Plant active ingredient composition for resisting hepatopathy attack and preparation method thereof

Abstract

The invention discloses an anti-hepatopathy plant active ingredient composition and a preparation method thereof, and belongs to the technical field of medicines. The composition is prepared from extracts of seven plants including radix gentianae, radix scutellariae and the like, vitamin B, taurine and VC according to a specific proportion, key components are subjected to ultrasonic-assisted extraction by adopting a deep eutectic solvent (choline chloride-lactic acid system), and a glycyrrhizic acid alkali extraction-resin purification process is matched, so that the yield of active components is remarkably increased, and the toxicity risk is reduced. The preparation method comprises the steps of step-by-step extraction, purification and drying, preferably, the ultrasonic power is 300-500W, and the spray drying temperature is 160-180 DEG C. The composition can be further added with nano-silver (20-50nm) or compounded with rice bran fatty alkanol, and experiments prove that the composition can significantly reduce liver injury markers and improve fatty degeneration, is suitable for preventing and treating liver injury, clearing liver fire, removing dampness, removing dryness, enhancing immunity, removing toxins in animal bodies, enhancing immunity, enhancing liver activity and reducing attack of liver diseases.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to a plant active ingredient composition for resisting liver disease attacks and a preparation method thereof. Background Art

[0002] The field of liver disease prevention and treatment has long faced three major technical bottlenecks: low efficiency of ingredient extraction, potential safety hazards of compound compatibility, and unclear multi-component synergistic mechanisms. Traditional plant compositions mostly use a single water extraction or alcohol extraction process, which makes it difficult to take into account the simultaneous and efficient extraction of active ingredients with large polarity differences (such as flavonoid glycosides and cyclopentadiene ether terpenes), and high-temperature and long-term treatment can easily lead to degradation of heat-sensitive components. Existing compound prescriptions often introduce medicinal materials containing aristolochic acid or anthraquinone components (such as Aristolochia guanylica and rhubarb) due to blind pursuit of efficacy, and long-term use poses a risk of liver and kidney damage. In addition, the simple mixing of vitamins and plant ingredients can easily lead to conflicts in physical and chemical properties (such as the Maillard reaction of taurine and phenolic substances), resulting in reduced bioavailability. How to construct an efficient, safe and multi-target synergistic plant composition has become a technical problem that needs to be solved in this field. Summary of the invention

[0003] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0004] In view of the above problems existing in the prior art, the inventor proposes the present invention.

[0005] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide a plant active ingredient composition for resisting liver disease attacks and a preparation method thereof.

[0006] In order to solve the above technical problems, the present invention provides the following technical solution: a plant active ingredient composition for resisting liver disease attacks, comprising the following raw materials in parts by weight:

[0007] 5-15 parts of gentian extract, 3-10 parts of liquorice extract, 5-12 parts of scutellaria extract, 3-8 parts of pulsatilla extract, 2-6 parts of akebia extract, 4-10 parts of plantain extract, 5-12 parts of isatis root extract, 0.1-0.5 parts of vitamin B complex, 1-3 parts of taurine, and 0.5-2 parts of vitamin C;

[0008] The plant extract is prepared by a combined water extraction-alcohol precipitation process, and the extraction solvent of gentian, scutellaria baicalensis and isatis root is a mixture of a low eutectic solvent choline chloride-lactic acid system (molar ratio 1:2) and water (volume ratio 1:1-3).

[0009] As a preferred embodiment of the plant active ingredient composition for preventing liver disease attacks according to the present invention, wherein: the vitamin B complex is composed of vitamin B1, B2, and B6 in a mass ratio of 1:0.8:1.2, and the mass ratio of taurine to vitamin C is 1.5:1 - 2.5:1.

[0010] As a preferred embodiment of the plant active ingredient composition for preventing liver disease attacks according to the present invention, wherein: it further contains 0.1 - 0.3 parts of grape seed proanthocyanidins or wolfberry leaf phenolic compounds.

[0011] As a preferred embodiment of the plant active ingredient composition for preventing liver disease attacks according to the present invention, wherein: 0.05 - 0.1% of silver nanoparticles with a particle size of 20 - 50 nm are added to the eutectic solvent system.

[0012] Use of the plant active ingredient composition for preventing liver disease attacks according to the present invention in the preparation of functional foods or drugs for preventing or treating alcoholic liver injury, drug-induced liver injury, or fatty liver.

[0013] As a preferred embodiment of the use of the plant active ingredient composition for preventing liver disease attacks according to the present invention, wherein: the composition is compounded and used with rice bran aliphatic alcohol in a mass ratio of 10:1 - 5:1.

[0014] A preferred embodiment of the preparation method of the plant active ingredient composition for preventing liver disease attacks includes the following steps:

[0015] (a) Mix gentian, scutellaria baicalensis, and isatis root in a mass ratio of 2:1.5:1, add a mixed solution of choline chloride - lactic acid eutectic solvent and water (material - liquid ratio 1:15 - 25), and perform ultrasonic - assisted extraction at 50 - 60 °C for 30 - 45 minutes, then filter to obtain the first extract;

[0016] (b) Mix pulsatilla chinensis, akebia quinata, and plantago asiatica in a mass ratio of 1:0.6:1.2, then perform reflux extraction with 70% ethanol twice, combine the filtrates and concentrate to a density of 1.1 - 1.2 g / cm 3 , to obtain the second extract;

[0017] (c) Extract licorice alone with a weakly alkaline aqueous solution with a pH of 8 - 9, and enrich the active ingredients through XAD7HP macroporous resin after centrifugation to obtain the third extract;

[0018] (d) Combine the extracts obtained in steps (a) - (c), add the vitamin B complex, taurine, and vitamin C, and obtain the finished product through spray drying.

[0019] As a preferred embodiment of the preparation method of the plant active ingredient composition for preventing liver disease attacks according to the present invention, in step (a), the ultrasonic power is 300-500 W and the frequency is 28-40 kHz.

[0020] As a preferred embodiment of the preparation method of the plant active ingredient composition for preventing liver disease attacks according to the present invention, in step (c), the eluent of the macroporous resin is a 50% ethanol solution, and the elution flow rate is 2-3 BV / h.

[0021] As a preferred embodiment of the preparation method of the plant active ingredient composition for preventing liver disease attacks according to the present invention, in step (d), the inlet air temperature for spray drying is 160-180 °C, and the outlet air temperature is 80-90 °C.

[0022] Beneficial effects of the present invention: The present invention systematically solves the above problems through three strategies of precise ingredient screening - innovative extraction process - collaborative system construction:

[0023] Efficient extraction: The combination of choline chloride-lactic acid deep eutectic solvent and ultrasonic technology is used to improve the yields of key components such as gentiopicroside and baicalin, and increase the polysaccharide retention rate; the selective enrichment of glycyrrhizic acid is achieved through the alkali extraction-resin purification process of licorice, avoiding anthraquinone and aristolochic acid components; the combination of gentiana, scutellaria, and isatis root inhibits the NF-κB inflammatory pathway and reduces the expression levels of TNF-α and IL-6; taurine and VC form a redox buffer system in a ratio of 1.5:1 - 2.5:1, increasing the SOD activity; pulsatilla and plantain seed promote bile acid secretion, accelerate the excretion of lipid toxins, and the combined diuretic effect of akebia quinata reduces the intrahepatic pressure. Finally, it realizes clearing the liver and purging fire, removing dampness and dryness, enhancing immunity, while removing toxins from the animal body, enhancing immunity, enhancing the activity of the liver, and reducing the onset of liver diseases. Detailed implementation manners

[0024] To make the above objects, features, and advantages of the present invention more obvious and understandable, the following detailed description of the specific implementation manners of the present invention is provided in conjunction with the examples of the specification.

[0025] In the following description, many specific details are set forth to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Those skilled in the art can make similar generalizations without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific examples disclosed below.

[0026] Secondly, the so-called "one embodiment" or "embodiment" herein refers to a specific feature, structure, or characteristic that can be included in at least one implementation manner of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it an embodiment that is separate or selectively exclusive of other embodiments.

[0027] Example 1

[0028] This example provides a composition of plant active ingredients for preventing liver disease attacks and a preparation method thereof.

[0029] Raw material ratio: 10 parts of gentian extract, 6 parts of licorice extract, 8 parts of scutellaria extract, 5 parts of pulsatilla extract, 4 parts of akebia quinata extract, 7 parts of plantain seed extract, 9 parts of isatis root extract, 0.3 part of vitamin B complex (B1:B2:B6 = 1:0.8:1.2), 2 parts of taurine, 1.2 parts of VC

[0030] Preparation process:

[0031] Gentian - Scutellaria - Isatis root extraction:

[0032] Mix the raw materials in a mass ratio of 2:1.5:1, add a mixed solution of choline chloride - lactic acid eutectic solvent (molar ratio 1:2) and water (volume ratio 1:2), and the material - liquid ratio is 1:20

[0033] Ultrasonic treatment (power 400W, frequency 35kHz) is carried out at 55°C for 40 minutes, and the extract is obtained by filtration

[0034] Pulsatilla - Akebia quinata - Plantain seed extraction:

[0035] Mix the raw materials in a ratio of 1:0.6:1.2, add 70% ethanol for reflux extraction 2 times (1.5 hours each time), and after combining the filtrates, concentrate under reduced pressure to a density of 1.15 g / cm 3

[0036] Licorice purification:

[0037] The raw materials are extracted with a NaHCO3 solution at pH 8.5 (material - liquid ratio 1:15), centrifuged, loaded onto an XAD7HP resin column, and eluted with a 50% ethanol solution at a flow rate of 2.5 BV / h, and the eluate is collected and concentrated;

[0038] Mixing and drying:

[0039] Combine the three parts of the extracts, and add vitamin B complex, taurine and VC

[0040] Spray drying (inlet air 170°C, outlet air 85°C) to obtain a light yellow powder

[0041] Effect verification:

[0042] Cell experiment: Using the AML - 12 hepatocyte model, after pre - treating with the composition (100 μg / mL), detect the index changes under ethanol - induced injury (200 mM):

[0043] Index Model group Treatment group Improvement rate Cell survival rate 58％ 82％ +41％ SOD activity (U / mg) 12.3 18.7 +52％ MDA (nmol / mg) 4.8 2.1 -56％

[0044] Furthermore, the hydrogen bond donor (lactic acid) and acceptor (chloride ion) in the choline chloride-lactic acid system can disrupt the cellulose structure of the plant cell wall and dissolve flavonoid components through π-π interactions.

[0045] The ultrasonic cavitation effect (400 W, 35 kHz) generates microjets that impact the cell wall, increasing the dissolution rate of gentiopicroside by 3 times (the result of kinetic model fitting).

[0046] Vitamins B1 / B2 / B6 are formulated in a ratio of 1:0.8:1.2 to form a "B vitamin metabolic cycle" in liver cells, promoting the regeneration of coenzyme NADPH.

[0047] The molar ratio of taurine (2 parts) to VC (1.2 parts) is 2:1, which is close to the glutathione redox potential (-240 mV) of cells, effectively scavenging hydroxyl radicals.

[0048] Example 2

[0049] This example provides a composition of plant active ingredients for preventing liver disease attacks and a preparation method.

[0050] Preparation improvement: Add 0.08% silver nanoparticles (particle size 30 nm) to the eutectic solvent in Step 1 of Example 1, and the remaining steps are the same.

[0051] Performance test:

[0052] Stability: The accelerated test (40 °C / RH 75%) shows that after 6 months, the flavonoid retention rate of the silver nanoparticle group is 89%, and that of the control group is 72%.

[0053] Bioavailability: After intragastric administration (200 mg / kg) to SD rats, the area under the plasma concentration-time curve (AUC 0-24 ) of the silver nanoparticle group is 1.8 times that of the normal group.

[0054] Antibacterial effect: The MIC value against common bacteria causing liver abscess (Klebsiella pneumoniae) decreased from 128 μg / mL to 64 μg / mL.

[0055] Furthermore, the surface plasmon resonance effect of silver nanoparticles: Silver particles with a particle size of 30 nm generate local surface plasmon resonance (LSPR) at a wavelength of 550 nm, promoting the photothermal conversion of components such as baicalin and increasing the cell membrane permeability (the transmembrane potential decreases by 25%).

[0056] Reactive oxygen species (ROS) regulation: Silver nanoparticles activate the Nrf2 pathway through slow release of Ag + and upregulate the expression level of HO-1 enzyme by 3.2 times (detected by qPCR), forming a cascade amplification effect with the antioxidant system of the composition.

[0057] Judging from the experimental data:

[0058] After intragastric administration in rats, Cmax increased from 12.3 μg / mL to 19.8 μg / mL, and T1 / 2 was extended to 6.8 h (4.2 h in the normal group);

[0059] The inhibition zone diameter against common pathogens of liver abscess (Klebsiella pneumoniae) expanded from 14 mm to 21 mm (paper disk diffusion method).

[0060] Example 3

[0061] This example provides a plant active ingredient composition for preventing liver disease attacks and a preparation method thereof.

[0062] Formula: 70 parts of basic composition, 10 parts of rice bran aliphatic alcohol, 15 parts of microcrystalline cellulose, 0.8 part of magnesium stearate, 4 parts of hypromellose

[0063] Preparation process:

[0064] The composition and rice bran alcohol are ball-milled and mixed (particle size D90 ≤ 50 μm)

[0065] After mixing with excipients, directly tabletted (tablet weight 500 mg, hardness 8 - 10 kp)

[0066] Animal experiment:

[0067] Model: NAFLD rats induced by high-fat diet (n = 10 / group), administration dose 150 mg / kg / day × 4 weeks

[0068] Results:

[0069]

[0070]

[0071] Furthermore, analysis of the synergistic mechanism:

[0072] Activation of the PPARα / γ dual pathway:

[0073] Rice bran alcohol (the main component is octacosanol) promotes fatty acid β-oxidation by activating PPARα (the activity of CPT1A enzyme increases by 80%);

[0074] Indirubin in Isatis indigotica Fort. activates PPARγ and enhances adiponectin secretion (serum adiponectin level increases from 5.6 μg / mL to 9.2 μg / mL).

[0075] Regulation of the gut-liver axis:

[0076] Akebia polysaccharide (molecular weight 8 - 10 kDa) in the composition promotes the proliferation of Bifidobacterium (16S rRNA sequencing shows a 15-fold increase in abundance) and inhibits the entry of endotoxin (LPS) into the liver;

[0077] Rice bran alkyl alcohol inhibits the intestinal FXR receptor and reduces the enterohepatic circulation load of bile acids.

[0078] Results of animal experiments:

[0079] Reversal of fatty liver: Oil red O staining of liver tissue in high-fat model rats shows that the lipid droplet area is reduced by 72%, and electron microscopy observation shows that the mitochondrial cristae structure is restored to integrity;

[0080] Improvement of glucose metabolism: Fasting blood glucose drops from 9.8 mmol / L to 6.2 mmol / L, and the insulin sensitivity index (ISI) is increased by 2.3 times.

[0081] In summary, the limitations of traditional extraction processes stem from a single understanding of the cell structure of medicinal materials and the polarity of components. The present invention creatively introduces a deep eutectic solvent (DES) system, and its hydrogen bond network characteristics can dissolve both polar (such as polysaccharides) and non-polar components (such as flavonoid glycosides) simultaneously. The dynamic viscosity of choline chloride-lactic acid DES (~200 cP at 25 °C) is higher than that of traditional solvents (such as ethanol viscosity ~1.2 cP), and stronger shear force is generated in the ultrasonic field, increasing the cell wall disintegration efficiency by 2 - 3 times. HPLC-MS analysis shows that this process increases the extraction rate of gentiopicroside in gentiana from 3.4% by the conventional method to 5.8%, and the polysaccharide molecular weight distribution (10 - 50 kDa) is more conducive to immunomodulatory effects.

[0082] Through molecular docking and metabolomics techniques, reveal the synergistic network of key components:

[0083] Gentiopicroside - baicalin complex: The two form a stable dimer through hydrophobic interaction and hydrogen bonding, and the binding energy (-8.7 kcal / mol) to the NF-κB p50 subunit is significantly higher than that of single components (-5.2 kcal / mol), explaining its anti-inflammatory synergistic effect;

[0084] Taurine - VC redox coupling: When the two are in a molar ratio of 1.5:1, the redox potential (Eh) is stabilized at +120 mV, forming a gradient with the cytoplasmic GSH / GSSG system (Eh -150 to -200 mV) to achieve directional scavenging of free radicals.

[0085] Nanosilver modification not only improves bioavailability, and its antibacterial effect highly coincides with the treatment requirements of liver diseases:

[0086] Nanosilver (30 nm) can be targeted to the liver through Kupffer cell phagocytosis and slowly release Ag in the acidic environment of lysosomes +, disrupt bacterial biofilms (CLSM shows an 80% loss of membrane integrity);

[0087] The rice bran alcohol compounding scheme breaks through the limitations of single plant compositions and realizes synchronous regulation of lipid and glucose metabolism through dual activation of the PPAR pathway. Preclinical studies show that its effect of reducing liver fibrosis scores (from F3 to F1) is better than that of single-agent preparations.

[0088] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention, and they should all be covered within the scope of the claims of the present invention.

Claims

1. A plant active ingredient composition for preventing liver disease attacks, characterized in that: It is composed of the following raw materials in parts by mass: Gentiana macrophylla extract 5 - 15 parts, licorice extract 3 - 10 parts, Scutellaria baicalensis extract 5 - 12 parts, Pulsatilla chinensis extract 3 - 8 parts, Akebia quinata extract 2 - 6 parts, Plantago asiatica extract 4 - 10 parts, Isatis indigotica extract 5 - 12 parts, vitamin B complex 0.1 - 0.5 part, taurine 1 - 3 parts, vitamin C 0.5 - 2 parts; Among them, the plant extract is prepared by a combined process of water extraction - alcohol precipitation, and the extraction solvents for Gentiana macrophylla, Scutellaria baicalensis, and Isatis indigotica are a eutectic solvent choline chloride - lactic acid system (molar ratio 1:2) and a mixed solution of water (volume ratio 1:1 - 3).

2. The plant active ingredient composition for preventing liver disease attacks according to claim 1, characterized in that The vitamin B complex is composed of vitamin B1, B2, and B6 in a mass ratio of 1:0.8:1.2, and the mass ratio of taurine to vitamin C is 1.5:1 - 2.5:

1.

3. The plant active ingredient composition for preventing liver disease attacks according to claim 1, characterized in that: It further contains 0.1 - 0.3 part of grape seed proanthocyanidins or phenolic compounds from wolfberry leaves.

4. A plant active ingredient composition for preventing liver disease attacks as described in claim 1, characterized in that: 0.05 - 0.1% of silver nanoparticles with a particle size of 20 - 50 nm are added to the eutectic solvent system.

5. Use of a plant active ingredient composition for preventing liver disease attacks as described in claim 1 in the preparation of a functional food or drug for preventing or treating alcoholic liver injury, drug - induced liver injury, or fatty liver.

6. The application according to claim 5, wherein: The composition is compounded and used with rice bran aliphatic alcohol in a mass ratio of 10:1 - 5:

1.

7. The preparation method of a plant active ingredient composition for preventing liver disease attacks as described in claim 1, characterized in that, It includes the following steps: (a) Mix Gentiana macrophylla, Scutellaria baicalensis, and Isatis indigotica in a mass ratio of 2:1.5:1, add a mixed solution of choline chloride - lactic acid eutectic solvent and water (material - liquid ratio 1:15 - 25), and perform ultrasonic - assisted extraction at 50 - 60 °C for 30 - 45 minutes, then filter to obtain the first extract; (b) Mix Pulsatilla chinensis, Akebia quinata, and Plantago asiatica in a mass ratio of 1:0.6:1.2, and then extract them twice by refluxing with 70% ethanol. Combine the filtrates and concentrate them to a density of 1.1 - 1.2 g / cm 3 , to obtain the second extract; (c) Extract licorice alone with a weakly alkaline aqueous solution with a pH of 8 - 9, and enrich the active ingredients through XAD7HP macroporous resin after centrifugation to obtain the third extract; (d) Combine the extracts obtained in steps (a) - (c), add vitamin B complex, taurine, and vitamin C, and prepare the finished product by spray drying.

8. The preparation method of a plant active ingredient composition for preventing liver disease attacks according to claim 7, characterized in that: In step (a), the ultrasonic power is 300 - 500 W, and the frequency is 28 - 40 kHz.

9. The method of using a composition of plant active ingredients for preventing liver disease attacks and its preparation method according to claim 7, characterized in that: In step (c), the eluent of the macroporous resin is a 50% ethanol solution, and the elution flow rate is 2 - 3 BV / h.

10. A plant active ingredient composition for preventing liver disease attacks and a preparation method thereof according to claim 7, characterized in that: In step (d), the inlet air temperature for spray drying is 160 - 180 °C, and the outlet air temperature is 80 - 90 °C.

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