Pharmaceutical composition for treating pancreatic cancer as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for treating pancreatic cancer as well as a preparation method and application thereof. The pharmaceutical composition is prepared from the following raw materials in percentage by weight: atractylol, eugenol, safflower red and pachymic acid, and can be prepared into various clinical common oral preparations. The pharmaceutical composition disclosed by the invention has the effects of reducing tumor cell proliferation, reducing toxicity and enhancing efficacy, and also has the effects of enhancing the immunity of a patient, prolonging the lifetime of the patient and improving the life quality of the patient, so that the pharmaceutical composition disclosed by the invention has a relatively good development and utilization prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a pharmaceutical composition for treating pancreatic cancer, a preparation method thereof, a pharmaceutical preparation and use. BACKGROUND

[0002] Pancreatic cancer is a common malignant tumor of the digestive tract in clinic, and its morbidity and mortality show a rising trend year by year, which seriously endangers the health of patients. Clinically, surgery, radiotherapy, chemotherapy and biological targeted drugs are mainly used for treatment. In the early stage of the disease, surgical treatment is the preferred and most effective method, and the 5-year survival rate of early pancreatic cancer patients after surgery can even reach 58%. However, due to insidious onset, more than half of the pancreatic cancer patients have lost the opportunity for surgery when they are found. For locally unresectable / metastatic pancreatic cancer, chemotherapy is the most important treatment, but once drug resistance occurs, the clinical efficacy is relatively limited. With the emergence of targeted drugs and immunotherapy, the treatment of many tumors has made great progress, but the treatment of pancreatic cancer has been slow. In summary, the overall treatment effect of pancreatic cancer is not ideal, and finding new treatment strategies to improve the prognosis of pancreatic cancer has always been an important issue in pancreatic cancer research. At present, how to more reasonably and effectively use existing treatment methods to improve efficacy and how to better balance efficacy and patient quality of life have become the focus of current research.

[0003] In recent years, the efficacy of traditional Chinese medicine in the treatment of various types of malignant tumors has been continuously verified, and its position in the multidisciplinary comprehensive treatment model of malignant tumors is also becoming more and more important. Various studies on the treatment of malignant tumors with traditional Chinese medicine are also being widely and deeply carried out, and the mechanisms by which traditional Chinese medicine exerts its effects are gradually being revealed. Traditional Chinese medicine believes that the pathogenesis of pancreatic cancer is related to dampness-heat toxin, phlegm and blood stasis, spleen and stomach qi deficiency, liver qi stagnation, and weakness of viscera, and the treatment principle is to regulate qi and induce defecation, clear heat and eliminate stagnation, promote blood circulation and resolve phlegm and stasis, and at the same time, tonify the spleen and invigorate the center, and eliminate evil. However, traditional Chinese medicine treatment of pancreatic cancer is generally less effective in directly killing cancer cells, and its main effect is in regulating yin and yang balance, enhancing the body's immunity, and improving the quality of life. In addition, traditional Chinese medicine decoction has a heavy taste and a large amount, which may stimulate the gastrointestinal tract and is inconvenient to carry, resulting in poor patient medication compliance, making it difficult for patients to adhere to long-term use. At the same time, due to the large number of herbs in traditional Chinese medicine prescriptions, the cost and safety risks are high, which is not conducive to effective quality control and large-scale production.

[0004] Therefore, it is imperative to develop a pharmaceutical composition for treating pancreatic cancer with good efficacy, small side effects and convenient administration by using modern pharmacological research methods to conduct in-depth research and screening on the active ingredient monomers of natural products in classical Chinese medicine prescriptions. SUMMARY

[0005] The present application aims to overcome the deficiencies in the prior art, and provides a safe, effective, convenient and economical pharmaceutical composition, and provides a new idea for the treatment of pancreatic cancer in clinical practice and the modernization of traditional Chinese medicine.

[0006] Specifically, the present application is realized through the following technical solutions:

[0007] In a first aspect, the present application provides a pharmaceutical composition for treating pancreatic cancer, which comprises atractylol, eugenol, crocin and pachymic acid.

[0008] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition comprises the following weight percentages of active ingredients: atractylol 20%-50%, eugenol 20%-50%, crocin 10%-25% and pachymic acid 10%-25%.

[0009] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition comprises the following weight percentages of active ingredients: atractylol 30%-40%, eugenol 30%-40%, crocin 10%-20% and pachymic acid 10%-20%.

[0010] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition comprises the following weight percentages of active ingredients: atractylol 35%, eugenol 35%, crocin 15% and pachymic acid 15%.

[0011] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition is made of the following weight percentages of active ingredients: atractylol 20%-50%, eugenol 20%-50%, crocin 10%-25% and pachymic acid 10%-25%.

[0012] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition is made of the following weight percentages of active ingredients: atractylol 30%-40%, eugenol 30%-40%, crocin 10%-20% and pachymic acid 10%-20%.

[0013] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition is made of the following weight percentages of active ingredients: atractylol 35%, eugenol 35%, crocin 15% and pachymic acid 15%.

[0014] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition has the effects of reducing tumor cell proliferation and reducing toxicity and enhancing efficacy.

[0015] The active ingredients atractylodin, eugenol, safflor yellow and pachymic acid used in the pharmaceutical composition of the present application can be extracted and separated from traditional Chinese medicinal materials containing the above active ingredients, such as plants of Atractylodes lancea, Lobelia chinensis, Bupleurum chinense and Pachyman, or purchased from commercial products.

[0016] In the second aspect, the present application provides a preparation method of the pharmaceutical composition of the first aspect, mixing the active ingredients of the first aspect.

[0017] In the third aspect, the present application provides a pharmaceutical preparation for treating pancreatic cancer, which comprises the pharmaceutical composition of the first aspect and a pharmaceutically acceptable carrier.

[0018] As an optional mode, in the above pharmaceutical preparation, the pharmaceutically acceptable carrier refers to a pharmaceutical carrier commonly used in the field of pharmaceutical preparation, which is selected from one or more of fillers, binders, disintegrants, lubricants, suspending agents, wetting agents, pigments, flavoring agents, solvents and surfactants.

[0019] As an optional mode, in the above pharmaceutical preparation, the pharmaceutical preparation is an oral preparation.

[0020] As an optional mode, in the above pharmaceutical preparation, the oral preparation is a tablet, a capsule, a granule or an oral liquid.

[0021] In the fourth aspect, the present application provides use of the pharmaceutical composition of the first aspect or the pharmaceutical preparation of the third aspect in the preparation of a medicament for treating pancreatic cancer.

[0022] Compared with the prior art, the present application has the following beneficial effects:

[0023] (1) The present application is based on the classical prescription for treating pancreatic cancer, and four natural product monomers are selected through a large number of pharmacological experiments, which are combined in a specific proportion to form a pharmaceutical composition for treating pancreatic cancer, thereby providing a new idea for modernization of traditional Chinese medicine.

[0024] (2) The pharmaceutical composition of the present application has the effects of reducing tumor cell proliferation, reducing toxicity and increasing efficacy, and also has the effects of enhancing the immunity of patients, prolonging the survival period of patients and improving the quality of life of patients.

[0025] (3) The natural product monomers used in the pharmaceutical composition of the present application are widely sourced and can be produced by mature methods, so that the pharmaceutical composition of the present application is convenient for industrialized mass production. In addition, the pharmaceutical composition of the present application is made into a conventional oral preparation by adding a pharmaceutically acceptable carrier, so that the drug treatment is stable, the efficacy is improved, and the drug is convenient to take. DETAILED DESCRIPTION

[0026] The pharmaceutical composition of the present application is screened on the basis of a classical formula for treating pancreatic cancer, which is a basic formula with Bupleurum, Radix Anemarrhena, Radix Curcumae, Fructus Aurantii, Radix Bistortae, Radix Paeoniae Alba, Rhizoma Atractylodis Macrocephalae, Poria, Radix Bupleuri, Herba Hedyotis Diffusae, Herba Lycopi, and Fructus Crataegi, etc. as the basic formula, and can be added or subtracted according to the symptoms. The classical formula has the effects of soothing liver and regulating qi, invigorating spleen and draining dampness, detoxifying and resisting cancer, and dissipating blood stasis and relieving pain.

[0027] In the in-depth research on the pharmacological mechanism of the classical formula for treating pancreatic cancer, the present inventors have found for the first time that the combination of natural product monomers atractylodes alcohol, syringic acid, safflower red and pachymic acid can significantly enhance the effect of treating pancreatic cancer through a large number of screening. On this basis, the present application is completed.

[0028] As used herein, the natural product monomers atractylodes alcohol, syringic acid, safflower red and pachymic acid in the pharmaceutical composition of the present application can be administered in the same pharmaceutical preparation, or in different pharmaceutical preparations. In the case of administration in different pharmaceutical preparations, the dosage forms of the natural product monomers atractylodes alcohol, syringic acid, safflower red and pachymic acid can be the same or different. Moreover, the natural product monomers atractylodes alcohol, syringic acid, safflower red and pachymic acid can be administered simultaneously or sequentially.

[0029] As used herein, "pharmaceutically acceptable carrier" refers to a pharmaceutical carrier conventional in the field of pharmaceutical preparations, which is selected from one or more of fillers, binders, disintegrants, lubricants, suspending agents, wetting agents, pigments, flavoring agents, solvents, and surfactants.

[0030] The fillers according to the present application include, but are not limited to, starch, microcrystalline cellulose, sucrose, dextrin, lactose, sugar powder, glucose, etc.; the lubricants include, but are not limited to, magnesium stearate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, poloxamer, etc.; the binders include, but are not limited to, water, ethanol, starch paste, sugar syrup, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, sodium alginate, polyvinylpyrrolidone, etc.; the disintegrants include, but are not limited to, starch effervescent mixture, i.e. sodium bicarbonate and citric acid, tartaric acid, low-substituted hydroxypropyl cellulose, etc.; the suspending agents include, but are not limited to, polysaccharides such as acacia, agar, alginic acid, cellulose ether and carboxymethyl chitin, etc.; the solvents include, but are not limited to, water, balanced salt solution, etc.

[0031] The pharmaceutical composition can be prepared into various solid oral preparations, liquid oral preparations, etc. The pharmaceutically acceptable oral solid preparations include ordinary tablets, dispersible tablets, enteric-coated tablets, granules, capsules, dripping pills, powders, etc., and the oral liquid preparations include oral liquids, emulsions, etc.

[0032] The above various dosage forms can be prepared according to the conventional processes in the field of pharmaceutical preparations.

[0033] In the above-mentioned medical uses, the time, number and frequency of administration of the composition of the present application are determined according to the specific diagnosis of the disease, which is within the skill of the person skilled in the art.

[0034] The treatment regimen for mice can be applied to humans, and the effective dose of a drug for humans can be calculated from the effective dose of the drug for mice, which is also easy for the person skilled in the art to achieve.

[0035] The present application will be further described below with reference to specific examples. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the scope of the present application.

[0036] If a specific technique or condition is not specified in the examples, it is performed according to the technique or condition described in the literature in the art or according to the product manual. If the manufacturer of the reagent or instrument is not specified, it is a conventional product that can be purchased through a regular channel.

[0037] The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples are commercially available products unless otherwise specified.

[0038] Preparation of the oral preparation of the pharmaceutical composition of the present application

[0039] 1. The following is an exemplary prescription:

[0040] Prescription 1: atractylodin 30 g, eugenol 40 g, safflower red 10 g, pachymic acid 20 g, total 100 g.

[0041] Prescription 2: atractylodin 35 g, eugenol 35 g, safflower red 15 g, pachymic acid 15 g, total 100 g.

[0042] Prescription 3: atractylodin 40 g, eugenol 30 g, safflower red 20 g, pachymic acid 10 g, total 100 g.

[0043] 2. The following is an exemplary dosage form:

[0044] Tablets: Take 100 g of each of prescriptions 1-3, 50 g of lactose, 15 g of compressible starch, 10 g of hydroxypropyl cellulose, 5 g of magnesium stearate, and an appropriate amount of 70% ethanol to make 1000 tablets.

[0045] Capsules: Take 100 g of each of prescriptions 1-3, 80 g of starch, 25 g of sodium carboxymethyl starch, 20 g of dextrin, and an appropriate amount of 70% ethanol to make 1000 capsules.

[0046] Effect Example:

[0047] Example 1: Inhibition of pancreatic cancer cell proliferation by the pharmaceutical composition of the present application1. Experimental materials

[0048] Human pancreatic cancer cells PANC1 cells were purchased from Shanghai Kanglang Biotechnology Co., Ltd.

[0049] Test drug: prescription 2 prepared in the preparation example part.

[0050] Control drug 1: atractylone 35 g, eugenol 35 g, safflower red 15 g, and pachymic acid 15 g.

[0051] Control drug 2: atractylol 35 g and pachymic acid 15 g.

[0052] Control drug 3: eugenol 35 g and safflower red 15 g.

[0053] The experimental drug was prepared by dissolving dimethyl sulfoxide (DMSO). The concentration of the test drug and control drug 1 was 100 μM, and the concentration of control drug 2 and control drug 3 was 50 μM.

[0054] The cells in the blank control group were given an equal volume of DMSO solution.

[0055] 2. Experimental method

[0056] Logarithmic growth phase human pancreatic cancer cells PANC1 cells were inoculated in a 96-well plate at 5000 cells per well with a volume of 200 μL / well. After 24 h of culture, the old culture medium was removed and fresh serum-free culture medium was added for starvation treatment for 6 h. After the starvation treatment was completed, 20 μL of experimental drug or DMSO was added, and the final product was placed in an incubator for 48 h. Four hours before the enzyme marker detection, 20 μL of MTT was added per well in the dark, and the product was placed in an incubator for 4 h. The culture medium was discarded, 150 μL of DMSO was added to each well in the dark, and the product was shaken on a shaker for 10 min to fully dissolve the crystals. The absorbance was measured at a wavelength of 490 nm, and the cell survival rate was calculated. Each group had 5 replicate wells, and the experiment was repeated three times.

[0057] The results were analyzed using SPSS 27.0 statistical software, and the data were expressed as mean ± standard deviation (x ± s). Single factor analysis of variance was used for comparison among multiple groups, t test was used for comparison between two groups, p < 0.05 indicated that the difference was statistically significant, and p < 0.01 indicated that the difference was significantly statistically significant.

[0058] Cell proliferation inhibition rate (%) = (1-OD 实验组 / OD 对照组 ) x 100%

[0059] 3. Experimental results

[0060] The cell proliferation inhibition results are shown in Table 1.

[0061] Table 1: Inhibitory effect of the pharmaceutical composition of the present invention on the proliferation of PANC1 cells (%) (x±s)

[0062] Group Inhibition of proliferation (%) Blank control -- Test drug (Formulation 2) 89.16±11.57 Control drug 1 58.72 ± 9.18 ** ]] Control drug 2 32.53 ± 6.25 *** ]] Control drug 3 25.08 ± 7.66 *** ]]

[0063] Note: Comparison of drugs under test ** p<0.01, *** p<0.001.

[0064] The experimental results in Table 1 show that the pharmaceutical composition of the present invention (test drug, formulation 2) can significantly inhibit the proliferation of PANC1 cells, with an inhibition rate of up to 89.16%. However, when some active ingredients in formulation 2 are replaced or deleted, its inhibitory effect on PANC1 cell proliferation significantly decreases. ** p<0.01, *** p<0.001 indicates that the inhibitory effect of the pharmaceutical composition of the present invention on the proliferation of PANC1 cells is closely related to the types of active ingredients in the formulation.

[0065] Furthermore, the experimental results comparing the test drug group, control drug group 2, and control drug group 3 also show that each component in the pharmaceutical composition of the present invention has a significant synergistic effect in inhibiting the proliferation of PANC1 cells.

[0066] Example 2: Pharmacodynamic study of the pharmaceutical composition of the present application for the treatment of pancreatic cancer

[0067] 1. Main experimental animals and materials

[0068] SPF-grade male BALB / c nude mice, 6-8 weeks old, weighing 20-22g, were purchased from Liaoning Changsheng Biotechnology Co., Ltd.

[0069] The human pancreatic cancer cell PANC1 cells were purchased from Shanghai Kanglang Biotechnology Co., Ltd.

[0070] 2. Experimental Methods

[0071] (1) Establishment of mouse xenografts

[0072] PANC1 cells from human pancreatic cancer cells in the logarithmic growth phase were prepared into a concentration of 2×10⁻⁶. 7 A suspension of tumor cells per mL was implanted subcutaneously into the axillary region of BALB / c nude mice (approximately 0.20 mL per mouse). After inoculation, the mice's mental state, diet, and defecation were observed regularly.

[0073] (2) Experimental grouping and drug administration

[0074] The major diameter (a) and minor diameter (b) of the tumor nodule were measured using vernier calipers, and the result was calculated using the formula V = a × b. 2×0.52 Calculate tumor volume, when subcutaneous transplantation tumor grows to 10mm 3 After that, the nude mice with consistent tumor size were randomly divided into 5 groups, 9 in each group, and the specific grouping is as follows:

[0075] Model group: with equal volume of normal saline, the administration method is once a day.

[0076] Positive control group: 5-fluorouracil (5-Fu), with 2mg / kg / day with equal volume of normal saline, once a day.

[0077] Test drug: prescription 2 prepared in the preparation example part.

[0078] Control drug 1: atractylone 35g, eugenol 35g, safflower red 15g, pachymic acid 15g.

[0079] Control drug 2: atractylol 35g, pachymic acid 15g.

[0080] Control drug 3: eugenol 35g, safflower red 15g.

[0081] Test drug group and control drug 1 group: with 5mg / kg / day with equal volume of normal saline, once a day. Control drug 2 group and control drug 3 group: with 2.5mg / kg / day with equal volume of normal saline, once a day.

[0082] After 21 days of continuous administration, the tumor mass was taken out 24 hours after the last administration, and the tumor weight was determined.

[0083] (3) Experimental data processing

[0084] The results were analyzed by SPSS 27.0 statistical software, and the data were expressed as mean ± standard deviation (x ± s). Single factor analysis of variance was used for comparison among multiple groups, t test was used for comparison between two groups, p<0.05 indicated that the difference was statistically significant, and p<0.01 indicated that the difference was significantly statistically significant.

[0085] Tumor inhibition rate (%) = (model group average tumor weight - drug group average tumor weight) / model group average weight weight × 100%

[0086] 3. Experimental results

[0087] The experimental results are shown in Table 2 below.

[0088] Table 2: Effect of the drug composition of the application on PANC1 mouse transplanted tumor (x ± s)

[0089] Group Tumor weight (g) Tumor inhibition rate (%) Model 2.24±0.91 --- Positive control 0.58 ± 0.13 *** ]] 74.11 Test drug (Formulation 2) 0.81 ± 0.24 *** ]] 63.84 Control drug 1 1.32 ± 0.67 * ]] 41.07 Control drug 2 1.95±0.59 12.95 Control drug 3 1.83±0.71 18.30

[0090] Note: compared with the model group, * p<0.05,*** p<0.001.

[0091] As can be seen from the experimental results in Table 2, the drug composition of the present invention (test drug, prescription 2) has an inhibitory effect on mouse pancreatic xenograft tumors comparable to that of the positive control drug 5-Fu, and its effect is significantly different from that of the model group. *** (p<0.001). However, when some active ingredients in Formula 2 were replaced or deleted, the effect of the pharmaceutical composition on inhibiting pancreatic tumors in mice was significantly weakened, indicating that the efficacy of the pharmaceutical composition of the present invention is closely related to the types of each medicinal ingredient in the formula.

[0092] Furthermore, the experimental results comparing the test drug group, control drug group 2, and control drug group 3 also show that the components in the pharmaceutical composition of the present invention have a significant synergistic effect in inhibiting mouse pancreatic xenograft tumors.

[0093] Furthermore, in the aforementioned animal experiments, no obvious signs of fear, anxiety, or refusal to eat were observed in the nude mice group treated with the pharmaceutical composition of the present invention. No significant adverse reactions were observed with the pharmaceutical composition of the present invention, and the experimental animals exhibited good tolerance to it.

[0094] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A pharmaceutical composition for treating pancreatic cancer, characterized by: The pharmaceutical composition comprises atractylol, eugenol, crocin and pachymic acid.

2. The pharmaceutical composition of claim 1, wherein: The pharmaceutical composition comprises the following weight percentages of active ingredients: atractylol 20-50%, eugenol 20-50%, crocin 10-25%, and pachymic acid 10-25%.

3. The pharmaceutical composition of claim 2, wherein: The pharmaceutical composition comprises the following weight percentages of active ingredients: atractylol 30-40%, eugenol 30-40%, crocin 10-20%, and pachymic acid 10-20%.

4. The pharmaceutical composition of claim 3, wherein: The pharmaceutical composition comprises the following weight percentages of active ingredients: atractylol 35%, eugenol 35%, crocin 15%, and pachymic acid 15%.

5. The pharmaceutical composition of claim 1, wherein: The pharmaceutical composition has the effects of reducing tumor cell proliferation and reducing toxicity and increasing efficacy.

6. A process for the preparation of a pharmaceutical composition as claimed in any one of claims 1 to 5, characterized in that: The various active ingredients of any one of claims 1 to 5 are mixed.

7. A pharmaceutical preparation for the treatment of pancreatic cancer, characterized by: The pharmaceutical preparation comprises the pharmaceutical composition of any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

8. The pharmaceutical preparation according to claim 7, characterized in that: The pharmaceutical preparation is an oral preparation.

9. The pharmaceutical preparation according to claim 8, characterized in that: The oral preparation is a tablet, a capsule, a granule, or an oral solution.

10. Use of the pharmaceutical composition of any one of claims 1 to 5 or the pharmaceutical preparation of any one of claims 7 to 9 in the manufacture of a medicament for treating pancreatic cancer.