Pharmaceutical composition for treating Parkinson's syndrome

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a pharmaceutical composition for treating Parkinson's syndrome. Researches show that the Antarctic fish protein and the multi-domain derivatives thereof have the functions of relieving PD pathology and protecting dopaminergic neurons.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and specifically relates to a pharmaceutical composition for treating Parkinson's syndrome. Background Art

[0002] Parkinson's disease (PD) is the second most common central nervous system degenerative disease in the elderly, second only to Alzheimer's disease. PD mainly occurs in middle-aged and elderly people. With the aging of the population, the incidence of PD is increasing. At present, the etiology of these diseases is unknown and incurable, posing a serious threat to human health and a huge economic burden. Its pathological feature is the aggregation of α-synuclein in multiple nervous systems and peripheral systems, but its distribution and type vary, and it may also be affected by other pathologies or cause similar manifestations. Before this, the disease was mainly considered a movement disorder, with typical features including resting tremor, bradykinesia, postural instability, and stiffness of the neck, trunk, and limbs. However, PD is now considered a disease with significant neuroinflammation and immune dysfunction. Almost all Parkinson's syndromes are affected by genetic factors. More than 100 genes or gene loci have been discovered, and most cases may be caused by the interaction of multiple common and rare genetic variants. These genetic variations involve pathogenic genes of early-onset and late-onset Parkinson's syndrome, such as PARKIN, PINK1, DJ-1, SNCA and LRRK2, as well as some susceptibility genes, such as GBA, TMEM175, GAK and HTRA2.

[0003] Genetic studies have revealed some unifying biological themes, including synaptic, lysosomal, mitochondrial, and immune-mediated mechanisms. There are interactions and regulation between these mechanisms, affecting the function and survival of neurons. For example, α-synuclein can affect synaptic release, mitochondrial respiration, lysosomal degradation, and inflammatory response, while LRRK2 can regulate lysosomal function, mitochondrial dynamics, autophagy, and immune signaling. Due to the heterogeneity of Parkinson's syndrome, more biomarkers need to be developed to diagnose, stratify, monitor, and predict disease progression and treatment response. At present, some biomarkers have been used in clinical or research, such as α-synuclein in cerebrospinal fluid, dopamine transporters in neuroimaging, and neuron-specific enolase. In addition, some targeted therapeutic strategies are also under development, such as inhibiting the aggregation of α-synuclein, improving mitochondrial function, activating lysosomal enzymes, and regulating the activity of LRRK2.

[0004] Many studies have shown that antifreeze proteins in Antarctic fish can protect plants and animals from low temperature stress, including protecting the integrity of cell membranes, regulating ion balance (including calcium and potassium ions), and regulating ROS levels. Studies have shown that antifreeze proteins in Antarctic fish can also maintain intracellular ATP levels and improve cell viability. ATP metabolism is also closely related to brain activity rate, and lower neuronal activity is associated with lower energy consumption. The inhibition of ion currents by antifreeze proteins has an impact on neuronal function, such as the generation of action potentials, synaptic transmission, and potential neuroprotection under conditions of excessive ion movement, such as cerebral hypoxia / ischemia and cold / reheating injury. Other studies have shown that AFP I has the ability to reversibly inhibit neural network activity without affecting the ability of neurons to survive, and can play a protective role in the low-temperature rewarming process of hippocampal neurons. The protective effect on cells may be related to the inhibitory effect of AFP I, which may also provide a neuroprotective mechanism-due to the lack of energy consumption, the intracellular ATP level is maintained, and neurons may be better able to survive the cold and rewarming process.

[0005] In our previous studies, we found that in the Antarctic fish genome, the PD pathogenic gene α-syn gene is adjacent to the AFPIII gene, indicating that there may be a certain regulatory relationship between AFPIII and α-syn. Summary of the invention

[0006] The present invention has been studied to show that Antarctic fish protein and its multi-domain derivatives have the functions of alleviating PD pathology and protecting dopaminergic neurons, thereby completing the present invention.

[0007] In a first aspect, the present invention provides a pharmaceutical composition for treating Parkinson's syndrome, the pharmaceutical composition comprising Antarctic fish protein and its derivatives, the Antarctic fish protein being selected from one or more of CYN-1, CYN-2, CYN-3 and CYN-4 proteins, the nucleotide sequences of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins being respectively as shown in SEQ ID NO: 1 to SEQ ID NO: 4, and the amino acid sequences thereof being respectively as shown in SEQ ID NO: 5 to SEQ ID NO: 8, wherein;

[0008] The nucleotide sequences of CYN-1 to CYN-4 are:

[0009] CYN-1 SEQ ID NO: 1

[0010] ATGAAGTCAGTTGTTTTAACTGGTTTGCTGTTCGTCCTCGCTTGTGTCGAGCACATGAGTACAGCCAACAAGGCGTCCGTGGTGGCCAACCAGCTGATCCCCGTAAATACTGCCCTGACTCTGATAATGATGAAGGCGAAGGCGGTCACCCCAATGGGCCTCCCCGCCGAGGAAATTCCCCGAATAATCGGAATGCAAGCGAACAAGGCAGTGCCGTTGGGCACAACCCTCATGCCAGACATGATCAAAAACTATGAGTAG

[0011] CYN-2SEQ ID NO:2

[0012] ATGAAGTCAGTTGTTTTAACTGGTTTGCTGTTCGTCCTCGCTTGTGTCGAGCACATGAGTACAGCCAACAAGGCGTCCGTGGTGGCCAACCAGCTGATCCCCGTAAATACTGCCCTGACTCTGATAATGATGAAGGCGAAGGCGGTCACCCCAATGGGCCTCCCCGCCGAGGAAATTCCCCGAATAATCGGAATGCAAGCGAACAAGGCAGTGCCGTTGGGCACAACCCTCATGCCAGACATGATCAAAAACTATGAGGATGTGACGACATGTCCAGGCTTTAAGTCCGTGGTGGCCAACCAGCTGATCCCCATAAATACTGCCCTGACTCTAGTGATGATGAAGGCGGAGGCGGTCAGCCCAAAGGGCCTCCCTGCCGAGGAAATCTCCAAACTAGTGGGAATGCAAGCGAACAAGGCAGTGTATCTGGACCAAACCCTCATGCCAGATATGATCAAAAACTATGAGTAG

[0013] CYN-3 SEQ ID NO:3

[0014] ATGAAGTCAGTTGTTTTAACTGGTTTGCTGTTCGTCCTCGCTTGTGTCGAGCACATGAGTACAGCCAACAAGGCGTCCGTGGTGGCCAACCAGCTGATCCCCGTAAATACTGCCCTGACTCTGATAATGATGAAGGCGAAGGCGGTCACCCCAATGGGCCTCCCCGCCGAGGAAATTCCCCGAATAATCGGAATGCAAGCGAACAAGGCAGTGCCGTTGGGCACAACCCTCATGCCAGACATGATCAAAAACTATGAGGATGTGACGACATGTCCAGGCTTTAAGTCCGTGGTGGCCAACCAGCTGATCCCCATAAATACTGCCCTGACTCTAGTGATGATGAAGGCGGAGGCGGTCAGCCCAAAGGGCCTCCCTGCCGAGGAAATCTCCAAACTAGTGGGAATGCAAGCGAACAAGGCAGTGTATCTGGACCAAACCCTCATGCCAGATATGATCAAAAACTATGAGGATGGGACGACATGTCCAGGCATTAAGTCCGTGGTGGCCAACCAGCTGATCCCCATAAATACTGCCCTGACTCTAGTGATGATGAAGGCGGAGGCGGTCAGCCCAAAGGGCCTCCCTGCCGAGGAACTCCCCAAACTAGTGGGAATGCAAGCGAACAAGGCAGTGTATCTGGACCAAACCCTCATGCCAGATATGATCAAAAACTATGAGTAG

[0015] CYN-4 SEQ ID NO:4

[0016] ATGAAGTCAGTTGTTTTAACTGGTTTGCTGTTCGTCCTCGCTTGTGTCGAGCACATGAGTACAGCCAACAAGGCGTCCGTGGTGGCCAACCAGCTGATCCCCGTAAATACTGCCCTGACTCTGATAATGATGAAGGCGAAGGCGGTCACCCCAATGGGCCTCCCCGCCGAGGAAATTCCCCGAATAATCGGAATGCAAGCGAACAAGGCAGTGCCGTTGGGCACAACCCTCATGCCAGACATGATCAAAAACTATGAGGATGTGACGACATGTCCAGGCTTTAAGTCCGTGGTGGCCAACCAGCTGATCCCCATAAATACTGCCCTGACTCTAGTGATGATGAAGGCGGAGGCGGTCAGCCCAAAGGGCCTCCCTGCCGAGGAAATCTCCAAACTAGTGGGAATGCAAGCGAACAAGGCAGTGTATCTGGACCAAACCCTCATGCCAGATATGATCAAAAACTATGAGGATGGGACGACATGTCCAGGCATTAAGTCCGTGGTGGCCAACCAGCTGATCCCCATAAATACTGCCCTGACTCTAGTGATGATGAAGGCGGAGGCGGTCAGCCCAAAGGGCCTCCCTGCCGAGGAACTCCCCAAACTAGTGGGAATGCAAGCGAACAAGGCAGTGTATCTGGACCAAACCCTCATGCCAGATATGATCAAAAACTATGAGGATGGGACGACATGTCCAGGCATTAAGTCCGTGGTGGCCAACCAGCTGATCCCCATAAATACTGCCCTGACTCTAGTGATGATGAAGGCGGAGGCGGTCAGCCCAAAGGGCCTCCCTGCCGAGGAACTCCCCAAACTAGTGGGAATGCAAGCGAACAAGGCAGTGTATCTGGACGAAACCCTCATGCCAGATATGATCAAAAACTATGAGTAG

[0017] The amino acid sequences of CYN-1 to CYN-4 are as follows:

[0018] CYN-1SEQ ID NO: 5

[0019] MKSVVLTGLLFVLACVEHMSTANKASVVANQLIPVNTALTLIMMKAKAVTPMGLPAEEIPRIIGMQANKAVPLGTTLMPDMIKNYE

[0020] CYN-2SEQ ID NO: 6

[0021] MKSVVLTGLLFVLACVEHMSTANKASVVANQLIPVNTALTLIMMKAKAVTPMGLPAEEIPRIIGMQANKAVPLGTTLMPDMIKNYEDVTTCPGFKSVVANQLIPINTALTLVMMKAEAVSPKGLPAEEISKLVGMQANKAVYLDQTLMPDMIKNYE

[0022] CYN-3SEQ ID NO: 7

[0023] MKSVVLTGLLFVLACVEHMSTANKASVVANQLIPVNTALTLIMMKAKAVTPMGLPAEEIPRIIGMQANKAVPLGTTLMPDMIKNYEDVTTPGFKSVVANQLIPINTALTLVMMKAEAVSPKGLPAEISKLVGMQANKAVYLDQTLMPDMIKNYEDGTTPPGIKSVVANQLIPINTALTLVMMKAEAVSPKGLPAEEIIPE

[0024] CYN-4SEQ ID NO: 8

[0025] MKSVVLTGLLFVLACVEHMSTANKASVVANQLIPVNTALTLIMMKAKAVTPMGLPAEEIPRIIGMQANKAVPLGTTLMPDMIKNYEDVTTPGFKSVVANQLIPINTALTLVMMKAEAVSPKGLPAEISKLVGMQANKAVYLDQTLMPDMIKNYEDGTTCPGIKKSVVANQLIPINTALTLVMMKAEAVSPKGLPAEEIIP

[0026] The derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins are those having at least 80% homology with the amino acid sequences of the proteins SEQ ID NO: 5-8, more preferably at least 85% homology; more preferably at least 90% homology; more preferably at least 95% homology; more preferably at least 97% homology; more preferably at least 98% homology; more preferably at least 99% homology.

[0027] Furthermore, the derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to the deletion, addition and / or mutation of 1-50 amino acids in the amino acid sequence SEQ ID NO: 5-8 of the proteins, and the deletion, addition and / or mutation sites are continuous or discontinuous. Furthermore, the deletion, addition and / or mutation of amino acids is 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, 1-5, 2, 3, 4 or 5.

[0028] Furthermore, the Antarctic fish protein is preferably CYN-4 protein.

[0029] Furthermore, the pharmaceutical composition also contains other active ingredients, which are drugs that are different from the Antarctic fish protein and can promote improvement of the body and / or treat the symptoms of Parkinson's syndrome.

[0030] Furthermore, the pharmaceutical composition can be prepared as sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, oral liquids, lozenges, granules, electuary preparations, pills, elixirs, suspensions, powders, wine preparations, preparations, drops, injections, powder injections, creams, sustained-release preparations, targeted agents, etc.

[0031] Furthermore, the administration of the pharmaceutical composition includes oral administration, injection, implantation, external application, spraying or inhalation.

[0032] In a second aspect, the present invention provides an application of an Antarctic fish protein in the preparation of a drug for treating Parkinson's syndrome, wherein the Antarctic fish protein is selected from one or more of CYN-1, CYN-2, CYN-3 and CYN-4 proteins, wherein the nucleotide sequences of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins are respectively shown in SEQ ID NO: 1 to SEQ ID NO: 4, and the amino acid sequences thereof are respectively shown in SEQ ID NO: 5 to SEQ ID NO: 8;

[0033] Among them, the derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to those having at least 80% homology with the amino acid sequences of SEQ ID NO: 5-8 of the proteins, and more preferably at least 85% homology; more preferably at least 90% homology; more preferably at least 95% homology; more preferably at least 97% homology; more preferably at least 98% homology; more preferably at least 99% homology.

[0034] Furthermore, the derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to the deletion, addition and / or mutation of 1-50 amino acids in the amino acid sequence SEQ ID NO: 5-8 of the proteins, and the deletion, addition and / or mutation sites are continuous or discontinuous. Furthermore, the deletion, addition and / or mutation of amino acids is 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, 1-5, 2, 3, 4 or 5.

[0035] Furthermore, the Antarctic fish protein and its derivatives can reduce the phosphorylation level and aggregation of α-Syn in patients with Parkinson's syndrome.

[0036] Furthermore, the Antarctic fish protein and its derivatives can reduce TH cell damage in patients with Parkinson's syndrome. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 :Antarctic fish CYN protein reduces the phosphorylation level and aggregation of α-Syn in A53T cells (A Expression plasmid map of Antarctic fish CYN protein; B Immunofluorescence staining of pSyn in A53T cells overexpressing Antarctic fish CYN; C Quantitative statistics of pSyn aggregation points in cells);

[0038] Figure 2 :Antarctic fish CYN-1 and CYN-4 proteins can improve the motor ability of α-Syn PFF-induced mouse PD model (A The construction mechanism of transgenic mice, B Antifreeze protein CYN-1 improves the motor ability of α-Syn PFF-injected mice, C Antifreeze protein CYN-4 improves the motor ability of α-Syn PFF-injected mice);

[0039] Figure 3 :Antarctic fish CYN-1 protein reduces the hyperphosphorylation level and aggregation of α-Syn in different brain regions of mice (A p_Syn immunostaining of brain sections 3 months after unilateral injection of PFF in striatum; B p_Syn intensity quantification of each brain region after immunofluorescence staining of mouse brain sections 3 months after unilateral injection of PFF in striatum (N=3-4 mice per group));

[0040] Figure 4 :Antarctic fish CYN-4 protein reduces the hyperphosphorylation level and aggregation of α-Syn in different brain regions of mice (A p_Syn immunostaining of brain sections 3 months after unilateral injection of PFF in striatum, B p_Syn intensity quantification of brain sections after immunofluorescence staining of mouse brain sections 3 months after unilateral injection of PFF in striatum (N=3-4 mice per group));

[0041] Figure 5 :Antarctic fish CYN protein reduces TH cell damage induced by α-Syn PFF (A TH immunofluorescence staining of SN-DA neurons in WT, CYN-1 and CYN-4 mice 3 months after α-Syn PFF injection; B TH positive neuron cell count). DETAILED DESCRIPTION

[0042] The Parkinson's syndrome involved in the present invention includes "resting tremor", which is often the first symptom, mostly starting from the distal part of one upper limb, and appears or is obvious when the body is in a static position. When the disease occurs, there is a "pill-rolling" action between the thumb and the flexed index finger.

[0043] The so-called "muscle rigidity" refers to the state of the patient's limbs being similar to the bending of soft lead pipes, which is called "lead pipe rigidity"; in patients with static tremors, intermittent pause-like tremors may occur, like the turning of gears, which is called "gear rigidity". In severe cases, patients may have special flexed positions or postures, and may even be unable to take care of themselves.

[0044] The so-called "bradykinesia" means that in the early stages it can be observed that the patient's fine finger movements are slow, such as unbuttoning or buttoning, tying shoelaces, etc., which is particularly obvious.

[0045] The "postural balance disorder" mentioned above refers to the occurrence in the middle and late stages of the disease, which manifests as difficulty in standing up and easy falling backwards. Sometimes after the patient takes a step, he walks faster and faster with very small steps and cannot stop in time, which is called forward gait or panic gait.

[0046] The “sensory impairment” mentioned above means that patients may experience decreased sense of smell in the early stages, and numbness and pain in the limbs in the middle and late stages of the disease.

[0047] The "sleep disorder" refers to the patient's frequent dreams at night, accompanied by loud shouting and dancing of the body.

[0048] The "autonomic dysfunction" refers to that the patient may have constipation, sweating, urination disorder, orthostatic hypotension, etc.

[0049] The "mental disorder" mentioned here means that about 50% of patients suffer from depression, often accompanied by anxiety. In the late stage of the disease, patients experience cognitive impairment, even dementia, and visual hallucinations, i.e., visual hallucinations.

[0050] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments, but this should not be construed as limiting the scope of the present invention to the following embodiments.

[0051] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0052] Example 1 Expression and pathological detection of Antarctic fish CYN protein in A53T cells

[0053] A53T is a point mutation in α-syn that is closely related to the pathogenesis and related functional impairments of Parkinson's disease. The A53T cell line is a stable genetic cell line created by transgenic technology that overexpresses human α-synuclein with a Parkinson's disease-associated mutation (A53T).

[0054] A53T cell line was cultured with DMEM medium, and the Antarctic fish CYN protein expression plasmid was transfected into A53T cells using lipo3000 transfection reagent (Invitrogen). 12 hours after transfection, in vitro synthesized a-Syn PFFs (5μg / mL) were added to the cells and treated for 48 hours. The level of α-syn phosphorylation (AB_2564891, Abcam) was detected by cell immunofluorescence and quantitatively analyzed.

[0055] First, the cells were washed 3 times with PBS, then fixed with 4% PFA for 15 minutes, and then washed twice with PBS. Next, 0.5% Triton was used for perforation for 15 minutes and blocked with 2% BSA for 1 hour. α-syn phosphorylation antibody (AB_2564891, Abcam) was added and incubated at room temperature for 1 hour, washed 3 times with PBS, and then incubated with secondary antibody (AlexaFluor594, Invitrogen) for 1 hour and Hoechst for nuclear staining for 5 minutes. Finally, the cells were washed 3 times with PBS and sealed for observation. Fluorescence images were taken using a confocal scanning microscope (LSM710, Carl Zeiss). All images were processed by Zeiss Zen software (Carl Zeiss), and the fluorescence intensity level was evaluated using ImageJ.

[0056] The results are as follows Figure 1 As shown, the Antarctic fish CYN protein is obtained by tandem duplication of different numbers of AFPIII domains (see Figure 1A), the Antarctic fish CYN protein expression plasmid was transfected into A53T cells using chemical transfection reagents, and treated with PFF for 48 hours to detect the phosphorylation level and aggregation of α-Syn. The results showed that the expression of Antarctic fish CYN can significantly reduce the phosphorylation level and aggregation of α-Syn ( Figure 1 BC), indicating that CYN protein can reduce the phosphorylation level of α-Syn in vitro and inhibit the aggregation of α-Syn.

[0057] Example 2 Establishment of Antarctic fish CYN gene-transfected mice and analysis of motor ability and pathological detection

[0058] To obtain mice expressing the Antarctic fish CYN protein, the CYN gene was inserted into the H11 locus of WT (C57BL6) mice by microinjection. These mice were cultured according to a standard feeding regimen. At 12 weeks of age, in vitro expressed α-syn PFFs were injected into the dorsal striatum of WT (C57BL6) and CYN transgenic heterozygous mice by stereotaxic brain localization injection (AP+0.8mm, ML+2.0mm, DV-4.5mm). After a single unilateral inoculation of α-syn PFFs into the striatum, WT and transgenic mice (N=7 and 10 mice) were evaluated for locomotion at 3 months (3mpi), including the rotarod fatigue test and the hanging test. The results were plotted and analyzed using GraphPad Prism 9.

[0059] Results: To further verify the effect of Antarctic fish CYN on Parkinson's disease in vivo, transgenic mice were constructed in which the Antarctic fish CYN gene was inserted into the H11 locus (see Figure 2 A). The transgenic heterozygous mice at 3 months old were injected with exogenously expressed PFFs through the striatum. After continuous feeding for 3 months, their motor ability was tested, including the rotarod fatigue test and the hanging test. The results showed that compared with WT mice, mice expressing Antarctic fish CYN-1 and CYN-4 proteins had significantly stronger fatigue resistance and hanging ability ( Figure 2 BC). This indicates that Antarctic fish CYN protein has a protective function on PD model mice.

[0060] Example 3 Establishment of mice transfected with the Antarctic fish CYN gene and analysis of motor ability and detection of pathological transformation

[0061] To confirm the pathological transformation, hyperphosphorylated α-syn (pSyn) deposition, a marker of Lewy bodies, was detected after a single inoculation of PFFs at 3 mpi. Briefly, whole brains were fixed with 4% PFA for 24 h after mouse perfusion and then dehydrated with 30% sucrose solution. Immunofluorescence (IF) was performed on 30 μm thick serial brain cryosections. Primary antibodies and working dilutions: p-α-syn ser129 (AB_2564891, Abcam) 1:500; Th (ab_10077691, Abcam) 1:1000. For immunofluorescence studies, sections labeled with primary antibodies were incubated with secondary antibodies conjugated to Alexa-fluor 488 or 594 (Invitrogen). Fluorescence images were obtained using a confocal scanning microscope (LSM710, Carl Zeiss). All images were processed by Zeiss Zen software (Carl Zeiss). ImageJ was used to assess the total number of TH-neurons and the level of hyperphosphorylated α-Syn (pSyn) deposition.

[0062] Results: Antarctic fish CYN protein can reduce the phosphorylation level and aggregation of α-Syn in different brain regions of mice. Pathological examination was performed 3 months after unilateral inoculation of PFF in the striatum. Immunofluorescence detection of the phosphorylation level of α-Syn showed that α-Syn hyperphosphorylation occurred in different brain regions of WT mice in the control group, and dense Lewy body-like aggregates were visible ( Figure 3 A). However, CYN-1 mice showed less α-Syn hyperphosphorylation pathology and fewer Lewy body-like aggregates ( Figure 3 AB), in contrast, CYN-4 mice showed less hyperphosphorylation of α-Syn ( Figure 4 AB). The above results indicate that Antarctic fish CYN protein reduces the phosphorylation level and aggregation of α-Syn in different brain regions of mice, thereby alleviating the pathological symptoms of PD.

[0063] Results: Antarctic fish CYN protein reduced α-Syn PFF-induced TH cell damage. Striatal PFF inoculation led to α-Syn pathology in dopamine (SN_DA) neurons in the substantia nigra region. In WT mice, α-Syn pathology in the substantia nigra region was accompanied by loss of TH-positive neurons ( Figure 5 A), indicating that the generation of intraneuronal α-Syn aggregates leads to DA neuron loss as previously reported. In contrast, α-Syn pathology and loss of TH-positive neurons were well alleviated in CYN-1 and CYN-4 mice. At the same time, compared with CYN-1 mice, CYN-4 mice had a stronger protective function on SN_DA neurons ( Figure 5 B).

[0064] The above results indicate that Antarctic fish CYN protein has a protective effect on the loss of TH-positive neurons caused by PFF inoculation in the striatum.

Claims

1. A pharmaceutical composition for treating Parkinson's syndrome, the pharmaceutical composition comprising Antarctic fish protein and its derivatives, the Antarctic fish protein being selected from one or more of CYN-1, CYN-2, CYN-3 and CYN-4 proteins, the nucleotide sequences of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins being respectively shown in SEQ ID NO: 1 to SEQ ID NO: 4, and the amino acid sequences thereof being respectively shown in SEQ ID NO: 5 to SEQ ID NO:

8.

2. The pharmaceutical composition according to claim 1, wherein The derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to those having at least 80% homology with the amino acid sequences of SEQ ID NOs: 5-8 of the proteins, and more preferably at least 85% homology; more preferably at least 90% homology; more preferably at least 95% homology; more preferably at least 97% homology; more preferably at least 98% homology; more preferably at least 99% homology.

3. The pharmaceutical composition according to claim 1 or 2, wherein The derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to the amino acid sequences of the proteins SEQ ID NOs: 5-8 in which 1-50 amino acids are deleted, added and / or mutated, and the deleted, added and / or mutated sites are continuous or discontinuous; further, the deleted, added and / or mutated amino acids are 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, 1-5, 2, 3, 4 or 5.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the Antarctic fish protein is preferably CYN-4 protein.

5. The pharmaceutical composition according to any one of claims 1 to 3, further comprising other active ingredients, wherein the other active ingredients are drugs different from the Antarctic fish protein and capable of promoting improvement of the body and / or treating symptoms of Parkinson's syndrome.

6. Use of an Antarctic fish protein in the preparation of a drug for treating Parkinson's syndrome, wherein the Antarctic fish protein is selected from one or more of CYN-1, CYN-2, CYN-3 and CYN-4 proteins, the nucleotide sequences of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins are respectively shown in SEQ ID NO: 1 to SEQ ID NO: 4, and the amino acid sequences thereof are respectively shown in SEQ ID NO: 5 to SEQ ID NO:

8.

7. The use according to claim 6, wherein: The derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to those having at least 80% homology with the amino acid sequences of SEQ ID NOs: 5-8 of the proteins, and more preferably at least 85% homology; more preferably at least 90% homology; more preferably at least 95% homology; more preferably at least 97% homology; more preferably at least 98% homology; more preferably at least 99% homology.

8. The use according to claim 6 or 7, wherein: The derivatives of the CYN-1, CYN-2, CYN-3 and CYN-4 proteins refer to the amino acid sequences of the proteins SEQ ID NOs: 5-8 in which 1-50 amino acids are deleted, added and / or mutated, and the deleted, added and / or mutated sites are continuous or discontinuous; further, the deleted, added and / or mutated amino acids are 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, 1-5, 2, 3, 4 or 5.

9. The use according to any one of claims 6 to 8, wherein the Antarctic fish protein is preferably CYN-4 protein.

10. The use according to any one of claims 6 to 8, wherein the Antarctic fish protein and its derivatives can reduce the phosphorylation level and aggregation of α-Syn in patients with Parkinson's syndrome and reduce TH cell damage.

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