Use of a pharmaceutical composition for the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis

Abstract

The application belongs to the technical field of medicine, and relates to application of a pharmaceutical composition in preparation of a medicine for treating and / or preventing rheumatoid arthritis. The application provides application of a pharmaceutical composition in preparation of a medicine for treating and / or preventing rheumatoid arthritis, wherein 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. The components of the pharmaceutical composition can effectively treat and / or prevent rheumatoid arthritis.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis. Background Technology

[0002] Rheumatoid arthritis (RA) is a prevalent autoimmune disease in humans, primarily characterized by non-suppurative inflammation of the joints and surrounding tissues. Studies show that RA can occur at any age, with a high incidence rate; typically, 1-2 out of every 100 people will have RA. In China, the incidence rate is 0.4%, with a large number of patients aged 30-50, and a 2-3 times higher incidence in women than men. If RA is not diagnosed and treated promptly, it can easily spread to other organs and joints. Studies indicate that the disability rate within 3 years for untreated RA patients is as high as 75%. Currently, there is no cure for rheumatoid arthritis. Clinically, the most commonly used treatments include four classes: nonsteroidal anti-inflammatory drugs (NSAIDs), glucocorticoids (GCs), DMARDs (diuretic rheumatoid arthritis drugs), and biologics. Nonsteroidal anti-inflammatory drugs (NSAIDs) (such as ibuprofen and celecoxib) are the most widely used in the treatment of rheumatoid arthritis. They are used to relieve pain in the early stages of the disease. Their mechanism of action is mainly to inhibit cyclooxygenase-2 (COX-2), thereby inhibiting prostaglandin production. However, these drugs cannot alter the disease progression or reduce joint damage; they are only used as adjunctive therapies. Furthermore, the inhibition of prostaglandin production can easily cause adverse reactions such as diarrhea, indigestion, headache, and nausea, and can also lead to kidney damage and cardiovascular disease. Glucocorticoids (such as dexamethasone, prednisolone, and dextrin) are widely used in the treatment of rheumatoid arthritis due to their very strong anti-inflammatory and immunomodulatory effects. However, these drugs cannot specifically target the pathological sites, and discontinuation or long-term use can lead to very serious side effects, such as insulin resistance, thinning of the skin, osteoporosis, hypertension, obesity, and inhibition of wound healing. While antirheumatic drugs such as methotrexate have long been considered first-line treatments, their effects are slow, taking 1 to 6 months to become apparent. They lack immediate analgesia and anti-inflammatory effects, have poor disease flare-up control, and long-term use can lead to side effects such as anorexia, nausea, vomiting, stomatitis, hair loss, leukopenia or thrombocytopenia, drug-induced pneumonia, and rashes. Biologics (anti-tumor necrosis factor, IL-1 receptor antagonists, IL-6 antagonists, β-cell and T-cell blockers, etc.) have gained recognition for their therapeutic effects in rheumatoid arthritis. However, these biologics are essentially immunosuppressants, focusing on blocking the activation of immune cells and the signaling pathways of related factors. Long-term systemic administration inevitably suppresses the body's normal immunity, increasing the likelihood of microbial infections and tumor development. Furthermore, their production technology is demanding, resulting in extremely high medical costs for patients, significantly limiting their clinical application. Currently, there is a need to develop safe and effective drugs for treating RA to alleviate the suffering of RA patients. Summary of the Invention

[0003] The purpose of this invention is to provide the use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis. The pharmaceutical composition of this invention can effectively prevent and / or treat rheumatoid arthritis, and has extremely high safety.

[0004] This invention provides the use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis, the pharmaceutical composition comprising the following components: ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract, and a pharmaceutically acceptable carrier.

[0005] 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%.

[0006] 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.

[0007] Preferably, the ginsenoside components include Rg1, Re, Rf, Rb1, Rc, Rd, and Rb2; the mass percentage of Rg1 in the ginseng ethanol-water extract is 2-3%, the mass percentage of Re in the ginseng ethanol-water extract is 1.0-2.5%, the mass percentage of Rf in the ginseng ethanol-water extract is 0.2-1.5%, the mass percentage of Rb1 in the ginseng ethanol-water extract is 0.5-1.5%, the mass percentage of Rc in the ginseng ethanol-water extract is 0.5-1.2%, the mass percentage of Rd in the ginseng ethanol-water extract is 0.5-0.8%, and the mass percentage of Rb2 in the ginseng ethanol-water extract is 0.3-0.8%.

[0008] Preferably, the ginseng ethanol-water extract is extracted from white ginseng, fresh ginseng, or red ginseng.

[0009] 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.

[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 carrier includes one or more of the following: solvent, buffer, coating, isotropic agent, wetting agent, emulsifier, preservative and antibacterial agent.

[0013] Preferably, the method for preparing the pharmaceutical composition includes 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] This invention provides the use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis (RA). The pharmaceutical composition of this invention can alleviate the inflammatory response of RA by reducing the levels of inflammatory factors and improving the body's antioxidant capacity. The pharmaceutical composition of this invention can be used as a pharmaceutical preparation for the treatment and / or prevention of rheumatoid arthritis. The composition of this invention is more stable and easier to store, and has significant application prospects. Experimental results show that the pharmaceutical composition of this invention can achieve effective treatment of rheumatoid arthritis. Attached Figure Description

[0016] 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.

[0017] Figure 1 The effect of the pharmaceutical compositions 1-3 provided by the present invention on rheumatoid arthritis in ICR rats, a model of rheumatoid arthritis. Detailed Implementation

[0018] This invention provides the use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis. The pharmaceutical composition comprises the following components: ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rg3, ginsenoside Rh4, ginsenoside Rk3, ginseng ethanol-water extract, and a pharmaceutically acceptable carrier. The composition of this invention exhibits extremely high safety; no LD50 was detected in acute toxicity tests. 50 In this invention, the diseases causing rheumatoid arthritis preferably include inflammatory diseases and / or autoimmune diseases. Specific combinations and proportions of the pharmaceutical compositions of this invention can achieve effective prevention and / or treatment of rheumatoid arthritis.

[0019] In this invention, the preferred mass percentage of ginsenoside Rk1 in the pharmaceutical composition is 5-12%, the preferred mass percentage of ginsenoside Rg5 is 5-12%, the preferred mass percentage of ginsenoside Rg3 is 5-12%, the preferred mass percentage of ginsenoside Rh4 is 3-8%, the preferred mass percentage of ginsenoside Rk3 is 2-6%, and the preferred mass percentage of ginseng ethanol-water extract is 50-80%.

[0020] In this invention, the ginsenoside Rk1 in the pharmaceutical composition has a mass percentage of 5-12%, preferably 5.3-10.8%. 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.

[0021]

[0022] In this invention, the mass percentage of ginsenoside Rg5 in the pharmaceutical composition is 5-12%, preferably 6.1-10.5%. 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.

[0023]

[0024] In this invention, the ginsenoside Rg3 in the pharmaceutical composition has a mass percentage of 5% to 12%, preferably 5.5% to 8.9%. 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.

[0025]

[0026] In this invention, the ginsenoside Rh4 in the pharmaceutical composition comprises 3-8% by mass, preferably 3.0-6.2%. 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.

[0027]

[0028] In this invention, the ginsenoside Rk3 in the pharmaceutical composition has a mass percentage of 2-6%, preferably 2.5-4.3%. 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.

[0029]

[0030] 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.

[0031] 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.

[0032] 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%.

[0033] In this invention, the ginseng ethanol-water extract is preferably extracted from white ginseng, fresh ginseng, or red ginseng.

[0034] 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.

[0035] 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.

[0036] 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:

[0037] (8-12), more preferably 1:10. In this invention, the heating method is preferably a water bath heating method. In this invention, the first filtrate is preferably stored in a refrigerator at 4°C.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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: solvent, buffer, coating, isotonist, wetting agent, emulsifier, preservative, and antibacterial agent. In this invention, the dosage form of the pharmaceutical composition preferably includes: tablets, powder, suspension, emulsion, capsules, granules, sugar-coated tablets, pills, liquid, medicated solution, or syrup.

[0043] In this invention, the method for preparing the pharmaceutical composition preferably includes the following steps:

[0044] 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.

[0045] To further illustrate the present invention, the application of a pharmaceutical composition provided by the present invention in the preparation of a medicament for the prevention and / or treatment of rheumatoid arthritis is described in detail below with reference to the accompanying drawings and embodiments, but these descriptions should not be construed as limiting the scope of protection of the present invention.

[0046] Example 1

[0047] Preparation of the composition of the present invention

[0048] Ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rh4, ginsenoside Rg3, ginsenoside Rh4, and ginsenoside Rk3 were all purchased from Shanghai Yuanye Biotechnology Co., Ltd. Their purity all met pharmaceutical standards. They were prepared according to the proportions shown in Tables 1 to 3 below.

[0049] Preparation method of ethanol-water extract:

[0050] 1. Dry the ginseng and grind it into powder;

[0051] 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.

[0052] 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.

[0053] 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.

[0054] 5. Combine the filtrates obtained in steps 2, 3 and 4, and use a rotary evaporator to evaporate most of the solvent;

[0055] 6. Freeze-dry the concentrated liquid, and grind the freeze-dried solid into powder.

[0056] The composition of the ginseng ethanol-water extract is shown in Table 4.

[0057] 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.

[0058] Table 1. Composition of the pharmaceutical composition

[0059]

[0060]

[0061] Table 2 Composition of the pharmaceutical composition

[0062]

[0063] Table 3. Composition of the pharmaceutical composition

[0064]

[0065]

[0066] Table 4 Ethanol-water extracts

[0067]

[0068] Example 2

[0069] Safety evaluation of the compositions of the present invention

[0070] The non-clinical safety evaluation of the compositions of the present invention was studied as follows:

[0071] 1. Acute oral toxicity test in mice

[0072] 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 composition of the present invention as >10.0 g / kg BW.

[0073] 2. Acute oral toxicity test in Beagle dogs

[0074] 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.

[0075] 3. Long-term toxicity of oral administration to rats

[0076] 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.

[0077] 4. Long-term toxicity test of oral administration in Beagle dogs

[0078] 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.

[0079] 5. General pharmacological tests

[0080] 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.

[0081] 6. Mutagenicity test

[0082] Chromosomal aberration assays in cultured mammalian cells (CHL), Ames assays, and mouse micronucleus assays showed that composition 1 of the present invention had no mutagenic effect.

[0083] Example 3

[0084] Pharmacodynamics

[0085] The composition of the present invention inhibits the development of rheumatoid arthritis in ICR mice.

[0086] Materials: ICR mice, compositions 1-3 and compositions 4-9 obtained in Example 1 of this invention.

[0087] Methods: Six- to eight-week-old ICR mice were randomly divided into 10 groups. Hair was removed from the right plantar surface of the mice, and the joints were disinfected. The normal control group was injected with physiological saline (0.02 mL / mouse), while the other groups were injected with 0.02 mL of complete Freund's adjuvant (CFA) into the right plantar surface, ideally with the appearance of obvious white blisters around the injection site, to induce inflammation and develop adjuvant-induced arthritis (AA). On day 15 after successful modeling, the model mice were injected with CFA again to better reflect the recurrent nature of clinical RA. Drug treatment began 20 days after modeling. Groups 1-3 and 4-9 of the drug compositions in Example 1 were administered the same volume of 0.9% physiological saline by gavage. The treatment period was 60 days. Various observation indicators of the mice were measured every 3 days. After the end of treatment, the mice were observed for another 12 days, with various indicators measured every 3 days.

[0088] Table 5. Effects on rheumatoid arthritis in ICR rats (x±s)

[0089]

[0090] Table 6. Effects on rheumatoid arthritis in ICR rats (x±s)

[0091]

[0092] Table 7. Effects on rheumatoid arthritis in ICR rats (x±s)

[0093]

[0094] The results are as follows Figure 1(Figure showing the effect of drug compositions 1-3 on rheumatoid arthritis in ICR rats) and Tables 5-7 indicate that compositions 1, 2, and 3 of this invention all have good therapeutic effects in the ICR mouse model of rheumatoid arthritis. Other drug compositions, compositions 4, 5, 6, 7, 8, and 9, showed no significant therapeutic effect. This indicates that the absence of any of the rare ginsenoside group components (ginsenoside Rk1, ginsenoside Rg5, ginsenoside Rk3, ginsenoside Rg3, and ginsenoside Rh4) in compositions 1-3 of this invention is necessary to achieve an effective therapeutic effect.

[0095] Example 4

[0096] Mr. Li, male, 63 years old, had suffered from rheumatoid arthritis for 8 years. After taking the drug of the present invention (the drug of composition 2 in Example 1) for 6 months, all his symptoms disappeared.

[0097] Mr. Tan, male, 56 years old, was diagnosed with rheumatoid arthritis at the University of Hong Kong-Shenzhen Hospital on February 3, 2023. After taking the drug of the present invention (the drug of composition 3 in Example 1) for 12 weeks starting on February 4, 2023, the patient's various physiological indicators tended to normalize, all symptoms disappeared, and his quality of life was significantly improved.

[0098] Mr. Mou, male, 65 years old, had been suffering from rheumatoid arthritis for 6 years. After taking the drug of the present invention (the drug of composition 1 in Example 1) for 10 weeks, the patient's joint function was significantly improved and his quality of life was enhanced.

[0099] Animal experiments showed that compositions 4-9 had no therapeutic effect, so they were not used in human trials.

[0100] 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. The use of a pharmaceutical composition in the preparation of a medicament for treating rheumatoid arthritis, said pharmaceutical composition comprising the following components in weight percentages: The ginsenoside Rk1 contains 5-12% by mass, ginsenoside Rg5 contains 5-12% by mass, ginsenoside Rg3 contains 5-12% by mass, ginsenoside Rh4 contains 3-8% by mass, ginsenoside Rk3 contains 2-6% by mass, ginsenoside ethanol-water extract contains 63-75% by mass, and pharmaceutically acceptable carrier contains 1-10% by mass. 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 application according to claim 1, characterized in that, The ginseng ethanol-water extract is composed of ginsenosides, ginseng polysaccharides, ginseng polypeptides, ginseng polyphenols, and ginseng flavonoids. The ginsenosides in the pharmaceutical composition have a mass percentage of 5-11.3%, the ginseng polysaccharides have a mass percentage of 20-30%, the ginseng polypeptides have a mass percentage of 22-32.7%, the ginseng polyphenols have a mass percentage of 1.0-2.0%, and the ginseng flavonoids have a mass percentage of 2.0-4.0%.

3. The application according to claim 2, characterized in that, The ginsenoside components are composed of Rg1, Re, Rf, Rb1, Rc, Rd, and Rb2; Rg1 has a mass percentage of 2-3% in the pharmaceutical composition, Re has a mass percentage of 1.0-2.5% in the pharmaceutical composition, Rf has a mass percentage of 0.2-1.5% in the pharmaceutical composition, Rb1 has a mass percentage of 0.5-1.5% in the pharmaceutical composition, Rc has a mass percentage of 0.5-1.2% in the pharmaceutical composition, Rd has a mass percentage of 0.5-0.8% in the pharmaceutical composition, and Rb2 has a mass percentage of 0.3-0.8% in the pharmaceutical composition.

4. The application according to claim 1, characterized in that, The ginseng ethanol-water extract is extracted from white ginseng, fresh ginseng, or red ginseng.

5. The application according to claim 1, characterized in that, The sources of the ginsenosides Rk1, Rg5, Rg3, Rh4, and Rk3 include natural extraction.

6. The application according to claim 5, characterized in that, The ginsenosides Rk1, Rg5, Rh4, Rg3, and Rk3 are extracted from wild ginseng, cultivated ginseng, American ginseng, Panax notoginseng, red ginseng, or white ginseng.

7. The application according to claim 1, characterized in that, The carrier includes one or more of the following: solvent, buffer, coating, isotropic agent, wetting agent, emulsifier, preservative and antibacterial agent.

8. The application according to claim 1, characterized in that, The method for preparing the pharmaceutical composition 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.

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