Use of ginkgolides in the preparation of a medicament for the prevention and / or treatment of Parkinson's disease

Abstract

The present application relates to the technical field of medicine, in particular to the use of ginkgolide in the preparation of a medicine for preventing and / or treating Parkinson's disease. The ginkgolide and the composition or extract thereof can improve the motor coordination of a Parkinson's disease model mouse, have a therapeutic effect on Parkinson's disease, and can be used in the preparation of a medicine for treating Parkinson's disease.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically to the use of ginkgolide lactone in the preparation of medicaments for the prevention and / or treatment of Parkinson's disease. Background Technology

[0002] Modern research indicates that the main active components in ginkgo leaves are flavonoids and ginkgolides. Ginkgo flavonoids include flavonol glycosides, biflavonoids, and catechins. Ginkgoolides include diterpenoid lactones and ginkgolides. Diterpenoid lactones include ginkgolides A, B, C, J, M, K, L, N, P, and Q.

[0003] Parkinson's disease is a serious neurodegenerative disorder that severely threatens the health and quality of life of middle-aged and elderly people. Clinically, it is characterized by resting tremor, bradykinesia, rigidity, postural instability, and speech impairment. Its incidence is increasing annually, and while there is no cure, it can be managed; therefore, early diagnosis is crucial. Among the various manifestations of Parkinson's disease, speech impairment is one of the typical early symptoms, affecting approximately 90% of Parkinson's patients to some degree.

[0004] Therefore, it is essential to develop new drugs for the prevention or treatment of Parkinson's disease. Summary of the Invention

[0005] The purpose of this invention is to provide its use in medicines for the prevention, treatment and / or adjunctive treatment of Parkinson's disease.

[0006] One or more embodiments of this application provide the use of ginkgolide lactone in the preparation of medicaments for the prevention and / or treatment of Parkinson's disease.

[0007] One or more embodiments of this application provide the use of pharmaceutical compositions comprising ginkgolide in the preparation of medicaments for the prevention, treatment and / or adjunctive treatment of Parkinson's disease.

[0008] One or more embodiments of this application provide the use of extracts containing ginkgolide lactones in the preparation of medicaments for the prevention, treatment and / or adjunctive treatment of Parkinson's disease.

[0009] In one or more embodiments, the ginkgolide comprises at least one of ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, and ginkgolide K, or is composed of one or more of the above components.

[0010] In one or more embodiments, the weight ratio of ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, and ginkgolide K is selected from one of the following:

[0011] Ginkgolide A: Ginkgolide B = (10–45): (5–40); for example, (10, 20, 30, or 45): (5, 10, 20, 30, or 40);

[0012] Ginkgolide A: Ginkgolide C = (10–45): (3–35); for example, (10, 20, 30, or 45): (3, 10, 20, 30, or 35);

[0013] Ginkgolide B: Ginkgolide C = (5–40): (3–35); for example, (5, 10, 20, 30, or 40): (3, 10, 20, 30, or 35);

[0014] Ginkgolide A: Ginkgolide B: Ginkgolide C = (10–45): (5–40): (3–35); for example, (10, 20, 30, or 45): (5, 10, 20, 30, or 40): (3, 10, 20, 30, or 35);

[0015] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide J = (10–45): (5–40): (3–35): (1–10); for example, (10, 20, 30, or 45): (5, 10, 20, 30, or 40): (3, 10, 20, 30, or 35): (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10);

[0016] Ginkgolide A: Ginkgolide K = (3-40): (0.01-5); for example, (3, 10, 20, 30, or 40): (0.01, 0.1, 1, 2, 3, 4, or 5);

[0017] Ginkgolide B: Ginkgolide K = (5-75): (0.01-5); for example, (5, 10, 20, 30, 40, 50, 60, 70, or 75): (0.01, 0.1, 1, 2, 3, 4, or 5);

[0018] Ginkgolide A: Ginkgolide B: Ginkgolide K = (3-40): (5-75): (0.01-5); for example, (3, 10, 20, 30, or 40): (5, 10, 20, 30, 40, 50, 60, 70, or 75): (0.01, 0.1, 1, 2, 3, 4, or 5).

[0019] In one or more embodiments, the ginkgolide further comprises ginkgolide.

[0020] In one or more embodiments, the ginkgolide is composed of one or more of ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, ginkgolide K, and ginkgolide.

[0021] In one or more embodiments, the mass ratio of ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, ginkgolide K, and ginkgolide is selected from one of the following:

[0022] Ginkgolide A: Ginkgolide B: Ginkgolide = (10–45): (5–40): (10–70); for example, (10, 20, 30, or 45): (5, 10, 20, 30, or 40): (10, 20, 30, or 40): (10, 20, 30, 40, 50, 60, or 70);

[0023] Ginkgolide A: Ginkgolide C: Ginkgolide = (10–45): (3–35): (10–70); for example, (10, 20, 30, or 45): (3, 10, 20, 30, or 35): (10, 20, 30, 40, 50, 60, or 70);

[0024] Ginkgolide B: Ginkgolide C: Ginkgolide = (5–40): (3–35): (10–70); for example, (5, 10, 20, 30, or 40): (3, 10, 20, 30, or 35): (10, 20, 30, 40, 50, 60, or 70);

[0025] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide = (10–45): (5–40): (3–35): (10–70); for example, (10, 20, 30, or 45): (5, 10, 20, 30, or 40): (3, 10, 20, 30, or 35): (10, 20, 30, 40, 50, 60, or 70):

[0026] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide J: Ginkgolide = (10–45): (5–40): (3–35): (1–10): (10–70); for example, (10, 20, 30, or 45): (5, 10, 20, 30, or 40): (3, 10, 20, 30, or 35): (10, 20, 30, 40, 50, 60, or 70):

[0027] Ginkgolide A: Ginkgolide K: Ginkgolide = (3-40): (0.01-5): (10-70); for example, (3, 10, 20, 30, or 40): (0.01, 0.1, 1, 2, 3, 4, or 5): (10, 20, 30, 40, 50, 60, or 70);

[0028] Ginkgolide B: Ginkgolide K: Ginkgolide = (5–75): (0.01–5): (10–70); for example, (5, 10, 20, 30, 40, 50, 60, 70, or 75): (0.01, 0.1, 1, 2, 3, 4, or 5): (10, 20, 30, 40, 50, 60, or 70):

[0029] Ginkgolide A: Ginkgolide B: Ginkgolide K: Ginkgolide = (3-40): (5-75): (0.01-5): (10-70); for example, (3, 10, 20, 30, or 40): (5, 10, 20, 30, 40, 50, 60, 70, or 75): (0.01, 0.1, 1, 2, 3, 4, or 5): (10, 20, 30, 40, 50, 60, or 70).

[0030] In one or more embodiments, the mass ratio of ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, ginkgolide K, and ginkgolide is selected from one of the following:

[0031] Ginkgolide A: Ginkgolide B: Ginkgolide = (20-30): (10-20): (35-50); for example, (20, 25, or 30): (10, 15, or 20): (35, 40, 45, or 50);

[0032] Ginkgolide A: Ginkgolide C: Ginkgolide = (20-30): (10-20): (35-50); for example, (20, 25, or 30): (10, 15, or 20): (35, 40, 45, or 50);

[0033] Ginkgolide B: Ginkgolide C: Ginkgolide = (10-20): (10-20): (35-50); for example, (10, 15, or 20): (10, 15, or 20): (35, 40, 45, or 50);

[0034] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide = (20-30): (10-20): (10-20):

[0035] (35–50); for example, (20, 25, or 30): (10, 15, or 20): (10, 15, or 20): (35, 40, 45, or 50):

[0036] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide = (10-20): (30-40): (3-10):

[0037] (35-50); for example, (10, 15, or 20): (30, 35, or 40): (3, 4, 5, 6, 7, 8, 9, or 10): (35, 40, 45, or 50);

[0038] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide = (10-15): (30-35): (5-10):

[0039] (40-50); for example, (10, 11, 12, 13, 14, or 15): (30, 31, 32, 33, 34, or 35): (5, 6, 7, 8, 9, or 10): (40, 41, 41, 43, 44, 45, 46, 47, 48, 49, or 50);

[0040] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide = (10–15): (30–35): (5–8): (45–50); for example, (10, 11, 12, 13, 14, or 15): (30, 31, 32, 33, 34, or 35): (5, 6, 7, or 8): (45, 46, 47, 48, 49, or 50):

[0041] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide = 12:34:6:48.

[0042] Ginkgolide A: Ginkgolide B: Ginkgolide C: Ginkgolide J: Ginkgolide = (20-30): (10-20): (10-20): (2-7): (35-50); for example, (20, 25, or 30): (10, 15, or 20): (10, 15, or 20): (2, 3, 4, 5, 6, or 7): (35, 40, 45, or 50);

[0043] Ginkgolide A: Ginkgolide K: Ginkgolide = (20-30): (0.01-5): (35-50); for example, (20, 25, or 30): (0.01, 0.1, 1, 2, 3, 4, or 5): (35, 40, 45, or 50);

[0044] Ginkgolide B: Ginkgolide K: Ginkgolide = (10-20): (0.01-5): (35-50); for example, (10, 15, or 20): (0.01, 0.1, 1, 2, 3, 4, or 5): (35, 40, 45, or 50);

[0045] Ginkgolide A: Ginkgolide B: Ginkgolide K: Ginkgolide = (20-30): (10-20): (0.01-5): (35-50); for example, (20, 25, or 30): (10, 15, or 20): (0.01, 0.1, 1, 2, 3, 4, or 5): (35, 40, 45, or 50).

[0046] In one or more embodiments, the dose of the ginkgolide lactone is 1 to 800 mg / kg / day (e.g., 100, 200, 300, 400, 500, 600, or 700 mg / kg / day).

[0047] In one or more embodiments, the dose of the ginkgolide lactone is 1 to 500 mg / kg / day.

[0048] One or more embodiments of this application have the following beneficial effects: Ginkgolide of the present invention can improve motor coordination in Parkinson's disease model mice, indicating that it has a preventive or therapeutic effect on Parkinson's disease, and can be used to prepare drugs for the prevention and / or treatment of Parkinson's disease, etc.

[0049] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0050] In one or more embodiments, the term "prevention" refers to preventing the occurrence of disease and / or preventing the recurrence of disease.

[0051] In one or more embodiments, "ginkgolic acid lactone" refers to ginkgolide compounds.

[0052] In one or more embodiments, ginkgolide lactones can be obtained by separation and purification using existing techniques, or they can be composed of corresponding ginkgolide lactone monomer compounds.

[0053] In one or more embodiments, when ginkgolide comprises two or more ginkgolide monomer compounds, it can be formed by combining the corresponding monomer compounds.

[0054] In one or more embodiments, the composition containing ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, ginkgolide K and / or ginkgolide can be obtained by directly purchasing commercially available ginkgolide injection, or by the method described in ZL200610103626.0 or ZL200610103625.6, or by combining monomeric compounds.

[0055] In one or more embodiments, the ginkgolide monomer compounds can be obtained by purchasing commercially available products or by separating and purifying them using existing methods.

[0056] In one or more embodiments, the ginkgolide monomer, specifically, may be, for example, ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide M, ginkgolide J, ginkgolide K, ginkgolide L, ginkgolide N, ginkgolide P, ginkgolide Q, or ginkgolide, etc.

[0057] In one or more embodiments, ginkgolides can be prepared by separation and purification using existing techniques. Attached Figure Description

[0058] Figure 1 The bar chart shows the time spent on the rotarod in each group of mice, where the ginkgolide group refers to composition A. Detailed Implementation

[0059] This specification provides a detailed description of specific embodiments. Those skilled in the art should recognize that the following embodiments are exemplary and should not be construed as limiting the invention. For those skilled in the art, various improvements and modifications can be made to the invention without departing from its principles; such improvements and modifications also fall within the scope of protection of the claims. The beneficial effects of the invention are illustrated below through specific examples.

[0060] Example 1

[0061] Efficacy evaluation of ginkgolide in a mouse model of chronic Parkinson's disease induced by MPTP

[0062] 1. Experimental Procedure

[0063] 1.1 Experimental Animals and Reagents

[0064] Male C57BL / 6J mice, 10-12 weeks old, 25-30g; purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0065] Composition A contains ginkgolides in a mass ratio of ginkgolide A: ginkgolide B: ginkgolide C: ginkgolide = 12:34:6:48.

[0066] 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP hydrochloride), purchased from Sigma, lot number S31504;

[0067] Probenecid, purchased from Aladdin, batch number H2117241.

[0068] 1.2 Establishment and administration of a chronic Parkinson's disease mouse model (PD)

[0069] Three days prior to group administration, all C57BL / 6J mice underwent rotarod training for three days to assess limb motor coordination, and mice with poor limb coordination were excluded. Mice were randomly divided into four groups of 10 mice each, based on body weight: a blank control group, a model group, a levodopa group (75 mg / kg per administration), a composition A group (150 mg / kg per administration), and a ginkgolide group (50 mg / kg per administration).

[0070] Except for the blank control group, mice in other groups were injected twice a week with MPTP (subcutaneous injection, 25 mg / kg) and Probenecid (intraperitoneal injection, 250 mg / kg), with a 30-minute interval between MPTP and Probenecid injections, for a total of 10 injections to establish a chronic PD model. Drug administration began at the same time as model establishment, with administration once daily by gavage for 9 weeks.

[0071] 1.3 Spinning Rod Behavior Test

[0072] At the end of week 9, a rotarod test was conducted to assess the motor coordination of the mice. The rotarod was approximately 2 cm in diameter. The mice were placed on the rotarod with their heads perpendicular to the long axis of the rotarod. The rotation speed of the rotarod was gradually increased from 10 rpm to 40 rpm over 5 minutes. Tests were conducted at 30-minute intervals, and the average value was recorded for three consecutive tests. The duration of rotarod testing for each group of mice was recorded to assess their motor coordination.

[0073] 1.4 Suspension Experiment

[0074] The suspension test was used to assess the coordination of limb movements in mice. The mice were suspended from both front paws on a fixed horizontal metal string, 25 cm above the ground. The suspension time was recorded. Failure was defined as only one paw grasping the string.

[0075] 1.5 Western blot analysis of tyrosine hydroxylase (TH) content in substantia nigra tissue

[0076] After the experiment, mice were euthanized, and their brains were removed on ice. Based on the mouse brain atlas, the substantia nigra was located, dissected, flash-frozen in liquid nitrogen, and stored at -80°C. Subsequent procedures, including protein extraction and SDS-PAGE gel electrophoresis, were performed to detect the expression of TH protein in the mouse substantia nigra.

[0077] 1.6 Data Analysis

[0078] SPSS 22.0 software was used for statistical analysis. Data are expressed as mean ± standard deviation. Differences among multiple groups were compared using one-way ANOVA:

[0079] Compared with the blank control group, #P<0.05, ##<0.01, ###<0.001;

[0080] Compared with the model group, *P<0.05, **P<0.01, ***<0.001.

[0081] 2. Experimental Results

[0082] 2.1 Results of the twirling rod behavioral test

[0083] The rotarod test in mice can be used to assess the coordination and balance abilities of mice, and the results are as follows: Figure 1 As shown. Compared with the blank control group, the model group mice had a significantly shorter dwell time on the stick (#P<0.05), indicating successful modeling. Compared with the model group, the mice in composition A group had a significantly longer dwell time on the stick (*P<0.05), which was statistically significant and superior to both the levodopa group (positive control) and the ginkgolide group, indicating that the composition containing ginkgolide in this application has a better effect on improving motor coordination in Parkinson's disease model mice.

[0084] 2.2 Suspension Test Results

[0085] Compared with the blank control group, the chronic PD model showed significant motor deficits.

[0086] 2.3 Quantitative analysis of tyrosine hydroxylase protein (TH)

[0087] To further verify the effect of ginkgolide on the damage of dopamine neurons in a Parkinson's disease model, the level of tyrosine hydroxylase protein in the substantia nigra of the brain was quantitatively analyzed using Western blot.

[0088] The results showed that, compared with the blank control group, the expression level of TH in the brain tissue of the model group was significantly reduced. Compared with the model group, composition A group increased the expression level of TH, and was superior to the levodopa group, indicating that the composition containing ginkgolide in this application has a protective effect against damage to dopamine neurons.

[0089] The above embodiments illustrate that the composition containing ginkgolide lactones of this application can improve the motor coordination of Parkinson's disease model mice, has a therapeutic or preventive effect on Parkinson's disease, and can be used to prepare drugs for the treatment or prevention of Parkinson's disease.

[0090] This specification provides a detailed description of specific embodiments. Those skilled in the art should recognize that the above embodiments are exemplary and should not be construed as limiting the invention. For those skilled in the art, various improvements and modifications can be made to the invention without departing from its principles or spirit, and the resulting technical solutions also fall within the scope of protection of the claims.

Claims

1. The use of ginkgolide as the sole active ingredient in the preparation of a medicament for the prevention and / or treatment of Parkinson's disease, wherein the ginkgolide is composed of ginkgolide A, ginkgolide B, ginkgolide C, and ginkgolide, and the weight ratio of ginkgolide A: ginkgolide B: ginkgolide C: ginkgolide = 12:34:6:48.

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