An anti-tumor N-acetyl-D-glucosamine compound wine and a preparation method thereof

Abstract

The application discloses an anti-tumor N-acetyl-D-glucosamine compound wine and a preparation method thereof, and belongs to the technical field of medicines. The anti-tumor N-acetyl-D-glucosamine compound wine provided by the application is characterized by comprising active ingredients and base wine; the active ingredients comprise N-acetyl-D-glucosamine, smilax riparian root, agrimony, oldenlandia, medlar and licorice. In the preparation, the traditional Chinese medicine components are first soaked and extracted by the base wine, then N-acetyl-D-glucosamine is added for dissolution, and the anti-tumor N-acetyl-D-glucosamine compound wine is obtained through aging and filling. The N-acetyl-D-glucosamine is compounded with traditional Chinese medicine such as smilax riparian root and agrimony for the first time, and the synergistic effect is exerted through the wine form. The in-vitro cell experiment and in-vivo animal experiment prove that the compound wine has a significant inhibitory effect on various tumor cells such as liver cancer, lung cancer and cervical cancer, and the effect is obviously better than that of the comparative examples; and the preparation process is simple and suitable for industrial production.

Description

Technical Field

[0001] This application belongs to the field of pharmaceutical technology, and in particular relates to an antitumor N-acetyl-D-glucosamine compound wine and its preparation method. Background Technology

[0002] Cancer is a major disease that seriously threatens human health, and the search for highly effective and low-toxicity anti-tumor drugs has always been a research hotspot. N-acetyl-D-glucosamine (NAG) is the basic structural unit of chitin and possesses various biological activities such as anti-inflammatory and immunomodulatory effects. Existing studies have shown that N-acetyl-D-glucosamine has certain anti-tumor activity, but its effect is limited when used alone, and a relatively high dose is required to achieve the desired inhibitory effect. In addition, although there are reports of adding N-acetyl-D-glucosamine to alcohol in existing technologies, these mainly focus on reducing alcohol-induced liver damage and improving intestinal function, and there is no research on its combination with specific traditional Chinese medicine components for anti-tumor purposes.

[0003] Traditional Chinese medicine (TCM) has unique advantages in adjuvant therapy for tumors. Actinidia chinensis root (root of the kiwifruit plant, Actinidia chinensis family) has the effects of clearing heat and detoxifying, dispelling wind and dampness, and promoting diuresis and stopping bleeding. Modern research shows that it contains triterpenoids, phenolic acids, and other components, which have a certain inhibitory effect on digestive tract tumors. Agrimonia pilosa (dried aerial parts of Agrimonia pilosa plant, Rosaceae family), also known as "toughening herb," ​​has the effects of astringing and stopping bleeding, treating malaria, stopping dysentery, detoxifying, and tonifying deficiency. Modern pharmacological studies have confirmed that it is rich in flavonoids and phenolic compounds, which can induce tumor cell apoptosis. Combining N-acetyl-D-glucosamine with Actinidia chinensis root, Agrimonia pilosa, and other TCM herbs with "strengthening the body's resistance and fighting cancer" properties may exert a synergistic anti-tumor effect through multiple targets and pathways. As a traditional TCM dosage form, medicinal wine has the characteristics of "unblocking blood vessels and promoting the efficacy of medicine," which can promote the absorption of active ingredients. Currently, there are no reports of using a combination of N-acetyl-D-glucosamine, Actinidia chinensis root, and Agrimonia pilosa to prepare an anti-tumor medicinal wine. Summary of the Invention

[0004] To overcome the aforementioned deficiencies in the prior art, this application provides an antitumor N-acetyl-D-glucosamine compound tincture and its preparation method.

[0005] To achieve the above-mentioned objectives, this application provides the following technical solution:

[0006] On the one hand, this application provides an anti-tumor N-acetyl-D-glucosamine compound wine preparation, including active ingredients and base wine;

[0007] The active ingredient is prepared from the following components in parts by weight: 5-20 parts of N-acetyl-D-glucosamine, 15-30 parts of Actinidia chinensis root, 10-25 parts of Agrimonia pilosa, 5-15 parts of Hedyotis diffusa, 5-10 parts of Lycium barbarum, and 3-8 parts of Glycyrrhiza uralensis.

[0008] Optionally, the active ingredient is prepared from a component comprising the following parts by weight: 10 parts N-acetyl-D-glucosamine, 20 parts Actinidia chinensis root, 15 parts Agrimonia pilosa, 10 parts Hedyotis diffusa, 8 parts Lycium barbarum, and 5 parts Glycyrrhiza uralensis.

[0009] Optionally, the base liquor includes baijiu or huangjiu;

[0010] The base liquor has an alcohol content of 35-60% vol.

[0011] Preferably, the base liquor has an alcohol content of 45% vol.

[0012] Optionally, the ratio of the active ingredient to the base wine is 1g:10~20mL.

[0013] Preferably, the ratio of the active ingredient to the base wine is 1g:15mL.

[0014] Secondly, this application provides a method for preparing the aforementioned antitumor N-acetyl-D-glucosamine compound tincture, comprising the following steps:

[0015] (1) The root of the Chinese herb, Agrimonia pilosa, Hedyotis diffusa, Lycium barbarum and Glycyrrhiza uralensis are crushed and sieved separately. The sieved components are collected and mixed to obtain a mixed powder of Chinese herbal medicines.

[0016] (2) Mix the Chinese herbal medicine powder with the base wine, seal and soak, filter, and take the filtrate to obtain the Chinese herbal medicine wine extract;

[0017] (3) Mix the Chinese herbal wine extract with N-acetyl-D-glucosamine, stir until completely dissolved, let stand and age, take the supernatant to obtain the anti-tumor N-acetyl-D-glucosamine compound wine.

[0018] Optionally, in step (1), the mesh size of the pulverized and sieved material is 20 to 50 mesh.

[0019] Preferably, in step (1), the pulverized and sieved material has a mesh size of 30.

[0020] Optionally, in step (2), the sealing and soaking time is 15 to 30 days;

[0021] During the sealed soaking process, the mixture is stirred once every 3 to 5 days, with a stirring speed of 30 to 60 rpm and a stirring time of 10 to 20 minutes each time.

[0022] The filtration is performed using a 150-250 mesh filter cloth.

[0023] Preferably, in step (2), the sealing and soaking time is 20 days;

[0024] During the sealed soaking process, the mixture is stirred once every 4 days, with a stirring speed of 50 rpm and a stirring time of 15 minutes each time.

[0025] The filtration is performed using a 200-mesh filter cloth.

[0026] Optionally, in step (3), the stirring speed is 60~120 rpm;

[0027] The temperature for static aging is 15~25℃;

[0028] The settling and aging period is 7 to 15 days.

[0029] Preferably, in step (3), the stirring speed is 90 rpm;

[0030] The temperature for static aging is 20°C;

[0031] The settling and aging period is 10 days.

[0032] Thirdly, this application provides the application of the above-mentioned antitumor N-acetyl-D-glucosamine compound wine, or the antitumor N-acetyl-D-glucosamine compound wine prepared by the above preparation method, in the preparation of drugs that inhibit the proliferation of tumor cells.

[0033] Optionally, the tumor cells include liver cancer cells, lung cancer cells, or cervical cancer cells.

[0034] Compared with the prior art, this application has the following advantages:

[0035] (1) This application provides an antitumor N-acetyl-D-glucosamine compound wine preparation, which for the first time combines N-acetyl-D-glucosamine with traditional Chinese medicines such as Actinidia chinensis root and Agrimonia pilosa. The synergistic effect of each component significantly improves the antitumor effect. Experimental results show that the inhibitory rate of the compound wine preparation of this application on human liver cancer cells HepG2, human lung cancer cells A549 and human cervical cancer cells HeLa is significantly higher than that of the control group, indicating that this application has established a reasonable compound compatibility relationship and produced a synergistic effect. Due to the synergistic effect, this application can achieve the ideal antitumor effect at a lower dose, which helps to reduce the potential side effects of N-acetyl-D-glucosamine. In addition, the use of wine preparation in this application can not only fully extract the effective components in traditional Chinese medicines, but also promote the absorption and utilization of active ingredients by taking advantage of the "potential effect" of wine, thereby improving bioavailability.

[0036] (2) The method for preparing an antitumor N-acetyl-D-glucosamine compound wine provided in this application adopts a room temperature extraction process, which avoids the destruction of active ingredients by high temperature, and is simple to operate and suitable for industrial production. Detailed Implementation

[0037] The present application is further illustrated below with reference to specific embodiments. The following descriptions are merely a few embodiments of the present application and are not intended to limit the present application in any way. Although the present application discloses preferred embodiments as follows, they are not intended to limit the present application. Any modifications or variations made by those skilled in the art without departing from the scope of the technical solution of the present application using the disclosed technical content are equivalent to equivalent implementation cases and all fall within the scope of the technical solution.

[0038] Unless otherwise specified, the raw materials used in the embodiments of this application are all purchased commercially and used directly without any special treatment.

[0039] Unless otherwise specified, the analytical methods in the embodiments all adopt conventional instrument or equipment settings and conventional analytical methods.

[0040] The N-acetyl-D-glucosamine used in the following examples and comparative examples was purchased from Zhejiang Lvchuang Biotechnology Co., Ltd., with a purity of 99%.

[0041] Example 1

[0042] A method for preparing an antitumor N-acetyl-D-glucosamine compound tincture, comprising the following steps:

[0043] (1) Weigh the following components by mass:

[0044] 5g N-acetyl-D-glucosamine, 15g Actinidia chinensis root, 10g Agrimonia pilosa, 5g Hedyotis diffusa, 5g Lycium barbarum, 3g Glycyrrhiza uralensis; 430mL of 35% vol baijiu as the base liquor.

[0045] (2) Wash the root of the Chinese pear, the agrimony, the white flower tongue grass, the wolfberry and the licorice to remove impurities, dry them at 50°C to constant weight, pulverize them separately and pass them through a 20-mesh sieve, collect the sieve-under-the-sieve components, mix them evenly, and obtain the Chinese herbal medicine mixed powder.

[0046] (3) Place the Chinese herbal medicine powder in a glass container, add base wine, seal and soak at room temperature for 15 days, stirring once every 3 days (each stirring speed is 30 rpm and each stirring time is 10 min), then filter with a 150 mesh filter cloth, take the filtrate, and obtain the Chinese herbal medicine wine extract.

[0047] (4) Mix the Chinese herbal wine extract with N-acetyl-D-glucosamine, stir at 60 rpm until completely dissolved, let stand and age at 15°C for 7 days, take the supernatant, fill it, sterilize at 80°C for 30 min, and obtain the anti-tumor N-acetyl-D-glucosamine compound wine.

[0048] Example 2

[0049] A method for preparing an antitumor N-acetyl-D-glucosamine compound tincture, comprising the following steps:

[0050] (1) Weigh the following components by mass:

[0051] 10g N-acetyl-D-glucosamine, 20g Actinidia chinensis root, 15g Agrimonia pilosa, 10g Hedyotis diffusa, 8g Lycium barbarum, 5g Glycyrrhiza uralensis; 1020mL of 45% vol baijiu (Chinese white liquor) as the base.

[0052] (2) Wash the root of the Chinese pear, the agrimony, the white flower tongue grass, the wolfberry and the licorice to remove impurities, dry them at 50°C to constant weight, pulverize them separately and pass them through a 30-mesh sieve, collect the sieve-under-the-sieve components, mix them evenly, and obtain the Chinese herbal medicine mixed powder.

[0053] (3) Place the mixed powder of Chinese medicine in a glass container, add base wine, seal and soak at room temperature for 20 days, stirring once every 4 days (stirring speed is 50 rpm and stirring time is 15 min each time), then filter with 200 mesh filter cloth, take the filtrate to obtain Chinese medicine wine extract;

[0054] (4) Mix the Chinese herbal wine extract with N-acetyl-D-glucosamine, stir at 90 rpm until completely dissolved, let stand and age at 20°C for 10 days, take the supernatant, fill it, sterilize at 80°C for 30 min, and obtain the anti-tumor N-acetyl-D-glucosamine compound wine.

[0055] Example 3

[0056] A method for preparing an antitumor N-acetyl-D-glucosamine compound tincture, comprising the following steps:

[0057] (1) Weigh the following components by mass:

[0058] 20g N-acetyl-D-glucosamine, 30g Actinidia chinensis root, 25g Agrimonia pilosa, 15g Hedyotis diffusa, 10g Lycium barbarum, 8g Glycyrrhiza uralensis; 2160mL of 60% vol baijiu as the base liquor.

[0059] (2) Wash the root of the Chinese pear, the agrimony, the white flower tongue grass, the wolfberry and the licorice to remove impurities, dry them at 50°C to constant weight, pulverize them separately and pass them through a 50-mesh sieve, collect the sieve-filled components, mix them evenly, and obtain the Chinese herbal medicine mixed powder.

[0060] (3) Place the Chinese herbal medicine powder in a glass container, add base wine, seal and soak at room temperature for 30 days, stirring once every 5 days (stirring speed is 60 rpm and stirring time is 20 min each time), then filter with a 250 mesh filter cloth, take the filtrate to obtain Chinese herbal medicine wine extract;

[0061] (4) Mix the Chinese herbal wine extract with N-acetyl-D-glucosamine, stir at 120 rpm until completely dissolved, let stand and age at 25°C for 15 days, take the supernatant, fill it, sterilize at 80°C for 30 min, and obtain the anti-tumor N-acetyl-D-glucosamine compound wine.

[0062] Comparative Example 1

[0063] NAG single-ingredient wine preparation (compared to Example 2):

[0064] Weigh 10g of N-acetyl-D-glucosamine and add it to 150mL of 45%vol liquor. Stir at 90rpm until completely dissolved. Let it stand and age at 20℃ for 10 days. Take the supernatant, fill it, and sterilize it at 80℃ for 30min to obtain NAG single-ingredient liquor.

[0065] Comparative Example 2

[0066] Traditional Chinese medicine compound wine preparation (compared to Example 2):

[0067] (1) Weigh the following components by mass:

[0068] 20g of *Actinidia chinensis* root, 15g of *Agrimonia pilosa*, 10g of *Hedyotis diffusa*, 8g of *Lycium barbarum*, and 5g of licorice; 870mL of 45% vol baijiu as the base liquor.

[0069] (2) Wash the root of the Chinese pear, the agrimony, the white flower tongue grass, the wolfberry and the licorice to remove impurities, dry them at 50°C to constant weight, pulverize them separately and pass them through a 30-mesh sieve, collect the sieve-under-the-sieve components, mix them evenly, and obtain the Chinese herbal medicine mixed powder.

[0070] (3) Place the mixed powder of Chinese medicine in a glass container, add base wine, seal and soak at room temperature for 20 days, stirring once every 4 days (stirring speed is 50 rpm and stirring time is 15 min each time), then filter with 200 mesh filter cloth, take the filtrate to obtain Chinese medicine wine extract;

[0071] (4) The Chinese herbal wine extract was left to stand at 20°C for 10 days, the supernatant was taken, bottled, and sterilized at 80°C for 30 minutes to obtain the Chinese herbal compound wine.

[0072] Comparative Example 3

[0073] The modified medicinal wine with altered active ingredients (compared to Example 2):

[0074] (1) Weigh the following components by mass:

[0075] 10g N-acetyl-D-glucosamine, 20g Astragalus membranaceus, 15g Ganoderma lucidum, 10g Hedyotis diffusa, 8g Lycium barbarum, 5g Glycyrrhiza uralensis; 1020mL of 45% vol baijiu (Chinese white liquor) as the base.

[0076] (2) Wash Astragalus membranaceus, Ganoderma lucidum, Hedyotis diffusa, Lycium barbarum and Glycyrrhiza uralensis to remove impurities, dry them at 50°C to constant weight, pulverize them separately and pass them through a 30-mesh sieve, collect the sieve-under-the-sieve components, mix them evenly, and obtain a mixed powder of Chinese medicine.

[0077] (3) Place the mixed powder of Chinese medicine in a glass container, add base wine, seal and soak at room temperature for 20 days, stirring once every 4 days (stirring speed is 50 rpm and stirring time is 15 min each time), then filter with 200 mesh filter cloth, take the filtrate to obtain Chinese medicine wine extract;

[0078] (4) Mix the Chinese herbal wine extract with N-acetyl-D-glucosamine, stir at 90 rpm until completely dissolved, let stand and age at 20°C for 10 days, take the supernatant, fill it, and sterilize at 80°C for 30 min to obtain the wine after changing the active ingredients of the Chinese herbal medicine.

[0079] Experimental Example 1

[0080] In vitro antitumor experiment

[0081] 1. Experimental materials

[0082] Cell lines: HepG2 human liver cancer cells, A549 human non-small cell lung cancer cells, and HeLa human cervical cancer cells were purchased from the Cell Bank of the Chinese Academy of Sciences, with catalog numbers THu 72, SCSP-503, and SCSP-504, respectively.

[0083] Test samples: the wines prepared in Examples 1-3 and Comparative Examples 1-3.

[0084] Positive control: 5-fluorouracil (5-FU, 10 μg / mL, purchased from MCE China).

[0085] Blank control: 45% vol spirits without any drugs (diluted with complete culture medium to the same alcohol concentration as the test sample).

[0086] 2. Experimental Methods

[0087] The MTT assay was used to detect cell proliferation inhibition rate. Cells in logarithmic growth phase were harvested and the cell density was adjusted to 5 × 10⁶ cells / year. 4 The sample was inoculated at 100 μL / mL into a 96-well plate. After 24 hours of incubation, the test sample was added (diluted with complete culture medium to a final concentration of 200 μg / mL (based on total crude drug, where the initial concentration of total crude drug was calculated according to the following formula: as in Example 1, total crude drug concentration = (5g + 15g + 10g + 5g + 5g + 3g) / 430mL = 0.1g / mL = 100mg / mL)). Six replicates were set for each concentration. After 48 hours of further incubation, 20 μL of 5mg / mL MTT solution was added to each well, and the plate was incubated for 4 hours. The supernatant was discarded, and 150 μL of DMSO was added. The plate was shaken for 10 minutes to completely dissolve the crystals. The absorbance (OD value) was measured at 490 nm using a microplate reader. The inhibition rate was calculated as follows: Inhibition rate (%) = (1 - OD value of experimental group / OD value of blank control group) × 100%.

[0088] The results are shown in Table 1. The blank control group showed good cell growth with OD values ​​between 0.8 and 0.9, indicating excellent cell condition and a stable and reliable experimental system.

[0089] Table 1 Comparison of inhibition rates of different samples on three types of tumor cells (%, n=6, x̄±s)

[0090] Grouping HepG2 cells A549 cells HeLa cells Example 1 69.37±3.42 64.51±3.52 60.84±3.62 Example 2 76.45±3.18 70.82±3.37 67.23±3.26 Example 3 71.96±3.28 66.93±3.41 63.17±3.39 Comparative Example 1 32.56±4.12 28.94±4.35 25.63±4.08 Comparative Example 2 38.74±3.65 35.26±3.78 32.41±3.85 Comparative Example 3 41.62±3.93 38.47±4.01 35.29±4.15 Positive control (5-FU) 85.42±2.86 81.37±3.02 79.84±3.26

[0091] As shown in Table 1, the compound medicinal wines prepared in Examples 1-3 of this application all exhibited significant inhibitory effects on the three types of tumor cells. Example 2 showed the best effect, with an inhibition rate of 76.45% against HepG2 cells, approaching the level of the positive control 5-FU. Comparative Example 1 (NAG alone) had a lower inhibition rate, only 32.56% against HepG2 cells. Comparative Example 2 (a compound traditional Chinese medicine formula, without NAG) had an inhibition rate (38.74%) higher than Comparative Example 1, but significantly lower than the examples, indicating that the traditional Chinese medicine components such as *Actinidia chinensis* root and *Agrimonia pilosa* themselves possess certain anti-tumor activity, but the effect is significantly enhanced when combined with NAG. Comparative Example 3 (with altered active ingredients of the traditional Chinese medicine) had an inhibition rate of 41.62%, higher than Comparative Examples 1 and 2, but still significantly lower than the examples, indicating that the anti-tumor effect of the compound medicinal wine of traditional Chinese medicine + NAG in this application is superior to other combinations of traditional Chinese medicine and NAG, demonstrating significant progress.

[0092] Experiment Example 2

[0093] Animal tumor suppression experiment

[0094] Experimental methods:

[0095] Establishment of H22 tumor-bearing mouse model: Well-grown H22 ascites tumor cells (purchased from Shanghai Huzhen Industrial Co., Ltd.) were collected, and the cell density was adjusted to 1×10⁻⁶. 7 0.2 mL of cyclophosphamide was administered subcutaneously to the right anterior axilla of Kunming mice (Beijing Vital River Laboratory Animal Technology Co., Ltd.). Twenty-four hours after inoculation, mice were randomly divided into seven groups of ten mice each: model control group (physiological saline), positive control group (cyclophosphamide, 20 mg / kg), Example 2 group (0.2 mL / 10 g gavage), Comparative Example 1 group (same volume gavage), Comparative Example 2 group (same volume gavage), and Comparative Example 3 group (same volume gavage). The mice were administered the drug for 14 consecutive days. Twenty-four hours after the last administration, the mice were sacrificed, the tumors were removed and weighed, and the tumor inhibition rate was calculated. Tumor inhibition rate (%) = (1 - average tumor weight of the administered group / average tumor weight of the model control group) × 100%. The results are shown in Table 2.

[0096] Table 2. Antitumor effects of different samples on H22 tumor-bearing mice (n=10, x̄±s)

[0097] Grouping Tumor weight (g) Tumor inhibition rate (%) Model control group 1.86±0.35 - Positive control group 0.48±0.12 74.19 Example 2 0.75±0.19 59.68 Comparative Example 1 1.42±0.31 23.66 Comparative Example 2 1.35±0.29 27.42 Comparative Example 3 1.28±0.27 31.18

[0098] As shown in Table 2, the in vivo tumor inhibition rate of Example 1 of this application reached 59.68%, which was significantly higher than that of Comparative Example 1 (23.66%), Comparative Example 2 (27.42%) and Comparative Example 3 (31.18%), consistent with the in vitro experimental results, further confirming the synergistic anti-tumor effect of the compound wine preparation of this application and its advantages over the prior art.

[0099] In summary, the N-acetyl-D-glucosamine compound tincture provided in this application has a simple preparation process and low cost. The resulting product has significant anti-tumor activity and can be used to prepare drugs or health foods for adjuvant treatment of liver cancer, lung cancer, cervical cancer, and other diseases, showing good prospects for industrial application.

[0100] The above description is merely a few embodiments of this application and is not intended to limit this application in any way. Although this application discloses preferred embodiments as described above, it is not intended to limit this application. Any changes or modifications made by those skilled in the art without departing from the scope of the technical solution of this application using the disclosed technical content are equivalent to equivalent implementation cases and fall within the scope of the technical solution.

Claims

1. A compound medicated wine containing N-acetyl-D-glucosamine for antitumor purposes, characterized in that, Including active ingredients and base wine; The active ingredient is prepared from the following components in parts by weight: 5-20 parts of N-acetyl-D-glucosamine, 15-30 parts of Actinidia chinensis root, 10-25 parts of Agrimonia pilosa, 5-15 parts of Hedyotis diffusa, 5-10 parts of Lycium barbarum, and 3-8 parts of Glycyrrhiza uralensis.

2. The antitumor N-acetyl-D-glucosamine compound tincture according to claim 1, characterized in that, The active ingredient is prepared from the following components in parts by weight: 10 parts N-acetyl-D-glucosamine, 20 parts Actinidia chinensis root, 15 parts Agrimonia pilosa, 10 parts Hedyotis diffusa, 8 parts Lycium barbarum, and 5 parts Glycyrrhiza uralensis.

3. The antitumor N-acetyl-D-glucosamine compound tincture according to claim 1, characterized in that, The base liquor includes baijiu or huangjiu; The base liquor has an alcohol content of 35-60% vol.

4. The antitumor N-acetyl-D-glucosamine compound tincture according to claim 1, characterized in that, The ratio of the active ingredient to the base wine is 1g:10~20mL.

5. A method for preparing an antitumor N-acetyl-D-glucosamine compound tincture according to any one of claims 1 to 4, characterized in that, Includes the following steps: (1) The root of the Chinese herb, Agrimonia pilosa, Hedyotis diffusa, Lycium barbarum and Glycyrrhiza uralensis are crushed and sieved separately. The sieved components are collected and mixed to obtain a mixed powder of Chinese herbal medicines. (2) Mix the Chinese herbal medicine powder with the base wine, seal and soak, filter, and take the filtrate to obtain the Chinese herbal medicine wine extract; (3) Mix the Chinese herbal wine extract with N-acetyl-D-glucosamine, stir until completely dissolved, let stand and age, take the supernatant to obtain the anti-tumor N-acetyl-D-glucosamine compound wine.

6. The method for preparing an antitumor N-acetyl-D-glucosamine compound tincture according to claim 5, characterized in that, In step (1), the mesh size of the pulverized and sieved material is 20 to 50 mesh.

7. The method for preparing an antitumor N-acetyl-D-glucosamine compound tincture according to claim 5, characterized in that, In step (2), the sealing and soaking time is 15 to 30 days; During the sealed soaking process, the mixture is stirred once every 3 to 5 days, with a stirring speed of 30 to 60 rpm and a stirring time of 10 to 20 minutes each time. The filtration is performed using a 150-250 mesh filter cloth.

8. The method for preparing an antitumor N-acetyl-D-glucosamine compound tincture according to claim 5, characterized in that, In step (3), the stirring speed is 60~120 rpm; The temperature for static aging is 15~25℃; The settling and aging period is 7 to 15 days.

9. The use of an antitumor N-acetyl-D-glucosamine compound tincture as described in any one of claims 1 to 4, or an antitumor N-acetyl-D-glucosamine compound tincture prepared by the preparation method described in any one of claims 5 to 8, in the preparation of a drug for inhibiting tumor cell proliferation.

10. The application according to claim 9, characterized in that, The tumor cells include liver cancer cells, lung cancer cells, or cervical cancer cells.