A pharmaceutical composition, a method for preparing the same and use thereof in the preparation of a medicament for preventing and / or treating cachexia

Abstract

The application belongs to the technical field of medicine, and relates to a kind of pharmaceutical composition and its preparation method and application in preparation of medicine for preventing and / or treating cachexia.The pharmaceutical composition includes the following components: ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract and pharmaceutically acceptable carrier.The components of the pharmaceutical composition can effectively improve the symptoms of cachexia.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a pharmaceutical composition, its preparation method, and its application in the preparation of drugs for the prevention and / or treatment of cachexia. Background Technology

[0002] Cachexia is a multifactorial syndrome characterized by persistent loss of skeletal muscle mass (with or without fat loss) that cannot be reversed by routine nutritional support. Clinically, the consensus definition of cachexia is a weight loss ≥5% or a body mass index (BMI) <20 kg / m². 2 Furthermore, traditional nutrition cannot completely reverse this effect.

[0003] Cancer cachexia is generally divided into three stages: pre-cachexia, cachexia, and treatment-resistant cachexia. Currently, there is no effective treatment for cancer cachexia. Common treatments include medication, nutritional support, and physical exercise, primarily aimed at stimulating appetite and increasing patient weight. Systemic drug therapy can usually reduce the tumor burden, but multiple studies have shown that chemotherapy can cause muscle atrophy in cancer patients, leading to a more severe cachexia state. Currently, there is a lack of safer drugs specifically for cachexia. Summary of the Invention

[0004] The purpose of this invention is to provide a pharmaceutical composition, a method for preparing the same, and its use in the preparation of remedies for the prevention and / or treatment of cachexia. The components of the pharmaceutical composition described in this invention can effectively improve the symptoms of cachexia.

[0005] The present invention provides a pharmaceutical composition comprising the following components: ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginsenoside ethanol-water extract, and a pharmaceutically acceptable carrier.

[0006] Preferably, in the pharmaceutical composition, the mass percentage of ginsenoside Rk1 is 5-12%, the mass percentage of ginsenoside Rg5 is 5-12%, the mass percentage of ginsenoside Rg3 is 5%-12%, the mass percentage of ginsenoside Rh4 is 3-8%, the mass percentage of ginsenoside Rk3 is 2-6%, and the mass percentage of ginsenoside ethanol-water extract is 50-80%.

[0007] Preferably, the ginseng ethanol-water extract comprises the following components in weight percentage: 5-11.3% ginsenosides, 20-30% ginseng polysaccharides, 22-32.7% ginseng polypeptides, 1.0-2.0% ginseng polyphenols, and 2.0-4.0% ginseng flavonoids;

[0008] The ginsenoside components include Rg1, Re, Rf, Rb1, Rc, Rd, and Rb2; Rg1 has a mass percentage of 2-3% in the ginseng ethanol-water extract, Re has a mass percentage of 1.0-2.5% in the ginseng ethanol-water extract, Rf has a mass percentage of 0.2-1.5% in the ginseng ethanol-water extract, Rb1 has a mass percentage of 0.5-1.5% in the ginseng ethanol-water extract, Rc has a mass percentage of 0.5-1.2% in the ginseng ethanol-water extract, Rd has a mass percentage of 0.5-0.8% in the ginseng ethanol-water extract, and Rb2 has a mass percentage of 0.3-0.8% in the ginseng ethanol-water extract.

[0009] Preferably, the ginseng ethanol-water extract is extracted from white ginseng, fresh ginseng, or red ginseng.

[0010] Preferably, the sources of ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4 and ginsenoside Rk3 include natural extraction.

[0011] Preferably, the ginsenosides Rk1, Rg5, Rh4, Rg3, and Rk3 are extracted from wild ginseng, cultivated ginseng, American ginseng, Panax notoginseng, red ginseng, or white ginseng.

[0012] Preferably, the extraction method of the ginseng ethanol-water extract includes the following steps: drying and pulverizing ginseng to obtain pulverized ginseng; mixing the pulverized ginseng with an ethanol aqueous solution with a volume percentage of 75%, heating at 55°C for 3 hours, and filtering to obtain a first filtrate and a first filter residue; mixing the first filter residue with an ethanol aqueous solution with a volume percentage of 45%, heating at 55°C for 3 hours, and filtering to obtain a second filtrate and a second filter residue; mixing the second filter residue with water, heating at 80°C for 3 hours, and filtering to obtain a third filtrate; combining the first filtrate, the second filtrate, and the third filtrate, evaporating the solvent, and freeze-drying to obtain the ginseng ethanol-water extract.

[0013] The present invention also provides a method for preparing the pharmaceutical composition described in the above technical solution, comprising the following steps:

[0014] Ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract, and a pharmaceutically acceptable carrier are mixed to obtain a pharmaceutical composition.

[0015] The present invention also provides the use of the pharmaceutical composition described above in the preparation of a medicament for the prevention and / or treatment of cachexia.

[0016] Preferably, the cachexia includes cachexia occurring in patients with advanced cancer.

[0017] This invention provides a pharmaceutical composition. The components of this pharmaceutical composition can effectively improve the symptoms of cachexia, effectively controlling the rapid weight loss in patients with advanced cancer, and can be used as a pharmaceutical preparation for the treatment and / or prevention of cachexia. The composition of this invention is more stable and easier to store, showing significant application prospects. Experimental results show that the pharmaceutical composition of this invention can significantly improve weight loss, effectively alleviate muscle atrophy caused by cachexia, effectively improve the reduction in gastrocnemius muscle weight, effectively improve the gap between muscle fibers and the cross-sectional area of ​​muscle bundles, effectively alleviate fat consumption, and effectively control and treat the development of cachexia caused by cancer. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is a graph showing the body weight-time curves of mice in each group during the pharmacodynamic test of Example 3 of the present invention;

[0020] Figure 2 This is a comparison chart of the weight of the gastrocnemius muscle in each group of mice during the pharmacodynamic test of Example 3 of the present invention;

[0021] Figure 3 HE staining images of the gastrocnemius muscle of mice in each group during the pharmacodynamic test of Example 3 of this invention;

[0022] Figure 4 These are photographs showing the comparison of adipose tissue in different groups of mice during the pharmacodynamic test in Example 3 of this invention.

[0023] Figure 5 This is a graph showing the protein expression of Fbxo32, Trim63, and MyoD in the gastrocnemius muscle of mice in each group during the pharmacodynamic test of Example 3 of the present invention. Detailed Implementation

[0024] This invention provides a pharmaceutical composition comprising the following components: ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginsenoside ethanol-water extract, and a pharmaceutically acceptable carrier. Cachexia is a multifactorial syndrome characterized by a persistent loss of skeletal muscle mass (with or without fat loss). This invention utilizes a combination of rare ginsenoside groups (a combination of ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, and ginsenoside Rk3) and ginsenoside ethanol-water extract. The composition of this invention exhibits extremely high safety; no LD50 was detected in acute toxicity tests. 50 Furthermore, the combination of the compositions described in this invention can significantly improve weight loss, effectively alleviate muscle atrophy caused by cachexia, effectively improve the reduction of gastrocnemius muscle weight, effectively improve the gap between muscle fibers and the cross-sectional area of ​​muscle bundles, effectively alleviate fat consumption, and effectively control and treat the development of cachexia caused by cancer.

[0025] In this invention, the ginsenoside Rk1 in the pharmaceutical composition comprises 5-12% by mass, preferably 4.5-11%. In this invention, ginsenoside Rk1, chemical name: 3β,6α,12β-trihydroxydammar-20(21),24-diene-6-O-β-D-glucopyranoside, molecular formula C 42 H 70 O 12 It has a molecular weight of 767.0, is odorless, and is a white powder. It is soluble in methanol and ethanol, slightly soluble in ethyl acetate, poorly soluble in water, and insoluble in chloroform and diethyl ether. CAS No.: 494753-69-4. Its structural formula is shown in Formula I.

[0026]

[0027] In this invention, the ginsenoside Rg5 in the pharmaceutical composition has a mass percentage of 5-12%, preferably 5.6-10.8%. In this invention, ginsenoside Rg5, chemical name: beta-D-Glucopyranoside,(3beta,12beta,20E)-12-hydroxydammara-20(22),24-dien-3-yl2-O-beta-D-glucopyranosyl, molecular formula C 42 H 70 O 12 Molecular weight: 767.0, odorless, white powder. Soluble in methanol and ethanol, slightly soluble in ethyl acetate, poorly soluble in water, insoluble in chloroform and diethyl ether. CAS number: 74964-14-0, structural formula as shown in Formula II.

[0028]

[0029] In this invention, the ginsenoside Rg3 in the pharmaceutical composition has a mass percentage of 5% to 12%, preferably 5.2% to 9.1%. In this invention, ginsenoside Rg3, chemically named 3-O-[β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl]-20(S)-protopanaxadiol, has the molecular formula: C 42 H 72 O 13 Molecular weight: 784.5, odorless, white powder. Soluble in methanol and ethanol, slightly soluble in ethyl acetate, poorly soluble in water, insoluble in chloroform and diethyl ether. CAS number: 14197-60-5. Structural formula as shown in Formula III.

[0030]

[0031] In this invention, the ginsenoside Rh4 in the pharmaceutical composition has a mass percentage of 3-8%, preferably 3.2-7.2%. In this invention, ginsenoside Rh4, scientifically known as 6-O-β-D-glucopyranosyl-20(-H2O)-trans-protopanaxatriol, has the molecular formula C2. 36 H 60 O8; Molecular weight: 620.4. Odorless, white powder. Soluble in methanol and ethanol, slightly soluble in ethyl acetate, poorly soluble in water, insoluble in chloroform and ether; CAS number: 174721-08-5, structural formula as shown in Formula IV.

[0032]

[0033] In this invention, the ginsenoside Rk3 in the pharmaceutical composition has a mass percentage of 2-6%, preferably 2.2-5.0%. In this invention, ginsenoside Rk3, scientifically known as Trihydroxydammar-20(21),24-diene-6-O-β-D-glucopyranoside, has the molecular formula C2. 36 H 60 O8; Molecular weight: 620.87, odorless, white powder. Soluble in methanol and ethanol, slightly soluble in ethyl acetate, poorly soluble in water, insoluble in chloroform and diethyl ether; CAS number: 364779-15-7, structural formula as shown in Formula V.

[0034]

[0035] In this invention, the ginseng ethanol-water extract in the pharmaceutical composition comprises 50-80% by mass, preferably 63-75%. In this invention, the ginseng ethanol-water extract preferably comprises the following components in mass percentage: 5-11.3% ginsenosides, 20-30% ginseng polysaccharides, 22-32.7% ginseng polypeptides, 1.0-2.0% ginseng polyphenols, and 2.0-4.0% ginseng flavonoids.

[0036] In this invention, the ginseng ethanol-water extract preferably comprises 5-11.3% ginsenosides, more preferably 7-9.3%, and more preferably 9%. In this invention, the ginsenosides preferably include Rg1, Re, Rf, Rb1, Rc, Rd, and Rb2; the mass percentage of Rg1 in the ginseng ethanol-water extract is 2-3%, preferably 2.4-2.7%, more preferably 2.6%; the mass percentage of Re in the ginseng ethanol-water extract is 1.0-2.5%, preferably 1.4-2.0%, more preferably 1.5%; the mass percentage of Rf in the ginseng ethanol-water extract is 0.2-1.5%, preferably 0.5-1.4%, more preferably 1.4%; and the mass percentage of Rb1 in the ginseng ethanol-water extract is... The ginseng ethanol-water extract contains 0.5-1.5% by mass, preferably 0.8-1.2%, more preferably 0.9% by mass; Rc contains 0.5-1.2% by mass, preferably 0.8-1.1%, more preferably 1.1% by mass; Rd contains 0.5-0.8% by mass, preferably 0.6-0.7%, more preferably 0.7% by mass; and Rb2 contains 0.3-0.8% by mass, preferably 0.5-0.8%, more preferably 0.8% by mass.

[0037] In this invention, the ginseng ethanol-water extract comprises 20-30% ginseng polysaccharides, preferably 29.3%. In this invention, the ginseng ethanol-water extract comprises 22-32.7% ginseng polypeptides, preferably 32.3%. In this invention, the ginseng ethanol-water extract comprises 1.0-2.0% ginseng polyphenols, preferably 1.3%. In this invention, the ginseng ethanol-water extract comprises 2.0-4.0% ginseng flavonoids, preferably 3.1%.

[0038] In this invention, the ginseng ethanol-water extract is preferably extracted from white ginseng, fresh ginseng, or red ginseng.

[0039] In this invention, the sources of ginsenosides Rk1, Rg5, Rg3, Rh4, and Rk3 are preferably natural extractions. In this invention, the ginsenosides Rk1, Rg5, Rh4, Rg3, and Rk3 are preferably extracted from wild ginseng, cultivated ginseng, American ginseng, Panax notoginseng, red ginseng, or white ginseng. In this invention, the extraction methods for ginsenosides Rk1, Rg5, Rg3, Rh4, and Rk3 preferably include one or more of the following: water extraction, organic solvent extraction, permeation, distillation, ultrasonic impregnation, extraction, and macroporous adsorption resin separation. In this invention, the ginsenosides Rk1, Rg5, and Rg3 are preferably obtained through the conversion of other diol-type ginsenosides. For example, ginsenosides Rk1 and Rg5 can be obtained by conversion of diol-type ginsenosides such as Rb1. In this invention, ginsenosides Rh4 and Rk3 are preferably obtained by conversion of other triol-type ginsenosides. For example, ginsenosides Rh4 and Rk3 can be obtained by conversion of triol-type ginsenoside Rg1. In this invention, the conversion method preferably includes enzymatic degradation. The sources of ginsenosides Rk1, Rg5, Rg3, Rh4, and Rk3 in this invention preferably also include direct purchase.

[0040] In this invention, the pharmaceutically acceptable carrier in the pharmaceutical composition preferably comprises 1-10% by mass. In this invention, the pharmaceutically acceptable carrier preferably includes one or more of the following: solvents, buffers, coatings, isotonics, wetting agents, emulsifiers, preservatives, and antibacterial agents.

[0041] In this invention, the dosage form of the pharmaceutical composition preferably includes: tablets, powders, suspensions, emulsions, capsules, granules, sugar-coated tablets, pills, liquids, medicated solutions, or syrups.

[0042] In this invention, the extraction method of the ginseng ethanol-water extract preferably includes the following steps: drying and pulverizing ginseng to obtain pulverized ginseng; mixing the pulverized ginseng with an ethanol aqueous solution with a volume percentage of 75%, heating at 55°C for 3 hours, and filtering to obtain a first filtrate and a first filter residue; mixing the first filter residue with an ethanol aqueous solution with a volume percentage of 45%, heating at 55°C for 3 hours, and filtering to obtain a second filtrate and a second filter residue; mixing the second filter residue with water, heating at 80°C for 3 hours, and filtering to obtain a third filtrate; combining the first filtrate, the second filtrate, and the third filtrate, evaporating the solvent, and freeze-drying to obtain the ginseng ethanol-water extract.

[0043] This invention involves drying and pulverizing ginseng to obtain pulverized ginseng. The invention does not specify any particular method for drying and pulverizing; conventional drying and pulverizing methods well-known to those skilled in the art can be used.

[0044] After obtaining the pulverized ginseng, the present invention mixes the pulverized ginseng with an ethanol aqueous solution containing 75% by volume, heats at 55°C for 3 hours, and filters to obtain a first filtrate and a first filter residue. In the present invention, the preferred volume ratio of the pulverized ginseng to the ethanol aqueous solution containing 75% by volume is 1:

[0045] (8-12), more preferably 1:10. In this invention, the heating method is preferably a water bath heating method.

[0046] In this invention, the first filtrate is preferably stored in a refrigerator at 4°C.

[0047] After obtaining the first filter residue, the present invention mixes the first filter residue with an aqueous ethanol solution of 45% by volume, heats at 55°C for 3 hours, and filters to obtain a second filtrate and a second filter residue. In the present invention, the volume ratio of the first filter residue to the aqueous ethanol solution of 45% by volume is preferably 1:(8-12), more preferably 1:10. In the present invention, the heating method is preferably a water bath heating method. In the present invention, the second filtrate is preferably stored in a refrigerator at 4°C.

[0048] After obtaining the second filter residue, the present invention mixes the second filter residue with water, heats it at 80°C for 3 hours, and filters it to obtain the third filtrate. In the present invention, the volume ratio of the second filter residue to water is preferably 1:(8-12), more preferably 1:10. In the present invention, the heating method is preferably a water bath heating method. In the present invention, the second filtrate is preferably stored in a refrigerator at 4°C.

[0049] After obtaining the first, second, and third filtrates, this invention combines the three filtrates, evaporates the solvent, and freeze-dries them to obtain ginseng ethanol-water extract. This invention does not have a specific limitation on the solvent evaporation method; conventional methods are acceptable, such as using a rotary evaporator to remove the solvent. After evaporating the solvent, this invention preferably freeze-dries the concentrate. This invention does not have a specific limitation on the freeze-drying method; conventional freeze-drying methods are acceptable. After freeze-drying, this invention preferably grinds the freeze-dried solid into powder.

[0050] The present invention also provides a method for preparing the pharmaceutical composition described in the above technical solution, comprising the following steps:

[0051] Ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract, and a pharmaceutically acceptable carrier are mixed to obtain a pharmaceutical composition.

[0052] This invention also provides the use of the pharmaceutical composition described above in the preparation of a medicament for the prevention and / or treatment of cachexia. In this invention, the disease causing the cachexia preferably includes malignant tumors. In this invention, the malignant tumors preferably include colorectal cancer, prostate cancer, kidney cancer, and lung cancer. In this invention, the cachexia includes cachexia occurring in patients with advanced cancer.

[0053] To further illustrate the present invention, a pharmaceutical composition and its preparation method, as well as its application in the preparation of medicaments for the prevention and / or treatment of cachexia, are described in detail below with reference to the accompanying drawings and embodiments. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0054] Example 1

[0055] Preparation of the pharmaceutical composition of the present invention.

[0056] Ginsenoside Rk1 was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0057] Ginsenoside Rg5 was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0058] Ginsenoside Rh4 was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0059] Ginsenoside Rg3 was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0060] Ginsenoside Rh4 was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0061] Ginsenoside Rk3 was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0062] Their purity all meets pharmaceutical standards.

[0063] Preparation method of ethanol-water extract:

[0064] 1. Dry the ginseng and grind it into powder;

[0065] 2. Heat ginseng powder with 10 times its volume of 75% ethanol in a 55°C water bath for 3 hours. Store the filtrate in a 4°C refrigerator.

[0066] 3. Heat the filter residue from the previous step with 10 times its volume of 45% ethanol in a 55°C water bath for 3 hours, and store the filtrate in a 4°C refrigerator.

[0067] 4. Heat the filter residue from the previous step with 10 times its volume of pure water in an 80°C water bath for 3 hours. Store the filtrate in a 4°C refrigerator and discard the filter residue.

[0068] 5. Combine the filtrates obtained in steps 2, 3 and 4, and use a rotary evaporator to evaporate most of the solvent;

[0069] 6. Freeze-dry the concentrated liquid, and grind the freeze-dried solid into powder.

[0070] Ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract, and dextrin (Shanghai Yuanye Biotechnology Co., Ltd.) were mixed to obtain a pharmaceutical composition.

[0071] The compositions of the present invention were prepared according to the proportions shown in Table 1.

[0072] Table 1. Composition of the pharmaceutical composition

[0073]

[0074]

[0075] The composition of ginseng ethanol-water extract is shown in Table 2.

[0076] Table 2 Composition of Ginseng Ethanol-Water Extract

[0077]

[0078] Example 2

[0079] Safety evaluation of the compositions of the present invention

[0080] The non-clinical safety evaluation of the compositions of the present invention is as follows:

[0081] 1. Acute oral toxicity test in mice

[0082] Mice were orally administered composition 1 of the present invention by gavage under the conditions of maximum dosing concentration and maximum dosing volume, and observed continuously for 14 days to obtain the maximum tolerated dose of the composition of the present invention >10.0 g / kg BW.

[0083] 2. Acute oral toxicity test in Beagle dogs

[0084] Composition 1 of the present invention was administered orally by gavage to Beagle dogs at the maximum dosing concentration and maximum dosing volume conditions, thereby obtaining a maximum tolerated dose of the composition of the present invention >2.0 g / kg BW.

[0085] 3. Long-term toxicity of oral administration to rats

[0086] A long-term toxicity test was conducted on SD rats using composition 1 of the present invention via oral gavage for three consecutive months, followed by a four-week recovery period after drug withdrawal. The results showed: ① General condition: During the administration and recovery periods, the animals' food and water intake were normal, their weight increased, their fur was smooth, and their behavior was normal; ② Hematological and blood biochemical indicators: After the administration and recovery periods, all hematological and blood biochemical indicators fluctuated within the normal range, with no obvious abnormalities observed; ③ Bone marrow and urinalysis indicators: After the administration and recovery periods, no obvious abnormalities were observed in the bone marrow and urinalysis indicators; ④ Histopathological indicators: After the administration and recovery periods, no obvious abnormalities were observed in the organs of the animals, and there were no significant differences in organ weight and organ coefficients compared to the control group. No obvious pathological changes were observed in any organ. The results indicate that no significant toxic reactions were observed with long-term administration to SD rats.

[0087] 4. Long-term toxicity test of oral administration in Beagle dogs

[0088] A long-term toxicity test was conducted on Beagle dogs using the composition 1 of the present invention via oral gavage for three months, followed by a four-week recovery period after drug withdrawal. The results showed: ① General condition: During the administration and recovery periods, food and water intake and body temperature were normal, weight increased, fur was smooth, and behavior was normal; ② Hematological and blood biochemical indicators: After the administration and recovery periods, all hematological and blood biochemical indicators fluctuated within the normal range, with no obvious abnormalities observed; ③ Electrocardiogram indicators: After the administration and recovery periods, all electrocardiogram indicators fluctuated within the normal range, with no obvious abnormalities observed; ④ Bone marrow and ophthalmological examination: After the administration and recovery periods, bone marrow cells... No abnormalities were observed in cells and their taxa; ophthalmic examination revealed clear vascular patterns without hemorrhage or exudation in the fundus of all groups of animals, no edema in the optic disc, and normal arteriovenous ratios; ⑤ Immunological and urinary / fecal indicators: After the end of administration and during the recovery period, all immunological and urinary / fecal indicators fluctuated within the normal range, with no significant abnormalities observed; ⑥ Histopathological indicators: After the end of administration and during the recovery period, no significant abnormalities were observed in any of the animals' organs grossly, and there were no significant differences in organ weight and organ coefficients compared to the control group. No significant pathological changes were observed in any organ. The results indicate that long-term administration to Beagle dogs did not result in significant toxic reactions.

[0089] 5. General pharmacological tests

[0090] Oral administration of composition 1 of the present invention to anesthetized Beagle dogs via gavage had no significant effect on their blood pressure (diastolic and systolic), heart rate, P wave, T wave, R wave, QRS interval, PR interval, QT interval, respiratory rate, or respiratory amplitude. The composition of the present invention also had no significant effect on the Irwin's behavioral test score and pole climbing test score in mice. This indicates that the composition of the present invention does not affect the central nervous system, cardiovascular system, or respiratory system of animals.

[0091] 6. Mutagenicity test

[0092] Chromosomal aberration assays, Ames assays, and mouse micronucleus assays in cultured mammalian cells (CHL) showed that composition 1 of the present invention had no mutagenic effect.

[0093] Example 3

[0094] Pharmacodynamics

[0095] The composition of the present invention inhibits the development of cancer cachexia in Balb / c mice.

[0096] Materials: Balb / c mice, and pharmaceutical compositions 1-3 obtained in Example 1 of this invention.

[0097] Methods: 6-8 week old Balb / c mice were subcutaneously injected with CT26 intestinal cancer cells. Once the tumors reached a sufficient size, they were harvested, and a CT26 tumor suspension was prepared. A second batch of 6-8 week old Balb / c mice were also subcutaneously injected with the CT26 tumor suspension. When a palpable subcutaneous tumor mass was obtained, the mice were administered drug compositions 1-3 from Example 1 of this invention via gavage. The mouse body weight and tumor size were measured daily. The tumor mass was calculated using the formula: Tumor mass (mg) = 0.52 × Tumor length (mm) × Tumor width. 2 (mm 2 The body weight of mice after tumor removal was calculated. A model group (subcutaneously injected with tumor cell suspension followed by gavage with drinking water) and a normal control group (injected with an equal volume of PBS solution as the tumor suspension, followed by gavage with drinking water) were also set up.

[0098] The results showed that the composition of the present invention significantly improved the weight loss of mice, such as... Figure 1 ,from Figure 1 As can be seen, over time, the body weight of the model group mice decreased significantly, while the body weight of mice using drug compositions 1-3 remained relatively stable; the composition of this invention effectively alleviates muscle atrophy caused by cachexia, such as Figure 2 and Figure 3 ,from Figure 2 It can be seen that the weight of the gastrocnemius muscle in the model group mice was significantly reduced compared with that in normal mice, while the weight reduction of the gastrocnemius muscle in mice was effectively improved after treatment with drug combinations 1-3. Figure 3 It can be seen that, compared with normal mice, the gaps between the gastrocnemius muscle fibers in the model group mice were significantly larger, and the cross-sectional area of ​​the muscle bundles was significantly reduced in the cross-section of the gastrocnemius muscle. Treatment with drug compositions 1-3 effectively improved both the gaps between muscle fibers and the cross-sectional area of ​​the muscle bundles. The drug composition of this invention can effectively alleviate fat consumption. Figure 4As can be seen, the epididymal fat of the model group mice was significantly depleted. After treatment with the drug composition 1-3, the depletion of epididymal fat in the mice was effectively controlled. Further comparison of the drug composition of the present invention with imperatorin (IMP) showed that, compared with IMP, the drug composition of the present invention better inhibited epididymal fat in mice. The drug composition of the present invention can effectively control and treat the development of cachexia caused by cancer, such as... Figure 5 ,from Figure 5 The results showed that the expression of two muscle tissue E3 ubiquitin ligases, Fbxo32 and Trim63, was significantly upregulated in the model group, indicating that strong protein degradation occurred in the gastrocnemius muscle of the model group mice. However, after administration of the drug composition of this invention (1-3), the expression of the two E3 ubiquitin ligases, Fbxo32 and Trim63, decreased to the level of the normal control group mice. MyoD is a myogenic differentiation factor that can promote the differentiation and generation of myocytes. Figure 5 It can be seen that the expression of MyoD in the model group was significantly reduced, while after treatment with the drug composition of the present invention 1-3, the expression of MyoD returned to the level of the normal control group mice.

[0099] Example 4

[0100] Drug stability test

[0101] Stability testing was performed on composition 2 from Example 1.

[0102] 1. Saponin extraction:

[0103] (1) Accurately weigh 10g of the test sample (composition 2) into a 50mL stoppered test tube, add 40mL of methanol, sonicate in an ultrasonic cleaner for 30min, filter, and extract the residue twice more. Combine the three filtrates and put them into a 250mL constant weight rotary evaporation flask. After concentration and drying under reduced pressure, defatt the evaporated saponins three times with petroleum ether, evaporate to dryness, transfer to a constant temperature drying oven (105℃) to dry to constant weight, and weigh.

[0104] (2) Weigh 20-30 mg of the sample obtained in (1), dissolve it in 50 mL of water, and then purify it by loading it onto a 50 mL pre-treated AB-8 macroporous adsorption resin column. After the saponins are completely adsorbed, wash away the unadsorbed components with 150-200 mL of water, and then elute with 150-200 mL of 70%-75% ethanol. After complete elution, evaporate to dryness and use it as the sample to be tested.

[0105] 2. Determination of various saponins:

[0106] (1) Chromatographic reference conditions:

[0107] a) Chromatographic column: C18 column (5μm, )

[0108] b) Mobile phase: Acetonitrile-water

[0109] c) Column temperature: 35℃

[0110] d) Flow rate: 1 mL / min

[0111] e) Detection wavelength: 203nm

[0112] f) Injection volume: 20 μL

[0113] g) The gradient elution procedure is shown in Table 3.

[0114] Table 3 Mobile phase elution procedure

[0115] Time / min Acetonitrile A / % Water (B) 0～20 20 80 20～31 20→30 80→70 31～35 30→33 70→67 35～45 33→47 67→53 45～50 47→55 53→45 50～75 55→85 45→15 75～85 85→100 15→0

[0116] (2) Plotting the standard curve

[0117] Using methanol as the solvent, prepare standard solutions of each saponin (ginsenoside Rk1, ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rh4, and ginsenoside Rk3) at a concentration of 2 mg / mL. Take 1 mL, 3 mL, 4 mL, 5 mL, and 6 mL of each solution and place them in 10 mL volumetric flasks, then dilute to the mark with methanol. Detect the solutions using high-performance liquid chromatography (HPLC), recording the peak areas. The injection volume is 20 μL. Plot a standard curve based on the peak area and the mass concentration of each saponin.

[0118] (3) Content determination

[0119] Dissolve the purified sample in methanol and transfer it to a 25 mL volumetric flask, then dilute to volume and filter through a 0.45 μm membrane. Inject 20 μL of the filtrate into the high-performance liquid chromatograph (HPLC) for detection. Record the peak area and compare it with the peak area of ​​the standard solution for quantification.

[0120] Table 4. Changes in saponin content and storage time of composition 2

[0121]

[0122] As shown in Table 4, the contents of the main saponins in Composition 2—ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, and ginsenoside Rk3—remained unchanged after three years of storage. Among them, ginsenoside Rk1 experienced the greatest loss rate after three years of storage, at only 1.4%. The results indicate that Composition 2 has strong stability.

[0123] Example 5

[0124] Mr. Hong, male, 63 years old, was a patient with advanced colorectal cancer. The tumor had metastasized to the liver and lungs. His body mass index (BMI) was 19.8. He started taking the drug of this invention (the drug of composition 2 in Example 1) in March 2018. After 2 months, he gained 3 kg in weight and his physical strength and energy were significantly improved.

[0125] Mr. Zhang, male, 78 years old, a patient with advanced prostate cancer, had a body mass index (BMI) of only 18.5. He started taking the drug of the present invention (the drug of composition 2 in Example 1) for 9 weeks on June 18, 2020. His BMI rose to 24, and his quality of life was significantly improved.

[0126] Ms. Xu, 89 years old, was a patient with mid-stage colorectal cancer that had metastasized to her liver. Her body mass index (BMI) was 16.2. She started taking the drug of this invention (the drug of composition 1 in Example 1) in March 2017. After six months, she gained 5 kg, her physical strength and energy improved significantly, and her appetite also increased significantly.

[0127] Ms. Liu, 55 years old, was an early-stage colorectal cancer patient with a body mass index (BMI) of 17.8. In the early stages of her illness, she experienced symptoms such as loss of appetite, nausea, vomiting, and abdominal pain. In March 2018, she began taking the drug of this invention (the drug of composition 3 in Example 1). Two months later, she gained 3 kilograms, her physical strength and energy improved significantly, her appetite returned, her nausea and vomiting disappeared, and her abdominal pain lessened.

[0128] Mr. Liang, male, 53 years old, was a patient with advanced colorectal cancer. During his illness, his body mass index (BMI) was only 17.2. He started taking the drug of this invention (the drug of composition 2 in Example 1) in April 2019. After 3 months, he gained 6 kg and his physical strength and energy were significantly improved.

[0129] Mr. Li, male, 70 years old, was an early-stage colorectal cancer patient with a body mass index (BMI) of 18.1. During his illness, he experienced abdominal distension and pain, significantly weakened digestive and absorptive functions, and significant weight loss. In June 2019, he began taking the drug of this invention (the drug of composition 3 in Example 1). Two months later, he gained 5 kg, his physical strength and energy improved significantly, and his abdominal pain disappeared.

[0130] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A pharmaceutical composition for the prevention and / or treatment of cachexia, characterized in that, The pharmaceutical composition comprises the following components: ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginsenoside ethanol-water extract, and a pharmaceutically acceptable carrier; wherein the mass percentage of ginsenoside Rk1 is 5-12%, the mass percentage of ginsenoside Rg5 is 5-12%, the mass percentage of ginsenoside Rg3 is 5%-12%, the mass percentage of ginsenoside Rh4 is 3-8%, the mass percentage of ginsenoside Rk3 is 2-6%, and the mass percentage of ginsenoside ethanol-water extract is 50-80%; wherein the ginsenoside ethanol-water extract comprises the following components in the following mass percentages: ginsenoside components 5-11.3%, ginsenoside polysaccharide components 20-30%, ginsenoside polypeptide components 22-32.7%, and ginsenoside polyphenol components. The ginsenosides consist of 1.0-2.0% ginseng and 2.0-4.0% ginseng flavonoids; the ginsenosides include Rg1, Re, Rf, Rb1, Rc, Rd, and Rb2; Rg1 has a mass percentage of 2-3% in the ginseng ethanol-water extract, Re has a mass percentage of 1.0-2.5% in the ginseng ethanol-water extract, Rf has a mass percentage of 0.2-1.5% in the ginseng ethanol-water extract, Rb1 has a mass percentage of 0.5-1.5% in the ginseng ethanol-water extract, Rc has a mass percentage of 0.5-1.2% in the ginseng ethanol-water extract, Rd has a mass percentage of 0.5-0.8% in the ginseng ethanol-water extract, and Rb2 has a mass percentage of 0.3-0.8% in the ginseng ethanol-water extract. The extraction method of the ginseng ethanol-water extract includes the following steps: drying and pulverizing ginseng to obtain pulverized ginseng; mixing the pulverized ginseng with a 75% (v / v) ethanol aqueous solution, heating at 55°C for 3 hours, and filtering to obtain a first filtrate and a first filter residue; mixing the first filter residue with a 45% (v / v) ethanol aqueous solution, heating at 55°C for 3 hours, and filtering to obtain a second filtrate and a second filter residue; mixing the second filter residue with water, heating at 80°C for 3 hours, and filtering to obtain a third filtrate; combining the first, second, and third filtrates, evaporating the solvent, and freeze-drying to obtain the ginseng ethanol-water extract.

2. The pharmaceutical composition for the prevention and / or treatment of cachexia according to claim 1, characterized in that, The sources of the ginsenosides Rk1, Rg5, Rg3, Rh4, and Rk3 include natural extraction.

3. A method for preparing the pharmaceutical composition for preventing and / or treating cachexia as described in claim 1 or 2, characterized in that, Includes the following steps: Ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract, and a pharmaceutically acceptable carrier are mixed to obtain a pharmaceutical composition for the prevention and / or treatment of cachexia.

4. Use of the pharmaceutical composition of claim 1 or 2 in the preparation of a medicament for the prevention and / or treatment of cachexia.

5. The application according to claim 4, characterized in that, The cachexia mentioned includes cachexia that occurs in patients with advanced cancer.

Images