.001

[0073] As shown in Table 3, the spleen index in the model group was significantly lower than that in the control group; and the spleen index was significantly higher in the low-dose and medium-dose administration groups compared with the model group. This indicates that the traditional Chinese medicine composition can significantly improve the spleen index in cyclophosphamide-immunosuppressed mice.

[0074] Table 4. Effects of traditional Chinese medicine composition on the number of white blood cells and neutrophils in mice.

[0075]

[0076] Note: Compared to the model group * P<0.05, ** P<0.01, *** P<0.001.

[0077] As shown in Table 4, compared with the control group, the number of white blood cells and neutrophils in the model group was significantly reduced, indicating successful modeling; compared with the model group, the number of white blood cells and neutrophils in the treatment group was significantly increased. This indicates that the traditional Chinese medicine composition can significantly increase the number of white blood cells and neutrophils in cyclophosphamide-immunosuppressed mice.

[0078] Depend on Figure 1 It can be seen that the spleen tissue in the control group was intact, with the white and red pulp clearly distinguishable, splenic corpuscles clearly visible, and cells evenly arranged. Compared with the control group, the spleen tissue in the model group was structurally disordered, with the white and red pulp indistinguishable and splenic corpuscles showing pathological changes. Compared with the model group, the drug-treated group improved the pathological damage to the spleen tissue, indicating that this traditional Chinese medicine composition can improve the pathological changes in the spleen tissue of cyclophosphamide-immunosuppressed mice.

[0079] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition, characterized in that, The raw materials include the following parts by weight: 1.5–90 parts monk fruit, 1–72 parts privet fruit, 0.5–54 parts ginseng leaf and 0.5–36 parts rose.

2. The composition according to claim 1, characterized in that, The raw materials include the following parts by weight: 3–60 parts monk fruit, 2–48 parts privet fruit, 1–36 parts ginseng leaves, and 1–12 parts rose petals.

3. The composition according to claim 1, characterized in that, It is made from the following ingredients in parts by weight: 9 parts monk fruit, 6 parts privet fruit, 3 parts ginseng leaves and 3 parts rose petals.

4. A method for preparing the composition according to any one of claims 1 to 3, characterized in that, include: Monk fruit, privet fruit, ginseng leaf and rose petals were mixed and extracted with water. The extract was then filtered and concentrated to obtain the composition.

5. The preparation method according to claim 4, characterized in that, The extraction method is water decoction; the water decoction is performed 2 to 3 times, and the decoction time is 0.5 to 1 hour each time.

6. The preparation method according to claim 5, characterized in that, In the decoction, the mass ratio of the total mass of the monk fruit, privet fruit, ginseng leaf and rose petals to the mass of water is 1:(8-10).

7. The preparation method according to any one of claims 4 to 6, characterized in that, Before the extraction, the process also includes soaking the monk fruit, privet fruit, ginseng leaves, and rose petals in water for 0.5 to 1 hour.

8. The use of the composition according to any one of claims 1 to 3 or the composition prepared by any one of claims 4 to 7 in the preparation of a medicament for the prevention and treatment of leukopenia.

9. The application according to claim 8, characterized in that, The prevention and treatment include: increasing the number of immune cells, increasing the immune organ index, and / or improving spleen tissue damage.

10. A drug for the prevention and treatment of leukopenia, characterized in that, It includes the composition according to any one of claims 1 to 3 or the composition prepared by any one of claims 4 to 7, and excipients.

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