A traditional Chinese medicine composition for treating vascular cognitive impairment, a preparation method and application thereof

Abstract

This invention discloses a traditional Chinese medicine composition for treating vascular cognitive impairment, its preparation method, and its application, belonging to the field of traditional Chinese medicine technology. It includes the following raw materials: stir-fried Atractylodes macrocephala, Fritillaria thunbergii, Ligusticum chuanxiong, Salvia miltiorrhiza, Curcuma longa, Acorus tatarinowii, Polygala tenuifolia, Cornus officinalis, Cistanche deserticola, Rehmannia glutinosa, Scrophularia ningpoensis, and borneol. Preparation method: First, the volatile oil is extracted by steam distillation and encapsulated with β-cyclodextrin; then, the lipid-soluble components in Salvia miltiorrhiza and Fritillaria thunbergii are extracted with ethanol; finally, the residue is combined with the remaining medicinal materials and decocted in water for extraction. Through the combination of Fritillaria thunbergii and borneol, the dual functions of "clearing inflammation + opening the blood-brain barrier" are achieved, increasing the brain concentration of lipid-soluble components such as tanshinone IIA. It significantly improves the learning and memory abilities of rats with vascular cognitive impairment, with efficacy superior to the Western medicine donepezil, and exhibits synergistic effects on multiple targets such as inhibiting NLRP3 inflammasomes and upregulating BDNF. This invention has a simple process, is suitable for industrial production, and has significant clinical application value.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine composition for treating vascular cognitive impairment, its preparation method and application, and more specifically, to a traditional Chinese medicine compound based on the treatment principle of "tonifying the kidney and resolving phlegm, opening the orifices and unblocking the collaterals," which specifically regulates the function of the neurovascular unit (NVU) to treat vascular dementia (VaD) and vascular mild cognitive impairment (VaMCI) caused by chronic cerebral hypoperfusion (CCH). Background Technology

[0002] Epidemiology and Pathological Mechanisms of Vascular Cognitive Impairment: Vascular cognitive impairment (VCI) is a syndrome ranging from mild cognitive impairment to dementia caused by cerebrovascular lesions, including vascular dementia (VaD) and vascular mild cognitive impairment (VaMCI). According to the World Health Organization, there are more than 50 million VCI patients worldwide, with approximately 10 million new cases each year. The prevalence in the elderly population is as high as 5-10%, making it the second leading cause of dementia after Alzheimer's disease (AD).

[0003] The core pathological mechanism of cerebral infarction (VCI) is chronic cerebral hypoperfusion (CCH), which is a long-term reduction in cerebral blood flow due to stenosis or occlusion of cerebral blood vessels. CCH triggers a series of pathological cascade reactions:

[0004] White matter damage: The deep white matter of the brain is extremely sensitive to ischemia and hypoxia. CCH leads to demyelination of white matter fibers and axonal damage, which manifests as leukoaraiosis on imaging.

[0005] Neurovascular Unit (NVU) Uncoupling: The NVU, composed of neurons, astrocytes, vascular endothelial cells, pericytes, and the basement membrane, is a functional unit that maintains brain homeostasis. CCH disrupts the coordination among the components of the NVU, leading to abnormal blood-brain barrier (BBB) ​​permeability and neurovascular coupling dysregulation.

[0006] Chronic neuroinflammation: Ischemia and hypoxia activate microglia, causing them to polarize towards the pro-inflammatory M1 type, releasing inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), forming a persistent aseptic inflammatory environment. Modern research has found that the NLRP3 inflammasome plays a key role in neuroinflammation in VCI.

[0007] Neuronal apoptosis and synapse loss: The above pathological changes eventually lead to neuronal apoptosis in key brain regions such as the hippocampus and cortex, resulting in decreased synaptic density and reduced expression of brain-derived neurotrophic factor (BDNF), manifesting as progressive memory loss, executive dysfunction, and abnormal mental behavior.

[0008] Limitations of existing Western medicine treatments: Currently, the main Western medicines used in clinical treatment of VCI include: (1) Cholinesterase inhibitors (such as donepezil and rivastigmine): By inhibiting acetylcholinesterase, they increase the concentration of acetylcholine in the synaptic cleft and improve cholinergic neurotransmission. However, these drugs can only temporarily improve cognitive symptoms and cannot prevent the progression of neuronal apoptosis and white matter damage. Long-term use has side effects such as gastrointestinal discomfort and bradycardia (Reference: Kavirajan H, Schneider LS. Efficacy and adverse effects of cholinesterase inhibitors and memantine in vascular dementia: a meta-analysis of randomised controlled trials. Lancet Neurol. 2007;6(9):782-792).

[0009] (2) NMDA receptor antagonists (such as memantine): reduce excitatory neurotoxicity by blocking overactivated NMDA receptors. However, the efficacy of this drug for VCI is limited and it has central side effects such as dizziness and drowsiness. Long-term use may lead to drug resistance (Reference: Dichgans M, Markus HS, Salloway S, et al. Donepezil in patients with subcortical vascular cognitive impairment: a randomised double-blindtrial in CADASIL. Lancet Neurol. 2008;7(4):310-318).

[0010] (3) Brain circulation improvers (such as nimodipine and ginkgo biloba extract): They work by dilating cerebral blood vessels and improving microcirculation, but they have no reversible effect on existing white matter damage and neuronal loss.

[0011] In summary, most existing Western medicines are symptomatic treatments, lacking a holistic intervention on the core pathological mechanisms of VCI—NVU functional uncoupling, chronic neuroinflammation, and lack of neurotrophic factors—resulting in unsatisfactory clinical efficacy.

[0012] The shortcomings of existing Chinese medicine patent technologies: In recent years, research on traditional Chinese medicine (TCM) treatment for ventricular intraepithelial neoplasia (VCI) has received increasing attention. A search of Chinese patent databases revealed several TCM patents related to VCI treatment, but all have certain limitations: (1) Patent CN108478718A “A Traditional Chinese Medicine Composition for Treating Vascular Dementia” (Publication Date: September 4, 2018) The patent discloses a compound formula for tonifying qi and promoting blood circulation, composed of ginseng, astragalus, salvia miltiorrhiza, chuanxiong, angelica sinensis, safflower, etc., prepared by traditional decoction method.

[0013] Problems identified: ① It focuses solely on "tonifying qi and promoting blood circulation," without specifically addressing the chronic inflammation ("phlegm and turbidity" in the brain of VCI patients, and lacks phlegm-resolving and nodule-dispersing drugs. ② It does not include drugs that open the orifices and awaken the mind, making it difficult to cross the blood-brain barrier and exert central effects. ③ The traditional decoction method results in low dissolution rates of fat-soluble components in Danshen (such as tanshinone IIA), leading to significant loss of effective ingredients.

[0014] (2) Patent CN109953907A "A Chinese medicine composition for treating cerebral ischemia and its preparation method" (publication date: July 2, 2019) discloses: This patent discloses a compound containing acorus tatarinowii, polygala tenuifolia, chuanxiong and other drugs that open the orifices and awaken the mind, which is prepared by traditional decoction method.

[0015] Problems: ① Although it contains orifice-opening herbs, the high-temperature decoction process causes a significant loss of volatile oils (such as β-asarone and tetramethylpyrazine) in Acorus tatarinowii and Ligusticum chuanxiong at 100℃, resulting in a substantial reduction in their orifice-opening function. ② The specific mechanism by which the volatile oil components exert their effects across the blood-brain barrier is not elucidated.

[0016] (3) Patent CN110638739A "Cerebroprotective Drug Composition Containing Borneol" (Publication Date: January 3, 2020) The patent discloses a combination of borneol, ginkgo leaf extract, and total saponins of Panax notoginseng for brain protection.

[0017] Problems identified: ① Borneol is used alone or in combination with Western medicine extracts, but not in combination with traditional Chinese medicine expectorants and blood-activating drugs, thus lacking a multi-target synergistic effect. ② Excessive borneol dosage (5-10 parts) may cause side effects such as central nervous system excitation. ③ The synergistic brain entry mechanism of borneol with other drug components has not been elucidated.

[0018] (4) Patent CN111184782A "A Traditional Chinese Medicine Composition for Tonifying the Kidney and Improving Intelligence" (Publication Date: May 22, 2020) The patent discloses a kidney-tonifying compound composed of Rehmannia glutinosa, Cornus officinalis, Cistanche deserticola, and Lycium barbarum.

[0019] Problems identified: ① It purely "tonifies the kidneys and replenishes essence," without considering the "symptoms" of VCI—phlegm and blood stasis obstructing the collaterals, and lacks phlegm-resolving and blood-activating drugs. ② It does not address the problem of the blood-brain barrier's difficulty in crossing, making it difficult for the large molecular components (such as polysaccharides) in kidney-tonifying drugs to enter the brain and exert their effects.

[0020] Technical bottlenecks of existing extraction processes: Besides the formulation and compatibility, the extraction process of traditional Chinese medicine preparations directly affects the retention of active ingredients and the efficacy of the medicine. Existing extraction processes for traditional Chinese medicine mainly have the following problems: (1) Traditional decoction method: Chinese herbal medicine formulas contain components with huge polarity differences, such as fat-soluble tanshinone IIA (log P=3.2), volatile β-asarone (boiling point 217℃), and water-soluble strychnine (log P=-1.5). The traditional "one-pot" decoction method can only effectively extract water-soluble components, the dissolution rate of fat-soluble components is less than 10%, and volatile components are almost completely lost with water vapor.

[0021] (2) Single alcohol extraction: Although it can extract fat-soluble components, ethanol will carry away volatile oil components and the extraction of water-soluble polysaccharide components is insufficient.

[0022] (3) Existing two-step method (alcohol extraction followed by water extraction): Some patents adopt the two-step method of "alcohol extraction followed by water extraction", but the volatile oil is not extracted separately, and the volatile components that open the mind and refresh the spirit will still be lost during the alcohol extraction process.

[0023] In summary, existing Chinese medicine patents have significant shortcomings in both formulation and extraction processes, making it difficult to achieve effective treatment for VCI.

[0024] Therefore, how to develop a traditional Chinese medicine composition for treating vascular cognitive impairment that targets and regulates the function of neurovascular units, as well as its preparation method and application, is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0025] Existing drugs for treating vascular cognitive impairment (VCI) and vascular dementia (VaD) have significant drawbacks: (1) Limitations of Western medicine: Commonly used cholinesterase inhibitors (such as donepezil) and NMDA receptor antagonists (such as memantine) can only temporarily improve neurotransmitter transmission and cannot reverse neuronal apoptosis and white matter damage caused by chronic cerebral hypoperfusion (CCH). They also have significant drug resistance and gastrointestinal side effects.

[0026] (2) Bottlenecks in Traditional Chinese Medicine: Most existing compound Chinese medicine formulas employ the traditional method of "decocting the entire formula in water." However, the core pathology of VCI lies in "phlegm and blood stasis obstructing the collaterals" and "kidney essence deficiency." The key active ingredients required for treatment exhibit vastly different physicochemical properties. Difficulty in penetrating the brain: Tanshinones (diterpenoid quinones) and Fritillaria thunbergii alkaloids (isosteroids), which have neuroprotective effects, are both highly lipid-soluble and poorly water-soluble, and have large molecular weights, making it difficult to cross the blood-brain barrier (BBB). Traditional decoction methods result in most of these components remaining in the dregs and being discarded, with a dissolution rate of less than 10%.

[0027] Volatile: Volatile oils with the effects of opening the orifices and refreshing the mind (such as β-asarone in Acorus tatarinowii and ligustrazine in Ligusticum chuanxiong) are very easy to escape with water vapor during high-temperature decoction, resulting in the complete loss of "medicinal aroma".

[0028] Single mechanism: Existing compound prescriptions mostly focus on tonifying the kidney or promoting blood circulation, lacking a holistic intervention to uncouple the core pathogenesis of VCI—the neurovascular unit (NVU) function, especially lacking specific means to clear the "micro-inflammatory environment (phlegm and turbidity) in the brain".

[0029] In view of this, the present invention provides a traditional Chinese medicine composition based on the method of "tonifying the kidney and resolving phlegm, opening the orifices and unblocking the meridians" and its stepwise targeted extraction process. Through specific formulation and process design, the present invention achieves "precise enrichment" and "targeted delivery" of active ingredients, solving the technical problems of severe loss of effective ingredients and difficulty in crossing the blood-brain barrier in traditional Chinese medicine.

[0030] In view of the above-mentioned deficiencies of the prior art, the present invention aims to provide a traditional Chinese medicine composition based on the treatment principle of "tonifying the kidney and resolving phlegm, opening the orifices and unblocking the meridians" and its stepwise targeted extraction process, to achieve the following technical objectives: By combining Fritillaria thunbergii and borneol, the inflammation of the brain nerves ("phlegm and turbidity") is cleared, and the blood-brain barrier is opened to promote the entry of drugs into the brain. By using a "kidney-tonifying herb group" (such as Rehmannia glutinosa, Cornus officinalis, and Cistanche deserticola), neurotrophic factors such as BDNF are upregulated, and damaged neurons are repaired. By using a "three-step targeted extraction process" (volatile oil distillation → alcohol extraction of fat-soluble components → water extraction of polysaccharides), the precise enrichment of components with different polarities is achieved, maximizing the retention of effective components; By synergistically targeting multiple targets (anti-inflammatory, antioxidant, neurotrophic, and improved circulation), a holistic intervention on the core pathological mechanisms of VCI can be achieved.

[0031] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A traditional Chinese medicine composition for treating vascular cognitive impairment, made from the following raw materials in parts by weight: 10-20 parts of stir-fried Atractylodes macrocephala, 10-20 parts of Fritillaria thunbergii, 10-20 parts of Ligusticum chuanxiong, 15-25 parts of Salvia miltiorrhiza, 5-15 parts of Curcuma longa, 10-20 parts of Acorus tatarinowii, 5-15 parts of Polygala tenuifolia, 15-25 parts of Cornus officinalis, 8-15 parts of Cistanche deserticola, 8-15 parts of Rehmannia glutinosa (processed), 8-15 parts of Scrophularia ningpoensis, and 0.5-2 parts of borneol.

[0032] Formula Mechanism: Among them, the principal drug (core treatment) is: 10-20 parts of stir-fried Atractylodes macrocephala and 10-20 parts of Fritillaria thunbergii. Assistant herbs (to aid circulation): Ligusticum chuanxiong 10-20 parts, Salvia miltiorrhiza 15-25 parts, Curcuma longa 5-15 parts, Acorus tatarinowii 10-20 parts; Adjuvant herbs (for nerve repair): Polygala tenuifolia 5-15 parts, Cornus officinalis 15-25 parts, Cistanche deserticola 8-15 parts, Rehmannia glutinosa 8-15 parts, Scrophularia ningpoensis 8-15 parts; Targeted drug: 0.5-2 parts of borneol.

[0033] Furthermore, the weight ratio of Fritillaria thunbergii to Salvia miltiorrhiza is 1:(1.2-1.8), and the weight of borneol is 0.8-1.5 parts.

[0034] The beneficial effects of adopting the above-mentioned further technical solution are as follows: This ratio optimizes the synergistic anti-inflammatory effect of fritillary acetylcholine A and tanshinone IIA in vivo. The dosage range of borneol is optimized based on the blood-brain barrier opening experiment. Below 0.5 parts, the opening effect is not obvious; above 2 parts, it may cause central nervous system excitation side effects; 0.8-1.5 parts yield the best effect.

[0035] Furthermore, it is made from the following raw materials in parts by weight: 15 parts stir-fried Atractylodes macrocephala, 15 parts Fritillaria thunbergii, 15 parts Ligusticum chuanxiong, 20 parts Salvia miltiorrhiza, 10 parts Curcuma longa, 15 parts Acorus tatarinowii, 10 parts Polygala tenuifolia, 20 parts Cornus officinalis, 12 parts Cistanche deserticola, 12 parts Rehmannia glutinosa, 12 parts Scrophularia ningpoensis, and 1 part Borneol.

[0036] Furthermore, in the traditional Chinese medicine composition, the content of tanshinone IIA is 1.68-3.12 mg / g, the content of β-asarone is 1.25-2.45 mg / g, and the content of strychnoside is 6.8-11.5 mg / g.

[0037] This invention also provides a method for preparing a traditional Chinese medicine composition for treating vascular cognitive impairment, comprising the following steps: (1) Extraction of volatile oil: Take Curcuma longa, Acorus tatarinowii and Ligusticum chuanxiong, crush them, add water and extract by steam distillation, collect the volatile oil, and use β-cyclodextrin to encapsulate it to obtain volatile oil inclusion complex; the aqueous solution after distillation is used as aromatic water for later use; the residue is drained to obtain residue I; (2) Alcohol extraction: Take Salvia miltiorrhiza and Fritillaria thunbergii, crush them, add 75%-85% ethanol solution, heat and extract at 78-82℃, combine the extracts, recover the ethanol under reduced pressure until there is no alcohol taste, concentrate to obtain alcohol extract; drain the residue to obtain residue II; (3) Water extraction: Mix the dregs I from step (1), the dregs II from step (2), stir-fried Atractylodes macrocephala, Polygala tenuifolia, Cornus officinalis, Cistanche deserticola, Rehmannia glutinosa and Scrophularia ningpoensis, add the aromatic water from step (1), and add purified water until the total solvent volume is 6-10 times the total weight of the medicinal materials and dregs in this step, decoct, combine the decoctions, and concentrate to obtain water extract; (4) Molding: Mix the alcohol extract from step (2) and the water extract from step (3) evenly, dry, granulate to obtain granules; mix the volatile oil inclusion complex from step (1) with borneol, add it evenly to the granules by spraying, and granulate to obtain the final product.

[0038] Further, in step (1), the powder is pulverized to 20-40 mesh, 6-10 times the amount of water is added, and steam distillation is used to extract for 3-5 hours. The weight ratio of β-cyclodextrin to volatile oil is (5-8):1. In step (2), take Salvia miltiorrhiza and Fritillaria thunbergii, pulverize them to 20-40 mesh, add 5-8 times the amount of ethanol solution, heat and extract 2-3 times, each extraction time is 1.0-2.0 hours; concentrate to 60℃ and measure the relative density to be 1.10-1.20 of the ethanol extract. In step (3), the water extract is decocted at 100℃ 2-3 times, with each decoction lasting 0.5-1.5 hours, and concentrated to a relative density of 1.15-1.25 at 60℃. In step (4), the product is dried at 60-80℃ until the moisture content is ≤5%, and then granulated to 10-20 mesh.

[0039] Preferably, the extraction time using steam distillation in step (1) is 4 hours.

[0040] Preferably, in step (1), the mass ratio of β-cyclodextrin to volatile oil is 6:1.

[0041] Preferably, the concentration of ethanol in step (2) is 80% (v / v).

[0042] Preferably, the extraction time in step (2) is 1.5 hours each time.

[0043] Preferably, the simmering time in step (3) is 1 hour each time.

[0044] Preferably, in step (1), the amount of water added is 8 times the weight of the medicinal material, in step (2) the amount of alcohol added is 6 times the weight of the medicinal material, and in step (3) the amount of water added is 8 times the weight of the medicinal material.

[0045] Preparation method mechanism: Core innovative process: Physicochemical property-guided "three-step targeted extraction method": This invention abandons the conventional "one-pot cooking" approach and, based on the differences in polarity and volatility of the active pharmaceutical ingredients, designs the following inseparable process flow: Step (1): Extraction of volatile neuroactive components (targeting Curcuma longa, Acorus tatarinowii, and Ligusticum chuanxiong): Procedure: Take Curcuma longa, Acorus tatarinowii, and Ligusticum chuanxiong, pulverize them to 20-40 mesh, add 6-10 times (preferably 8 times) of water, and extract by steam distillation for 3-5 hours (preferably 4 hours). Collect the volatile oil (yield approximately 1.2%), and encapsulate it with β-cyclodextrin at a ratio of cyclodextrin:volatile oil = (5-8):1 (preferably 6:1, by weight). The distilled aqueous solution is used as aromatic water for later use. Drain the residue to obtain residue I.

[0046] Principle: The core components of these three herbs—β-Asarone, Ligustrazine, and Turmerone—are all volatile small molecules. Traditional decoction would cause them to be rapidly lost through evaporation with steam at 100°C. This step utilizes the principle of oil-water separation to enrich them in the volatile oil phase.

[0047] Special treatment: The collected volatile oils must be immediately solidified using β-cyclodextrin inclusion complex technology to prevent secondary volatilization during formulation storage. The hydrophobic cavities of β-cyclodextrin can encapsulate the volatile oil molecules, forming stable inclusion complexes.

[0048] Step (2): Alcohol precipitation extraction of lipid-soluble anti-inflammatory / vascular protective components (for Danshen and Fritillaria thunbergii): Procedure: Take Salvia miltiorrhiza and Fritillaria thunbergii, pulverize them to 20-40 mesh, add 5-8 times (preferably 6 times) of 75%-85% (v / v) ethanol (preferably 80%), and heat to extract 2-3 times at ethanol reflux temperature (78-82℃), each time for 1.0-2.0 hours (preferably 1.5 hours). Combine the extracts, recover the ethanol under reduced pressure until there is no alcohol odor, and concentrate to an alcohol extract with a relative density of 1.10-1.20 (measured at 60℃, preferably 1.15); drain the residue to obtain Residue II.

[0049] Principle: Tanshinone IIA and Tanshinone I in Salvia miltiorrhiza are extremely lipophilic molecules (logP>3) and have very low solubility in water; Peimine A and Peiminine B in Fritillaria thunbergii are also alkaloids, and their alcohol solubility is much better than their water solubility.

[0050] Key parameters: Ethanol concentration is limited to the range of 75%-85%. Studies have shown that ethanol concentrations below 60% lead to the dissolution of a large number of water-soluble impurities (such as starch and pectin), hindering the desorption of fat-soluble components; concentrations above 90% lead to cell wall dehydration and shrinkage, which is not conducive to solvent penetration. This concentration range is the "golden window" for the extraction of this component.

[0051] Step (3): Decoction extraction of water-soluble neurotrophic components (for tonifying medicinal materials and residues from previous treatments): Procedure: Mix dregs I and II with the remaining medicinal materials (fried Atractylodes macrocephala, Polygala tenuifolia, Cornus officinalis, Cistanche deserticola, Rehmannia glutinosa, and Scrophularia ningpoensis), add the aromatic water from step (1) and 6-10 times (preferably 8 times) of water, and decoct at 100°C 2-3 times, each time for 0.5-1.5 hours (preferably 1 hour). Combine the decoctions and concentrate to a water extract with a relative density of 1.15-1.25 (measured at 60°C, preferably 1.20).

[0052] Principle: The effective components of tonifying drugs are mostly phenylethanoid glycosides (such as echinacoside and verbascoside), iridoid glycosides (such as strychnoside), and polysaccharides (such as Rehmannia glutinosa polysaccharides). These components are highly polar, heat-resistant, and have the highest extraction efficiency through water extraction. By co-decoctioning the residue from the previous step, the advantage of cell wall disruption during alcohol extraction can be further utilized to dissolve the remaining water-soluble components.

[0053] Step (4): Molding: Mix the alcohol extract from step (2) and the water extract from step (3) evenly, dry them at 60-80℃ (preferably 70℃) until the water content is ≤5%, and granulate them to 10-20 mesh (preferably 15 mesh); mix the volatile oil inclusion complex from step (1) with borneol, and then add it evenly to the granules by spraying, and granulate to obtain the final product.

[0054] The present invention also provides the application of the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method in the preparation of a drug for treating vascular cognitive impairment, wherein the vascular cognitive impairment includes vascular dementia and vascular mild cognitive impairment, and the vascular cognitive impairment is caused by chronic cerebral hypoperfusion.

[0055] Furthermore, the drug formulation is one of granules, capsules, tablets, pills, or oral liquid; the drug is prepared into a unit formulation, and the effective extract contained in each unit is equivalent to 18-28 grams of raw medicinal material.

[0056] Furthermore, when the preparation is in the form of granules, each bag contains 10-15 grams, which is equivalent to 18-28 grams of raw medicinal materials per bag.

[0057] Furthermore, the drug is administered orally, 2-3 times daily, 10-15 grams each time, equivalent to 18-28 grams of raw medicinal materials, for 4-12 weeks.

[0058] Furthermore, the vascular cognitive impairment is caused by chronic cerebral hypoperfusion due to stenosis or occlusion of both common carotid arteries, and patients exhibit progressive memory loss, executive dysfunction, inattention, and abnormal mental and behavioral characteristics.

[0059] Furthermore, the traditional Chinese medicine composition synergistically regulates the function of the neurovascular unit, exerting its therapeutic effect through the following mechanisms: Borneol transiently and pulsatilely opens the blood-brain barrier by temporarily downregulating the expression of tight junction proteins Claudin-5 and Occludin, promoting the entry of lipid-soluble drug components into the brain. At the same time, the kidney-tonifying and blood-activating herbs in the formula (such as Rehmannia glutinosa, Cornus officinalis, and Ligusticum chuanxiong) can promote the repair of cerebral microvascular endothelial cells after drug absorption, restore and maintain the long-term integrity of the blood-brain barrier, thereby blocking the continuous infiltration of peripheral inflammatory factors. Fritillary acetylene from Fritillaria thunbergii specifically inhibits the assembly and activation of the NLRP3 inflammasome, blocking the neuroinflammatory cascade. The phenylethanoid glycosides in Cornus officinalis, Rehmannia glutinosa, and Cistanche deserticola activate the PI3K / Akt / CREB signaling pathway, upregulate the expression of brain-derived neurotrophic factor (BDNF), and promote neuronal survival and synaptic plasticity. Danshen and Chuanxiong improve cerebral microcirculation, increase cerebral blood flow, and provide blood oxygen support for nerve repair.

[0060] Formulation mechanism and drug-drug interaction: This invention, based on the holistic view of the "neurovascular unit (NVU)," constructs a rigorous "principal, assistant, adjuvant, and guide" pharmacological network: The first mechanism: reversible opening of the blood-brain barrier (BBB) ​​mediated by borneol (“biological key” effect). Dynamic opening and closing mechanism: This 'opening' is not a destructive, continuous opening, but rather a 'time window' opening that matches the pharmacokinetic characteristics. Borneol is rapidly absorbed after administration (reaching peak concentration in about 1-2 hours) and opens the blood-brain barrier, guiding active ingredients such as tanshinone IIA into the brain; as borneol is rapidly metabolized, the blood-brain barrier closes again under the nourishment of tonifying drugs, achieving targeted delivery while avoiding the risk of long-term exposure to brain tissue.

[0061] Technical challenge: Large molecule drugs such as tanshinone IIA and fritillary acetonide A have difficulty passing through tight junctions in the body's biological barrier (BBB).

[0062] This invention represents a breakthrough: Borneol (whose main component is dextrorotatory borneol) acts as an "adjuvant" and possesses a unique bidirectional regulatory effect. Modern pharmacology has confirmed that borneol can reversibly open the blood-brain barrier by downregulating the expression of tight junction proteins (such as Claudin-5 and Occludin) and inhibiting the efflux pump function of P-glycoprotein (P-gp).

[0063] Synergistic effect: Borneol acts as a "key," opening a pathway to brain tissue for other active ingredients in the formula that are difficult to penetrate. Experimental data shows that after being combined with borneol, the distribution concentration of tanshinone IIA in the hippocampus increased by more than 3.7 times.

[0064] The second mechanism: the anti-inflammatory blocking effect of Fritillaria thunbergii and Atractylodes macrocephala ("scavenger effect"). Pathological correspondence: VCI patients have persistent aseptic inflammation in their brains, abnormal activation of microglia (M1 polarization), and release of factors such as TNF-α and IL-1β, which is considered to be "phlegm-toxin-induced disease" in traditional Chinese medicine.

[0065] Pharmacological mechanism: Fritillary acetylcholine A from Fritillaria thunbergii has been shown to specifically inhibit phosphorylation of the NF-κB signaling pathway, blocking the assembly of the NLRP3 inflammasome, thereby inhibiting the transformation of microglia into the pro-inflammatory M1 phenotype. Atractylodes macrocephala (fried) reduces peripheral LPS (lipopolysaccharide) entering the brain by regulating the gut microbiota (brain-gut axis), cutting off the source of inflammation. The combined use of these two herbs clears "phlegm (inflammation)" from the brain and protects the integrity of white matter fibers.

[0066] The third mechanism: the nerve remodeling effect of kidney-tonifying and blood-activating drugs ("fertilization effect"). Pathological correspondence: Chronic ischemia leads to neuronal apoptosis and synapse loss, i.e., "empty marrow sea".

[0067] Pharmacological mechanism: Rehmannia glutinosa and Cistanche deserticola are rich in neurotrophic factor mimics (such as echinacoside), which can activate the PI3K / Akt / CREB signaling pathway and significantly upregulate the expression of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF). Ligusticum chuanxiong and Salvia miltiorrhiza provide blood oxygen support for nerve regeneration by improving microcirculation. The two work synergistically to promote the proliferation and differentiation of neural stem cells in the hippocampus and repair damaged memory networks.

[0068] In summary, the formulation and process of this invention are related as a "lock and key." The stepwise extraction process preserves both the "key" (volatile oil) and the "ammunition" (lipid-soluble components). Borneol in the formulation guides the "key" through the blood-brain barrier, while Fritillaria thunbergii and kidney-tonifying herbs act as the "ammunition" to enter the brain, clear inflammation, and repair nerves. This three-pronged strategy of "process preservation + borneol guidance + compound treatment" constitutes the fundamental inventiveness that distinguishes this invention from existing technologies.

[0069] The beneficial effects of this invention are: Compared with the prior art, the present invention has the following significant advantages: (1) Ingredient advantages brought about by process innovation: This invention employs a three-step targeted extraction process: "volatile oil distillation → alcohol extraction of fat-soluble components → water extraction of polysaccharides," achieving precise enrichment of components with different polarities. Experimental data show that: The content of tanshinone IIA in the preparation was 16 times higher than that of the traditional decoction method (2.45 mg / g vs 0.15 mg / g). The β-asarone content increased 23 times (1.88 mg / g vs 0.08 mg / g); The yield of volatile oils increased to 1.2%, while the yield of traditional processes was almost zero; (2) The efficacy advantages brought about by innovative drug combination: This invention is the first to use the "Zhejiang Fritillaria-Borneol" drug pair, which achieves the dual functions of "clearing inflammation + opening the barrier": Animal experiments showed that it increased the brain concentration of lipid-soluble components such as tanshinone IIA by 3.7 times (45.6 ng / g vs 12.4 ng / g). Fritillaria thunbergii specifically inhibits the NLRP3 inflammasome, reducing TNF-α levels in the hippocampus by 62% (72.3 pg / mg vs 189.5 pg / mg); (3) Therapeutic advantages: Morris water maze test results showed that the cognitive improvement effect of the composition of this invention was superior to that of the positive control drug donepezil: Escape latency: 25.6±3.1 s (this invention) vs 28.4±3.5 s (donepezil), P<0.05; Number of platform crossings: 4.8±0.9 times (this invention) vs 4.2±0.8 times (donepezil); (4) Mechanism advantages: This invention achieves synergistic effects across multiple targets: anti-inflammatory, antioxidant, and neurotrophic. Hippocampal tissue showed a 62% reduction in TNF-α and a 63% reduction in IL-6. The lipid peroxidation product MDA decreased by 54%; BDNF protein expression recovered to 88% of normal. (5) Security advantages: The compositions of this invention are all medicinal and edible herbs or commonly used traditional Chinese medicines in clinical practice, and no obvious toxic side effects have been observed with long-term administration. Acute toxicity test: Maximum tolerated dose (MTD) > 100 times the clinically equivalent dose; Long-term administration (90 days): Liver and kidney function indicators were normal, and no abnormalities were found in the pathological examination of major organs; (6) Advantages of industrialization: The process and equipment of this invention are simple, suitable for large-scale production, and the product quality is stable and controllable. The RSD of tanshinone IIA content was <5% in 3 batches of samples; β-Asaurophene content RSD < 4%; The volatile oil yield RSD is less than 6%.

[0070] In summary, this invention, through the combination of Fritillaria thunbergii and borneol, achieves the dual functions of "clearing inflammation and opening the blood-brain barrier," increasing the brain concentration of lipid-soluble components such as tanshinone IIA by 3.7 times. Animal experiments show that this composition significantly improves the learning and memory abilities of rats with vascular cognitive impairment, with efficacy superior to the Western medicine donepezil, and its mechanism of action involves synergistic action targeting multiple targets, including inhibiting the NLRP3 inflammasome and upregulating BDNF. This invention has a simple process, is suitable for industrial production, and has significant clinical application value.

[0071] Compared with existing patents, this application has the following advantages: (1) Patent CN108478718A “A Traditional Chinese Medicine Composition for Treating Vascular Dementia” (Publication Date: September 4, 2018) The patent discloses a compound formula for tonifying qi and promoting blood circulation, composed of ginseng, astragalus, salvia miltiorrhiza, chuanxiong, angelica sinensis, safflower, etc., prepared by traditional decoction method.

[0072] The difference between this invention and the present invention is that the present invention introduces Fritillaria thunbergii to specifically inhibit the NLRP3 inflammasome, clearing "phlegm and turbidity" (neuritis) in the brain; it is combined with borneol to open the blood-brain barrier and promote drug entry into the brain; and it adopts a three-step targeted extraction process, which increases the content of tanshinone IIA by 16 times compared with the water decoction method.

[0073] (2) Patent CN109953907A "A Chinese medicine composition for treating cerebral ischemia and its preparation method" (publication date: July 2, 2019) discloses: This patent discloses a compound containing acorus tatarinowii, polygala tenuifolia, chuanxiong and other drugs that open the orifices and awaken the mind, which is prepared by traditional decoction method.

[0074] The difference between this invention and the present invention is that the present invention uses steam distillation to extract volatile oil separately and uses β-cyclodextrin to encapsulate and prevent volatilization, thereby increasing the yield of volatile oil to 1.2% (compared to nearly 0% by the traditional decoction method); it also clearly proposes that borneol activates the molecular mechanism of BBB by downregulating Claudin-5 and Occludin.

[0075] (3) Patent CN110638739A "Cerebroprotective Drug Composition Containing Borneol" (Publication Date: January 3, 2020) The patent discloses a combination of borneol, ginkgo leaf extract, and total saponins of Panax notoginseng for brain protection.

[0076] The difference between this invention and the present invention is that the dosage of borneol is precisely controlled at 0.5-2 parts (preferably 1 part), which can both open the BBB and avoid side effects; borneol is combined with specific drugs such as Fritillaria thunbergii and Salvia miltiorrhiza to achieve a synergistic brain-entry effect of "key (borneol) + ammunition (lipid-soluble anti-inflammatory components)," which has not been revealed in the prior art.

[0077] (4) Patent CN111184782A "A Traditional Chinese Medicine Composition for Tonifying the Kidney and Improving Intelligence" (Publication Date: May 22, 2020) The patent discloses a kidney-tonifying compound composed of Rehmannia glutinosa, Cornus officinalis, Cistanche deserticola, and Lycium barbarum.

[0078] The difference between this invention and the present invention is that the present invention adopts a "treating both the symptoms and the root cause" strategy, which has both Fritillaria thunbergii and borneol to "treat the symptoms" (resolve phlegm, open the orifices, and reduce inflammation and enter the brain), and Rehmannia glutinosa and Cornus officinalis to "treat the root cause" (tonify the kidneys and replenish essence, and regulate BDNF). Furthermore, by using borneol to open the BBB, the effective components of the kidney-tonifying medicine can also enter the brain. Attached Figure Description

[0079] Figure 1 Bar chart showing the escape latency period for each experimental group in the Morris water maze test.

[0080] Figure 2 This is a schematic diagram illustrating the mechanism of action of the traditional Chinese medicine composition of the present invention.

[0081] Figure 3 This is a comparison of HPLC fingerprints of formulations produced using different processes.

[0082] Figure 4 The bar chart shows the relative expression levels of BDNF, NLRP3, and Claudin-5 proteins in the hippocampus of rats in the sham-operated group, model group, Example 1 group, and Comparative Example 2 group. Detailed Implementation

[0083] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0084] Example 1 A method for preparing a traditional Chinese medicine composition for treating vascular cognitive impairment includes the following steps: Weigh out the following ingredients: 150g of stir-fried Atractylodes macrocephala, 150g of Fritillaria thunbergii, 150g of Ligusticum chuanxiong, 200g of Salvia miltiorrhiza, 100g of Curcuma longa, 150g of Acorus tatarinowii, 100g of Polygala tenuifolia, 200g of Cornus officinalis, 120g of Cistanche deserticola, 120g of Rehmannia glutinosa (processed), 120g of Scrophularia ningpoensis, and 10g of Borneol; (1) Extraction of volatile oils: Take Curcuma longa, Acorus tatarinowii, and Ligusticum chuanxiong, grind them to 20 mesh, add 8 times the amount of water (3200 mL in total), and extract by steam distillation for 4 hours. Collect 4.8 g of volatile oil (yield of about 1.2%), and encapsulate it with 28.8 g of β-cyclodextrin to obtain 32.5 g of volatile oil inclusion complex. Use about 3000 mL of the distilled aqueous solution as aromatic water for later use. Drain the residue to obtain residue I. (2) Alcohol extraction: Take Salvia miltiorrhiza and Fritillaria thunbergii, grind them to 20 mesh, add 6 times the amount of 80% (v / v) ethanol solution, heat and extract twice at reflux temperature of 80℃, each extraction time is 1.5 hours; combine the extracts, recover the ethanol under reduced pressure until there is no alcohol odor, concentrate to 60℃ to obtain an alcohol extract with a relative density of 1.15 of about 280g; drain the residue to obtain residue II; (3) Water extraction: Mix the dregs I from step (1), the dregs II from step (2), stir-fried Atractylodes macrocephala, Polygala tenuifolia, Cornus officinalis, Cistanche deserticola, Rehmannia glutinosa, and Scrophularia ningpoensis. Add about 3000 mL of aromatic water from step (1) and add about 4200 mL of water. Decoction twice at 100°C for 1 hour each time. Combine the decoctions and concentrate to about 900 g of water extract with a relative density of 1.20 at 60°C. (4) Molding: Mix 280g of alcohol extract from step (2) and 900g of water extract from step (3) evenly, dry at 70°C to a moisture content of 4.5%, granulate to 15 mesh, and obtain approximately 800g of granules; mix 33.6g of volatile oil inclusion complex from step (1) with 10g of borneol, and then evenly add the mixture to the granules by spraying, and granulate to obtain approximately 850g of the product.

[0085] The contents of tanshinone IIA and strychnine were determined by HPLC, the contents of β-asarone were determined by GC, and the moisture content was determined by drying method (moisture determination method of Chinese Pharmacopoeia). The yield of volatile oil was calculated by volume / weight method. The contents of tanshinone IIA, β-asarone, and strychnine in this traditional Chinese medicine composition were 2.45±0.12 mg / g, 1.88±0.09 mg / g, 9.8±0.5 mg / g, moisture content: 4.5%, and volatile oil yield: 1.2%.

[0086] Example 2 1. Prescription: Weigh out the following ingredients: 100g of stir-fried Atractylodes macrocephala, 100g of Fritillaria thunbergii, 100g of Ligusticum chuanxiong, 150g of Salvia miltiorrhiza, 50g of Curcuma longa, 100g of Acorus tatarinowii, 50g of Polygala tenuifolia, 150g of Cornus officinalis, 80g of Cistanche deserticola, 80g of Rehmannia glutinosa (processed), 80g of Scrophularia ningpoensis, and 5g of Borneol. 2. Preparation: (1) Extraction of volatile oils: Take 50g of Curcuma longa, 100g of Acorus tatarinowii, and 100g of Ligusticum chuanxiong, grind them to 20 mesh, add 8 times the amount of water (2000mL in total), and extract by steam distillation for 4 hours. Collect about 3.0g of volatile oil, encapsulate it with 18.0g of β-cyclodextrin, and dry it to obtain about 20.5g of volatile oil inclusion complex. Use about 1800mL of the distilled aqueous solution as aromatic water for later use. Drain the residue to obtain residue I. (2) Alcohol extraction: Take 150g of Salvia miltiorrhiza and 100g of Fritillaria thunbergii, grind them to 20 mesh, add 6 times the amount of 80% (v / v) ethanol solution (total 1500mL), and heat to extract twice at reflux temperature of 80℃, each extraction time being 1.5 hours; combine the extracts, recover the ethanol under reduced pressure until there is no alcohol odor, concentrate to about 200g of ethanol extract with a relative density of 1.15 measured at 60℃; drain the residue to obtain residue II; (3) Water extraction: Mix the dregs I from step (1), the dregs II from step (2), 100g of stir-fried Atractylodes macrocephala, 50g of Polygala tenuifolia, 150g of Cornus officinalis, 80g of Cistanche deserticola, 80g of Rehmannia glutinosa, and 80g of Scrophularia ningpoensis. Add about 1800mL of aromatic water from step (1) and add about 2600mL of water. Decoction twice at 100℃, each time for 1 hour. Combine the decoctions and concentrate to about 550g of water extract with a relative density of 1.20 measured at 60℃. (4) Molding: Mix 200g of alcohol extract from step (2) and 550g of water extract from step (3) evenly, dry at 70°C to a moisture content of 4.5%, granulate to 15 mesh, and obtain approximately 450g of granules; mix 20.5g of volatile oil inclusion complex from step (1) with 5g of borneol, and then evenly add the mixture to the granules by spraying, and granulate to obtain approximately 475g of the product.

[0087] 3. Quality Inspection: Tanshinone IIA content: 1.68±0.08 mg / g; β-Asaurophene content: 1.25±0.06 mg / g; Strychnoside content: 6.8±0.4 mg / g; 4. Efficacy testing: A VCI rat model (n=12) was established using the 2-VO method. The Morris water maze test was performed 30 days after drug administration. Escape latency: 29.8 ± 3.5 s; Number of platform crossings: 3.9 ± 0.7 times; It was significantly improved compared to the model group (56.8±5.4 s) (P<0.01), but slightly inferior to Example 1 (25.6±3.1 s).

[0088] 5. Conclusion: This demonstrates that the lower limit formulation of the present invention still has significant therapeutic effects, but the therapeutic effects are slightly lower than those of the formulation in Example 1.

[0089] Example 3 1. Prescription: 200g of stir-fried Atractylodes macrocephala, 200g of Fritillaria thunbergii, 200g of Ligusticum chuanxiong, 250g of Salvia miltiorrhiza, 150g of Curcuma longa, 200g of Acorus tatarinowii, 150g of Polygala tenuifolia, 250g of Cornus officinalis, 150g of Cistanche deserticola, 150g of Rehmannia glutinosa (processed), 150g of Scrophularia ningpoensis, and 20g of Borneol.

[0090] 2. Preparation: The process flow and process parameters are the same as in Example 1, except that the dosage of other raw materials, except for the medicinal materials, is adjusted according to the proportions in Example 1.

[0091] 3. Quality Inspection: Tanshinone IIA content: 3.12±0.15 mg / g; β-Asaurophene content: 2.45±0.12 mg / g; Strychnoside content: 11.5±0.6 mg / g; 4. Efficacy testing: Escape latency: 24.1 ± 3.2 s; Number of platform crossings: 5.1 ± 0.9 times; There was no significant difference from Example 1 (25.6 ± 3.1 s) (P > 0.05).

[0092] 5. Cost Analysis: Raw material costs increased by about 30%, but the therapeutic effect did not improve significantly.

[0093] 6. Conclusion: This demonstrates that Example 1 is the most cost-effective solution, and that excessively increasing the dosage of medicinal materials cannot further improve the therapeutic effect.

[0094] Comparative Example 1: Traditional whole-formula decoction process (process comparison): 1. Prescription: Same as Example 1.

[0095] 2. Preparation: All 11 medicinal materials except borneol were mixed, pulverized, and passed through a 20-mesh sieve. They were then decocted twice with 8 times the amount of water, each time for 1.5 hours. The filtrate was concentrated to a relative density of 1.10-1.20 (measured at 60℃) to form a paste. Borneol was dissolved in 5 times the amount of 95% ethanol solution and then sprayed into the paste.

[0096] 3. Quality Inspection: Tanshinone IIA content: 0.15±0.03 mg / g (only 6% of that in Example 1); β-Asauroyl ether content: 0.08±0.01 mg / g (only 4% of that in Example 1); Strychnoside content: 9.2 ± 0.5 mg / g (equivalent to Example 1); 4. Efficacy testing: Escape latency: 45.2 ± 4.8 s; Number of platform crossings: 2.1 ± 0.6 times; The therapeutic effect was significantly weaker than that of Example 1 (P<0.01).

[0097] 5. Conclusion: Traditional decoction methods result in significant loss of fat-soluble and volatile components, greatly reducing the efficacy of the medicine.

[0098] Comparative Example 2: Group lacking "symptomatic treatment" drugs (Formula comparison - lacking Fritillaria thunbergii and borneol): 1. Prescription: Based on Example 1, remove 150g of Fritillaria thunbergii and 10g of borneol, while keeping the other ingredients and dosages unchanged.

[0099] 2. Preparation: The process flow and parameters are the same as in Example 1, except that the dosage of other raw materials is adjusted according to the proportions in Example 1. (The borneol encapsulation step is omitted because there is no borneol; and only salvia miltiorrhiza is extracted with alcohol because there is no fritillaria thunbergii).

[0100] 3. Quality Inspection: Tanshinone IIA content: 2.38±0.11 mg / g (the content of the preparation is similar to that in Example 1); Intracranial tanshinone IIA concentration: 12.4 ± 1.8 ng / g (only 27% of that in Example 1); 4. Efficacy testing: Escape latency: 38.5 ± 4.2 s; Number of platform crossings: 2.8 ± 0.7 times; Hippocampal TNF-α: 158.4±13.5 pg / mg (poor inflammation control); 5. Conclusion: After removing Fritillaria thunbergii and borneol, although the content of tanshinone IIA in the preparation was normal, the concentration in the brain decreased significantly (due to the lack of borneol to activate the BBB), and neuroinflammation was not effectively controlled (due to the lack of Fritillaria thunbergii inhibiting NLRP3), resulting in a significant decrease in efficacy. This demonstrates that the "Fritillaria thunbergii-borneol" drug pair plays a core role in this invention.

[0101] Comparative Example 3: Group lacking "root cause treatment" herbs (Formula comparison - lacking Rehmannia glutinosa and Cornus officinalis): 1. Prescription: Based on Example 1, remove 120g of Rehmannia glutinosa and 200g of Cornus officinalis.

[0102] 2. Preparation: The process flow and process parameters are the same as in Example 1, except that the dosage of other raw materials, except for the medicinal materials, is adjusted according to the proportions in Example 1.

[0103] 3. Efficacy testing: Escape latency: 32.1 ± 3.8 s; Number of platform crossings: 3.5 ± 0.8 times; Hippocampal BDNF protein expression: 0.45±0.05 (only recovered to 45% of normal); 4. Conclusion: Although the inflammation was controlled after removing the kidney-tonifying medication, the upregulation of neurotrophic factor (BDNF) was insufficient, weakening nerve repair capacity and resulting in a less effective treatment compared to the full-formula approach. This demonstrates the crucial role of "kidney-tonifying and essence-replenishing" medications in nerve regeneration.

[0104] Comparative Example 4: Reversing the Extraction Order (Process Comparison - First Alcohol Extraction, Then Volatile Oil Extraction): 1. Prescription: Same as Example 1.

[0105] 2. Preparation: First, all medicinal materials (except borneol) are pulverized and passed through a 20-mesh sieve. Then, 6 times the amount of 80% ethanol is added for extraction twice, each time for 1.5 hours, and the extract is concentrated to a relative density of 1.15 (measured at 60℃). Then, add 8 times the amount of water to the alcohol-extracted residue and steam distill for 4 hours to extract the volatile oil. The remaining dregs were extracted twice with 8 times the amount of water, and concentrated to a water extract with a relative density of 1.20 (measured at 60℃). Combine the alcohol extract and water extract, add the volatile oil inclusion complex and borneol, and shape.

[0106] 3. Quality Inspection: Volatile oil yield: 0.3% (only 25% of that in Example 1); β-Asauroyl ether content: 0.45±0.06 mg / g (only 24% of that in Example 1); Tanshinone IIA content: 2.38 ± 0.12 mg / g (equivalent to Example 1); 4. Cause Analysis: During the alcohol extraction process, ethanol carries away volatile oil components, causing a significant decrease in the yield of volatile oil during subsequent distillation.

[0107] 5. Conclusion: This proves that the process design of "first extracting volatile oil" in this invention is necessary, and the order of steps cannot be reversed.

[0108] Experimental Example 1: Pharmacodynamic and mechanistic study of the composition of the present invention in a rat model of vascular cognitive impairment: 1. Experimental Materials and Model Construction: Animals: SPF grade SD rats, male, weighing 250-280g.

[0109] Modeling method (2-VO method): A chronic cerebral hypoperfusion (CCH) model was prepared by bilateral common carotid artery permanent ligation. This model can simulate white matter damage and cognitive decline caused by long-term reduction in cerebral blood flow in clinical VCI patients and is an internationally recognized standard model for VCI.

[0110] Procedure: Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (350 mg / kg). A midline incision was made in the neck, and the bilateral common carotid arteries were separated and permanently ligated with 4-0 silk sutures. In the sham-operated group, only the blood vessels were separated without ligation. Postoperatively, penicillin was administered intramuscularly to prevent infection.

[0111] Grouping (n=15 / group): Sham surgery group: only blood vessels were separated without ligation; Model group: After modeling, physiological saline was administered by gavage; Positive control group (Donepezil): given donepezil hydrochloride (0.5 mg / kg / d); Example 1 group (the present invention): The drug prepared in Example 1 was administered (equivalent to 9.0 g / kg / d of crude drug); Comparative Example 1 (conventional process): The drug prepared in Comparative Example 1 was administered (at the same dosage as above). Comparative Example 2 (missing standard group): Administered the drug prepared in Comparative Example 2; Comparative Example 3 (missing this group): Administered the drug prepared in Comparative Example 3; Dosing cycle: Dosing began in the 4th week after modeling and continued for 30 days via gavage.

[0112] 2. Detection indicators and methods: 2.1 Morris Water Maze (MWM): Assess spatial learning and memory abilities. The water maze is 150 cm in diameter and the water temperature is 22±1℃.

[0113] Positioning and navigation test (days 26-30): The platform is positioned in the third quadrant, 1.5 cm below the water surface. Training is conducted 4 times a day, and the escape latency (time from entering the water to finding the platform) is recorded.

[0114] Space exploration experiment (day 31): Remove the platform and record the number of times the platform is traversed within 60 seconds and the time spent in the target quadrant.

[0115] 2.2 HPLC content determination: The contents of tanshinone IIA and β-asarone in the brain tissue of rats after administration of the drug preparations in each group were determined to directly verify the advantages of the process and the guiding effect of borneol.

[0116] Determination of formulation content: Accurately weigh 1.0 g of sample powder, add 25 mL of methanol, extract by ultrasonication for 30 minutes, centrifuge, filter the supernatant through a 0.22 μm filter membrane, and determine by HPLC.

[0117] Brain tissue content determination: 100 mg of hippocampal tissue was taken, 1 mL of methanol was added, homogenized, extracted by ultrasonication, centrifuged, and the supernatant was filtered through a 0.22 μm filter membrane and determined by HPLC.

[0118] Chromatographic conditions: Agilent C18 column (4.6 × 250 mm, 5 μm); mobile phase methanol-water (75:25); flow rate 1.0 mL / min; detection wavelength 270 nm; column temperature 30℃.

[0119] 2.3 ELISA and Biochemical Detection: Two hours after the last gavage administration (when the blood concentrations of borneol and tanshinone IIA, etc., reach their peak), rats were decapitated, and their brains were harvested. Hippocampal tissue was isolated, and tissue homogenate (physiological saline, 1:9, w / v) was prepared. ELISA kits (Beijing Solarbio Science & Technology Co., Ltd.) were used for detection. Inflammatory factors: TNF-α, IL-1β, IL-6; Oxidative stress indicators: SOD (superoxide dismutase), MDA (malondialdehyde); 2.4 Western Blot: The expression of BDNF (brain-derived neurotrophic factor), p-Akt (phosphorylated protein kinase B), NLRP3 (inflammasome), and Claudin-5 (tight junction protein) in the hippocampus was detected.

[0120] Total protein was extracted, and its concentration was determined by the BCA method. SDS-PAGE electrophoresis, followed by transfer to a PVDF membrane; 5% skim milk powder sealed for 1 hour; Primary antibody incubation overnight (4℃): anti-BDNF (1:1000), anti-NLRP3 (1:1000), anti-Claudin-5 (1:1000), anti-β-actin (1:2000); Secondary antibody incubation for 1 hour (room temperature); ECL color development, gel imaging system for taking pictures, and ImageJ software for analyzing grayscale values; 2.5 HE staining: Observe the morphology and survival of neurons in the CA1 region of the hippocampus.

[0121] Rats were perfused and fixed (4% paraformaldehyde); Brain tissue was harvested, embedded in paraffin, and sectioned (5μm). HE staining, mounting with neutral resin; Observed under an optical microscope (400×); 2.6 Statistical methods: All data are expressed as mean ± standard deviation (SD). Mean ± s. Analyses were performed using SPSS 26.0 statistical software. One-way ANOVA was used for comparisons among multiple groups, Levene's test was used for homogeneity of variance, and LSD-t test was used for pairwise comparisons between groups. P < 0.05 was considered statistically significant.

[0122] Significance labeling rules: - # P<0.05 vs Sham group - ## P<0.01 vs Sham group - P<0.05 vs Model group - P<0.01 vs Model group - △ P<0.05 vs Donepezil group - △ △ P<0.01 vs Donepezil group 3. Detailed analysis of experimental results: 3.1 Verification of Process Advantages: Comparison of Active Ingredient Content and Brain Distribution: To visually demonstrate the necessity of "stepwise extraction" and "borneol guidance," we first conducted a chemical composition analysis.

[0123] Table 1: Content of key components and their distribution in the brain after administration in formulations produced by different processes ( ±s, n=6)

[0124] Note: Compared with Example 1, ##P<0.01; compared with Example 1 (brain concentration) P<0.01, △△P<0.01.

[0125] In-depth analysis: Extraction efficiency: The tanshinone IIA content in the preparation of Example 1 (stepwise extraction) was 16 times that of Comparative Example 1 (water decoction), and the β-asarone content was 23 times that of Comparative Example 1. This irrefutably demonstrates that the traditional water decoction process leads to the catastrophic loss of fat-soluble and volatile components, while the process of the present invention successfully preserves these key components.

[0126] Guide effect: The tanshinone content in the preparations of Example 1 and Comparative Example 2 was the same, but the concentration of tanshinone in the brain of rats in Example 1 (containing borneol) was 3.7 times that in Comparative Example 2 (without borneol). This directly confirms the "drug-guide" function of borneol in opening the blood-brain barrier and promoting drug entry into the brain.

[0127] 3.2 Behavioral testing: Improvement in cognitive function: The results of the Morris Water Maze test on day 5 are as follows: Table 2: Results of the Morris water maze test ( ±s, n=12)

[0128] Note: Compared with the Sham group, ##P<0.01; compared with the Model group. P<0.05, P<0.01; compared with the Donepezil group, △P<0.05.

[0129] In-depth analysis: Advantages of this invention: The latency period in Example 1 group was the shortest, even better than that of the positive control drug donepezil (P<0.05). This indicates that the traditional Chinese medicine compound, through multi-target intervention, has a more comprehensive cognitive improvement ability than Western medicine that simply increases acetylcholine levels.

[0130] Disadvantages of the flavor-deficient group: The effect of Comparative Example 2 (deficient Fritillaria thunbergii and borneol) was significantly worse than that of Example 1, which proved the decisive role of the "symptomatic treatment" drug in improving VCI (especially clearing intracranial inflammation and unblocking microcirculation).

[0131] 3.3 Molecular Mechanism Verification: Anti-inflammatory, Antioxidant, and Neuroprotective Effects Table 3: Results of biochemical and protein expression detection in hippocampal tissue ( ±s, n=8)

[0132] Note: Compared with the Sham group, ##P<0.01; compared with the Model group. P<0.05, P<0.01.

[0133] Key Data Interpretation: Anti-inflammatory mechanism (TNF-α, IL-6): The levels of inflammatory factors in Example 1 group were the lowest, close to those in the sham-operated group. In contrast, the inflammation levels in Comparative Example 2 group (lacking Fritillaria thunbergii and borneol) were extremely high, almost indistinguishable from the model group. This directly confirms that the "nodule-dispersing" effect of Fritillaria thunbergii is actually an "anti-inflammatory" effect; without the guidance of Fritillaria thunbergii and borneol, the compound formula cannot inhibit neuroinflammation in the brain.

[0134] Neurotrophic factor (BDNF): In Example 1 group, BDNF expression recovered to 88% of normal. In contrast, in Comparative Group 3 (lacking kidney-tonifying drugs), although inflammation was somewhat controlled, BDNF levels only recovered to 45%. This confirms that kidney-tonifying drugs such as Rehmannia glutinosa and Cornus officinalis are the core material basis for promoting the synthesis of neurotrophic factors and repairing neurons.

[0135] Oxidative stress (MDA, SOD): The first group significantly reduced lipid peroxidation product MDA and increased the activity of antioxidant enzyme SOD, suggesting that it has a strong ability to resist oxidative stress and protect mitochondrial function.

[0136] 3.4 Western Blot protein expression analysis: Table 4: Relative expression levels of key proteins in hippocampal tissue ( ±s, n=6)

[0137] Note: Compared with the Sham group, ##P<0.01; compared with the Model group. P<0.05, P<0.01; compared with the Donepezil group, △P<0.05, △△P<0.01.

[0138] In-depth analysis: NLRP3 inflammasome: The expression of NLRP3 was significantly inhibited in the first example group, confirming the anti-inflammatory mechanism of Fritillaria thunbergii.

[0139] Claudin-5 tight junction protein: In Example 1, Claudin-5 was significantly downregulated (0.58) 2 hours after administration, accurately capturing the pharmacokinetic characteristic of borneol's 'instantaneous opening' of the BBB, confirming the role of the biological key; while in Comparative Example 2 (without borneol), Claudin-5 had recovered to a higher level (0.95) at this time, which, although repairing the pathological barrier leakage, also prevented the drug from entering the brain. This indicates that the composition of the present invention, through the active and moderate downregulation of tight junction proteins by borneol within a specific time window at the beginning of medication, is a key prerequisite for achieving precise drug delivery to the brain.

[0140] BDNF and p-Akt: In Example 1, BDNF and p-Akt were significantly upregulated, activating the neurotrophic pathway.

[0141] 4. Experimental Conclusion: This study, through rigorous pharmaceutical and pharmacodynamic comparisons, draws the following conclusions: Innovative Process: The stepwise targeted extraction process of this invention is a prerequisite for therapeutic efficacy. Traditional decoction methods (Comparative Example 1) lead to the complete loss of key components such as tanshinone and volatile oils, resulting in a loss of efficacy. This process maximizes the retention of active ingredients.

[0142] Scientific basis of the formulation: "Fritillaria thunbergii-borneol" is the "vanguard" of this formula. Borneol opens up the barrier, while Fritillaria thunbergii clears inflammation (phlegm toxins). Both are indispensable (as confirmed in Comparative Example 2). "Kidney-tonifying herbs" are the "logistical support" of this formula, providing neurotrophic support such as BDNF and repairing damaged nerves (as confirmed in Comparative Example 3).

[0143] Overall therapeutic effect: The composition of the present invention significantly improves the cognitive function of VCI rats through the synergistic effect of multiple targets such as "anti-inflammatory, antioxidant and neurotrophic". Its efficacy is superior to that of single Western medicine and traditional process preparations, and it has extremely high clinical application value.

[0144] Experimental Example 2: Acute Toxicity Test 1. Experimental objective: Evaluate the acute toxicity of the compositions of the present invention, and determine the maximum tolerated dose (MTD) and safety range.

[0145] 2. Experimental materials: Animals: Kunming mice, SPF grade, half male and half female, weighing 18-22g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0146] Test drug: The composition of the present invention prepared in Example 1 was formulated into a suspension with 0.5% sodium carboxymethyl cellulose (CMC-Na).

[0147] 3. Experimental Methods: Grouping and drug administration (n=10 / group, half male and half female): Blank control group: administered 0.5% CMC-Na solution by gavage; Low-dose group: 10 times the clinically equivalent dose (equivalent to 90 g / kg of raw drug). Medium dose group: 30 times the clinically equivalent dose (equivalent to 270 g / kg of raw drug); High-dose group: 50 times the clinically equivalent dose (equivalent to 450 g / kg of raw drug); Extremely high dose group: 100 times the clinically equivalent dose (equivalent to 900 g / kg of raw drug). Note: Clinical equivalent dose is calculated based on body surface area. A human (60kg) taking 90g of raw herb per day is equivalent to a mouse taking 9g / kg.

[0148] Administration method: single oral gavage, gavage volume 20 mL / kg.

[0149] Observation indicators: General condition: After administration, mice were observed for 14 consecutive days, and their activity, coat color, food intake, water intake, defecation, and respiration were recorded.

[0150] Weight changes: Weigh yourself before administration and on days 1, 3, 7, and 14 after administration.

[0151] Death details: Record the time of death and the number of deaths.

[0152] Dissection and observation: On day 14, surviving mice were sacrificed and the appearance of major organs such as heart, liver, spleen, lungs and kidneys was observed with the naked eye.

[0153] 4. Experimental Results: Table 5: Results of Acute Toxicity Tests (n=10)

[0154] Anatomical observation: No obvious abnormalities were observed in the appearance of the main organs such as heart, liver, spleen, lungs and kidneys of mice in each dose group, and no pathological changes such as congestion, hemorrhage or swelling were found.

[0155] 5. Conclusion: The maximum tolerated dose (MTD) of the composition of this invention is >100 times the clinically equivalent dose (equivalent to 900 g / kg of crude drug), indicating good safety and a wide safety margin. Even at extremely high doses, only transient reduction in activity occurred, with no deaths, proving that the composition of this invention is safe and reliable for clinical application.

[0156] Experimental Example 3: Effects of Borneol on Blood-Brain Barrier Permeability and Verification of the Brain Entry Mechanism of Tanshinone IIA: 1. Experimental objective: To verify whether borneol increases blood-brain barrier permeability by downregulating tight junction proteins, thereby promoting the entry of tanshinone IIA into the brain, this study provides direct experimental evidence for the innovative combination of "Zhejiang fritillary bulb-borneol".

[0157] 2. Experimental materials: Animals: Male SD rats, weighing 250±20g.

[0158] Drugs: Pure tanshinone IIA (purity ≥98%, Chengdu Mansite Biotechnology Co., Ltd.); Natural borneol (purity ≥98%, Sichuan Senke Traditional Chinese Medicine Pieces Co., Ltd.); 3. Experimental Methods: Grouping and drug administration (n=8 / group): Control group: administered 0.5% CMC-Na solution by gavage; Tanshinone IIA group: Pure tanshinone IIA (20 mg / kg, prepared as a suspension with 0.5% CMC-Na) was administered by gavage. Tanshinone IIA + Borneol group: Tanshinone IIA (20 mg / kg) + Borneol (5 mg / kg) were administered by gavage. Administration and sampling: Two hours after administration, rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (350 mg / kg), decapitated and brain removed. Hippocampal tissue was quickly separated, and part of it was used for HPLC to determine the concentration of tanshinone IIA and part of it was used for Western blotting to detect tight junction proteins.

[0159] Testing indicators: (1) HPLC determination of tanshinone IIA concentration in the brain: Take 100 mg of hippocampal tissue, add 1 mL of methanol, homogenize, extract by sonication for 30 minutes, centrifuge (12000 rpm, 10 minutes), filter the supernatant through a 0.22 μm filter membrane, and determine by HPLC.

[0160] The chromatographic conditions were the same as in Example 1.

[0161] (2) Western Blot detection of tight junction proteins: The expression of Claudin-5 and Occludin proteins in the hippocampus was detected.

[0162] The operation method is the same as in Experiment 1.

[0163] (3) Immunofluorescence observation of BBB integrity (supplementary experiment): Obtain frozen sections of brain tissue (10 μm); Rabbit anti-Claudin-5 primary antibody (1:200) incubated overnight; Incubate with FITC-labeled secondary antibody (1:500) for 1 hour; DAPI counterstaining of cell nuclei; Observation using a laser confocal microscope; 4. Experimental Results: Table 6: Tanshinone IIA concentration and tight junction protein expression in the brains of rats in each group ( ±s, n=8)

[0164] Note: Compared with the control group P<0.01; compared with the tanshinone IIA group, ΔΔP<0.01.

[0165] In-depth analysis: Brain penetration efficiency: After being combined with borneol, the intracerebral concentration of tanshinone IIA increased from 8.5 ng / g to 31.2 ng / g, an increase of 3.7 times (P<0.01). This directly proves the "guiding" effect of borneol.

[0166] BBB opening mechanism: Claudin-5 expression was downregulated by 42% (P<0.01) and Occludin expression was downregulated by 38% (P<0.01) in the borneol group. Claudin-5 and Occludin are key proteins of the BBB tight junction, and their downregulation means that the tight junction is relaxed and the BBB permeability is increased.

[0167] Reversibility: When tested again 24 hours after administration, the expression of Claudin-5 and Occludin had basically returned to normal, indicating that the opening of the BBB by borneol is reversible and will not cause permanent damage.

[0168] Immunofluorescence results: In the control group and the tanshinone IIA group, the fluorescence intensity of Claudin-5 around the hippocampal microvessels was high and the distribution was continuous, indicating that the BBB was tight and intact.

[0169] In the tanshinone IIA + borneol group: the fluorescence intensity of Claudin-5 was significantly reduced and the distribution was discontinuous, indicating that the tight junctions of the BBB were loose.

[0170] 5. Conclusion: This experiment directly confirms that borneol reversibly opens the blood-brain barrier by downregulating the expression of tight junction proteins Claudin-5 and Occludin, thereby increasing the intracerebral concentration of tanshinone IIA by 3.7 times. This result provides a solid experimental basis for the inventive combination of borneol and Fritillaria thunbergii, revealing the molecular mechanism of the synergistic brain entry of the "key (borneol) + ammunition (tanshinone IIA)" of this invention.

[0171] Test Example 4: Long-term toxicity test (90 days): 1. Experimental objective: To evaluate the safety of long-term administration of the compositions of the present invention.

[0172] 2. Experimental materials: Animals: SD rats, SPF grade, half male and half female, weighing 200±20g.

[0173] Test drug: The composition of the present invention prepared in Example 1.

[0174] 3. Experimental Methods: Grouping and drug administration (n=20 / group, half male and half female): Control group: administered 0.5% CMC-Na solution by gavage; Low-dose group: 1 times the clinically equivalent dose (equivalent to 9 g / kg / d of raw drug). Medium dose group: 3 times the clinically equivalent dose (equivalent to 27 g / kg / d of raw drug). High-dose group: 9 times the clinically equivalent dose (equivalent to 81 g / kg / d of raw drug). Administration cycle: 90 consecutive days by gavage, once daily.

[0175] Observation indicators: General status: Daily observation record; Weight: Weigh yourself once a week; Food intake: Recorded once a week; Complete blood count: Tests were performed before drug administration, 45 days after drug administration, and 90 days after drug administration (WBC, RBC, HGB, PLT, etc.). Liver and kidney function: measured before administration, 45 days after administration, and 90 days after administration (ALT, AST, BUN, Cr, etc.); Pathological examination: Rats were sacrificed 90 days after administration of the drug, and organs such as heart, liver, spleen, lung, kidney, and brain were collected, fixed in 10% formaldehyde, embedded in paraffin, stained with hematoxylin and eosin (HE), and observed under a light microscope.

[0176] 4. Experimental Results: Table 7: Key Indicators of Long-Term Toxicity Tests ( (±s, n=20)

[0177] Blood routine test results: WBC, RBC, HGB, PLT and other indicators in each dose group were within the normal range and there was no significant difference compared with the control group (P>0.05).

[0178] Pathological examination results: No obvious pathological changes were found in the major organs such as heart, liver, spleen, lung, kidney and brain of rats in each dose group. The liver cell structure was clear, the glomeruli and renal tubules were normal, the myocardial fibers were neatly arranged, and the alveolar structure was intact.

[0179] 5. Conclusion: When the composition of this invention was administered continuously for 90 days, even at 9 times the clinically equivalent dose, the rats maintained good general condition, normal weight gain, normal liver and kidney function indicators, and no abnormalities were found in the pathological examination of major organs. This demonstrates that the composition of this invention has good long-term safety with no obvious toxic side effects and is suitable for long-term clinical application.

[0180] Quality Standards: The compositions of this invention shall meet the following quality requirements: 1. Characteristics: This product consists of brownish-brown granules; it has a slightly fragrant odor and a bitter, slightly spicy taste.

[0181] 2. Identification: (1) Thin-layer chromatography identification: Identification of Danshen: Take 2g of the powder, add 20mL of ethanol, extract by ultrasonication for 30 minutes, filter, and concentrate the filtrate to 1mL as the test solution. Separately, take tanshinone IIA reference standard, add ethanol to prepare a 1mg / mL solution as the reference solution. Perform thin-layer chromatography (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia). Apply 5μL of each of the above two solutions to the same silica gel G thin-layer plate, using petroleum ether-ethyl acetate (9:1) as the developing solvent. Develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference standard.

[0182] Identification of Fritillaria thunbergii: Take 2g of the powder, moisten with 2mL of ammonia solution, add 20mL of chloroform, extract by sonication for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1mL of chloroform to prepare the test solution. Separately, prepare a 1mg / mL solution of fritillary acetate reference standard in chloroform to prepare the reference solution. Perform thin-layer chromatography, applying 5μL of each of the above two solutions separately to the same silica gel G thin-layer plate. Develop using chloroform-methanol-concentrated ammonia solution (8:2:0.1), remove, air dry, and spray with potassium bismuth iodide solution. The test sample chromatogram should show spots of the same color at the corresponding positions as the reference standard chromatogram.

[0183] Identification of Borneol: Take 1g of the powder, add 10mL of petroleum ether, extract by sonication for 15 minutes, filter, and use the filtrate as the test solution. Separately, take borneol reference standard, add petroleum ether to prepare a 1mg / mL solution, and use this as the reference solution. Perform thin-layer chromatography, applying 5μL of each of the above two solutions separately to the same silica gel G thin-layer plate. Use cyclohexane-ethyl acetate (9:1) as the developing solvent, develop, remove, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference standard.

[0184] (2) Microscopic identification: Take a small amount of the powder, add water and slide it. Microscopic examination reveals: numerous starch granules, single granules are round, oval or irregular in shape; calcium oxalate clusters are 20-80 μm in diameter; cork cells are brownish-yellow; thin-walled cells around fiber bundles contain calcium oxalate crystals.

[0185] 3. Inspection: (1) Moisture content: not more than 9.0% (General Chapter 0832, Method II, Chinese Pharmacopoeia 2020 Edition); (2) Content variation: It should comply with the regulations (General Chapter 0942 of the 2020 edition of the Chinese Pharmacopoeia); (3) Microbial limits: should comply with the regulations (General Chapters 1105, 1106, 1107, and 1108 of the 2020 edition of the Chinese Pharmacopoeia); 4. Content determination: (1) Determination of tanshinone IIA content (HPLC method): Chromatographic conditions: Chromatographic column: Octadecylsilane bonded silica column (4.6 mm × 250 mm, 5 μm); Mobile phase: methanol-water (75:25); Detection wavelength: 270nm; Flow rate: 1.0 mL / min; Column temperature: 30℃; Injection volume: 10 μL; Assay method: Accurately weigh approximately 1.0 g of the powder and place it in a stoppered conical flask. Accurately add 25 mL of methanol, seal tightly, and weigh. Sonicate (250 W, 40 kHz) for 30 minutes, cool, and weigh again. Make up the lost weight with methanol, shake well, filter, and collect the filtrate. Determine the content under the above chromatographic conditions using the external standard method, calculating the content based on peak area.

[0186] Limit: Each 1g must contain no less than 1.5mg of tanshinone IIA (C19H18O3).

[0187] (2) Determination of β-asarone content (GC method): Chromatographic conditions: Chromatographic column: polyethylene glycol (PEG-20M) capillary column (30m×0.32mm×0.25μm); Programmed temperature rise: Initial temperature 80℃, hold for 2 minutes, then increase to 180℃ at 10℃ / min, hold for 5 minutes; Inlet temperature: 250℃; Detector temperature: 250℃ (FID); Carrier gas: Nitrogen; Flow rate: 1.0 mL / min; Flow split ratio: 10:1; Injection volume: 1 μL; Assay method: Accurately weigh approximately 0.5 g of the powder and place it in a stoppered conical flask. Accurately add 10 mL of diethyl ether, seal tightly, weigh, sonicate for 30 minutes, cool, weigh again, replenish the lost weight with diethyl ether, shake well, filter, and collect the filtrate. Determine the content under the above chromatographic conditions using the external standard method based on peak area.

[0188] Limit: Each 1g shall contain not less than 1.0mg of β-asarone (C12H16O3).

[0189] (3) Determination of strychnine content (HPLC method): Chromatographic conditions: Chromatographic column: Octadecylsilane bonded silica column (4.6 mm × 250 mm, 5 μm); Mobile phase: Acetonitrile-water (12:88); Detection wavelength: 238nm; Flow rate: 1.0 mL / min; Column temperature: 30℃; Injection volume: 10 μL; Assay method: Accurately weigh approximately 1.0 g of the powder and place it in a stoppered conical flask. Accurately add 25 mL of methanol, seal tightly, weigh, sonicate for 30 minutes, cool, weigh again, replenish the lost weight with methanol, shake well, filter, and collect the filtrate. Determine the content under the above chromatographic conditions using the external standard method based on peak area.

[0190] Limit: Each 1g must contain not less than 6.0mg of strychnine (C17H26O10).

[0191] 5. Fingerprint pattern: The fingerprint chromatogram of this product was established using HPLC. Chromatographic conditions: Chromatographic column: Octadecylsilane bonded silica column (4.6 mm × 250 mm, 5 μm); Mobile phase: Acetonitrile (A) - 0.1% phosphoric acid aqueous solution (B), gradient elution (0-10 min, 10%-20% A; 10-30 min, 20%-50% A; 30-50 min, 50%-80% A); Detection wavelength: 280nm; Flow rate: 1.0 mL / min; Column temperature: 30℃; The fingerprint spectrum of this product should have more than 15 common peaks, and the similarity should not be less than 0.90.

[0192] 6. Indications: It nourishes the kidneys, resolves phlegm, and clears the orifices and meridians. It is used for vascular cognitive impairment, characterized by memory loss, slow reaction, dizziness, headache, numbness of limbs, etc., which belong to the syndrome of kidney essence deficiency and phlegm and blood stasis obstructing the meridians.

[0193] 7. Usage and dosage: Dissolve in boiling water and drink. Take 10-15g each time, 2-3 times a day, for 4-12 weeks, or as directed by your doctor.

[0194] 8. Specifications: Each bag contains 15g (equivalent to 27.6g of raw medicinal materials).

[0195] 9. Storage: Seal and store in a cool, dry place.

[0196] 10. Validity period: 24 months.

[0197] in conclusion: This invention provides a traditional Chinese medicine composition for treating vascular cognitive impairment, characterized by an innovative "Zhejiang Fritillaria-Borneol" compatibility strategy and a matching "stepwise targeted extraction process".

[0198] This invention not only perfects the system of "treating dementia from the perspective of phlegm, blood stasis, and kidney deficiency" in traditional Chinese medicine theory; Furthermore, at the level of modern medicine, experimental data have confirmed the molecular mechanisms by which it increases the rate of fat-soluble components entering the brain, inhibits NLRP3-mediated neuroinflammation, and upregulates the BDNF / TrkB signaling pathway.

[0199] Comparative experiments have fully demonstrated that the technical solution of the present invention is significantly superior to the prior art in terms of component retention rate and biological effect, possessing outstanding substantive features and significant progress.

[0200] This invention provides a safe and effective novel traditional Chinese medicine preparation for the treatment of vascular cognitive impairment, which has important clinical application value and broad market prospects.

[0201] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A traditional Chinese medicine composition for treating vascular cognitive impairment, characterized in that, Made from the following parts by weight of active pharmaceutical ingredient: 10-20 parts of stir-fried Atractylodes macrocephala, 10-20 parts of Fritillaria thunbergii, 10-20 parts of Ligusticum chuanxiong, 15-25 parts of Salvia miltiorrhiza, 5-15 parts of Curcuma longa, 10-20 parts of Acorus tatarinowii, 5-15 parts of Polygala tenuifolia, 15-25 parts of Cornus officinalis, 8-15 parts of Cistanche deserticola, 8-15 parts of Rehmannia glutinosa (processed), 8-15 parts of Scrophularia ningpoensis, and 0.5-2 parts of borneol.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The weight ratio of Fritillaria thunbergii to Salvia miltiorrhiza is 1:(1.2-1.8), and the weight of borneol is 0.8-1.5 parts.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, It is made from the following raw materials in parts by weight: 15 parts stir-fried Atractylodes macrocephala, 15 parts Fritillaria thunbergii, 15 parts Ligusticum chuanxiong, 20 parts Salvia miltiorrhiza, 10 parts Curcuma longa, 15 parts Acorus tatarinowii, 10 parts Polygala tenuifolia, 20 parts Cornus officinalis, 12 parts Cistanche deserticola, 12 parts Rehmannia glutinosa, 12 parts Scrophularia ningpoensis, and 1 part Borneol.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, In the aforementioned traditional Chinese medicine composition, the content of tanshinone IIA is 1.68-3.12 mg / g, the content of β-asarone is 1.25-2.45 mg / g, and the content of strychnoside is 6.8-11.5 mg / g.

5. A method for preparing a traditional Chinese medicine composition for treating vascular cognitive impairment as described in any one of claims 1-4, characterized in that, Includes the following steps: (1) Extraction of volatile oil: Take Curcuma longa, Acorus tatarinowii and Ligusticum chuanxiong, crush them, add water and extract by steam distillation, collect the volatile oil, and use β-cyclodextrin to encapsulate it to obtain volatile oil inclusion complex; the aqueous solution after distillation is used as aromatic water for later use; the residue is drained to obtain residue I; (2) Alcohol extraction: Take Salvia miltiorrhiza and Fritillaria thunbergii, crush them, add 75%-85% ethanol solution, heat and extract at 78-82℃, combine the extracts, recover the ethanol under reduced pressure until there is no alcohol taste, concentrate to obtain alcohol extract; drain the residue to obtain residue II; (3) Water extraction: Mix the dregs I from step (1), the dregs II from step (2), stir-fried Atractylodes macrocephala, Polygala tenuifolia, Cornus officinalis, Cistanche deserticola, Rehmannia glutinosa and Scrophularia ningpoensis, add the aromatic water from step (1), and add purified water until the total solvent volume is 6-10 times the total weight of the medicinal materials and dregs in this step, decoct, combine the decoctions, and concentrate to obtain water extract; (4) Molding: Mix the alcohol extract from step (2) and the water extract from step (3) evenly, dry, granulate to obtain granules; mix the volatile oil inclusion complex from step (1) with borneol, add it evenly to the granules by spraying, and granulate to obtain the final product.

6. The preparation method according to claim 5, characterized in that, In step (1), the powder is pulverized to 20-40 mesh, 6-10 times the amount of water is added, and steam distillation is used to extract for 3-5 hours. The weight ratio of β-cyclodextrin to volatile oil is (5-8):

1. In step (2), take Salvia miltiorrhiza and Fritillaria thunbergii, pulverize them to 20-40 mesh, add 5-8 times the amount of ethanol solution, heat and extract 2-3 times, each extraction time is 1.0-2.0 hours; concentrate to 60℃ and measure the relative density to be 1.10-1.20 of the ethanol extract. In step (3), the water extract is decocted at 100℃ 2-3 times, with each decoction lasting 0.5-1.5 hours, and concentrated to a relative density of 1.15-1.25 at 60℃. In step (4), the product is dried at 60-80℃ until the moisture content is ≤5%, and then granulated to 10-20 mesh.

7. The use of a traditional Chinese medicine composition according to any one of claims 1-4 or a traditional Chinese medicine composition prepared by the preparation method according to any one of claims 5-6 in the preparation of a drug for treating vascular cognitive impairment, characterized in that, The vascular cognitive impairment includes vascular dementia and vascular mild cognitive impairment, which is caused by chronic cerebral hypoperfusion.

8. The application according to claim 7, characterized in that, The drug is prepared as one of granules, capsules, tablets, pills, or oral liquid; The drug is prepared into unit formulations, and each formulation unit contains 18-28 grams of effective extract equivalent to crude drug.

9. The application according to claim 7, characterized in that, The vascular cognitive impairment is caused by chronic cerebral hypoperfusion due to stenosis or occlusion of both common carotid arteries. Patients exhibit progressive memory loss, executive dysfunction, inattention, and abnormal mental and behavioral characteristics.

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