Use of rubidium chloride in the manufacture of a medicament for the treatment of parkinsonism
By using a drug prepared with rubidium chloride, the motor coordination and brain tissue damage of Parkinson's disease mice were improved, α-synuclein expression was reduced, and dopamine neuron function was enhanced, thus overcoming the shortcomings of existing treatment methods.
Patent Information
- Application Number
- CN202310819479.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Priority Date
- 2023-02-27
- Filing Date
- 2023-07-06
- Publication Date
- 2026-02-27
- Estimated Expiration
- 2043-07-06
AI Technical Summary
Currently, there are no effective drugs targeting the pathogenesis of Parkinson's disease, and existing treatments have failed to effectively improve movement disorders and brain tissue damage.
Rubidium chloride is used as the active ingredient and prepared into a pharmaceutically permissible dosage form. It is administered by gavage at a dose of not less than 0.5 mg/kg·d for the treatment of Parkinson's syndrome.
It significantly improved MPTP-induced motor dysfunction in mice, protected the substantia nigra and striatum structures, reduced α-synuclein expression, and enhanced dopamine neuron function.
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Figure CN116832057B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of medicine. More particularly, the present application relates to the use of rubidium chloride in the preparation of a medicament for treating Parkinson's syndrome. BACKGROUND
[0002] Parkinson's disease (PD) is a common neurodegenerative disease, its clinical manifestations mainly include resting tremor, bradykinesia, muscle rigidity and postural gait disorder, and patients can also have depression, constipation and sleep disorders and other non-motor symptoms. The diagnosis of Parkinson's disease mainly relies on the history, clinical symptoms and signs, and the general auxiliary examination has no abnormal change. Its pathogenesis is not yet fully understood, and there is no drug for treating its pathogenesis. Therefore, finding a drug that can target the research of its pathogenesis has become a hot spot of domestic and foreign research. SUMMARY
[0003] The present application provides the use of rubidium chloride in the preparation of a medicament for treating Parkinson's syndrome, rubidium chloride has a protective effect on MPTP-induced Parkinson's syndrome in mice, and provides a theoretical basis for preventing and treating Parkinson's syndrome.
[0004] In order to achieve these objects and other advantages in accordance with the present application, a use of rubidium chloride in the preparation of a medicament for treating Parkinson's syndrome is provided.
[0005] Preferably, the medicament contains a therapeutically effective amount of rubidium chloride and a pharmaceutically acceptable carrier.
[0006] Preferably, the pharmaceutically acceptable carrier includes diluents, solubilizers, latent solvents, disintegrants, dispersants, lubricants, flavoring agents, antioxidants, binders, absorbents, humectants, buffers, cross-linking agents.
[0007] Preferably, the medicament is prepared into a pharmaceutically acceptable dosage form.
[0008] Preferably, the medicament is prepared into a dosage form for intragastric administration.
[0009] Preferably, the dosage of rubidium chloride is not less than 0.5 mg / kg·d.
[0010] The present application at least includes the following beneficial effects:
[0011] MPTP modeling mice occurred dyskinesia, coordination ability decreased, rubidium chloride administration can significantly improve the mouse dyskinesia, improve motor coordination. At the same time, HE staining to observe the mouse brain structure, model group of brain structure and striatum damage, rubidium chloride administration can significantly improve the brain tissue structure and striatum damage. MPTP modeling α-synuclein significantly increased, rubidium chloride can significantly reduce the expression of α-synuclein. It is proved that rubidium chloride has protective effect on MPTP induced Parkinson's disease mice.
[0012] Other advantages, objects, and features of the present application will be apparent from the following specification and appended claims, and will be understood by those skilled in the art. BRIEF DESCRIPTION OF DRAWINGS
[0013] Figure 1 The effect of rubidium chloride on MPTP mouse parkinsonism score;
[0014] Figure 2 The effect of rubidium chloride on MPTP mouse open field test score;
[0015] Figure 3 The effect of rubidium chloride on MPTP mouse pole climbing test score;
[0016] Figure 4 The effect of rubidium chloride on MPTP mouse hanging test score;
[0017] Figure 5 The effect of rubidium chloride on MPTP mouse swimming test score;
[0018] Figure 6 The effect of rubidium chloride on MPTP mouse 5 behavioral score addition statistical results;
[0019] Figure 7 The effect of rubidium chloride on MPTP mouse brain tissue substantia nigra;
[0020] Figure 8 The effect of rubidium chloride on MPTP mouse brain tissue striatum;
[0021] Figure 9 The effect of rubidium chloride on MPTP mouse brain tissue α-synuclein;
[0022] Figure 10 The effect of rubidium chloride on DAT mRNA expression;
[0023] Figure 11 The effect of rubidium chloride on TH mRNA expression. DETAILED DESCRIPTION
[0024] The application will be further described in details below with reference to the accompanying drawings so that those skilled in the art can implement the application according to the description.
[0025] It should be understood that the terms such as "have", "contain" and "include" used herein do not exclude the presence or addition of one or more other elements or combinations thereof.
[0026] It should be noted that the experimental methods in the following embodiments are all conventional methods, and the reagents and materials are commercially available unless otherwise specified.
[0027] Use of rubidium chloride in the preparation of a medicament for treating Parkinson's syndrome.
[0028] The medicament contains a therapeutically effective amount of rubidium chloride and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier includes diluents, solubilizers, latent solvents, disintegrants, dispersants, lubricants, flavoring agents, antioxidants, binders, absorbents, humectants, buffers, cross-linking agents.
[0029] The medicament is prepared into a pharmaceutically acceptable dosage form. The dosage form includes pills, tablets, powders, capsules, granules, powders, dripping pills, dripping agents, sprays, injections, suspensions, gels, suppositories.
[0030] The medicament is prepared into a dosage form for gavage administration. The dosage form for gavage administration includes but is not limited to solutions, suspensions, solid preparations.
[0031] The administration dose of rubidium chloride is not less than 0.5 mg / kg·d. The pharmaceutical preparation of the application is used in the form of unit weight per dose.
[0032] 1 Experimental materials and methods
[0033] MPTP (purity ≥98%, sigma company), rubidium chloride (RbCl) (purity ≥98%, maklin company), compound levodopa tablets (shanghai roche pharmaceutical co., ltd.).
[0034] 1.1 Experimental animals
[0035] C57 / BL6J mice, 60 male, 7-8w old, 25-30g. The mice were adaptively fed for three days, all the mice were subjected to three open field experiments, and the mice with abnormal spontaneous behavior were excluded.
[0036] 1.2 Animal modeling
[0037] Randomly take 50 mice to give MPTP solution, intraperitoneal injection of 30 mg / kg, once a day, for 7 days, on the 3rd day and the 7th day, behavioral experiment detection and record score, according to the score of behavioral experiment, after reaching 7.5 points, it is judged that the modeling is successful, and the grouping administration is started.
[0038] 1.3 animal grouping
[0039] The mice were divided into 6 groups: blank control group (normal saline 30 mg / kg), model control group (MTPT 30 mg / kg), RbCl low dose group (7.5 mg / kg), RbCl medium dose group (15 mg / kg), RbCl high dose group (30 mg / kg), positive control group (compound levodopa 30 mg / kg), and MPTP solvent was normal saline. The above groups were respectively simplified as Control, MPTP, MPTP+RbCl 7.5mg / kg, MPTP+RbCl 15mg / kg, MPTP+RbCl 30mg / kg, MPTP+Dopamine 30mg / kg. Figures 1-11
[0040] 1.4 animal administration
[0041] After the successful modeling of the mice, the body weight was weighed and recorded every day, and the mice were gavaged according to the body weight of 0.1 mL / 10 g for 14 days, once a day, and the tremor behavior of the animals was observed every two days. On the 7th day and the 14th day of administration, behavioral experiment detection and record score were carried out. The next day, 30 mg / kg modeling agent was given to maintain the symptoms of PD.
[0042] 1.5 behavioral experiment
[0043] Behavioral detection includes: tremor paralysis, open field experiment, pole climbing experiment, suspension experiment and swimming experiment.
[0044] (1) tremor paralysis
[0045] Observe whether the mice appear to stand on their tails, stand on their hair, drool, tremor, paralysis, death and other conditions within 1 hour after administration, and score according to the mouse performance: 0 points, similar to normal mice, without any symptoms; 1 point, stand on their hair, arch their back, intermittent small tremor, but move freely; 2 points, frequent swallowing, frequent tremor, hind limbs open, tail tremor, and movement gradually limited; 3 points, drooling, persistent tremor, limbs stiff, and movement limited; 4 points, death due to whole body paralysis.
[0046] (2) open field experiment (to judge the activity)
[0047] The open field is made of 50 cm x 50 cm x 50 cm square box of acrylic material, evenly divided into 9 grids, the mice are put into the middle grid of the open field, the number of times of crossing into the adjacent grid is recorded, and the observation lasts for 2 min. The scoring standard is: 0 points, crossing frequency > 40 times; 1 point, crossing frequency is 31-40 times; 2 points, crossing frequency is 21-30 times; 3 points, crossing frequency < 21 times.
[0048] (3) Pole climbing experiment (judging the state of the forelimbs)
[0049] A softwood ball with a diameter of 25 cm is fixed on the top end of a wooden pole with a length of 50 cm and a diameter of 1 cm, and gauze is wrapped around the wooden pole to prevent slipping. The mouse to be tested is placed on the ball, and the time for the mouse to climb from the ball to the bottom is recorded. The scoring standard is: 0 points, pole climbing time < 4.01 s; 1 point, pole climbing time is 4.01-8.0 s; 2 points, pole climbing time is 8.01-12.0 s; 3 points, pole climbing time > 12.00 s.
[0050] (4) Suspension experiment (judging the gripping state of the hind paws)
[0051] The mouse to be tested is inverted and suspended, with both forepaws placed at the midpoint of a horizontal line with a length of 30 cm and a height of 25 cm. Then the mouse is released, and the gripping condition is recorded. The scoring standard is: 0 points, both hind paws can grip the line; hind paws repeatedly grip the line, intermittently can grip, can be scored as 0.5 points; 1 point, one hind paw can grip the line; 2 points, both hind paws cannot grip the line.
[0052] (5) Swimming experiment (judging the coordination ability of the mouse)
[0053] The mouse to be tested is placed in a water tank with a size of 40 cm x 25 cm x 16 cm, the water depth is 15 cm, and the water temperature is (27±2)℃. The floating time of the mouse within 3 min is recorded. The scoring standard is: 0 points, floating 0-30 s; 1 point, floating 31-90 s; 2 points, floating 91-150 s; 3 points, floating 151-180 s; 4 points, choking during floating.
[0054] 1.6 Sampling and detection of related indicators
[0055] (1) 1 day after the last administration, brain tissue is taken for HE staining of the substantia nigra and striatum;
[0056] (2) Immunohistochemical detection of α-synuclein expression in brain tissue;
[0057] (3) qRT-PCR detection of dopamine transporter (DAT) and tyrosine hydroxylase (TH) mRNA expression in mouse brain tissue.
[0058] 2 Experimental results
[0059] 2.1 Effect of RbCl on MPTP mouse tremor paralysis score
[0060] As shown in Figure 1 ### P < 0.001 vs Control; * P < 0.05, *** P < 0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP modeling, mice appeared piloerection, arching, intermittent tremor, salivation, limb stiffness, reduced activity, after administration, RbCl low dose group (7.5 mg / kg), RbCl middle dose group (15 mg / kg), RbCl high dose group (30 mg / kg) can significantly improve the mouse paralysis phenomenon, and RbCl high dose group (30 mg / kg) effect is most significant.
[0061] 2.2 Open field test
[0062] As shown in Figure 2 ### P < 0.001 vs Control; * P < 0.05, ** P < 0.01, *** P < 0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP modeling, open field test score was significantly increased, after administration, RbCl low dose group (7.5 mg / kg), RbCl middle dose group (15 mg / kg), RbCl high dose group (30 mg / kg) can significantly reduce the score, and RbCl high dose group (30 mg / kg) effect is most significant.
[0063] 2.3 Pole climbing test
[0064] As shown in Figure 3 ### P < 0.001 vs Control; * P < 0.05, ** P < 0.01, *** P < 0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP modeling, pole climbing test score was significantly increased, after administration, RbCl low dose group (7.5 mg / kg), RbCl middle dose group (15 mg / kg), RbCl high dose group (30 mg / kg) can significantly reduce the score, and RbCl high dose group (30 mg / kg) effect is most significant.
[0065] 2.4 Hanging test
[0066] As shown in Figure 4 ### P<0.001 vs Control; * P<0.05, ** P<0.01, *** P<0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP, the score of the suspension experiment was significantly increased, after administration, the RbCl low dose group (7.5 mg / kg), the RbCl middle dose group (15 mg / kg), and the RbCl high dose group (30 mg / kg) could significantly reduce the score, and the RbCl high dose group (30 mg / kg) was the most significant.
[0067] 2.5 Swimming experiment
[0068] As Figure 5 shown, ### P<0.001 vs Control; * P<0.05, ** P<0.01, *** P<0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP, the score of the swimming experiment was significantly increased, after administration, the RbCl low dose group (7.5 mg / kg), the RbCl middle dose group (15 mg / kg), and the RbCl high dose group (30 mg / kg) could significantly reduce the score, and the RbCl high dose group (30 mg / kg) was the most significant.
[0069] 2.6 Comprehensive statistics
[0070] The scores of the five behavioral tests of the test mice were added, and the results are shown in Figure 6 ### P<0.001 vs Control; *** P<0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP, the score of the model control group mice was significantly increased, after administration, the RbCl low dose group (7.5 mg / kg), the RbCl middle dose group (15 mg / kg), and the RbCl high dose group (30 mg / kg) could significantly reduce the score, and the RbCl high dose group (30 mg / kg) was the most significant, indicating that RbCl had an improving effect on the behavioral changes of mice with PD.
[0071] 2.7 Effect of RbCl on the structure of the substantia nigra and striatum of the brain tissue of MPTP mice
[0072] Figure 7 A, B, C, D, E, F represent blank control group, model control group (MPTP 30mg / kg), positive control group (compound levodopa 30mg / kg), RbCl low-dose group (7.5mg / kg), RbCl medium-dose group (15mg / kg), RbCl high-dose group (30mg / kg) respectively, Figure 8 A, B, C, D, E, F represent blank control group, model control group (MPTP 30mg / kg), positive control group (compound levodopa 30mg / kg), RbCl low-dose group (7.5mg / kg), RbCl medium-dose group (15mg / kg), RbCl high-dose group (30mg / kg) respectively. HE staining results showed that compared with the blank control group, after intraperitoneal injection of MPTP modeling, the structure of mouse brain substantia nigra and striatum was damaged, and after administration, RbCl low-dose group (7.5mg / kg), RbCl medium-dose group (15mg / kg), RbCl high-dose group (30mg / kg) could significantly improve the damage of substantia nigra and striatum structure.
[0073] 2.8 Effect of RbCl on α-synuclein in brain tissue of MPTP mice
[0074] Figure 9 A, B, C, D, E, F represent blank control group, model control group (MPTP 30mg / kg), positive control group (compound levodopa 30mg / kg), RbCl low-dose group (7.5mg / kg), RbCl medium-dose group (15mg / kg), RbCl high-dose group (30mg / kg) respectively. α-synuclein is a soluble protein expressed in the presynaptic and perikaryon of the central nervous system, which is closely related to the pathogenesis and dysfunction of Parkinson's disease. Immunohistochemical results showed that compared with the blank control group, after intraperitoneal injection of MPTP modeling, the expression of α-synuclein in mouse brain tissue was significantly increased, and after administration, RbCl low-dose group (7.5mg / kg), RbCl medium-dose group (15mg / kg), RbCl high-dose group (30mg / kg) could significantly reduce the expression of α-synuclein. It is proved that RbCl can improve Parkinson's disease by reducing α-synuclein.
[0075] 2.9 Effect of RbCl on the expression of dopamine transporter (DAT) and tyrosine hydroxylase (TH) in brain tissue of MPTP mice
[0076] As shown in Figure 10 ### P<0.001 vs Control; * P<0.05, ** P<0.01, *** P<0.001 vs MPTP, as Figure 11 shown, ### P<0.001 vs Control; ** P<0.01, *** P<0.001 vs MPTP, compared with the blank control group, after intraperitoneal injection of MPTP, DAT level and TH mRNA level were significantly reduced, after administration, RbCl low dose group (7.5 mg / kg), RbCl middle dose group (15 mg / kg), RbCl high dose group (30 mg / kg) can significantly increase the DAT and TH mRNA levels of mouse brain tissue. It shows that RbCl has protective effect on MPTP-induced Parkinson's disease mouse model of substantia nigra dopamine neurons.
[0077] 3 Conclusion
[0078] The experimental results found that RbCl has protective effect on MPTP-induced Parkinson's disease in mice, can improve the behavior change, and brain injury of substantia nigra and striatum. Its mechanism may be through reducing the expression of α-synuclein, increasing DAT and TH to play a protective effect on dopamine neurons related.
[0079] The number of devices and the scale of processing described herein are intended to be illustrative of aspects of the present application. Applications, modifications and variations of the present application that are obvious to those of ordinary skill in the art are intended to be within the scope of the present application.
[0080] Although embodiments of the present application have been disclosed as above, it is not limited to the use listed in the specification and embodiments, it can be fully applied to various fields suitable for the present application, and additional modifications can be easily realized by those skilled in the art, therefore, the present application is not limited to specific details and the figures shown and described herein, without departing from the general concept defined by the claims and the equivalent scope.
Claims
1. The use of rubidium chloride as the sole active ingredient in the preparation of a drug for treating Parkinson's syndrome, characterized in that, The drug is formulated into a dosage form for oral administration, the dosage form including solutions, suspensions, and solid dosage forms.
2. The use as described in claim 1, characterized in that, The drug contains a therapeutically effective amount of rubidium chloride and a pharmaceutically acceptable carrier.
3. The use as described in claim 2, characterized in that, Pharmaceutically acceptable carriers include diluents, solubilizers, cosolvents, disintegrants, dispersants, lubricants, flavoring agents, antioxidants, binders, absorbents, humectants, buffers, and crosslinking agents.
4. The use as described in claim 1, characterized in that, The dosage of rubidium chloride is not less than 0.5 mg / kg•d.