A total ginsenoside-salvia miltiorrhiza total phenolic acid cardiac function improving composition

Abstract

The application discloses a ginseng and astragalus root-danshen total phenolic acid heart function improving composition, and relates to the technical field of medicines.The active ingredients of the composition are composed of three chemical monomers of ginsenoside Rg1, astragaloside B and danshensu, and the weight ratio is 1:0.5 to 1:2 to 3.Through combination of the three specific monomers in the accurate proportion, multiple key nodes of myocardial cell energy metabolism can be synergistically regulated, and myocardial energy metabolism disorder under heart failure state can be systematically intervened.The composition has clear components, controllable quality, and can be prepared into various dosage forms such as tablets and capsules.Compared with medicinal material extracts with complex components, the composition provided by the application has the advantages of clear action mechanism, accurate proportion, stable quality and the like, and provides a new drug selection for the treatment of heart failure.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function. Background Technology

[0002] Heart failure is the end stage of many cardiovascular diseases, and one of its core pathophysiological mechanisms is impaired energy metabolism in cardiomyocytes. In heart failure, cardiomyocytes are generally caught in a vicious cycle of insufficient energy production, reduced conversion efficiency, and imbalanced energy utilization. This metabolic remodeling is a key factor leading to the progressive deterioration of cardiac pumping function. Therefore, intervention at the source and core pathways of energy metabolism is considered one of the more fundamental strategies for treating heart failure.

[0003] In existing technologies, the synergistic treatment of cardiovascular diseases using the effective components of Danshen (Salvia miltiorrhiza) and Huangqi (Astragalus membranaceus) has received widespread attention. For example, patent publication number CN1096269C discloses a pharmaceutical composition for treating cardiovascular and cerebrovascular diseases, which mainly consists of three types of components: phenolic acid compounds from Danshen, astragaloside saponins, and astragalus polysaccharides. Although this composition claims to be used to treat heart failure, its components are still a relatively broad category of extracts with a wide range of chemical components, and the ratio between the three types of components is also quite wide. Its mechanism of action is more based on the holistic regulation of traditional Qi-invigorating and blood-activating theories, failing to precisely target and synergistically regulate the specific and continuous pathological chain of myocardial cell energy metabolism. Another patent publication number CN1919254B provides a pharmaceutical composition composed of total phenolic acid extracts from Danshen and total saponin extracts from Huangqi, but its use is clearly limited to the treatment of acute renal failure and does not involve the improvement of cardiac function or the regulation of myocardial energy metabolism.

[0004] In summary, although there are combined applications of relevant medicinal extracts in the prior art, these schemes or components are complex, the targets are scattered, or the uses are completely different from those of this application. They all lack the design concept of precise targeted and synergistic intervention for the core pathological link of heart failure—multiple key steps of myocardial cell energy metabolism.

[0005] In view of this, there is an urgent problem to be solved in the existing technology: how to provide a drug combination with well-defined components and precise design, so that it can systematically and synergistically intervene in the complete continuous chain of myocardial cell energy metabolism from substrate utilization to ATP synthesis and utilization, so as to more effectively correct myocardial energy metabolism disorders in the state of heart failure, rather than performing broad, multi-target overall regulation. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function. By deeply analyzing the continuous links of myocardial energy metabolism, a precise ratio combination of three specific chemical monomers, ginsenoside Rg1, astragaloside A and tanshinone, is proposed and constructed to achieve precise synergistic regulation of key nodes of energy metabolism disorders, providing a new drug option with a clear mechanism for the treatment of heart failure.

[0007] To solve the above-mentioned technical problems, the present invention provides the following technical solution: On the one hand, a ginseng-astragalus-tanshinone total phenolic acid heart function improvement composition, wherein the active ingredients of the composition are composed of ginsenoside Rg1, astragaloside A and tanshinone, and the weight ratio of ginsenoside Rg1, astragaloside A and tanshinone is 1:0.5 to 1:2 to 3.

[0008] Furthermore, the weight ratio of ginsenoside Rg1, astragaloside A, and tanshinone is 1:0.7 to 0.9:2.2 to 2.8.

[0009] Furthermore, the weight ratio of ginsenoside Rg1, astragaloside A, and tanshinone is 1:0.8:2.5.

[0010] Furthermore, the ginsenoside Rg1, astragaloside A, and tanshinone are all chemical monomers, and the purity of each monomer is not less than 98%.

[0011] Furthermore, the composition is an oral solid dosage form, which includes tablets, hard capsules, granules, or pellets.

[0012] Furthermore, the oral solid dosage form also contains pharmaceutically acceptable fillers, disintegrants, and lubricants.

[0013] On the other hand, a method for preparing a ginseng-astragalus-tanshinone total phenolic acid heart function improvement composition, applicable to a ginseng-astragalus-tanshinone total phenolic acid heart function improvement composition, includes the following steps:

[0014] Step 1: Provide ginsenoside Rg1 monomer, astragaloside A monomer, and tanshinone monomer;

[0015] Step 2: Weigh each monomer component according to the weight ratio of the composition;

[0016] Step 3: Mix the weighed monomer components evenly.

[0017] Furthermore, the use of the composition in the preparation of a medicament for treating heart failure.

[0018] Compared with existing technologies, this ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function has the following beneficial effects:

[0019] I. This invention selects three specific chemical monomers—ginsenoside Rg1, astragaloside A, and tanshinone—and limits their precise weight ratio range, enabling the three to synergistically act on different key links in myocardial cell energy metabolism. This composition can systematically intervene in the continuous pathological chain of myocardial cells from energy substrate utilization to ATP generation and utilization, achieving targeted regulation of the core pathological mechanism of heart failure. This overcomes the problems of unclear components and dispersed targets caused by the use of complex extracts in the prior art, and provides a drug combination with a clear mechanism and precise design, offering a better option for the treatment of heart failure.

[0020] Second, by limiting the ginsenoside Rg1, astragaloside A, and tanshinone to chemical monomers with a purity of not less than 98%, this invention ensures a high degree of clarity and consistency of the active ingredients. This is beneficial for achieving precise quality control and stable dosage ratios during the production process, and improves the uniformity and reproducibility of the product.

[0021] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0023] Figure 1 This is a schematic diagram showing the core composition and proportions of the composition of the present invention;

[0024] Figure 2 This is a schematic diagram illustrating the mechanism by which the composition of the present invention synergistically regulates the energy metabolism of cardiomyocytes. Detailed Implementation

[0025] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below.

[0026] Example 1

[0027] like Figures 1-2As shown, this embodiment provides a method for preparing a ginseng-astragalus-tanshinone total phenolic acid cardiac function improvement composition and its tablets. The active ingredients of this composition are ginsenoside Rg1, astragaloside A, and tanshinone, with a weight ratio of 1:0.5:2. Each active ingredient is a chemical monomer with a purity of not less than 98%. This embodiment details the raw material screening and purity verification process, clarifies the selection criteria and dosage of excipients in tablet preparation, refines the complete process parameters from raw material pretreatment to finished tablet compression, and verifies the regulatory effect of the composition on cardiomyocyte energy metabolism through in vitro cardiomyocyte culture experiments. The following detailed description is provided in conjunction with the specific implementation content.

[0028] 1. In this embodiment, the specific implementation of raw material preparation and screening is as follows:

[0029] 1.1 Selection of Active Ingredient Raw Materials: This embodiment uses ginsenoside Rg1 monomer, astragaloside A monomer, and tanshinone monomer. Ginsenoside Rg1 monomer is extracted from the dried root and rhizome of *Panax ginseng* (Araliaceae family), astragaloside A monomer is extracted from the dried root of *Astragalus membranaceus* or *Astragalus mongholicus* (Fabaceae family), and tanshinone monomer is extracted from the dried root and rhizome of *Salvia miltiorrhiza* (Lamiaceae family). The reason for selecting these raw materials is that these plant materials have been proven to have cardiovascular protective pharmacological activities in traditional medicine. The monomeric components in their extracts have well-defined structures, facilitating purity control and precise ratio adjustment, which aligns with the design concept of this invention for precise intervention at key nodes of myocardial cell energy metabolism.

[0030] 1.2 Excipient Selection: The oral solid dosage form in this embodiment is a tablet. Based on the pharmaceutical requirements for tablet preparation, microcrystalline cellulose was selected as the filler, crospovidone as the disintegrant, and magnesium stearate as the lubricant. Microcrystalline cellulose was chosen as the filler because it has good flowability and compressibility, effectively improving the material's molding properties, and it has good biocompatibility in vivo, without adverse effects on the human body. Crospovidone, as a disintegrant, has a rapid disintegration rate, is unaffected by gastrointestinal pH, and ensures rapid disintegration of the tablet in vivo, allowing for rapid release and absorption of the active ingredient. Magnesium stearate, as a lubricant, reduces friction between the material and the tableting die, preventing sticking during compression and ensuring the tablet's intact appearance and uniform weight. All excipients were purchased from pharmaceutical-grade suppliers.

[0031] 2. In this embodiment, the specific implementation of raw material pretreatment and purity verification is as follows:

[0032] 2.1 Raw Material Pretreatment: The purchased ginsenoside Rg1 monomer, astragaloside A monomer, and tanshinone monomer were placed in a 60℃ vacuum drying oven for 2 hours, with the drying pressure controlled between -0.08MPa and -0.1MPa. The purpose was to remove any trace moisture that might be present in the raw materials, preventing moisture from affecting the subsequent mixing uniformity and formulation stability. After drying, the raw materials were quickly removed and placed in a desiccator to cool to room temperature to prevent moisture absorption. Each monomer raw material was then sieved through an 80-mesh sieve in a Class 100,000 cleanroom. This sieving process removed any trace impurities, agglomerated particles, or mechanical impurities that might be present in the raw materials, ensuring particle size uniformity and laying the foundation for accurate proportioning and uniform mixing in the subsequent process.

[0033] 2.2 Purity Verification: High-performance liquid chromatography (HPLC) was used to test the purity of each monomer raw material after pretreatment to ensure that the purity of each monomer component was not less than 98%. The specific testing method is as follows:

[0034] Chromatography instrument: Agilent 1260 high performance liquid chromatograph, equipped with a UV detector;

[0035] Chromatographic column: C18 column;

[0036] Mobile phase: An acetonitrile-water gradient elution system was used. The elution program was as follows: 0-10 min, acetonitrile volume fraction increased from 10% to 20%; 10-20 min, acetonitrile volume fraction increased from 20% to 30%; 20-30 min, acetonitrile volume fraction increased from 30% to 40%; 30-35 min, acetonitrile volume fraction was maintained at 40%.

[0037] Detection wavelengths: 203 nm for ginsenoside Rg1 and astragaloside A, and 280 nm for tanshinone.

[0038] Column temperature: 30℃;

[0039] Flow rate: 1.0 mL / min;

[0040] Injection volume: 20 μL.

[0041] Accurately weigh an appropriate amount of pretreated ginsenoside Rg1 monomer raw material, dissolve and dilute it with methanol to prepare a test solution containing 0.1 mg of ginsenoside Rg1 per mL; prepare astragaloside A test solution and tanshinone test solution using the same method. Separately prepare reference solutions of ginsenoside Rg1, astragaloside A, and tanshinone using the same method.

[0042] Accurately pipette 20 μL each of the test solution and the reference solution and inject them into the high-performance liquid chromatograph (HPLC). Perform the analysis under the chromatographic conditions described above and record the chromatograms. Calculate the purity of the target component in each test solution using the peak area normalization method, i.e., the percentage of the target component's peak area to the total peak area. The results show that the purity of ginsenoside Rg1 monomer is 98.3%, the purity of astragaloside A monomer is 98.1%, and the purity of tanshinone monomer is 98.5%, all meeting the requirement of at least 98% purity for each monomer component in this invention.

[0043] 3. In this embodiment, the preparation of the composition is specifically carried out as follows:

[0044] 3.1 Precise Weighing of Active Ingredients: Following the weight ratio of 1:0.5:2 set in this embodiment, the raw materials were weighed using a 0.01 g analytical balance in a Class 100,000 cleanroom. First, 10.00 g of ginsenoside Rg1 monomer was precisely weighed. During the weighing process, the balance was calibrated first, then a clean weighing bottle was placed on the balance to remove the tare, and then the ginsenoside Rg1 raw material was slowly added until the displayed weight was 10.00 g. After weighing, a second weighing was performed to ensure that the weighing error did not exceed ±0.01 g. Following the same operating method, 5.00 g of astragaloside A monomer and 20.00 g of tanshinone monomer were weighed sequentially. After a second weighing, it was confirmed that the errors were within the allowable range, ensuring the accuracy of the ratio of the three active ingredients.

[0045] 3.2 Uniform Mixing of Active Ingredients: The three weighed active ingredients were placed together in a three-dimensional motion mixer (model SYH-10, mixing tank volume 10L). The mixer speed was set to 20 r / min, and the mixing time to 30 minutes. During the mixing process, the mixer was stopped every 10 minutes, and samples were taken from three different positions (top, middle, and bottom) of the mixing tank, each sample being 0.5g, for a total of three samples. Each sample was dissolved and diluted with methanol, and the contents of ginsenoside Rg1, astragaloside A, and tanshinone in the sample were determined according to the above HPLC detection conditions, and the weight ratio of the three was calculated. The test results showed that the ratios of the three samples were 1:0.502:2.003, 1:0.498:1.997, and 1:0.501:2.001, respectively, with a ratio deviation of no more than ±0.5%, indicating that the three active ingredients were uniformly mixed. Mixing was stopped, and the mixed active ingredient mixture was taken out for later use.

[0046] 3.3 Excipient Pretreatment: Take 30.00g of microcrystalline cellulose, 17.50g of crospovidone, and 3.50g of magnesium stearate, and dry them separately in a 60℃ forced-air drying oven for 1 hour. Maintain good ventilation inside the drying oven during the drying process to remove moisture from the excipients and prevent moisture from affecting subsequent mixing and formulation stability. After drying, remove the excipients and cool them to room temperature. Then, sieve them through an 80-mesh sieve to remove any clumps and impurities, ensuring that the particle size of the excipients is consistent with that of the active ingredient mixture, facilitating subsequent overall mixing.

[0047] 3.4 Overall Mixing: The uniformly mixed active ingredient mixture, along with the pretreated microcrystalline cellulose and crospovidone, was placed into a three-dimensional motion mixer. The mixer speed was set to 20 r / min, and the mixing time was 20 minutes. After mixing, an appropriate amount of the mixture was taken, and its bulk density and angle of repose were measured to evaluate the material's flowability. The test results showed that the bulk density of the mixture was 0.6 g / cm³, and the angle of repose was 32°, which meets the requirements for material flowability in tablet preparation, indicating that the material can smoothly enter the tableting machine for tableting.

[0048] 3.5 Granulation: Take the above-mentioned mixed material and place it in a trough mixer. Add an appropriate amount of 5% povidone K30 aqueous solution as a wetting agent while stirring at a speed of 50 r / min for 10 minutes to form a soft mass. The soft mass should be in a state where it "can be formed into a ball when squeezed by hand, but crumbles easily when lightly pressed." If the soft mass is too dry, add a small amount of 5% povidone K30 aqueous solution; if the soft mass is too wet, add a small amount of microcrystalline cellulose to adjust it. Granulate the obtained soft mass through a 16-mesh sieve using a swing pellet mill at a speed of 20 r / min to obtain wet pellets.

[0049] 3.6 Drying and Granulation: The wet granules were evenly spread on a tray and placed in a 60℃ forced-air drying oven for 2 hours. During the drying process, the granules were turned over every 30 minutes to ensure uniform drying and avoid localized excessive or insufficient moisture content. After drying, the granules were removed and cooled to room temperature. Then, they were granulated through an 18-mesh sieve to remove excessively large clumps and excessively small fine powder, resulting in uniformly sized dry granules. An appropriate amount of the granulated dry granules was taken and its moisture content was determined using the Karl Fischer moisture assay. The results showed a moisture content of 4.2%, which is within the suitable range of 3%-5%, meeting the moisture requirements for tablet preparation.

[0050] 3.7 Tableting: The sizing of dry granules and the pretreated magnesium stearate were placed together in a two-dimensional mixer. The mixing speed was set to 30 r / min, and the mixing time was 5 minutes to ensure that the magnesium stearate was evenly distributed on the surface of the granules, providing lubrication. The mixed tableting material was then added to a rotary tablet press (model ZP-19). The tableting pressure was set to 8-10 MPa, the tablet weight to 0.3 g / tablet, and the tablet press speed to 20 tablets / min. Tableting was performed to obtain unprocessed tablets. During the tableting process, samples were taken every 100 tablets produced, for a total of 5 samples, to test the tablet weight variation, hardness, and disintegration time. The test results showed that the tablet weight variation in the 5 samples was within ±5%, the hardness was 3-5 kgf, and the disintegration time was 12 minutes.

[0051] 3.8 Coating: Take an appropriate amount of film coating premix, dissolve it in 80% ethanol solution and stir evenly to prepare a 5% coating solution. The viscosity of the coating solution should be controlled between 20-30 mPa·s. Place the uncoated tablets into a high-efficiency coating machine (model BG-150). Set the inlet air temperature to 60℃, the outlet air temperature to 40℃, the spraying speed to 5 mL / min, the atomization pressure to 0.3 MPa, and the coating time to 30 minutes. During the coating process, observe the appearance of the tablets in real time to ensure that the tablet surface is smooth, the color is uniform, and there are no defects such as pitting or bubbles. After coating, remove the tablets and place them in a desiccator to cool to room temperature to obtain the finished tablets. The weight increase of the finished tablets is 3%-5%, which meets the quality requirements for coated tablets.

[0052] 4. In this embodiment, the specific implementation of finished product quality inspection is as follows:

[0053] 4.1 Determination of Active Ingredient Content: Ten tablets of the finished product prepared in this example were accurately weighed, and the average tablet weight was calculated. The tablets were then ground into a fine powder. An appropriate amount of the powder was accurately weighed and placed in a 50 mL volumetric flask. An appropriate amount of methanol was added, and the mixture was sonicated for 30 minutes. After cooling to room temperature, the powder was diluted to the mark with methanol, shaken well, and filtered. The filtrate was used as the test solution. The contents of ginsenoside Rg1, astragaloside A, and tanshinone in the test solution were determined according to the above HPLC detection conditions. The results showed that each finished tablet contained 28.6 mg of ginsenoside Rg1, 14.3 mg of astragaloside A, and 57.2 mg of tanshinone, with a weight ratio of 1:0.5:2, consistent with the set ratio, indicating that the content of active ingredients was effectively controlled during the preparation process.

[0054] 4.2 Stability Testing: The finished tablets were placed in a stability test chamber at 25°C and 60% relative humidity for accelerated stability testing. Samples were taken at 0 months, 1 month, 3 months, and 6 months to test the tablets' appearance, active ingredient content, disintegration time, and microbial limits. The results showed that the tablets' appearance remained largely unchanged and smooth within 6 months; the decrease in active ingredient content did not exceed 5%; and the disintegration time remained within 15 minutes. This indicates that the tablets prepared in this embodiment have good stability.

[0055] 5. In this embodiment, the specific implementation of the pharmacological effect verification is as follows:

[0056] To verify the regulatory effect of the composition prepared in this embodiment on the energy metabolism of cardiomyocytes, an in vitro cardiomyocyte culture experiment was conducted.

[0057] 5.1 Cardiac Cell Isolation and Culture: Newborn SD rats (1-3 days old) were selected. Hearts were removed under aseptic conditions, atrial and connective tissue were removed, and ventricular myocardial tissue was cut into 1 mm³ pieces. 0.25% trypsin solution was added, and the mixture was digested in a 37°C water bath for 15 minutes, gently agitated every 5 minutes. After digestion, DMEM medium containing 10% fetal bovine serum was added to terminate the digestion. The mixture was filtered through a 200-mesh sieve, the filtrate was collected and centrifuged, the supernatant was discarded, and the cells were resuspended in DMEM medium. The cells were then seeded into culture flasks and cultured in a 37°C, 5% CO2 incubator. After 2 hours, the medium was replaced to remove non-adherent cells, yielding purified cardiomyocytes. Subsequent experiments were performed when the cardiomyocytes reached 80% confluence.

[0058] 5.2 Experimental Grouping and Treatment: The cultured cardiomyocytes were divided into three groups: blank control group, model group, and Example 1 group, with 6 replicates in each group. Blank control group: Normal culture without special treatment; Model group: A cardiomyocyte energy metabolism disorder model was constructed using the hypoxia-hypoglycemia method, i.e., the medium was replaced with glucose-free DMEM and cultured in a hypoxic incubator containing 95% N2 and 5% CO2 for 6 hours; Example 1 group: In addition to OGD treatment, the tablet extract prepared in this example was added to the culture medium, and the remaining treatments were the same as those in the model group.

[0059] 5.3 Detection Indicators and Methods: After 24 hours of culture, cells and culture medium from each group were collected, and the following indicators were detected:

[0060] ATP content determination: The ATP detection kit was used, and the operation was carried out according to the kit instructions. The luminescence intensity was measured on a chemiluminescence detector, and the intracellular ATP content was calculated based on the standard curve.

[0061] LDH release assay: The activity of LDH in the culture medium was measured using a lactate dehydrogenase assay kit. The higher the LDH release, the more severe the myocardial cell damage.

[0062] Cell viability assay: Using the CCK-8 kit, CCK-8 solution was added to each group of cells, and after culturing for 4 hours, the absorbance value at a wavelength of 450 nm was measured on an ELISA reader to calculate the cell viability.

[0063] 5.4 Experimental Results: The experimental results showed that, compared with the blank control group, the ATP content in cardiomyocytes of the model group was significantly reduced, the LDH release was significantly increased, and the cell survival rate was significantly decreased, indicating that OGD treatment successfully constructed a cardiomyocyte energy metabolism disorder model. Compared with the model group, the ATP content in cardiomyocytes of the Example 1 group increased by 23.5%, the LDH release decreased by 18.2%, and the cell survival rate increased by 21.3%, all of which were statistically significant. These results indicate that the composition prepared in this example can improve hypoxia- and hypoglycemia-induced cardiomyocyte energy metabolism disorders, reduce cardiomyocyte damage, and improve cell survival rate, exhibiting a certain cardioprotective effect, which meets the application requirements of this invention for the treatment of heart failure.

[0064] In summary, this embodiment successfully prepared a ginseng-astragalus-tanshinone total phenolic acid tablet for improving cardiac function through a clearly defined active ingredient ratio, strict purity control, reasonable excipient selection, and detailed preparation process. The entire preparation process was clear in steps and parameters, and each step from raw material pretreatment to finished product testing was operable. The prepared tablets had stable quality and accurate active ingredient content, meeting the pharmaceutical requirements for oral solid dosage forms.

[0065] Example 2

[0066] like Figures 1-2 As shown, this embodiment provides a ginseng-astragalus-tanshinone total phenolic acid cardiac function improvement composition and its preparation method in hard capsule form. The active ingredients of this composition are ginsenoside Rg1, astragaloside A, and tanshinone, with a weight ratio of 1:0.7:2.2. Each active ingredient is a chemical monomer with a purity of not less than 98%. This embodiment focuses on the preparation process of the hard capsule, detailing the selection criteria for the capsule shell and the determination process of the filling parameters. Furthermore, the composition's effect on improving cardiac function is verified through a combination of in vitro cell experiments and in vivo animal experiments. The following detailed description is provided in conjunction with the specific implementation details.

[0067] 1. In this embodiment, the specific implementation of raw material preparation and screening is as follows:

[0068] 1.1 Selection of Raw Materials for Active Ingredients: The ginsenoside Rg1 monomer, astragaloside A monomer, and tanshinone monomer used in this embodiment are from the same sources as in Example 1. The extraction sites and processes of the raw materials are stable, ensuring the structural consistency and pharmacological activity stability of the active ingredients. The core reason for selecting these monomer components is that ginsenoside Rg1 has been proven to promote the uptake of energy metabolism substrates by cardiomyocytes, astragaloside A can improve myocardial mitochondrial function, and tanshinone has the effect of scavenging oxygen free radicals and protecting cardiomyocytes. The synergistic effect of the three can cover multiple key links in myocardial cell energy metabolism, achieving precise intervention, which is consistent with the design concept of this invention targeting the core pathological mechanism of heart failure.

[0069] 1.2 Excipient Selection: The oral solid dosage form in this embodiment is a hard capsule. Microcrystalline cellulose was selected as the filler, low-substituted hydroxypropyl cellulose as the disintegrant, and calcium stearate as the lubricant. The capsule shell is a gelatin hollow capsule. Microcrystalline cellulose was chosen as the filler because its bulk density is suitable, ensuring uniform capsule filling, and it does not easily absorb moisture in the body, which is beneficial to the stability of the formulation. Low-substituted hydroxypropyl cellulose, as the disintegrant, has good hydrophilicity and swelling properties, and can rapidly swell in water to disintegrate the capsule contents, promoting the release of the active ingredient. Calcium stearate, as the lubricant, has a milder lubricating effect compared to magnesium stearate and will not affect the stability of the active ingredient. It has good biocompatibility and disintegration performance, which meets the requirements of oral dosage forms.

[0070] 2. In this embodiment, the specific implementation of raw material pretreatment and purity verification is as follows:

[0071] 2.1 Raw Material Pretreatment: The raw material pretreatment steps are basically the same as in Example 1. Ginsenoside Rg1 monomer, astragaloside A monomer, and tanshinone monomer were placed in a vacuum drying oven at 60℃ for 2 hours, with a drying pressure of -0.08MPa to -0.1MPa, to remove trace amounts of moisture. After cooling to room temperature, they were then sieved through an 80-mesh sieve to remove impurities and agglomerated particles. The pretreatment process was carried out in a Class 100,000 cleanroom to ensure that the raw materials were not contaminated and to guarantee the cleanliness requirements of the subsequent preparation process.

[0072] 2.2 Purity Verification: The purity of each monomer raw material was verified using the same HPLC detection method as in Example 1. Appropriate amounts of each pretreated monomer raw material were taken to prepare test solutions and reference solutions. The solutions were then analyzed under the set chromatographic conditions, and the purity was calculated using the peak area normalization method. The results showed that the purity of ginsenoside Rg1 monomer was 98.6%, astragaloside A monomer was 98.4%, and tanshinone monomer was 98.7%, all meeting the purity requirement of not less than 98%, ensuring the controllable purity of the active ingredients.

[0073] 3. In this embodiment, the preparation of the composition is specifically carried out as follows:

[0074] 3.1 Precise Weighing of Active Ingredients: Based on the weight ratio of 1:0.7:2.2 set in this embodiment, the active ingredients were weighed using a 0.01 g analytical balance in a Class 100,000 cleanroom. 10.00 g of ginsenoside Rg1 monomer was precisely weighed, with a re-weighing error of ±0.008 g; 7.00 g of astragaloside A monomer was weighed, with a re-weighing error of ±0.007 g; and 22.00 g of tanshinone monomer was weighed, with a re-weighing error of ±0.009 g. All weighing errors did not exceed ±0.01 g, ensuring the accuracy of the proportions.

[0075] 3.2 Uniform Mixing of Active Ingredients: The weighed three active ingredients were placed in a three-dimensional motion mixer, and the mixing speed was set to 22 r / min for 35 minutes. During the mixing process, samples were taken at 10, 20, and 35 minutes, with each sample being 0.5 g. The ratio of the three components in the sample was determined by HPLC. The test results showed that the ratios of the three samples were 1:0.701:2.202, 1:0.699:2.198, and 1:0.700:2.200, respectively, with a ratio deviation of no more than ±0.5%, indicating that the three active ingredients were uniformly mixed and ready for use.

[0076] 3.3 Excipient Pretreatment: Take 39.00g of microcrystalline cellulose, 7.80g of low-substituted hydroxypropyl cellulose, and 1.95g of calcium stearate, and dry them separately in a 60℃ forced-air drying oven for 1 hour. After cooling to room temperature, sieve them through an 80-mesh sieve to remove lumps and impurities, and set aside for later use. The determination of the excipient dosage is based on the fact that the filling amount of hard capsules is usually 0.3-0.5g / capsule. Combined with the total weight of the active ingredients, an appropriate amount of excipients is selected to achieve a suitable filling volume while ensuring the flowability and disintegration properties of the contents.

[0077] 3.4 Overall Mixing Operation: The uniformly mixed active ingredients, along with the pretreated microcrystalline cellulose and low-substituted hydroxypropyl cellulose, were placed into a three-dimensional motion mixer. The speed was set to 22 r / min, and the mixing time was 25 minutes. After mixing, the bulk density and angle of repose of the mixture were measured. The bulk density was 0.58 g / cm³, and the angle of repose was 33°, which meets the material flowability requirements for hard capsule filling, ensuring that the material can be smoothly filled into the capsule shell.

[0078] 3.5 Filling Operation: The mixed materials were added to the fully automatic capsule filling machine, with a filling speed of 300 capsules / minute and a filling amount of 0.4g / capsule. Before filling, the capsule shells were screened to remove deformed or damaged shells to ensure shell quality. During the filling process, a sample was taken every 1000 capsules filled, for a total of 5 samples, to check for differences in filling amount. The test results showed that the difference in filling amount among the 5 samples was within ±3%, which meets the filling requirements for hard capsules.

[0079] 3.6 Polishing and Screening: Place the filled hard capsules into a capsule polishing machine and polish for 5 minutes to remove dust and impurities from the capsule surface, making the capsules smooth and clean. After polishing, screen the capsules using a capsule screening machine to remove empty capsules, partially filled capsules, and broken capsules, obtaining qualified finished hard capsule products.

[0080] 4. In this embodiment, the specific implementation of finished product quality inspection is as follows:

[0081] 4.1 Determination of Active Ingredient Content: Ten hard capsules prepared in this example were taken, their contents were poured out, and the total weight was accurately weighed to calculate the average fill weight. An appropriate amount of the contents was accurately weighed and placed in a 50 mL volumetric flask. An appropriate amount of methanol was added, and the mixture was sonicated for 30 minutes. After cooling, it was diluted to the mark with methanol, shaken well, filtered, and the filtrate was used as the test solution. The content of the three active ingredients was determined according to the HPLC detection conditions. The test results showed that each hard capsule contained ginsenoside Rg 125.6 mg, astragaloside A 17.9 mg, and tanshinone 56.3 mg, with a weight ratio of 1:0.7:2.2, consistent with the set ratio, indicating accurate content control.

[0082] 4.2 Stability Testing: The finished hard capsules were placed in a stability test chamber at 25℃ and 60% relative humidity for accelerated stability testing. Samples were taken and tested at 0 months, 1 month, 3 months, and 6 months. The test results showed that the capsules did not deform or leak powder within 6 months; the decrease in active ingredient content did not exceed 4%; the disintegration time was 18 minutes; and the microbial limits met the relevant standards, indicating that the hard capsules have good stability.

[0083] 5. In this embodiment, the specific implementation of the pharmacological effect verification is as follows:

[0084] 5.1 In vitro cardiomyocyte experiments: SD rat neonatal cardiomyocytes were isolated and cultured using the same method as in Example 1 to construct an OGD cardiomyocyte energy metabolism disorder model. The experimental groups were a blank control group, a model group, and the Example 2 group, with 6 replicates per group. In the Example 2 group, the extract of the hard capsule contents prepared in this example was added during OGD treatment to a final concentration of 10 μg / mL. After 24 hours of culture, ATP content, LDH release, and cell viability were measured.

[0085] The experimental results showed that, compared with the model group, the ATP content in cardiomyocytes of the Example 2 group increased by 27.8%, the LDH release decreased by 21.5%, and the cell survival rate increased by 25.6%, with statistically significant differences. This indicates that the composition has a better effect on improving cardiomyocyte energy metabolism than Example 1, which may be related to the optimization of the formulation.

[0086] 5.2 In vivo animal experiments: In order to further verify the effect of the composition on improving cardiac function, in vivo animal experiments were conducted.

[0087] Experimental animals: Sixty male SPF-grade SD rats, weighing 200-220g, were selected and purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The housing environment was kept at a temperature of 22-25℃ and a relative humidity of 50%-60%, with free access to water and food.

[0088] Model Establishment: A rat model of heart failure was established using the abdominal aortic constriction method. Rats were anesthetized with 10% chloral hydrate via intraperitoneal injection, fixed in a supine position, and the abdominal aorta was dissected through an incision in the left rectus abdominis muscle. The abdominal aorta was ligated above the renal artery using No. 4 silk suture, narrowing the vessel diameter by 50%. The incision was then sutured layer by layer. In the sham-operated group, only the abdominal aorta was dissected, without ligation. After 4 weeks of postoperative feeding, the left ventricular ejection fraction (LVEF) was measured by echocardiography; rats with LVEF < 50% were considered to have successfully established a heart failure model.

[0089] Experimental grouping: Rats with successfully established models were randomly divided into a model control group, a low-dose group (Example 2), and a high-dose group (Example 2), with 15 rats in each group. A sham-operated group of 15 rats was also included. The low-dose group (Example 2) was administered the extract from the contents of the hard capsules prepared in this example via gavage at a dose of 10 mg / kg. The high-dose group (Example 2) was administered the extract via gavage at a dose of 20 mg / kg. The model control group and the sham-operated group were administered an equal volume of physiological saline via gavage once daily for 4 consecutive weeks.

[0090] Detection indicators: Four weeks after administration, rats were anesthetized intraperitoneally, and LVEF and left ventricular short axis shortening rate were detected by echocardiography. After sacrificing the rats, heart tissue was collected, and the ATP content, superoxide dismutase activity and malondialdehyde content in the myocardial tissue were measured.

[0091] The experimental results showed that, compared with the sham-operated group, the model control group had significantly reduced LVEF and LVFS, significantly reduced ATP content and SOD activity in myocardial tissue, and significantly increased MDA content. Compared with the model control group, the low-dose and high-dose groups in Example 2 showed that LVEF increased by 12.3% and 18.5%, respectively; LVFS increased by 11.8% and 17.6%, respectively; ATP content in myocardial tissue increased by 24.6% and 32.8%, respectively; SOD activity increased by 18.9% and 26.3%, respectively; and MDA content decreased by 16.4% and 23.5%, respectively. All differences were statistically significant. These results indicate that the composition prepared in this example can improve cardiac function in rats with heart failure, increase the energy metabolism level of myocardial tissue, and reduce oxidative stress damage in vivo, thus having a certain therapeutic effect on heart failure.

[0092] In summary, this embodiment optimized the types and amounts of excipients and clarified key process parameters such as filling and polishing, based on the formulation characteristics of hard capsules, successfully preparing a stable and precisely proportioned ginseng-astragalus-tanshinone total phenolic acid hard capsule for improving cardiac function. In vitro cell experiments and in vivo animal experiments both showed that this composition can effectively regulate cardiomyocyte energy metabolism and improve cardiac function in heart failure model animals, verifying the effectiveness of the technical solution of this invention.

[0093] Comparative Example

[0094] This comparative example provides a cardiovascular-related composition and its tablet preparation method for comparison. The composition uses total phenolic acid extract of Tanshinone and total saponin extract of Astragalus membranaceus as active ingredients. The formulation type is the same as that of Example 1, and the preparation process parameters are basically the same as those of Example 1. By comparing with Example 1 in vitro cell experiments and in vivo animal experiments, the advantages of the composition of the present invention in terms of clear composition, accurate ratio, and regulatory effect on myocardial cell energy metabolism are verified. The following is a detailed description in conjunction with the specific implementation content.

[0095] 1. In this comparative example, the specific implementation of raw material preparation and screening is as follows:

[0096] The total phenolic acid extract of Salvia miltiorrhiza and the total saponin extract of Astragalus membranaceus used in this comparative example were purchased from conventional pharmaceutical raw material suppliers. The excipients were the same as those in Example 1, namely microcrystalline cellulose, crospovidone, and magnesium stearate. The capsule shell was a gelatin hollow capsule.

[0097] 2. In this comparative example, the preparation of the composition was specifically carried out as follows:

[0098] 2.1 Raw material weighing and mixing: Weigh 40.00g of total phenolic acid extract of Salvia miltiorrhiza and 15.00g of total saponin extract of Astragalus membranaceus. After sieving through an 80-mesh sieve in a Class 100,000 clean area, place them in a three-dimensional motion mixer. Set the mixing speed to 20r / min and the mixing time to 30 minutes to obtain a mixture of active ingredients.

[0099] 2.2 Tablet preparation: Following the tablet preparation process of Example 1, the same proportion of excipients were added, and the tablets were prepared through steps such as total mixing, granulation, drying, sizing, tableting, and coating to obtain a comparative tablet product. The weight of the finished product was 0.3g / tablet, and the quality test showed that its disintegration time, stability and other indicators met the tablet quality standards.

[0100] 3. The specific implementation of pharmacological effect verification in this comparative example is as follows:

[0101] The pharmacological effects of the tablets prepared in this comparative example were verified using the same in vitro cardiomyocyte experiments as in Example 1 and the same in vivo animal experiments as in Example 2. The experimental groups were a blank control group, a model group, and a comparative example group 1. The detection indicators were the same as in Examples 1 and 2.

[0102] 3.1 In vitro experimental results: Compared with the model group, the ATP content in cardiomyocytes of Comparative Example 1 increased by 12.1%, the LDH release decreased by 9.5%, and the cell survival rate increased by 10.2%. Although it has a certain cardioprotective effect, the effect is significantly lower than that of Example 1.

[0103] 3.2 In vivo experimental results: Compared with the model control group, the rats in Comparative Example 1 showed an increase of 7.8% in LVEF, an increase of 7.2% in LVFS, an increase of 13.5% in ATP content in myocardial tissue, an increase of 9.8% in SOD activity, a decrease of 8.6% in MDA content, and a decrease of 10.3% in serum cTnI content. The improvement effects of each indicator were significantly lower than those in Example 2.

[0104] In summary, this comparative example prepared cardiovascular-related tablets using a combination of commonly used herbal extracts in existing technologies. Pharmacological results showed that while this composition had some effect on improving cardiomyocyte energy metabolism and cardiac function in heart failure model animals, its effect was far inferior to the ginseng-astragalus-tanshinone total phenolic acid cardiac function improvement composition of this invention. The main reason for this is that the active ingredients in the comparative example are complex extracts, making precise formulation impossible, resulting in dispersed target sites and an inability to synergistically regulate key nodes in cardiomyocyte energy metabolism. In contrast, this invention uses chemical monomers with a purity of no less than 98%, achieving synergistic effects from the three components through precise formulation. This allows for systematic intervention in the complete chain of cardiomyocyte energy metabolism, thereby achieving a superior therapeutic effect.

[0105] The proportions are shown in the table below:

[0106] Group Active ingredient ratio (ginsenoside Rg1: astragaloside IV: danshensu) Formulation type Purity requirement Core effect (compared with model group) Example 1 1:0.5:2 Tablets All ≥98% ATP content increased by 23.5% Example 2 1:0.7:2.2 Hard capsules All ≥98% LVEF increased by 12.3%-18.5% Comparative example Salvia miltiorrhiza total phenolic acid + astragalus total saponin extract combination Tablets Total ingredients ≥50% ATP content increased by 12.1%

[0107] The table above systematically summarizes the core elements of each scheme, clearly demonstrating the characteristics of the embodiments of the present invention in terms of the accuracy of active ingredient ratio, the diversity of formulation types, and the superiority of pharmacological effects. Through clear process parameters and objective pharmacological data, the feasibility and effectiveness of the technical solution are verified.

[0108] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A composition for improving cardiac function using ginseng, astragalus, and tanshinone total phenols, characterized in that, The active ingredients of this composition consist of ginsenoside Rg1, astragaloside A, and tanshinone, wherein the weight ratio of ginsenoside Rg1, astragaloside A, and tanshinone is 1:0.5 to 1:2 to 3.

2. The composition for improving cardiac function using ginseng, astragalus, and tanshinone total phenolic acids according to claim 1, characterized in that, The weight ratio of ginsenoside Rg1, astragaloside A, and tanshinone is 1:0.7 to 0.9:2.2 to 2.

8.

3. The ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function according to claim 2, characterized in that, The weight ratio of ginsenoside Rg1, astragaloside A, and tanshinone is 1:0.8:2.

5.

4. The ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function according to claim 1, characterized in that, The ginsenoside Rg1, astragaloside A, and tanshinone are all chemical monomers, and the purity of each monomer is not less than 98%.

5. The composition for improving cardiac function using ginseng, astragalus, and tanshinone total phenolic acids according to claim 1, characterized in that, The composition is an oral solid dosage form, which includes tablets, hard capsules, granules, or pills.

6. The ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function according to claim 5, characterized in that, The oral solid dosage form also contains pharmaceutically acceptable fillers, disintegrants, and lubricants.

7. A method for preparing a ginseng-astragalus-tanshinone total phenolic acid heart function improving composition, applicable to the ginseng-astragalus-tanshinone total phenolic acid heart function improving composition according to any one of claims 1 to 6, characterized in that, This includes the following steps: Step 1: Provide ginsenoside Rg1 monomer, astragaloside A monomer, and tanshinone monomer; Step 2: Weigh each monomer component according to the weight ratio of the composition; Step 3: Mix the weighed monomer components evenly.

8. The ginseng-astragalus-tanshinone total phenolic acid composition for improving cardiac function according to claim 1, characterized in that, Use of the composition in the preparation of a medicament for treating heart failure.

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