Application of UK-356618 in preparation of product for preventing and / or treating Parkinson's disease

Abstract

The invention relates to application of UK-356618 in preparation of a product for preventing and / or treating Parkinson's disease, and belongs to the technical field of biological medicine. The invention provides an application of UK-356618 in preparation of a product for preventing and / or treating Parkinson's disease. In-vitro experiments find that the UK-356618 has the effects of preventing dopaminergic neuron apoptosis of the Parkinson's disease, improving dopamine activity, protecting neuron synaptic integrity and reducing aggregation of phosphorylated alpha-synuclein, so that the risk of the Parkinson's disease caused by reduction of dopamine activity of brain cells is improved, and the application of the UK-356618 in treatment of the Parkinson's disease is realized. It is prompted that UK-356618 is expected to be developed into a novel product for preventing and / or treating Parkinson's disease.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more particularly to the use of UK-356618 in the preparation of products for the prevention and / or treatment of Parkinson's disease. Background Technology

[0002] Parkinson's disease (PD) is the second most common neurodegenerative disease in the world. Currently, there is no effective treatment to prevent PD or stop its progression. Both medication and surgery can only improve symptoms; they cannot stop the disease's progression or cure it.

[0003] Current Parkinson's medications only relieve symptoms by supplementing dopamine, but cannot prevent neuronal death and have no neuroprotective effect. Furthermore, their efficacy decreases as the disease progresses, leading to "on-off," end-of-dose, and freezing phenomena. Long-term use of levodopa can easily induce dyskinesia, hallucinations, hypotension, and other side effects. Moreover, there are significant individual differences and poor compliance, resulting in a narrow therapeutic window. Therefore, it is still a symptomatic treatment that does not address the root cause.

[0004] Therefore, there is an urgent need to develop a compound with relatively few side effects for treating Parkinson's disease. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide the use of UK-356618 in the preparation of products for the prevention and / or treatment of Parkinson's disease.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides the use of UK-356618 in the preparation of products for the prevention and / or treatment of Parkinson's disease.

[0007] Secondly, the present invention provides the application of UK-356618 in the preparation of nerve protection products.

[0008] Thirdly, the present invention provides the application of UK-356618 in the preparation of products that reduce α-synuclein buildup.

[0009] As a preferred embodiment of the application described in this invention, the products for preventing and / or treating Parkinson's disease and / or protecting nerves and / or reducing α-synuclein buildup include at least one of pharmaceuticals, daily chemical products, food, and health products.

[0010] As a preferred embodiment of the application described in this invention, the dosage forms of the products for preventing and / or treating Parkinson's disease and / or protecting nerves and / or reducing α-synuclein buildup include, but are not limited to, at least one of solid dosage forms, liquid dosage forms, semi-solid dosage forms and gaseous dosage forms.

[0011] As a preferred embodiment of the application described in this invention, when the dosage form of the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is a liquid dosage form, the content of UK-356618 in the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is 5-10 nmol / mL.

[0012] As a preferred embodiment of the application described in this invention, when the dosage form of the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is a liquid dosage form, the content of UK-356618 in the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is 6 nmol / mL.

[0013] As a preferred embodiment of the application described in this invention, the products for preventing and / or treating Parkinson's disease and / or protecting nerves and / or reducing α-synuclein buildup also contain excipients.

[0014] As a preferred embodiment of the application described in this invention, the excipients include, but are not limited to, at least one of fillers, binders, lubricants, antioxidants, disintegrants, preservatives, fragrances, colorants, and solvents.

[0015] As a preferred embodiment of the application described in this invention, the filler includes, but is not limited to, at least one of starch, dextrin, sucrose, hydroxybenzoic acid cellulose and mannitol.

[0016] As a preferred embodiment of the application described in this invention, the adhesive includes, but is not limited to, at least one of water, gelatin, starch paste, and polyethylene glycol.

[0017] As a preferred embodiment of the application described in this invention, the lubricant includes, but is not limited to, at least one of gum arabic, polyethylene glycol, and magnesium stearate.

[0018] As a preferred embodiment of the application described in this invention, the antioxidant includes, but is not limited to, at least one of sulfites, vitamin C, and vitamin E.

[0019] As a preferred embodiment of the application described in this invention, the disintegrant includes, but is not limited to, at least one of crospovidone, sodium carboxymethyl starch, and crospovidone carboxymethyl cellulose.

[0020] As a preferred embodiment of the application described in this invention, the preservative includes, but is not limited to, at least one of benzoic acid, sodium benzoate, propylene glycol, ethyl acetate, and sorbic acid.

[0021] As a preferred embodiment of the application described in this invention, the fragrance includes, but is not limited to, at least one of ethyl acetate, menthol, vanillin, ethyl maltol, and benzyl alcohol.

[0022] In a preferred embodiment of the application described in this invention, the colorant includes, but is not limited to, at least one of sucrose, sorbitol, mannitol, orange peel oil, carmine, and malic acid. As a preferred embodiment of the application described in this invention, the solvent includes, but is not limited to, at least one of water, ethanol, glycerol, and polysorbate 80.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention, through in vitro experiments, has shown that UK-356618 can prevent apoptosis of dopaminergic neurons in Parkinson's disease, increase dopamine activity, protect the integrity of neuronal synapses, and reduce the aggregation of phosphorylated α-synuclein, thereby improving the risk of Parkinson's disease by reducing dopamine activity in brain cells. This suggests that UK-356618 has the potential to be developed into a novel product for the prevention and / or treatment of Parkinson's disease. Attached Figure Description

[0024] Figure 1 In Example 1 of this invention, the HT-22 mouse hippocampal neuron cell line was used instead of primary mouse midbrain neurons for rotenone toxicity testing. The figure shows cells marked with "". "" indicates that there is a statistically significant difference between this group and the Control group (p<0.0002); Figure 2 In Example 1 of this invention, the HT-22 mouse hippocampal neuron cell line was used to replace primary mouse midbrain neurons for concentration gradient toxicity testing of UK-356618. The figure is marked with " "" indicates a statistically significant difference between this group and the Control group (p<0.0021), marked with " "" indicates that there is a statistically significant difference between this group and the Control group (p<0.0001); Figure 3 This invention, in Example 1, describes the effect of HT-22 mouse hippocampal neuron cell line, used as a substitute for primary mouse midbrain neurons, on improving cell viability through pretreatment with different concentrations of UK-356618. The text includes a tagline indicating that the cell viability is improved. "" indicates a statistically significant difference between this group and the Control group (p<0.0021), marked with " "" indicates that there is a statistically significant difference between this group and the Control group (p<0.0001).

[0025] Figure 4The figures show the survival status (A) and fluorescence statistics (B) of neurons in different treatment groups in Example 1 of this invention, marked with " "" indicates that there is a statistically significant difference between this group and the Control group (p<0.0001); Figure 5 This figure shows the synaptic integrity and phosphorylated α-synuclein accumulation of neurons in different treatment groups in Example 1 of this invention. A is a fluorescence-stained micrograph, B is the statistical result of synaptic integrity for each group, and C is the statistical result of α-synuclein accumulation. The figures are marked with "". "" indicates a statistically significant difference between this group and the Control group (p<0.0002), marked with " "" indicates that there is a statistically significant difference between this group and the Control group (p<0.0001). Detailed Implementation

[0026] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0027] Unless otherwise specified, all other materials and reagents used in the examples, comparative examples, and effect examples are commercially available.

[0028] The UK-356618 described in this invention can be obtained from various commercial channels. In the following examples, the UK-356618 was purchased from MedChemExpress, with a purity of 98%, catalog number HY-107394, and CAS number 230961-08-7.

[0029] Cell viability assay kits are available from various commercial sources. The cell viability assay kit used in the following examples was purchased from Thermo Fisher Scientific, catalog number L3224.

[0030] Example 1 To investigate the effects of UK-356618 on Parkinson's disease, a Parkinson's disease cell model was constructed and co-cultured with UK-356618. The specific protocol is as follows: 1. Construct a mouse model of Parkinson's disease by neuronal transmission.

[0031] Brain tissue was isolated from C57 suckling mice aged 0-3 days by decapitation. The brain tissue was immersed in D-Hanks balanced solution, and the meninges were peeled off (removing fibroblasts and vascular membranes), separating the cerebral cortex and midbrain tissue. The midbrain tissue was digested with trypsin to form a neuronal suspension, which was then cultured for 7 days in Neurobasal-A Medium (10888022, Gibco); 4v / v% B-27 supplement (50×) (17504044, Gibco); 0.5v / v% L-Glutamine (100×) (25030081, Gibco); 1v / v% 100× Pen Strep (15140-122, Gibco) (containing no cytokines required for glial cell growth to inhibit glial cell growth), yielding brain neurons. On day 7, filamentous synaptic structures were observed around the cell bodies of the neurons, preparing for the next stage of experiments.

[0032] Since neurons are terminally differentiated cells and do not proliferate significantly, directly measuring the drug's effect on neuronal activity is not feasible. Furthermore, digesting and reseeding mature primary neurons would severely damage their health and function, as pancreatic enzymes would directly kill a large number of neurons, leading to unreliable experimental data. Therefore, HT-22 (mouse hippocampal neuronal cell line) was used for preliminary experiments to explore drug toxicity.

[0033] HT-22 cells were treated with different concentrations of rotenone to establish rotenone's IC50. 50 The curve, the result is as follows Figure 1 As shown, the optimal PD modeling time and treatment concentration are 8 h and 4 μmol / mL.

[0034] Meanwhile, in the preliminary experiments, neuronal culture medium was used to prepare the drug, and after obtaining different concentrations of the drug, 1 mL of the drug was added to each well for 24 hours during cell culture. Cell viability was then measured according to the kit instructions. The results are as follows: Figure 2 As shown, HT-22 cells treated with UK-356618 at concentrations of 3-1000 nmol / mL all exhibited good cell viability, indicating that UK-356618 has high safety. Subsequently, UK-356618 concentrations of 3, 6, and 12 nmol / mL were selected.

[0035] Subsequently, HT-22 cells were pretreated with 3, 6, or 12 nmol / mL UK-356618 for 2 h, followed by incubation with 4 μmol / mL rotenone and the same concentration of UK-356618 for 8 h. Cell viability was measured, and it was found that compared with the PD model exposed to rotenone, HT-22 cells pretreated with 6 nmol / mL UK356618 and those co-incubated with rotenone showed the highest viability. Figure 3 The results showed significant differences compared to other treatment groups. Therefore, 6 nmol / mL UK356618 was subsequently selected for validation on neurons.

[0036] First, mouse midbrain neurons were pre-incubated with UK-356618 (6 nmol / mL) for 2 hours, followed by 24 hours of culture with 400 μmol / mL rotenone to establish a mouse midbrain neuronal Parkinson's disease model. UK-356618 (6 nmol / mL) was added during the rotenone treatment. Separate control groups were established: a pre-incubation group treated only with UK-356618, a rotenone-only treatment group, and a blank control group (no UK-356618 or rotenone treatment). After a second 24-hour culture with UK-356618, Calcein-AM / PI staining was used to visualize neuronal apoptosis. Furthermore, MAP2 and phosphorylated α-synuclein immunofluorescence staining was used to visualize neuronal morphology and α-synuclein accumulation. All experiments were performed according to the instructions of the fluorescent staining kit. Results are shown in [Figure number missing]. Figure 4-5 .

[0037] like Figure 4-5 As shown, the addition of UK-356618 can effectively improve the apoptosis of dopaminergic neurons in neuronal cells, enhance dopamine activity, protect the synaptic integrity of neurons, and reduce the aggregation of phosphorylated α-synuclein. This indicates that UK-356618 has the effect of preventing further lesions of Parkinson's disease and can be used in the preparation of products for the prevention and / or treatment of Parkinson's disease.

[0038] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. Application of UK-356618 in the preparation of products for the prevention and / or treatment of Parkinson's disease.

2. Application of UK-356618 in the preparation of nerve protection products.

3. Application of UK-356618 in the preparation of products that reduce α-synuclein buildup.

4. The application as described in any one of claims 1-3, characterized in that, The products for the prevention and / or treatment of Parkinson's disease and / or the products for the protection of nerves and / or the products for reducing α-synuclein buildup include at least one of pharmaceuticals, daily chemical products, food and health products.

5. The application as described in any one of claims 1-3, characterized in that, The dosage forms of the products for the prevention and / or treatment of Parkinson's disease and / or the products for the protection of nerves and / or the products for the reduction of α-synuclein buildup include at least one of solid dosage forms, liquid dosage forms, semi-solid dosage forms and gaseous dosage forms.

6. The application as described in claim 5, characterized in that, When the dosage form of the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is a liquid dosage form, the content of UK-356618 in the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is 5-10 nmol / mL.

7. The application as described in claim 6, characterized in that, When the dosage form of the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is a liquid dosage form, the content of UK-356618 in the product for preventing and / or treating Parkinson's disease and / or the product for protecting nerves and / or the product for reducing α-synuclein buildup is 6 nmol / mL.

8. The application as described in any one of claims 1-3, characterized in that, The products for the prevention and / or treatment of Parkinson's disease and / or the products for the protection of nerves and / or the products for the reduction of α-synuclein buildup also contain excipients.

9. The application as described in claim 8, characterized in that, The excipients include, but are not limited to, at least one of fillers, binders, lubricants, antioxidants, disintegrants, preservatives, fragrances, colorants, and solvents.

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