Application of songorine in treatment of Parkinson's disease

Abstract

The invention relates to the technical field of medicines, in particular to application of songorine in treatment of Parkinson's disease. The C20-type diterpenoid alkaloid songnin provided by the invention can obviously and continuously improve dyskinesia caused by dopaminergic neuron injury in MPTP-induced Parkinson's disease model mice, and the onset dose of songnin is obviously lower than that of clinically common positive medicine, namely medoxolane, so that the songnin shows a stronger drug effect and can be used for preparing drugs for treating Parkinson's disease. The compound can be developed into a novel efficient anti-Parkinson disease treatment medicine.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of Songguoling in the treatment of Parkinson's disease. Background Technology

[0002] Parkinson's disease (PD) is a common neurodegenerative disease characterized by the progressive degeneration and loss of dopaminergic neurons in the substantia nigra pars compacta of the midbrain. Its clinical manifestations mainly include motor symptoms such as resting tremor, bradykinesia, rigidity, and postural instability, and are often accompanied by a variety of non-motor symptoms such as depression, anxiety, sleep disorders, and autonomic dysfunction.

[0003] Currently, the core pathological feature of Parkinson's disease (PD) is the selective and progressive loss of dopaminergic neurons in the substantia nigra pars compacta of the midbrain, leading to a significant reduction in dopamine levels in the striatum and consequently causing typical motor dysfunction. Existing clinical treatments mainly involve levodopa and its compound preparations, dopamine receptor agonists, monoamine oxidase B inhibitors, and catechol-O-methyltransferase inhibitors. Their basic mechanism of action is to supplement or enhance dopamine function in the central nervous system, thereby improving the patient's motor symptoms. These drugs usually have some efficacy in the early or middle stages of the disease, but long-term use can easily lead to adverse reactions such as fluctuating efficacy, dyskinesia, and neuropsychiatric abnormalities. Furthermore, they are difficult to effectively delay or block the neurodegenerative process of the disease, and the overall treatment effect remains significantly limited. Summary of the Invention

[0004] This invention covers the following technical solution: the application of Songguoling in the preparation of drugs for treating Parkinson's disease.

[0005] Experimental results show that the C provided by this invention 20 The diterpenoid alkaloid songguoling significantly and persistently improved motor dysfunction caused by dopaminergic neuron damage in MPTP-induced Parkinson's disease model mice. In terms of efficacy comparison, the onset dose of songguoling was significantly lower than that of the commonly used clinical positive control drug levodopa, and it showed stronger efficacy, suggesting its potential as a novel and highly effective anti-Parkinson's disease treatment. Attached Figure Description

[0006] Reference will now be made to detailed embodiments of the present invention, one or more of which are described below. Each example is provided for explanation and not for limitation of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made to the invention without departing from its scope or spirit. For example, features described or illustrated as part of one embodiment may be used in another embodiment to produce further embodiments.

[0007] Unless otherwise stated, all terms used to disclose this invention (including technical and scientific terms) should be understood as having the meaning commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein is for the purpose of describing specific embodiments only and is not intended to limit the scope of protection of this invention. Unless the context clearly defines otherwise, the scientific and technical terms used herein, as well as terms and laboratory procedures in related fields such as neurobiology, animal behavior, physiology, psychology, medicinal chemistry, and pharmacology, are all conventional terms and standard methods well-known and widely used in the art. To facilitate understanding of the technical solutions of this invention, some related terms are further defined and explained below.

[0008] The terms “containing,” “comprising,” and “including” as used in this invention are synonyms and are inclusive or open-ended, not excluding additional, uncited members, elements, or method steps.

[0009] In this invention, the numerical range represented by endpoints includes all numerical values ​​and fractions contained within that range, as well as the endpoints mentioned.

[0010] Furthermore, in describing representative embodiments of the invention, this specification may present the methods and / or processes of the invention as a specific sequence of steps. However, the method or process should not be limited to the specific order of the steps described herein, to the extent that the method or process does not depend on the specific order of the steps presented herein. As will be understood by those skilled in the art, other sequences of steps are also possible. Therefore, the specific order of steps presented in the specification should not be construed as a limitation of the claims. Additionally, the claims relating to the methods and / or processes of the invention should not be limited to the execution of their steps in the order they are written, and those skilled in the art will readily recognize that the sequence can be changed while still remaining within the spirit and scope of the invention.

[0011] This invention relates to concentration values, which include fluctuations within a certain range. For example, fluctuations are allowed within a corresponding precision range. For instance, 2% can fluctuate within ±0.1%. For larger values ​​or values ​​that do not require overly precise control, even greater fluctuations are permitted.

[0012] As used in this invention, unless otherwise stated, the singular forms of the articles “a,” “an,” and “the” include plural referents.

[0013] In this invention, the terms "multiple" or "various" are used unless otherwise specified, referring to a quantity of 2 or more.

[0014] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0015] In this invention, terms such as "preferred," "better," "more suitable," and "ideal" merely describe implementation methods or embodiments with better effects and should be understood not to limit the scope of protection of this invention. In this invention, terms such as "optionally," "optionally," and "optional" mean that something is optional, that is, selected from either "with" or "without" a parallel solution. If multiple "optional" statements appear in a technical solution, unless otherwise specified and without contradiction or mutual constraint, each "optional" statement is independent.

[0016] In this description, the term "Songorine" refers to a C-type compound derived from the traditional Chinese medicine Aconitum carmichaelii. 20 This is a type of diterpenoid alkaloid, with CAS number 509-18-2 and molecular formula C2. 22 H 29 NO4, molecular weight 371.43. The Songguoling described in this invention includes Songguoling extracted from natural sources, as well as structurally identical or substantially equivalent compounds obtained through chemical synthesis, semi-synthesis, or biotransformation. It also includes possible stereoisomers, tautomers, solvates, hydrates, and pharmaceutically acceptable salts thereof, all of which are included within the scope of protection of this invention. The pharmaceutically acceptable salts include addition salts formed from inorganic or organic acids, such as hydrochlorides, sulfates, phosphates, methanesulfonates, tartrates, citrates, oxalates, etc., and salts formed from inorganic or organic bases. These salts exhibit good biological compatibility and are suitable for pharmaceutical preparation and clinical application.

[0017] In this invention, the term "treatment" and related terms include "treatment", "therapeutic intervention", "for treatment", etc., which refers to the partial or complete relief, improvement, inhibition, delay or control of clinical symptoms, pathological state or disease progression in patients or suspected patients with Parkinson's disease by administering songguoling or its pharmaceutical composition, and includes the reduction or prevention of the risk of disease recurrence or aggravation.

[0018] In this invention, the term "Parkinson's disease" refers to a neurodegenerative disease characterized by progressive degeneration of dopaminergic neurons in the substantia nigra of the midbrain and dysfunction of striatal dopaminergic neurons, with motor symptoms such as bradykinesia, resting tremor, rigidity, and postural instability, as well as non-motor symptoms such as sleep disturbances, cognitive impairment, autonomic dysfunction, and mood disorders as its main pathological basis. This term includes not only typical idiopathic Parkinson's disease, but also hereditary Parkinson's disease, secondary Parkinsonian syndromes, and related Parkinson's spectrum diseases. Furthermore, "Parkinson's disease" includes patient states at different stages of disease progression, such as early, intermediate, and late stages, or different stages from stage I to stage V according to the Hoehn-Yahr classification; it also includes Parkinson's disease patients with different clinical subtypes, such as tremor-dominant, rigid-bradykinesic, and mixed subtypes. Furthermore, the term "Parkinson's disease" also includes complex clinical manifestations accompanied by varying degrees of cognitive impairment, behavioral abnormalities, or other systemic dysfunctions, as well as different disease stages such as stable, progressive, or fluctuating phases. Any disease state that meets the above pathological characteristics, clinical manifestations, and classification features should be understood as being included within the scope of protection of Parkinson's disease as described in this invention.

[0019] In this invention, the term "idiopathic Parkinson's disease" refers to a type of Parkinson's disease with an unclear etiology and no clear genetic defects or exogenous damaging factors, characterized by the spontaneous degeneration of dopaminergic neurons.

[0020] In this specification, the term "effective dose" or "therapeutic effective dose" refers to the dosage of songguoling or its pharmaceutical composition that can induce the desired anti-Parkinson's biological effect or therapeutic effect without producing unacceptable toxic side effects. The effective dose may be adjusted according to factors such as the subject's age, weight, disease severity, route of administration, frequency of administration, and individual differences, and shall be reasonably determined by those skilled in the art based on conventional medical and pharmacological knowledge.

[0021] In this invention, the terms "subject" and "patient" are used interchangeably and refer to a mammalian individual, including preferred humans and non-human primates, as well as common laboratory animals (such as rabbits, rats, mice, dogs, monkeys, etc.). The term specifically refers to an individual to whom the compounds, compositions, or formulations of this invention may be administered. Suitable subjects can be determined through conventional screening methods and are typically individuals diagnosed with a relevant disease or pathological condition requiring treatment, relief, prevention, or improvement.

[0022] This invention relates to the use of sine oxaliplatin in the preparation of a medicament for the treatment of Parkinson's disease.

[0023] According to the present invention, Songguoling is a type of C20 Diterpenoid alkaloids, acting on the central nervous system, significantly improve Parkinson's disease-related dopaminergic neuronal damage and motor dysfunction. In a specific embodiment, a subacute Parkinson's disease mouse model was established using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The efficacy of Songguoling was systematically evaluated using behavioral tests such as the rotarod test, pole climbing test, and open field test. The results showed that after administration of Songguoling, the model animals' motor coordination, spontaneous activity, and exploratory behavior were significantly improved. Simultaneously, the degree of damage to dopaminergic neurons in the substantia nigra pars compacta was significantly reduced, and the number and expression level of TH-positive neurons significantly increased, demonstrating a good neuroprotective effect.

[0024] Furthermore, under different dosage conditions, Songguoling consistently produced stable therapeutic effects in Parkinson's disease model animals, and even at lower dosage levels, it exhibited efficacy superior to or equivalent to commonly used clinical positive control drugs, indicating that Songguoling possesses high pharmacodynamic activity and good application potential. This invention, through systematic pharmacodynamic experiments, verified the basis of Songguoling's effects in improving dopaminergic nerve function damage, alleviating motor disorders, and related behavioral deficits, thus confirming its suitability for preparing drugs for treating Parkinson's disease. This provides reliable experimental evidence and technical support for the development of novel anti-Parkinson's disease therapeutic agents.

[0025] In some further embodiments of the present invention, the Songguoling is prepared as a drug for treating Parkinson's disease, suitable for patients with Parkinson's disease of different etiologies and clinical characteristics, including but not limited to idiopathic Parkinson's disease, hereditary Parkinson's disease, and secondary Parkinsonian syndrome. In some embodiments of the present invention, the Parkinson's disease is idiopathic Parkinson's disease.

[0026] In some embodiments, the sinecuronium is combined with pharmaceutically acceptable excipients to form a pharmaceutical composition.

[0027] Pharmaceutical compositions can be formulated using methods known to those skilled in the art. For example, they can be administered parenterally as a sterile solution or suspension of water or a pharmaceutically acceptable solution other than water. For example, suitable combinations of pharmacologically acceptable carriers or media, specifically sterile water, physiological saline, vegetable oils, emulsifiers, suspending agents, surfactants, stabilizers, flavoring agents, excipients, mediators, preservatives, binders, etc. It is considered to be formulated by mixing in a commonly accepted unit dosage form required for pharmaceutical administration. The amount of the active ingredient in these formulations is set to obtain an appropriate volume within the indicated range.

[0028] Sterile compositions for injection can be formulated using excipients such as distilled water for injection, following conventional formulation methods.

[0029] Examples of aqueous solutions for injection include isotonic solutions containing, for example, physiological saline, lactose, glucose, and other supplements (such as D-sorbitol, D-mannose, D-mannitol, and sodium chloride). Appropriate dissolving aids can be used in combination, such as alcohols (ethanol, etc.), polyols (propylene glycol, polyethylene glycol, etc.), and nonionic surfactants (polysorbate 80(TM), HCO-50, etc.).

[0030] As an oily liquid, sesame oil and soybean oil can be examples, and benzyl benzoate and / or benzyl alcohol can be used in combination as a solubilizing agent. The composition can be mixed with buffers (e.g., phosphate buffer solutions and sodium acetate buffer solutions), analgesics (e.g., procaine hydrochloride), stabilizers (e.g., benzyl alcohol and phenol), and antioxidants. The prepared injection solution is typically packaged in appropriate ampoules.

[0031] In some embodiments, the pharmaceutical composition is an oral formulation, an injectable formulation, a mucosal administration formulation, or a sustained-release formulation. This adapts to different clinical application scenarios and patient needs. The oral formulation is suitable for long-term maintenance therapy or outpatient use, the injectable formulation is suitable for rapid onset of action or intervention during acute exacerbations, the mucosal administration formulation is suitable for improving absorption rate and bioavailability, and the sustained-release or controlled-release formulation is suitable for prolonging the duration of drug action and reducing the frequency of administration. By rationally selecting the formulation type and prescription composition, the release behavior and pharmacokinetic characteristics of songguoling in vivo can be further optimized, thereby improving its therapeutic effect on Parkinson's disease.

[0032] In some embodiments, the oral formulation is a tablet, capsule, granule, pill, oral liquid, suspension, or drop. Tablets and capsules can be prepared by wet granulation, dry granulation, or direct compression, and can be coated with a film or enteric coating as needed. Granules and pills can be obtained by extrusion spheronization, spray drying, or coating molding. Oral liquids and suspensions can be prepared by dissolving or dispersing sine in a suitable solvent system. Drops can be obtained by melt dripping or cold forming. These oral formulations have the advantages of convenient administration and high patient compliance, making them suitable for routine treatment of Parkinson's disease.

[0033] In some embodiments, the injectable formulation is an injectable solution or a lyophilized powder for injection. The injectable solution can be prepared by dissolving succinate in sterile water for injection or a solution containing a suitable buffer system and solubilizer, followed by sterile filtration and aseptic filling. The lyophilized powder for injection can be prepared by first preparing a succinate solution, then aseptically dispensing and freeze-drying it to form a solid formulation, which is then reconstituted and injected before use. By preparing the injectable formulation, succinate can rapidly enter the systemic circulation, thereby facilitating a faster onset of action in the treatment of Parkinson's disease.

[0034] In some embodiments, the mucosal administration formulation is a sublingual tablet, an orally disintegrating tablet, a nasal spray, or an oral spray. The sublingual tablet and orally disintegrating tablet can be rapidly disintegrated or dissolved in the oral cavity by controlling the excipient composition and tableting process, thereby promoting the absorption of Songguoling via the oral mucosa. The nasal spray and oral spray can be prepared by formulating Songguoling into a solution or a microdispersion system and using a spray device to achieve quantitative administration. Mucosal administration can partially avoid the first-pass effect, increase the drug absorption rate, and facilitate the rapid anti-Parkinson's effect of Songguoling.

[0035] In some embodiments, the sustained-release formulation is a sustained-release tablet, sustained-release capsule, or microsphere formulation. The sustained-release tablet or capsule can be designed using a sustained-release matrix material, coating material, or multilayer structure to allow for the slow release of sinecurin in vivo. The microsphere formulation can be obtained through processes such as emulsification solvent evaporation, spray drying, or phase separation, and the controlled-release effect can be achieved by adjusting the properties of the carrier material. By preparing sustained-release or controlled-release formulations, the effective duration of action of sinecurin in vivo can be prolonged, the frequency of dosing can be reduced, and patient compliance and treatment stability can be improved.

[0036] In some embodiments, the subject of the drug is a mammal.

[0037] In some embodiments, the subject of the drug is a primate.

[0038] In some embodiments, the subject of the drug is a human being.

[0039] The present invention also relates to a method for treating Parkinson's disease, the method comprising administering to a subject with Parkinson's disease an effective amount of sinecuronium or a pharmaceutical composition as described above.

[0040] In some embodiments of the present invention, the dosage of Songguoling can be adjusted according to the subject's age, weight, disease severity, route of administration, and individual differences. In animal experiments, Songguoling, administered at a dose range of 0.3–3 mg / kg, significantly improved motor dysfunction and dopaminergic neuron damage in Parkinson's disease model animals, with a more stable and significant efficacy observed in the 1–3 mg / kg dose range. In clinical applications, based on the above animal experimental results and pharmacokinetic conversion relationships, the recommended dosage of Songguoling is 0.01–1 mg / kg / day, preferably 0.05–0.5 mg / kg / day. The specific dosing regimen can be appropriately adjusted according to individual patient differences and treatment response. Songguoling can be administered as a single dose or in divided doses, with a dosing frequency of once daily or two to three times daily, to ensure therapeutic efficacy while reducing the risk of adverse reactions, thereby achieving safe and effective long-term medication management.

[0041] In some embodiments of the present invention, the Songguoling can be administered to subjects via various routes of administration, including but not limited to oral administration, injection administration, and transmucosal administration. Oral administration can be in the form of tablets, capsules, granules, oral solutions, or suspensions, suitable for long-term maintenance therapy; injection administration can be administered intravenously, intramuscularly, or subcutaneously, suitable for clinical scenarios requiring rapid relief of Parkinson's symptoms; transmucosal administration can be achieved through sublingual tablets, orally disintegrating tablets, or nasal sprays to improve absorption rate and bioavailability. Those skilled in the art can rationally select the route of administration and frequency of administration according to specific clinical needs.

[0042] In some embodiments of the present invention, Songguoling can be used alone as an active drug for the treatment of Parkinson's disease to fully exert its anti-Parkinson's activity. In this embodiment, Songguoling can be administered to the subject as the sole primary therapeutic ingredient, thereby achieving a significant anti-Parkinson's effect at a lower dose and reducing the risk of adverse reactions associated with the combined use of multiple drugs.

[0043] In another embodiment of the invention, Songguoling can also be used in combination with other anti-Parkinson's drugs to further improve the therapeutic effect or improve the therapeutic response of specific patient groups. The other anti-Parkinson's drugs may be selected from levodopa, carbidopa, benserazide, entacapone, tocapone, octopcapone, rasagiline, selegiline, safenamide, amantadine, pramipexole, ropinirole, rotigotine, apomorphine, levopraprazole, piribedil, bromocriptine, cabergoline, lisuva, trihexyphenidyl, biperiden, propiconamide, droxidopa, itratheline, caffeine, coenzyme Q10, alpha-lipoic acid, ginkgo biloba extract, and nilotinib. In the combination therapy embodiment, Songguoling can be administered simultaneously, at different times, or alternately with the above-mentioned drugs, and the specific dosing regimen can be adjusted according to the patient's condition and clinical experience.

[0044] In a further embodiment, the treatment method can be implemented using a phased or periodic dosing regimen. For example, during the acute phase, intensive treatment can be administered at a higher frequency or via injection, and after symptom relief, maintenance treatment can be switched to oral formulations to consolidate the therapeutic effect and reduce the risk of relapse. By rationally designing the dosing regimen and combination therapy strategy, Songguoling can exert a stable and continuous anti-Parkinson's effect at different stages of treatment, thereby improving the overall treatment effect and the patient's quality of life.

[0045] The embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the technical content of the present invention, rather than to limit the protection scope of the present invention. If specific experimental conditions are not clearly defined in the following examples, the guidance given in the specification of the present invention shall be preferentially referred to, or the experiments may also be carried out according to the generally recognized experimental manuals or conventional experimental conditions in the art, or other known experimental methods in the art, or according to the conditions recommended by relevant reagent or instrument manufacturers. In specific examples, for the measurement parameters of raw material components, if there is no special description, slight deviations within the weighing accuracy range are allowed; for parameters such as temperature and time, reasonable deviations caused by instrument detection accuracy or operation accuracy are also allowed.

[0046] Example 1 This example discloses the effect of the C-type diterpenoid alkaloid songorine on MPTP-induced subacute Parkinson's model mice, which is specifically verified by the following process: 20 1. Experimental materials and instruments MPTP, Apetaldine B, and Madopar are all commercially available products; songorine and total alkaloids of Aconitum carmichaeli Debx. are prepared in the laboratory; TH antibody (rabbit anti-mouse, Abcam, ab137869); DAB chromogenic kit (Beijing Zhongshan Jinqiao).

[0047] Climbing pole device (wooden pole length 50 cm, diameter 1 cm, with a wooden ball fixed at the top); rotarod (Zhongshi Technology Co., Ltd.); open field box (50 cm×50 cm×40 cm, VideoTrack real-time analysis by Zhongshi Technology Co., Ltd.).

[0048] 2. Experimental animals 8-week-old male C57BL / 6N mice, weighing 25–30 g, SPF grade, purchased from Beijing Vital River, license number: SYXK (Beijing) 2023-0001. Feeding environment: temperature 23±2 °C, humidity 50±10%, 12 h / 12 h light-dark cycle, free access to food and water. After 3 days of adaptive feeding, the experiment was started, with free access to water.

[0049] 3. Experimental grouping and model establishment 120 mice were randomly divided into 8 groups (n = 15) according to body weight: ① Blank group (normal saline) ② Model group (MPTP + normal saline) ③ Madopar group (MPTP + Madopar 50 mg / kg) ④ Low-dose songorine group (MPTP + songorine 0.3 mg / kg) ⑤ Medium-dose group of Songguoling (MPTP + Songguoling 1 mg / kg) ⑥ High-dose Songguoling group (MPTP + Songguoling 3 mg / kg) ⑦ Apetaldine B group (MPTP+Apetaldine B 10 mg / kg) ⑧ Total alkaloids of Aconitum carmichaelii group (MPTP + total alkaloids of Aconitum carmichaelii 10 mg / kg) Modeling method: Except for the blank group, the other groups were injected intraperitoneally with MPTP 30 mg / kg (0.1 mL / 10 g) daily from day 1 to day 5, while the blank control group was given an equal volume of physiological saline.

[0050] 4. Dosing regimen Starting from day 2 after the first MPTP injection (days 2-15), the drug was administered once daily by gavage for 14 consecutive days, at a volume of 0.1 mL / 10 g. The blank control group and the MPTP model group were given normal saline.

[0051] 5. Behavioral testing Complete the following sequentially 1 hour after the last dose: ① Rotating bar experiment: linear acceleration at 4–40 rpm for 5 min, record the dwell time, and take the average value; ② Pole climbing experiment: Record the time to turn around and the total time (s) to climb to the bottom of the pole, and take the average of 3 times; ③ Open field experiment: Explore freely for 5 minutes, and record the total distance traveled, the time spent in the central area, and the number of times the central area was crossed.

[0052] 6. TH staining and quantification After behavioral testing, four mice were randomly selected from each of the blank control group, model group, levodopa group, and low, medium, and high dose groups of sine trimethoprim. After deep anesthesia, the mice were fixed by cardiac perfusion with 4% paraformaldehyde. The brains were then removed and fixed for another 24 hours. After dehydration with 30% sucrose, the brains were prepared into coronal frozen sections (30 μm thick) covering the substantia nigra region.

[0053] After washing with PBS and blocking with serum, the slides were incubated with mouse anti-TH primary antibody at 4°C for 48 hours; subsequently, they were incubated sequentially with biotin-labeled secondary antibody and ABC complex, followed by DAB staining. Finally, the slides were mounted, dehydrated, cleared, and sealed.

[0054] The stained sections were scanned in their entirety using a 3D Histech Pannoramic MIDI II fully automated digital slicing system. The scanned images were analyzed using the accompanying QuantCenter software: the region of interest in the substantia nigra was manually delineated, and the DAB signal was separated via color deconvolution. Then, Image-Pro Plus 6.0 image analysis software was used to count and analyze the TH-positive neurons.

[0055] 7. Statistical processing Data are expressed as mean ± standard error (Mean ± SD). Experimental results were analyzed using GraphpadPrism 10.0 software. One-way ANOVA was first performed among multiple groups, followed by statistical testing using Tukey's multiple comparisons test. P A value <0.05 is considered statistically significant.

[0056] 8. Experimental Results As shown in Table 1, compared with the model group, all dose groups of Songguoling (0.3, 1, and 3 mg / kg) significantly prolonged the time spent on the rotarod in MPTP-induced Parkinson's disease model mice, demonstrating a significant improvement in motor function. Among them, the high-dose group of Songguoling (3 mg / kg) showed significantly better improvement than the treatment groups treated with first-line clinical drugs such as Madopar (50 mg / kg), Apetaldine B (10 mg / kg), and Aconitum carmichaelii total alkaloids (10 mg / kg).

[0057] Table 1. Effects of Song Guoling on Rotor Duration in MPTP-Induced Parkinson's Disease Model Mice

[0058] (Compared to the blank group, # P <0.05; compared with the model group, P <0.001, P <0.0001; compared with the Madopar group, @ P <0.05) As shown in Table 2, compared with the model group, all dose groups of Songguoling (0.3, 1, and 3 mg / kg) significantly shortened the pole descent time in MPTP-induced Parkinson's disease model mice, indicating that it has a significant effect on improving motor function in PD mice. Among them, the high-dose group of Songguoling (3 mg / kg) showed significantly better improvement than the treatment groups of the first-line clinical positive control drugs, Madopar (50 mg / kg), Apetaldine B (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg).

[0059] Table 2. Effects of Song Guoling on pole climbing and descent time in MPTP-induced Parkinson's disease model mice

[0060] (Compared to the blank group, #### P <0.0001; compared with the model group, P <0.05, P <0.01, P <0.0001; compared with the Madopar group, @ P <0.05) As shown in Table 3, compared with the model control group, all dose groups of Songguoling (0.3, 1, 3 mg / kg) significantly increased the total distance of movement in the open field test of MPTP-induced Parkinson's disease model mice, indicating that it can effectively improve the autonomous exploration behavior and overall motor level of model mice. Among them, the efficacy of the high-dose group of Songguoling was significantly better than that of the positive control drugs Madopar (50 mg / kg), Apetaldine B (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg), further confirming the ameliorative effect of Songguoling on Parkinson's disease-related behavioral deficits.

[0061] Table 3. Effects of Song Guoling on the total distance traveled in the open field experiment of MPTP-induced Parkinson's disease model mice.

[0062] (Compared to the blank group, ## P <0.01; compared with the model group, P <0.01, P <0.001, P <0.0001) In open field experiments, central zone distance is a key indicator reflecting the animal's exploratory behavior and anxiety state. Table 4 shows that the central zone distance of the model group mice was significantly shorter than that of the control group, indicating that MPTP induction successfully induced behavioral changes in mice. Compared with the model group, the positive control drug levodopa group and all doses of succinate (0.3, 1, and 3 mg / kg) significantly increased the central zone distance of mice. The high-dose succinate group (3 mg / kg) showed the most significant effect, significantly superior to the positive control drug levodopa (50 mg / kg), apetaldine B (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg) treatment groups.

[0063] The results indicate that Songguoling can improve the exploratory behavior and reduce anxiety-like symptoms in MPTP-induced Parkinson's disease model mice in a dose-dependent manner.

[0064] Table 4. Effects of Song Guoling on central zone distance in MPTP-induced Parkinson's disease model mice during open field experiments.

[0065] (Compared to the blank group, # P <0.05; compared with the model group, P <0.01, P <0.001, P <0.0001) As shown in Table 5, compared with the model group, all dose groups of Songguoling (0.3, 1, and 3 mg / kg) significantly increased the number of times MPTP-induced PD model mice entered the central region in the open field experiment, indicating that it had an ameliorative effect on the exploratory behavior and anxiety-like state of the model mice. Among them, the high-dose group of Songguoling (3 mg / kg) had the most significant effect, which was significantly better than the positive control drugs Madopar (50 mg / kg), Apetaldine B (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg), and the dose-dependent effect was better.

[0066] Table 5. Effect of Song Guoling on the number of central region entries in MPTP-induced Parkinson's disease model mice during the open field experiment.

[0067] (Compared to the blank group, # P <0.05; compared with the model group, P <0.01, P <0.001, P <0.0001) like Figure 1 As shown, changes in dopaminergic neurons in the substantia nigra pars compacta of MPTP-induced Parkinson's disease model mice were observed using TH immunohistochemical staining. The results showed that the number of TH-positive neurons in the substantia nigra pars compacta of the MPTP model group was significantly reduced compared to the control group, indicating significant damage to dopaminergic neurons. Compared to the model group, the number of TH-positive neurons in each dose group of levodopa and sine trimethoprim increased to varying degrees. Among them, each dose group of sine trimethoprim (0.3, 1, and 3 mg / kg) showed a significant protective effect, and the high-dose group (3 mg / kg) was more effective than the levodopa group (50 mg / kg), suggesting that the protective effect of sine trimethoprim on TH neurons is dose-dependent and superior to positive control drugs at high doses. Further image analysis showed that sine trimethoprim treatment significantly increased the mean optical density of TH-positive neurons and reduced neuronal morphological atrophy.

[0068] The above results indicate that Songguoling can significantly reduce MPTP-induced damage to dopaminergic neurons in the substantia nigra, demonstrating a clear neuroprotective effect. Its effect is superior to that of Madopar at high doses.

[0069] In summary, the present invention describes a C 20 The diterpenoid alkaloid songguoling has a significant anti-Parkinson's disease effect, with efficacy significantly superior to that of Madopar, a first-line clinical drug for Parkinson's disease. Moreover, its effective dose is much lower than that of positive control drugs, making it a potential candidate for new anti-Parkinson's disease drugs.

[0070] Application Example 1 This invention discloses a capsule formulation using Songguoling as the active pharmaceutical ingredient, the components of which are as follows: Songguoling 30.0mg 6.0g of starch Sodium metabisulfite 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 pills.

[0071] The specific preparation process is as follows: Mix Songguoling, starch, and sodium metabisulfite evenly, add anhydrous ethanol to make a soft material, pass it through a 24-mesh sieve to make granules, dry it, add magnesium stearate, mix well, and fill into capsules.

[0072] Application Example 2 This invention discloses a granule formulation using the compound succinate as a raw material, the components of which are as follows: Songguoling 25.0mg 6.0g of starch Sodium bisulfite 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 bags.

[0073] The specific preparation process is as follows: After mixing Songguoling with starch and sodium bisulfite, anhydrous ethanol is added to form a soft material. The material is then passed through a 24-mesh sieve to form granules, dried, and then magnesium stearate is added. The mixture is then mixed and packaged.

[0074] Application Example 3 This invention discloses an oral liquid using the compound Songguoling as a raw material, the components of which are as follows: Songguoling 20.0mg 3.0g sucrose Sodium bisulfite 0.2g Methylparaben 0.2g Sodium bicarbonate 0.5g 1000.0 mL of water for injection Make 100 pieces.

[0075] The specific preparation process is as follows: After the above components are mixed, they can be dispensed using conventional oral liquid preparation methods.

[0076] Application Example 4 This invention discloses an injection formulation using the compound succinate as the active pharmaceutical ingredient, the components of which are as follows: Songguoling 25.0mg Vitamin C 0.2g Sodium chloride 6.0g Sodium bicarbonate 0.5g 1000.0 mL of water for injection Make 100 pieces.

[0077] The specific preparation process is as follows: After the above components are mixed, 100 vials can be obtained by using conventional injection preparation methods.

[0078] Application Example 5 This invention discloses a tablet using the compounds songguoling and levodopa as active pharmaceutical ingredients, the composition of which is as follows: Songguoling 20.0mg Levodopa 60.0mg Hydroxypropyl methylcellulose 18.0g Talc powder 0.4g Lactose 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 pieces.

[0079] The specific preparation process is as follows: Mix Songguoling, L-DOPA, hydroxypropyl methylcellulose, talc, lactose, and magnesium stearate evenly, add anhydrous ethanol to make a soft mass, pass it through a 24-mesh sieve to make granules, dry it, add magnesium stearate, mix well, and compress into tablets.

[0080] Application Example 6 This invention discloses a capsule formulation using the compounds sineb and levodopa as active pharmaceutical ingredients, the components of which are as follows: Songguoling 3.5mg Madopar 300.0mg 6.0g of starch Sodium metabisulfite 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 pills.

[0081] The specific preparation process is as follows: Take succinate, benzyl hydrazine and sodium metabisulfite, mix them, add anhydrous ethanol to make a soft material, pass it through a 24-mesh sieve to make granules, dry it, add magnesium stearate, mix well, and fill it into capsules.

[0082] Application Example 7 This invention discloses an injection using the compounds songguoling and carbidopa as active pharmaceutical ingredients, the components of which are as follows: Songguoling 5.0mg Carbidopa 20.0mg Vitamin C 0.2g Sodium chloride 6.0g Sodium bicarbonate 0.5g 1000.0 mL of water for injection Make 100 pieces.

[0083] The specific preparation process is as follows: After the above components are mixed, 100 vials can be obtained by using conventional injection preparation methods.

[0084] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. Application of Songguoling in the preparation of drugs for the treatment of Parkinson's disease.

2. The application according to claim 1, wherein the Parkinson's disease is idiopathic Parkinson's disease.

3. The application according to claim 1, wherein the sinecurin is combined with pharmaceutically acceptable excipients to form a pharmaceutical composition.

4. The application according to claim 3, wherein the pharmaceutical composition is an oral formulation, an injectable formulation, a mucosal administration formulation, or a sustained-release formulation.

5. The application according to claim 4, wherein the oral preparation is a tablet, capsule, granule, pill, oral liquid, suspension, or drop.

6. The application according to claim 4, wherein the injectable formulation is an injection solution or a lyophilized powder for injection.

7. The application according to claim 4, wherein the transmucosal administration formulation is a sublingual tablet, an orally disintegrating tablet, a nasal spray, or an oral spray.

8. The application according to claim 4, wherein the sustained-release formulation is a sustained-release tablet, sustained-release capsule, or microsphere formulation.

9. The application according to any one of claims 1 to 8, wherein the subject of the drug is a mammal.

10. The application according to claim 9, wherein the subject of the drug is a human.

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