A traditional Chinese medicine extract for treating pancreatic cancer, and a preparation method and application thereof

Abstract

The application provides a traditional Chinese medicine extract for treating pancreatic cancer and a preparation method and application thereof, and belongs to the technical field of biological medicine. The traditional Chinese medicine extract comprises the following steps: obtaining white peony root powder by stir-frying, obtaining atractylodes powder by stir-frying atractylodes with wheat bran, obtaining radix astragali powder by wine-frying, then mixing the white peony root powder, anemarrhena asphodeloides powder, codonopsis powder, atractylodes powder, radix astragali powder, citrus peel powder, magnolia officinalis powder, gigeriae gall powder, ginger powder, chuanxiong powder, cassia twig powder, safflower powder and peach kernel powder to obtain mixed powder, mixing the mixed powder with an ethanol aqueous solution, refluxing extraction, filtering, collecting the extract, concentrating, drying, and obtaining the traditional Chinese medicine extract; the traditional Chinese medicine extract can significantly inhibit the activity, migration ability of pancreatic cancer cells and the growth of pancreatic cancer, and research shows that the tumor killing effect of the traditional Chinese medicine extract with a high dose is better than that of a positive drug. The preparation method is simple and easy to implement, and is suitable for industrial production.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to a traditional Chinese medicine extract for treating pancreatic cancer, its preparation method, and its application. Background Technology

[0002] Pancreatic cancer often develops insidiously, with few or no early symptoms, and is often diagnosed at an advanced stage. Early symptoms of pancreatic cancer may include upper abdominal discomfort, dull pain, and indigestion. Abdominal pain is the most common initial symptom. Other gastrointestinal symptoms such as loss of appetite, nausea, vomiting, and weight loss are also clinical manifestations of pancreatic cancer.

[0003] Currently, surgical resection is the best treatment for pancreatic cancer. However, due to the rapid progression of pancreatic cancer, most patients are diagnosed at an advanced stage and have lost the opportunity for surgery. For patients with inoperable pancreatic cancer, the main treatment is chemotherapy. However, patients easily develop resistance to chemotherapy drugs, so most patients are not sensitive to chemotherapy, and chemotherapy is prone to adverse reactions, making it difficult for patients to tolerate and hindering the overall treatment effect of pancreatic cancer. Traditional Chinese medicine (TCM) focuses on a holistic approach, using syndrome differentiation and treatment to treat pancreatic cancer systemically, not just focusing on the lesion itself, thereby improving the symptoms of pancreatic cancer. Traditional Chinese medicine (TCM) classifies pancreatic cancer under categories such as "jaundice," "abdominal masses," and "hidden masses," with complex etiologies and pathogenesis. Spleen and stomach weakness and liver qi stagnation are key pathogenic factors. Emotional distress leads to liver qi stagnation and spleen deficiency, resulting in impaired spleen function. External invasion of damp-heat and cold toxins further damages spleen and stomach function; liver qi stagnation leads to qi and blood stasis; improper diet disrupts spleen function, causing spleen deficiency and internal retention of dampness and phlegm, which then transforms into heat; prolonged blood stasis and phlegm accumulation damage the pancreatic vessels, leading to cancer. Although researchers are currently dedicated to developing drugs for pancreatic cancer treatment, the incidence rate of pancreatic cancer is increasing year by year. Therefore, based on the fundamental theories of TCM and the characteristics of Chinese herbal medicine, selecting an effective Chinese herbal extract for treating pancreatic cancer remains a pressing technical challenge. Summary of the Invention

[0004] Therefore, the purpose of this invention is to provide a traditional Chinese medicine extract for treating pancreatic cancer, its preparation method, and its application.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a method for preparing a traditional Chinese medicine extract for treating pancreatic cancer, comprising the following steps: The herbal extract comprises the following raw materials in parts by weight: White peony root 5-12 parts, Anemarrhena rhizome 5-12 parts, Codonopsis root 4-8 parts, Atractylodes rhizome 4-8 parts, Astragalus root 4-8 parts, Tangerine peel 4-8 parts, Magnolia bark 4-8 ​​parts, Chicken gizzard lining 4-8 parts, Fresh ginger 4-8 parts, Ligusticum chuanxiong rhizome 2-4 parts, Cinnamon twig 2-4 parts, Safflower 4-8 parts, and Peach kernel 4-8 parts; (1) Stir-fry the white peony slices in a hot pan, cool them down to obtain stir-fried white peony, grind them, and sieve them to obtain white peony powder; (2) Mix Atractylodes macrocephala slices with wheat bran, stir-fry the resulting Atractylodes macrocephala and wheat bran mixture in a hot pan, cool it down to obtain stir-fried Atractylodes macrocephala, grind it, and sieve it to obtain Atractylodes macrocephala powder; (3) Mix the astragalus slices with rice wine and let them soak. Then stir-fry them in a hot pan and cool them down to obtain wine-roasted astragalus. Grind the astragalus and sieve it to obtain astragalus powder. (4) Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius and Prunus persica were ground and sieved to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder and Prunus persica powder. (5) Weigh out the following powders: white peony powder, anemarrhena powder, codonopsis powder, atractylodes powder, astragalus powder, tangerine peel powder, magnolia bark powder, chicken gizzard powder, ginger powder, chuanxiong powder, cinnamon twig powder, safflower powder, and peach kernel powder respectively, mix them together to obtain a mixed powder, mix the mixed powder with an ethanol aqueous solution, reflux to extract, filter, collect the extract, concentrate and dry to obtain the Chinese herbal extract.

[0006] Preferably, in step (1), the stir-frying is carried out at 150~200℃ for 10~20 minutes; the sieve mesh size is 40~60 mesh; and the temperature is lowered to 20~25℃.

[0007] Preferably, in step (2), the mass ratio of the Atractylodes macrocephala slices to wheat bran is 8~11:1; the stir-frying is carried out at 160~210℃ for 15~25 minutes; the sieve mesh size is 40~60 mesh; and the temperature is lowered to 20~25℃.

[0008] Preferably, in step (3), the mass-to-volume ratio of the Astragalus membranaceus slices to the rice wine is 4~6:1g / mL; the stir-frying is carried out at 100~120℃ for 8~12 minutes; the sieve mesh size is 40~60 mesh; and the temperature is lowered to 20~25℃.

[0009] Preferably, in step (5), the mass-to-volume ratio of the mixed powder to the ethanol aqueous solution is 1:8~12 g / mL; and the volume percentage of the ethanol aqueous solution is 30%~50%.

[0010] Preferably, the extraction is performed 1 to 3 times, and each extraction takes 0.5 to 2.5 hours.

[0011] This invention provides a traditional Chinese medicine extract prepared by the above-described preparation method.

[0012] This invention provides an application of the above-mentioned traditional Chinese medicine extract in the preparation of drugs for the prevention and treatment of pancreatic cancer.

[0013] Preferably, the herbal extract can inhibit the growth and migration of pancreatic cancer cells.

[0014] Preferably, the herbal extract accounts for 20% to 90% of the mass of the drug.

[0015] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a traditional Chinese medicine extract for treating pancreatic cancer, its preparation method, and its application. For the first time, this invention utilizes a mixture of raw materials including white peony root, anemarrhena rhizome, codonopsis root, atractylodes rhizome, astragalus root, tangerine peel, magnolia bark, chicken gizzard lining, ginger, chuanxiong rhizome, cinnamon twig, safflower, and peach kernel. The resulting extract significantly inhibits the activity, migration ability, and growth of pancreatic cancer cells. Furthermore, studies have shown that high doses of the extract are more effective at killing tumors than positive control drugs. This invention provides a new option and theoretical basis for the clinical treatment of pancreatic cancer. The preparation method is simple and easy to implement, suitable for industrial production. Attached Figure Description

[0016] Figure 1 The results show the effects of different treatment groups on the viability of pancreatic cancer cells; Figure 2 The results show the effects of different treatment groups on the migration ability of pancreatic cancer cells; Figure 3 The results show the effects of different treatment groups on pancreatic cancer growth. Detailed Implementation

[0017] This invention provides a method for preparing a traditional Chinese medicine extract for treating pancreatic cancer, comprising the following steps: The herbal extract comprises the following raw materials in parts by weight: White peony root 5-12 parts, Anemarrhena rhizome 5-12 parts, Codonopsis root 4-8 parts, Atractylodes rhizome 4-8 parts, Astragalus root 4-8 parts, Tangerine peel 4-8 parts, Magnolia bark 4-8 ​​parts, Chicken gizzard lining 4-8 parts, Fresh ginger 4-8 parts, Ligusticum chuanxiong rhizome 2-4 parts, Cinnamon twig 2-4 parts, Safflower 4-8 parts, and Peach kernel 4-8 parts; (1) Stir-fry the white peony slices in a hot pan, cool them down to obtain stir-fried white peony, grind them, and sieve them to obtain white peony powder; (2) Mix Atractylodes macrocephala slices with wheat bran, stir-fry the resulting Atractylodes macrocephala and wheat bran mixture in a hot pan, cool it down to obtain stir-fried Atractylodes macrocephala, grind it, and sieve it to obtain Atractylodes macrocephala powder; (3) Mix the astragalus slices with rice wine and let them soak. Then stir-fry them in a hot pan and cool them down to obtain wine-roasted astragalus. Grind the astragalus and sieve it to obtain astragalus powder. (4) Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius and Prunus persica were ground and sieved to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder and Prunus persica powder. (5) Weigh out the following powders: white peony powder, anemarrhena powder, codonopsis powder, atractylodes powder, astragalus powder, tangerine peel powder, magnolia bark powder, chicken gizzard powder, ginger powder, chuanxiong powder, cinnamon twig powder, safflower powder, and peach kernel powder respectively, mix them together to obtain a mixed powder, mix the mixed powder with an ethanol aqueous solution, reflux to extract, filter, collect the extract, concentrate and dry to obtain the Chinese herbal extract.

[0018] In this invention, as a preferred embodiment, the herbal extract comprises the following raw materials in parts by weight: The herbal extract is preferably made from the following ingredients in parts by weight: 6-10 parts white peony root, 6-10 parts anemarrhena rhizome, 5-7 parts codonopsis root, 5-7 parts atractylodes rhizome, 5-7 parts astragalus root, 5-7 parts tangerine peel, 5-7 parts magnolia bark, 5-7 parts chicken gizzard lining, 5-7 parts fresh ginger, 2.5-3.5 parts chuanxiong rhizome, 2.5-3.5 parts cinnamon twig, 5-7 parts safflower, and 5-7 parts peach kernel. This invention does not have any special limitations on the source of white peony root, anemarrhena rhizome, codonopsis root, atractylodes rhizome, astragalus root, tangerine peel, magnolia bark, chicken gizzard lining, ginger, chuanxiong rhizome, cinnamon twig, safflower, and peach kernel; any commercially available products known in the art can be used.

[0019] In this invention, the white peony root is the root of *Paeonia lactiflora* Pall., a plant belonging to the genus *Paeonia* of the Ranunculaceae family; it has the effects of nourishing blood and regulating menstruation, astringing yin and stopping sweating, softening the liver and relieving pain, and calming liver yang. In this invention, the anemarrhena rhizome is a heat-clearing medicinal material, the dried rhizome of *Anemarrhena asphodeloides* Bge., a plant belonging to the genus *Anemarrhena* of the Liliaceae family; it has the effects of clearing heat and purging fire, nourishing yin and moistening dryness. In this invention, the codonopsis root is the dried root of *Codonopsis pilosula*, a plant belonging to the Campanulaceae family; it has the effects of strengthening the spleen and lungs, nourishing blood and promoting body fluids. In this invention, the atractylodes rhizome is the dried rhizome of *Atractylodes macrocephala*, a plant belonging to the Asteraceae family; it has the effects of strengthening the spleen and replenishing qi, drying dampness and promoting diuresis. The astragalus root is the dried root of *Astragalus membranaceus*, a plant belonging to the genus *Astragalus* of the Fabaceae family; it has the effects of replenishing qi and raising yang, consolidating the exterior and stopping sweating, promoting diuresis and reducing swelling, promoting body fluids and nourishing blood, promoting circulation and relieving pain, promoting pus drainage and detoxification, and promoting wound healing and tissue regeneration. The tangerine peel mentioned above is the dried, mature peel of *Citrus reticulata* Blanco, a plant belonging to the Rutaceae family. It has the effects of regulating qi and strengthening the spleen, drying dampness and resolving phlegm. The magnolia bark mentioned above is the dried bark, root bark, and branch bark of *Magnolia officinalis*, a plant belonging to the Magnoliaceae family. It has the effects of drying dampness and resolving phlegm, lowering qi and relieving fullness, and is mainly used to treat dampness stagnation in the middle jiao, epigastric fullness and vomiting, food stagnation and qi stagnation, abdominal distension and constipation, and phlegm-dampness cough. The chicken gizzard lining mentioned above is a digestive medicine, the inner wall of the gizzard of the domestic chicken (*Gallus gallus domesticus* Brisson), a pheasant family animal. It has the effects of strengthening the stomach and promoting digestion, astringing essence and stopping seminal emission, and clearing urinary tract stones and dissolving stones. The ginger mentioned above is the fresh rhizome of *Zingiber officinale* Rosc., a plant belonging to the Zingiberaceae family. It has the effects of relieving exterior syndromes and dispersing cold, warming the middle jiao and stopping vomiting, resolving phlegm and stopping cough, and detoxifying fish and crab poisoning. The *Ligusticum chuanxiong* mentioned above is the dried rhizome of *Ligusticum chuanxiong* Hort., a plant belonging to the genus *Ligustrum* of the family Apiaceae. It has the effects of promoting blood circulation, regulating qi, dispelling wind, and relieving pain. The *Cinnamomum cassia* twig mentioned above is the tender branch of *Cinnamomum cassia* Presl, a plant belonging to the genus *Cinnamomum* of the family Lauraceae. It has the effects of inducing sweating and relieving muscle tension, warming and unblocking the meridians, assisting yang qi, and calming and descending qi. The *Carthamus tinctorius* mentioned above is a blood-activating and menstruation-regulating medicinal material, the dried flower of *Carthamus tinctorius* L., a plant belonging to the genus *Carthamus* of the family Asteraceae. It has the effects of promoting blood circulation, regulating menstruation, dispersing blood stasis, and relieving pain. The *Prunus persica* kernel mentioned above has the effects of promoting blood circulation, removing blood stasis, moistening the intestines and relieving constipation, relieving cough and asthma. It is neutral in nature, bitter and sweet in taste, and enters the heart, liver, and large intestine meridians. This invention has found that the herbal extract prepared from the above raw materials has a significant inhibitory effect on the activity, migration ability, and growth of pancreatic cancer cells.

[0020] In this invention, white peony root slices are stir-fried in a hot pan, cooled, and then ground and sieved to obtain white peony root powder. The stir-frying is performed at 150-200℃ for 10-20 minutes, more preferably at 160-190℃ for 12-18 minutes, and even more preferably at 180℃ for 15 minutes. The sieve mesh size is preferably 40-60 mesh, more preferably 45-55 mesh, and even more preferably 50 mesh. The cooling temperature is 20-25℃, more preferably 21-24℃, and even more preferably 22℃. The thickness of the white peony root slices is preferably 1.5-2.5 mm, more preferably 1.6-2.4 mm, and even more preferably 2 mm. This invention, through the above-described white peony root processing method, can obtain active ingredients that effectively prevent and treat pancreatic cancer.

[0021] In this invention, Atractylodes macrocephala slices are mixed with wheat bran, and the resulting Atractylodes macrocephala-wheat bran mixture is stir-fried in a hot pan, cooled, and then obtained stir-fried Atractylodes macrocephala. After grinding, it is sieved to obtain Atractylodes macrocephala powder. The preferred mass ratio of Atractylodes macrocephala slices to wheat bran is 8-11:1, more preferably 9-10:1, and even more preferably 10:1. The stir-frying is preferably carried out at 160-210℃ for 15-25 minutes, more preferably at 170-205℃ for 17-23 minutes, and even more preferably at 200℃ for 20 minutes. The preferred mesh size for sieving is 40-60 mesh, more preferably 45-55 mesh, and even more preferably 50 mesh. The cooling temperature is 20-25℃, more preferably 21-24℃, and even more preferably 22℃. The thickness of the Atractylodes macrocephala slices is preferably 1.5-2.5 mm, more preferably 1.6-2.4 mm, and even more preferably 2 mm. This invention, through the above-described Atractylodes macrocephala processing method, can obtain active ingredients that effectively prevent and treat pancreatic cancer.

[0022] In this invention, astragalus slices are mixed with rice wine and simmered, then stir-fried in a hot pan, cooled, and processed to obtain wine-processed astragalus. The astragalus is then ground and sieved to obtain astragalus powder. The preferred mass-to-volume ratio of astragalus slices to rice wine is 4-6:1 g / mL, more preferably 4.5-5.5:1 g / mL, and even more preferably 5:1 g / mL. The stir-frying is preferably carried out at 100-120℃ for 8-12 minutes, more preferably at 105-115℃ for 9-11 minutes, and even more preferably at 110℃ for 10 minutes. The sieve mesh size is 40-60 mesh, more preferably 45-55 mesh, and even more preferably 50 mesh. The cooling temperature is 20-25℃, more preferably 21-24℃, and even more preferably 22℃. The thickness of the astragalus slices is preferably 1.5-2.5 mm, more preferably 1.6-2.4 mm, and even more preferably 2 mm. The present invention can obtain active ingredients that are effective in preventing and treating pancreatic cancer through the above-mentioned processing method of wine-processed Astragalus membranaceus.

[0023] In this invention, Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius, and Prunus persica kernel are ground and sieved to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder, and Prunus persica kernel powder. The sieve mesh size is 50-70 mesh, more preferably 55-65 mesh, and more preferably 60 mesh.

[0024] In this invention, white peony root powder, anemarrhena rhizome powder, codonopsis root powder, atractylodes rhizome powder, astragalus root powder, tangerine peel powder, magnolia bark powder, chicken gizzard lining powder, ginger powder, chuanxiong rhizome powder, cinnamon twig powder, safflower powder, and peach kernel powder are weighed and mixed to obtain a mixed powder. The mixed powder is then soaked in an ethanol-water solution, refluxed for extraction, filtered, and the extract is collected, concentrated, and dried to obtain a traditional Chinese medicine extract. The mass-to-volume ratio of the mixed powder to the ethanol-water solution is preferably 1:8~12 g / mL, more preferably 1:9~11 g / mL, and even more preferably 1:10 g / mL; the volume percentage of the ethanol-water solution is preferably 30%~50%, more preferably 35%~45%, and even more preferably 40%. The volume percentage refers to the percentage of anhydrous ethanol volume to the total volume of the ethanol-water solution. The extraction is performed 1~3 times, such as 2 times, with each extraction lasting 0.5~2.5 hours, more preferably 1~2 hours, and even more preferably 1, 1.5, or 2 hours. This invention does not impose any particular limitation on the methods of concentration and drying; any method known in the art can be used, such as vacuum concentration, and air drying, oven drying, or freeze drying. By using the above methods to extract the mixed powder, this invention can obtain active ingredients that significantly prevent and treat pancreatic cancer.

[0025] This invention provides a traditional Chinese medicine extract prepared by the above-described preparation method.

[0026] This invention provides an application of the above-mentioned traditional Chinese medicine extract in the preparation of drugs for the prevention and treatment of pancreatic cancer.

[0027] In this invention, the traditional Chinese medicine extract can inhibit the growth and migration of pancreatic cancer cells. The traditional Chinese medicine extract preferably accounts for 20% to 90% of the mass of the drug. In this invention, the drug also includes pharmaceutically acceptable excipients, such as excipients, flavoring agents, antioxidants, or surfactants. This invention allows the traditional Chinese medicine extract to be used in combination with other drugs for treating pancreatic cancer for prevention and treatment, or the traditional Chinese medicine extract to be used as the sole active ingredient for prevention and treatment of pancreatic cancer. The pancreatic cancer cells include pancreatic cancer BxPC-3.

[0028] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0029] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0030] In the following embodiments, data analysis was performed using GraphPad software. Experimental data were expressed as mean ± standard deviation and normality was tested. One-way ANOVA was used for comparisons among multiple groups. P < 0.05 indicated that the difference was statistically significant, and P > 0.05 indicated that the difference was not statistically significant.

[0031] Example 1 A method for preparing a traditional Chinese medicine extract for treating pancreatic cancer, comprising the following steps: The herbal extract is made from the following raw materials: White peony root 9g, Anemarrhena rhizome 9g, Codonopsis root 6g, Atractylodes rhizome 6g, Astragalus root 6g, Tangerine peel 6g, Magnolia bark 6g, Chicken gizzard lining 6g, Fresh ginger 6g, Ligusticum chuanxiong rhizome 3g, Cinnamon twig 3g, Safflower 6g, and Peach kernel 6g.

[0032] (1) Cut the white peony into 2mm thick slices, stir-fry in a hot pan at 180℃ for 15 minutes, let it cool to room temperature (22℃), grind the stir-fried white peony, and pass it through a 50-mesh sieve to obtain white peony powder; (2) Cut Atractylodes macrocephala into 2mm thick slices. Mix the slices with wheat bran at a mass ratio of 10:1. Stir-fry the resulting Atractylodes macrocephala and wheat bran mixture in a hot pan at 200℃ for 20 minutes. Let it cool to room temperature (22℃). Grind the stir-fried Atractylodes macrocephala and pass it through a 50-mesh sieve to obtain Atractylodes macrocephala powder. (3) Cut Astragalus membranaceus into 2mm thick slices. Mix 100g of Astragalus membranaceus slices with 20mL of rice wine and let it soak for 60min. Then stir-fry it in a hot pan at 110℃ for 10min. Let it cool to room temperature (22℃). Grind the wine-processed Astragalus membranaceus and pass it through a 50-mesh sieve to obtain Astragalus membranaceus powder. (4) Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius and Prunus persica were ground separately and passed through a 60-mesh sieve to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder and Prunus persica powder. (5) Weigh out 9g of white peony powder, 9g of anemarrhena powder, 6g of codonopsis powder, 6g of atractylodes powder, 6g of astragalus powder, 6g of tangerine peel powder, 6g of magnolia bark powder, 6g of chicken gizzard powder, 6g of ginger powder, 3g of chuanxiong powder, 3g of cinnamon twig powder, 6g of safflower powder and 6g of peach kernel powder respectively, mix them to obtain a mixed powder, mix 78g of the mixed powder with 780mL of 40% ethanol aqueous solution and soak for 30min, then reflux extract the mixed solution for 1.5h, filter and collect the first extract and the first filter residue, mix the first filter residue with 40% ethanol aqueous solution at a mass-volume ratio of 1:10g / mL, reflux extract for 1h, filter and collect the second extract, mix the first extract and the second extract to obtain the total extract, concentrate under reduced pressure to 1 / 5 of the original volume, dry and obtain the Chinese herbal extract.

[0033] Example 2 A method for preparing a traditional Chinese medicine extract for treating pancreatic cancer, comprising the following steps: The herbal extract is made from the following raw materials: White peony root 5g, Anemarrhena rhizome 5g, Codonopsis root 4g, Atractylodes rhizome 4g, Astragalus root 4g, Tangerine peel 4g, Magnolia bark 4g, Chicken gizzard lining 4g, Fresh ginger 4g, Ligusticum chuanxiong rhizome 2g, Cinnamon twig 2g, Safflower 4g, and Peach kernel 4g.

[0034] (1) Cut the white peony into 1.5 mm thick slices, stir-fry in a hot pan at 150°C for 20 min, let it cool to room temperature (22°C), grind the stir-fried white peony, and pass it through a 40 mesh sieve to obtain white peony powder; (2) Cut Atractylodes macrocephala into 1.5 mm thick slices. Mix the slices with wheat bran at a mass ratio of 8:1. Stir-fry the resulting Atractylodes macrocephala and wheat bran mixture in a hot pan at 160°C for 25 min. Let it cool to room temperature (22°C). Grind the stir-fried Atractylodes macrocephala and pass it through a 40-mesh sieve to obtain Atractylodes macrocephala powder. (3) Cut Astragalus membranaceus into 1.5mm thick slices. Mix 80g of Astragalus membranaceus slices with 20mL of rice wine and let it soak for 50min. Then stir-fry it in a hot pan at 100℃ for 12min. Let it cool to room temperature (22℃). Grind the wine-processed Astragalus membranaceus and pass it through a 40-mesh sieve to obtain Astragalus membranaceus powder. (4) Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius and Prunus persica were ground separately and passed through a 70-mesh sieve to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder and Prunus persica powder. (5) Weigh out 5g of white peony powder, 5g of anemarrhena powder, 4g of codonopsis powder, 4g of atractylodes powder, 4g of astragalus powder, 4g of tangerine peel powder, 4g of magnolia bark powder, 4g of chicken gizzard powder, 4g of ginger powder, 2g of chuanxiong powder, 2g of cinnamon twig powder, 4g of safflower powder and 4g of peach kernel powder respectively, mix them to obtain a mixed powder, mix 50g of the mixed powder with 600mL of 50% ethanol aqueous solution and soak for 25min, then reflux extract the resulting mixed solution for 1h, filter and collect the first extract and the first filter residue, mix the first filter residue with 50% ethanol aqueous solution at a mass-volume ratio of 1:12g / mL, reflux extract for 50min, filter and collect the second extract, mix the first extract and the second extract to obtain the total extract, concentrate under reduced pressure to 1 / 5 of the original volume, dry and obtain the Chinese herbal extract.

[0035] Example 3 A method for preparing a traditional Chinese medicine extract for treating pancreatic cancer, comprising the following steps: The herbal extract is made from the following raw materials: White peony root 12g, Anemarrhena rhizome 12g, Codonopsis root 8g, Atractylodes rhizome 8g, Astragalus root 8g, Tangerine peel 8g, Magnolia bark 8g, Chicken gizzard lining 8g, Fresh ginger 8g, Ligusticum chuanxiong rhizome 4g, Cinnamon twig 4g, Safflower 8g, and Peach kernel 8g.

[0036] (1) Cut the white peony into 2.5 mm thick slices, stir-fry in a hot pan at 200°C for 10 min, let it cool to room temperature (22°C), grind the stir-fried white peony and pass it through a 60 mesh sieve to obtain white peony powder; (2) Cut Atractylodes macrocephala into 2.5 mm thick slices. Mix the slices with wheat bran at a mass ratio of 11:1. Stir-fry the resulting Atractylodes macrocephala and wheat bran mixture in a hot pan at 210°C for 15 min. Let it cool to room temperature (22°C). Grind the stir-fried Atractylodes macrocephala and pass it through a 60-mesh sieve to obtain Atractylodes macrocephala powder. (3) Cut Astragalus membranaceus into 2.5 mm thick slices. Mix 120 g of Astragalus membranaceus slices with 20 mL of rice wine and let it soak for 70 min. Then stir-fry it in a hot pan at 120 °C for 8 min. Let it cool to room temperature (22 °C). Grind the wine-processed Astragalus membranaceus and pass it through a 60 mesh sieve to obtain Astragalus membranaceus powder. (4) Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius and Prunus persica were ground separately and passed through a 50-mesh sieve to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder and Prunus persica powder. (5) Weigh out 12g of white peony powder, 12g of anemarrhena powder, 8g of codonopsis powder, 8g of atractylodes powder, 8g of astragalus powder, 8g of tangerine peel powder, 8g of magnolia bark powder, 8g of chicken gizzard powder, 8g of ginger powder, 4g of chuanxiong powder, 4g of cinnamon twig powder, 8g of safflower powder and 8g of peach kernel powder respectively, mix them to obtain a mixed powder, mix 104g of the mixed powder with 832mL of 30% ethanol aqueous solution and soak for 35min, then reflux extract the mixed solution for 2h, filter and collect the first extract and the first filter residue, mix the first filter residue with 30% ethanol aqueous solution at a mass-volume ratio of 1:8g / mL, reflux extract for 1.5h, filter and collect the second extract, mix the first extract and the second extract to obtain the total extract, concentrate under reduced pressure to 1 / 5 of the original volume, dry and obtain the Chinese herbal extract.

[0037] Experimental Example 1 The therapeutic effect of the traditional Chinese medicine extract prepared in Example 1 on pancreatic cancer (1) Effect of the traditional Chinese medicine extract prepared in Example 1 on the in vitro activity of pancreatic cancer cells BxPC-3 Pancreatic cancer cell viability assay: BxPC-3 pancreatic cancer cells were cultured in a cell incubator containing 10% fetal bovine serum and RPMI-1640 medium at 37°C and 5% CO2. A suspension of BxPC-3 cells in the logarithmic growth phase was collected and cultured at a concentration of 2 × 10⁻⁶ cells / mL. 4 BxPC-3 cells were seeded at a concentration of 100 μL / mL into 96-well plates and cultured overnight. The cells were then randomly divided into five groups: a blank control group, a low-dose traditional Chinese medicine extract group (low-dose group), a medium-dose traditional Chinese medicine extract group (medium-dose group), a high-dose traditional Chinese medicine extract group (high-dose group), and a positive control group (positive control group). Each group had six replicates. The blank control group was cultured for 72 h with the same volume of PBS as the drug-treated groups. The low-dose traditional Chinese medicine extract group was cultured with a traditional Chinese medicine extract solution (PBS prepared with PBS) at a final concentration of 10 μg / mL. The medium-dose group was cultured with a PBS solution (prepared with PBS) to a final concentration of 20 μg / mL for 72 h; the high-dose group was cultured with a PBS solution (prepared with PBS) to a final concentration of 40 μg / mL for 72 h; the positive control group was cultured with 5-fluorouracil to a final concentration of 40 μg / mL for 72 h, after which the supernatant was discarded, 20 μL of 5 mg / mL MTT solution was added to each well, and the cells were incubated for another 4 h. The supernatant was carefully removed by pipetting, 100 μL of DMSO was added to each well, and the cells were shaken for 15 min. The absorbance of each well was measured using a microplate reader (490 nm wavelength), and the tumor cell viability was calculated. Tumor cell viability (%) = OD 样品处理组 ÷OD 空白对照组×100%. Calculate the tumor inhibition rate, inhibition rate (%) = (OD) / (D). 空白对照组 -OD 样品处理组 )÷OD 空白对照组 ×100%, the results are shown in Table 1.

[0038] Table 1. Results of the effects of different doses of traditional Chinese medicine extracts on pancreatic cancer activity.

[0039] Note: Different letters indicate significant differences between groups, p≤0.0001.

[0040] Figure 1 The results in Table 1 show that, compared with the blank control group, the pancreatic cancer cell viability was significantly reduced in the low-dose group, medium-dose group, high-dose group, and positive control group in a dose-dependent manner. Furthermore, the pancreatic cancer cell viability in the high-dose group was significantly lower than that in the positive control group. Therefore, the herbal extract of this invention can effectively kill pancreatic cancer cells.

[0041] Scratch assay to detect cell migration ability: BxPC-3 pancreatic cancer cells in logarithmic growth phase were subjected to a scratch assay at a concentration of 5 × 10⁻⁶ cells / year. 5 After seeding BxPC-3 cells at a concentration of 10 μg / mL into 6-well plates and culturing overnight, the cells were randomly divided into five groups after scoring the center of the plate with a pipette tip: blank control group, low-dose traditional Chinese medicine extract group (denoted as low-dose), medium-dose traditional Chinese medicine extract group (denoted as medium-dose), high-dose traditional Chinese medicine extract group (denoted as high-dose), and positive control group (denoted as positive control), with 3 replicates per group. The blank control group was cultured for 48 h with the same volume of PBS as the drug-treated groups. The low-dose traditional Chinese medicine extract group was cultured with a traditional Chinese medicine extract solution (prepared with PBS) at a final concentration of 10 μg / mL. The cells were cultured for 48 hours. The medium-dose herbal extract group was cultured for 48 hours with a herbal extract solution (prepared with PBS) at a final concentration of 20 μg / mL. The high-dose herbal extract group was cultured for 48 hours with a herbal extract solution (prepared with PBS) at a final concentration of 40 μg / mL. The positive control group was cultured for 48 hours with 5-fluorouracil at a final concentration of 40 μg / mL. The cell scratch interval was measured at 0 h and 48 h. The relative scratch area ratio was calculated as: (Area of ​​cell scratch in each group after 48 h) ÷ (Area of ​​cell scratch in each group at 0 h) × 100%.

[0042] Figure 2 The results in Table 2 show that, compared with the blank control group, the relative scratch area ratios of the low-dose group, medium-dose group, high-dose group, and positive drug group were significantly increased, and the relative scratch area ratio of the high-dose group was significantly higher than that of the positive drug group. Therefore, the traditional Chinese medicine extract prepared in this invention can effectively inhibit the migration ability of pancreatic cancer.

[0043] Table 2. Effects of different groups on the migration ability of pancreatic cancer cells.

[0044] Note: Different letters indicate significant differences between groups, p < 0.01.

[0045] (1) Effect of the traditional Chinese medicine extract prepared in Example 1 on the killing of pancreatic cancer in vivo 1. Materials 1.1 Animals BALB / c nude mice, 4-6 weeks old, each weighing 18-20g, were kept at a temperature of 24±1℃ and a humidity of 50±10%, with regular alternation of light and dark, and free access to food and water. They were acclimatized for 1 week.

[0046] 1.2 Construction of a mouse model of pancreatic cancer 0.2 mL of logarithmic growth phase pancreatic cancer cells BxPC-3 were administered at a dose of 5 × 10⁻⁶. 6 Cell suspension at a density of [number] cells / mL was subcutaneously injected into each BALB / c nude mouse. After 9 days, round masses appeared at the injection site, indicating that the pancreatic cancer mouse model was successfully established. The pancreatic cancer model mice were randomly divided into five groups of 10 mice each: model group, low-dose traditional Chinese medicine extract group (low-dose group), medium-dose traditional Chinese medicine extract group (medium-dose group), high-dose traditional Chinese medicine extract group (high-dose group), and positive drug group (positive drug). The pancreatic cancer model mice in the model group received no drug treatment and were administered physiological saline by gavage once daily for 30 consecutive days. The low-dose traditional Chinese medicine extract group received 5 g / kg of the traditional Chinese medicine extract prepared in Example 1 by gavage once daily for 30 consecutive days. The medium-dose group received 10 g / kg of the traditional Chinese medicine extract prepared in Example 1 by gavage once daily for 30 consecutive days. The high-dose group received 20 g / kg of the traditional Chinese medicine extract prepared in Example 1 by gavage once daily for 30 consecutive days. The positive control group received 0.03 g / kg of 5-fluorouracil by gavage once daily for 30 consecutive days. After the experiment, the body weight of each group of mice was measured, and then the mice were sacrificed, and subcutaneous tumors were dissected. The tumor inhibition rate was calculated as follows: Tumor inhibition rate (%) = (Tumor mass of model group - Tumor mass of experimental group) ÷ Average tumor mass of model group × 100%.

[0047] Table 3. Results of tumor inhibition rates after treatment in different groups.

[0048] Note: Different letters in Table 3 indicate significant differences between groups, p < 0.01.

[0049] Figure 3 The results in Table 3 indicate that, compared with the model group, the low-dose group, medium-dose group, high-dose group, and positive drug group can significantly inhibit the growth of pancreatic cancer tumors, and the tumor inhibition rate of the high-dose group is higher than that of the positive drug group.

[0050] Table 4. Results of mouse body weight after different treatment groups

[0051] The results in Table 4 show that there was no significant difference in body weight among the groups of mice, indicating that the use of the herbal extract of the present invention had no effect on the body weight of pancreatic cancer model mice.

[0052] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made 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 method for preparing a traditional Chinese medicine extract for treating pancreatic cancer, characterized in that, Includes the following steps: The herbal extract comprises the following raw materials in parts by weight: White peony root 5-12 parts, Anemarrhena rhizome 5-12 parts, Codonopsis root 4-8 parts, Atractylodes rhizome 4-8 parts, Astragalus root 4-8 parts, Tangerine peel 4-8 parts, Magnolia bark 4-8 ​​parts, Chicken gizzard lining 4-8 parts, Fresh ginger 4-8 parts, Ligusticum chuanxiong rhizome 2-4 parts, Cinnamon twig 2-4 parts, Safflower 4-8 parts, and Peach kernel 4-8 parts; (1) Stir-fry the white peony slices in a hot pan, cool them down to obtain stir-fried white peony, grind them, and sieve them to obtain white peony powder; (2) Mix Atractylodes macrocephala slices with wheat bran, stir-fry the resulting Atractylodes macrocephala and wheat bran mixture in a hot pan, cool it down to obtain stir-fried Atractylodes macrocephala, grind it, and sieve it to obtain Atractylodes macrocephala powder; (3) Mix the astragalus slices with rice wine and let them soak. Then stir-fry them in a hot pan and cool them down to obtain wine-roasted astragalus. Grind the astragalus and sieve it to obtain astragalus powder. (4) Anemarrhena asphodeloides, Codonopsis pilosula, Citrus reticulata peel, Magnolia officinalis, chicken gizzard lining, ginger, Ligusticum chuanxiong, Cinnamomum cassia, Carthamus tinctorius and Prunus persica were ground and sieved to obtain Anemarrhena asphodeloides powder, Codonopsis pilosula powder, Citrus reticulata peel powder, Magnolia officinalis powder, chicken gizzard lining powder, ginger powder, Ligusticum chuanxiong powder, Cinnamomum cassia powder, Carthamus tinctorius powder and Prunus persica powder. (5) Weigh out the following powders: white peony powder, anemarrhena powder, codonopsis powder, atractylodes powder, astragalus powder, tangerine peel powder, magnolia bark powder, chicken gizzard powder, ginger powder, chuanxiong powder, cinnamon twig powder, safflower powder, and peach kernel powder respectively, mix them together to obtain a mixed powder, mix the mixed powder with an ethanol aqueous solution, reflux to extract, filter, collect the extract, concentrate and dry to obtain the Chinese herbal extract.

2. The preparation method according to claim 1, characterized in that, In step (1), the stir-frying is carried out at 150~200℃ for 10~20 minutes; the sieve mesh size is 40~60 mesh; and the temperature is lowered to 20~25℃.

3. The preparation method according to claim 1, characterized in that, In step (2), the mass ratio of Atractylodes macrocephala slices to wheat bran is 8~11:1; the stir-frying is carried out at 160~210℃ for 15~25 minutes; the sieve mesh size is 40~60 mesh; and the temperature is lowered to 20~25℃.

4. The preparation method according to claim 1, characterized in that, In step (3), the mass-to-volume ratio of Astragalus membranaceus slices to rice wine is 4~6:1g / mL; the stir-frying is carried out at 100~120℃ for 8~12 minutes; the sieve mesh size is 40~60 mesh; and the temperature is lowered to 20~25℃.

5. The preparation method according to claim 1, characterized in that, In step (5), the mass-to-volume ratio of the mixed powder to the ethanol aqueous solution is 1:8~12 g / mL; the volume percentage of the ethanol aqueous solution is 30%~50%.

6. The preparation method according to claim 1 or 5, characterized in that, The extraction is performed 1 to 3 times, with each extraction lasting 0.5 to 2.5 hours.

7. The traditional Chinese medicine extract prepared by the preparation method according to any one of claims 1 to 6.

8. The application of the traditional Chinese medicine extract according to claim 7 in the preparation of drugs for the prevention and treatment of pancreatic cancer.

9. The application according to claim 8, characterized in that, The herbal extract described above can inhibit the growth and migration of pancreatic cancer cells.

10. The application according to claim 8, characterized in that, The herbal extract accounts for 20% to 90% of the mass of the drug.

Images