A chondroitin sulfate oligosaccharide composition and its application in preparing Parkinson's disease treatment drugs

Abstract

The present invention belongs to the field of biomedicine, and specifically relates to a composition of chondroitin sulfate oligosaccharide and its application in preparing a drug for treating Parkinson's disease; the chondroitin sulfate oligosaccharide is a chondroitin sulfate octose with the sulfate group at position 6 removed and the sulfate group at position 4 specifically retained; the chondroitin sulfate oligosaccharide oral preparation contains N A composition of sodium ‑(8‑[2‑hydroxybenzoyl]‑amino) caprylate (SNAC) and chondroitin sulfate octasaccharide retaining the sulfate group at position 4. N Sodium ‑(8‑[2‑hydroxybenzoyl]‑amino) caprylate can effectively promote the oral absorption of chondroitin sulfate and can be used as an oral absorption enhancer; the chondroitin sulfate oligosaccharide has neuroprotective activity and can be used to treat Parkinson's disease.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to a pharmaceutical composition comprising chondroitin sulfate oligosaccharides and an application thereof in preparing a drug for treating Parkinson's disease. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance understanding of the overall background of the invention and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] Parkinson's disease is a neurodegenerative disorder that primarily affects middle-aged and elderly individuals. The primary pathological change is the selective degeneration of dopaminergic neurons in the substantia nigra. Parkinson's disease is second only to Alzheimer's disease in incidence, affecting approximately 2-3% of people aged 65 and older. Clinical manifestations include resting tremor, bradykinesia, muscle rigidity, postural instability, and gait abnormalities. Current treatments primarily focus on symptom relief, and no therapeutic intervention can reverse the progression of Parkinson's disease. Dopamine and levodopa remain the gold standard treatments; however, long-term use reduces their effectiveness and produces side effects. Therefore, there is an urgent need to develop more effective neuroprotective and disease-modifying therapies.

[0004] In recent years, glycosaminoglycans (GAGs) and their binding proteins have attracted widespread attention from both academia and the pharmaceutical industry. GAGs are a class of linear anionic polysaccharides composed of repeating disaccharide units. They are ubiquitous on cell surfaces and in the extracellular matrix, playing important roles in regulating biological and physiological processes. Due to their negative charge, GAGs can interact with various proteins and exert a wide range of biological activities, including anti-inflammatory, anti-tumor, antiviral, and neuroprotective effects. GAGs can be classified into heparin (HP), heparan sulfate (HS), and chondroitin sulfate (CS), depending on the disaccharide unit composition and the degree of sulfation. Chondroitin sulfate is the most prevalent GAG in the brain's extracellular matrix. However, due to its large molecular weight, high negative charge, hydrophilicity, and instability under acidic conditions, its oral bioavailability is low, insufficient to provide the desired clinical therapeutic effect. Therefore, finding effective strategies to improve the oral bioavailability of chondroitin sulfate is of great importance. Summary of the Invention

[0005] In previous research (CN119285810B), the inventors provided a chondroitin sulfate oligosaccharide desulfurization product that has the ability to promote the repair of chronic hepatitis and anti-inflammatory activity. It can be used for oral treatment of non-alcoholic fatty liver disease. When combined with deoxycholethamine, it can significantly improve oral bioavailability. The present invention found that the above compound not only has anti-inflammatory activity, but also has anti-Parkinson's activity. In order to further expand the Parkinson's therapeutic activity of the above compound and improve the bioavailability of the oral preparation, the present invention combined it with N The combined use of sodium (8-[2-hydroxybenzoyl]-amino) caprylate (SNAC) effectively improves the bioavailability of oral preparations and the therapeutic effect of Parkinson's disease. Based on the above technical effects, the present invention provides the following technical solutions:

[0006] In a first aspect, a pharmaceutical composition is provided, comprising an active dose of the chondroitin sulfate oligosaccharide according to the first aspect; the structure of the chondroitin sulfate oligosaccharide is shown in the following formula (I):

[0007]

[0008] Formula (I);

[0009] In formula (I), R is SO3H or H.

[0010] The chondroitin sulfate oligosaccharide represented by formula (I) is uronic acid and N -A linear polysaccharide composed of repeated disaccharide units of acetylgalactosamine, with a degree of polymerization of 8. The octasaccharide is specifically removed N -The 6-position sulfate group of acetylgalactosamine is retained, and only the 4-position sulfate group is retained, named CS-dp8-de6S. The preparation method thereof comprises the following steps:

[0011] (1) Enzymatic hydrolysis of chondroitin sulfate into low molecular weight chondroitin sulfate;

[0012] (2) The low molecular weight chondroitin sulfate prepared in step (1) is separated by chromatography to obtain chondroitin sulfate octasaccharide;

[0013] (3) The chondroitin sulfate octasaccharide prepared in step (2) was subjected to H + Ion exchange column, adding pyridine to the product of the column to obtain chondroitin sulfate octasaccharide pyridinium salt, and mixing it with N -methyl- N -trimethylsilyltrifluoroacetamide (MSTFA) was stirred at 90~110℃ for 10~14 h to obtain the above-mentioned CS-dp8-de6S.

[0014] The specific operations of step (1) are as follows:

[0015] Chondroitin sulfate was dissolved in a partial enzymatic hydrolysis buffer, and chondroitin sulfate ABC enzyme was added and incubated at 35-39°C for 7-9 hours for enzymatic hydrolysis. After completion, the reaction system was inactivated at high temperature, and the supernatant was retained by centrifugation to obtain low molecular weight chondroitin sulfate.

[0016] Furthermore, the partial enzymatic hydrolysis buffer is a mixture of sodium acetate and calcium acetate, wherein the molar ratio of sodium acetate to calcium acetate is 1.8-2.2:0.02-0.04.

[0017] Furthermore, the dosage ratio of the chondroitin sulfate, partial enzymatic hydrolysis buffer, and chondroitin sulfate ABC enzyme is 1:28-32 mL:0.7-0.8 IU.

[0018] Furthermore, the purpose of the high-temperature inactivation is to inactivate the chondroitin sulfate ABC enzyme in the reaction system, and the method usually adopted is high-temperature heating, such as heating by water bath or microwave.

[0019] Furthermore, the centrifugation parameters are as follows: high-speed centrifugation at 12,000 to 16,000 rpm for 12 to 17 min.

[0020] In step (2), the mobile phase for the chromatographic separation is a 0.2 M aqueous solution of ammonium bicarbonate, the chromatographic column is P10, and the retention time of the chromatographic separation of the chondroitin sulfate octasaccharide is 380-425 min.

[0021] The specific operation of step (3) is as follows: dissolve the above chondroitin sulfate octasaccharide in water to a concentration of 0.2-0.3 g / mL, + An ion exchange column was used, and pyridine was added to the eluate received by the column until the pH of the eluate was 7. After thorough stirring, the eluate was concentrated and dried to obtain chondroitin sulfate octasaccharide pyridinium salt. After redissolved in pyridine, the salt was stirred with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) at 90-110°C for 11-13 h to obtain the above-mentioned CS-dp8-de6S.

[0022] Furthermore, the process + The eluent for the ion exchange column was water.

[0023] Furthermore, the dosage ratio of the chondroitin sulfate octasaccharide pyridinium salt to MSTFA is 1 g: 0.8-1.2 mL.

[0024] Furthermore, CS-dp8-de6S was isolated as follows: the solution system after the reaction of chondroitin sulfate octasaccharide pyridinium salt and MSTFA was dialyzed against water for 22 to 26 hours, and the components in the dialysis bag were freeze-dried to obtain CS-dp8-de6S.

[0025] The "active dose" referred to in the first aspect, according to the general understanding in the field, can be the drug dose for the purpose of prevention, improvement, treatment or auxiliary treatment. After clarifying the therapeutic activity of the above-mentioned ingredients, the dose belongs to the technical content that can be routinely inferred based on the general technology mastered by those skilled in the art.

[0026] In addition to the CS-dp8-de6S described in the first aspect, the above composition also includes other auxiliary ingredients. The feasible types of the other auxiliary ingredients are absorption enhancers, carriers, diluents, adhesives, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, antibacterial agents or buffers, or a combination of at least two. In one embodiment verified by the present invention, the other auxiliary ingredients are absorption enhancers, specifically: N -(8-[2-hydroxybenzoyl]-amino) sodium caprylate (SNAC). In this embodiment, the mass ratio of CS-dp8-de6S to SNAC is 2-4:1.

[0027] In a second aspect, the present invention provides the use of the chondroitin sulfate oligosaccharide and pharmaceutical composition described in the first aspect in the preparation of a drug for treating Parkinson's disease.

[0028] Existing studies have shown that low-molecular-weight chondroitin sulfate can protect dopaminergic neurons in the MPTP-induced PD model. Low-molecular-weight chondroitin sulfate is a highly complex polysaccharide mixture. The present invention provides a chondroitin sulfate oligosaccharide with a specific structure. From the perspective of pharmaceutical formulation development, the research results of the present invention are more conducive to drug quality control. Those skilled in the art can obtain the corresponding compound based on the preparation method provided by the present invention, or they can prepare it through chemical synthesis or other methods.

[0029] On the other hand, Parkinson's disease is a chronic disease requiring long-term medication, making oral formulations more suitable for patients. However, since chondroitin sulfate is a large molecule with low bioavailability, to improve the oral bioavailability of this compound, the present invention designed a co-administration of CS-dp8-de6S with ingredients that promote its absorption. According to the present research results, the oral administration of a drug combination of CS-dp8-de6S and SNAC can effectively increase the expression of tyrosine hydroxylase in the brain tissue of Parkinson's disease model mice, thereby increasing brain dopamine content and improving Parkinson's symptoms.

[0030] Depending on the form of drug administration to the affected area, the dosage form of the Parkinson's disease treatment drug includes but is not limited to intramuscular injection, subcutaneous injection, intravenous injection, oral preparation, or interventional preparation ingested by surgical means. In one embodiment verified by the present invention, the above-mentioned pharmaceutical composition is in an oral form, specifically, an oral tablet.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] 1. The pharmaceutical composition of SNAC and CS-dp8-de6S provided by the present invention effectively promotes the oral absorption of chondroitin sulfate octasaccharide and greatly improves the oral bioavailability of chondroitin sulfate octasaccharide.

[0033] 2. The pharmaceutical composition provided by the present invention has good neuroprotective activity and can be used for oral treatment of Parkinson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0035] Figure 1 This is the P10 chromatogram of the chondroitin sulfate octasaccharide prepared in Example 1;

[0036] Figure 2 This is the HSQC NMR spectrum of the chondroitin sulfate octasaccharide prepared in Example 1;

[0037] Figure 3 HSQC NMR spectrum of CS-dp8-de6S prepared in Example 1;

[0038] Figure 4 The blood concentration curves of CS-dp8-de6S after subcutaneous injection and oral administration in Example 5;

[0039] Figure 5 This is the immunohistochemical staining image of tyrosine hydroxylase in Example 5;

[0040] in, Figure 5 Middle A shows the immunohistochemical staining results of substantia nigra sections of brain tissue of mice in the normal group;

[0041] Figure 5 Middle B is the immunohistochemical staining results of substantia nigra sections of brain tissue of mice in the model group;

[0042] Figure 5 Middle C is the immunohistochemical staining results of substantia nigra sections of brain tissue of mice in the drug-treated group;

[0043] Figure 6 This is a statistical analysis graph of the average optical density of Example 5. DETAILED DESCRIPTION

[0044] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0045] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof.

[0046] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0047] Example 1

[0048] In this embodiment, a chondroitin sulfate oligosaccharide is provided, named CS-dp8-de6S, and its structure is shown in the following formula:

[0049]

[0050] Formula (I);

[0051] In formula (I), R is SO3H or H.

[0052] This embodiment also provides a method for preparing the above-mentioned CS-dp8-de6S, comprising the following steps:

[0053] 1. Preparation of low molecular weight chondroitin sulfate by enzymatic hydrolysis

[0054] Weigh 1 g of chondroitin sulfate and dissolve it in 30 mL of partial enzymatic hydrolysis buffer. Add 0.75 IU of chondroitin sulfate ABC enzyme and incubate at 37°C for 8 h. After enzymatic hydrolysis, boil in a 100°C water bath to inactivate the chondroitin sulfate ABC enzyme. Centrifuge at 14,000 rpm for 15 min, collect the supernatant, and lyophilize to obtain low-molecular-weight chondroitin sulfate.

[0055] The aforementioned partial enzymatic hydrolysis buffer was prepared by mixing 0.1 mol / L sodium acetate (adjusted to pH 7.0 with acetic acid) and 0.01 mol / L calcium acetate in a 20:3 volume ratio. 0.1 mol / L sodium acetate was prepared as follows: 0.2461 g sodium acetate was dissolved in 30 mL of water and the pH was adjusted to 7.0 with acetic acid. 0.01 mol / L calcium acetate was prepared as follows: 26.43 mg calcium acetate was dissolved in 15 mL of water.

[0056] 2. Chromatographic separation to obtain chondroitin sulfate octasaccharide

[0057] Mobile phase: 0.2 M ammonium bicarbonate aqueous solution, ultrasonically dissolve, filter to remove impurities, and ultrasonically remove bubbles.

[0058] Chromatographic column: P10; loading method: Add packing material to a beaker and soak it in 20% ethanol. Replace the 20% ethanol with high-purity water. Filter the packing material to remove bubbles and pour it into the column. First, compact the packing material with water, then fill it with 0.2 M ammonium bicarbonate mobile phase to compact the packing material. Sample loading is then possible.

[0059] Chromatographic separation: 300 mg of the obtained low molecular weight chondroitin sulfate was dissolved in water and separated by high performance liquid chromatography to obtain chondroitin sulfate octasaccharide. The results were characterized as follows: Figure 1 、 Figure 2 shown.

[0060] 3. Preparation of CS-dp8-de6S by directional desulfation of the sulfate group

[0061] Weigh 5 g of the above chondroitin sulfate octasaccharide and dissolve it in 20 mL of deionized water. + Ion exchange column. Add pyridine to the eluate collected after column flow until the solution reaches pH 7. Stir at room temperature for 1 hour, concentrate under vacuum, and lyophilize to obtain chondroitin sulfate octasaccharide pyridinium salt. Weigh 1 g of chondroitin sulfate octasaccharide pyridinium salt and dissolve it in 6 mL of anhydrous pyridine. Add 1 mL of MSTFA. Stir at 100°C for 12 hours, then dilute the solution with 5 mL of purified water. Dialyze the solution against water for 24 hours and lyophilize to obtain the desulfurized product of chondroitin sulfate octasaccharide, namely CS-dp8-de6S.

[0062] Characterization: 15 mg of the above CS-dp8-de6S was weighed, dissolved in deuterium oxide, and lyophilized three times to replace the exchangeable protons. The lyophilized sample was dissolved in 200 µL of deuterium oxide, vortexed, centrifuged, and the supernatant was analyzed by NMR HSQC spectroscopy. The results are shown in Figure 2. Figure 3 shown.

[0063] Example 2

[0064] In this example, another method for preparing CS-dp8-de6S is provided, which differs from Example 1 in that:

[0065] 1. Preparation of low molecular weight chondroitin sulfate by enzymatic hydrolysis

[0066] Weigh 1 g of chondroitin sulfate and dissolve it in 28 mL of partial enzymatic hydrolysis buffer. Add 0.7 IU of chondroitin sulfate ABC enzyme and incubate at 35°C for 9 h. After enzymatic hydrolysis, boil in a 100°C water bath to inactivate the chondroitin sulfate ABC enzyme. Centrifuge at 12,000 rpm for 17 min, collect the supernatant, and lyophilize to obtain low-molecular-weight chondroitin sulfate.

[0067] The above-mentioned partial enzymatic hydrolysis buffer was prepared by mixing 0.1 mol / L sodium acetate (adjusted to pH 7.0 with acetic acid) and 0.01 mol / L calcium acetate in a volume ratio of 18:2.

[0068] 2. Chromatographic separation to obtain chondroitin sulfate octasaccharide

[0069] Same as Example 1.

[0070] 3. Preparation of CS-dp8-de6S by directional desulfation of the sulfate group

[0071] Weigh 4 g of the above chondroitin sulfate octasaccharide and dissolve it in 20 mL of deionized water. + Ion exchange column. Add pyridine to the eluate collected after column flow until the solution reaches pH 7. Stir at room temperature for 1 hour, concentrate under vacuum, and lyophilize to obtain chondroitin sulfate octasaccharide pyridinium salt. Weigh 1 g of chondroitin sulfate octasaccharide pyridinium salt and dissolve it in 6 mL of anhydrous pyridine. Add 0.8 mL of MSTFA. Stir at 90°C for 13 hours, then dilute the solution with 5 mL of purified water. Dialyze the solution against water for 24 hours and lyophilize to obtain the desulfurized product of chondroitin sulfate octasaccharide, namely CS-dp8-de6S.

[0072] Example 3

[0073] In this example, another method for preparing CS-dp8-de6S is provided, which differs from Example 1 in that:

[0074] 1. Preparation of low molecular weight chondroitin sulfate by enzymatic hydrolysis

[0075] Weigh 1 g of chondroitin sulfate and dissolve it in 32 mL of partial enzymatic hydrolysis buffer. Add 0.8 IU of chondroitin sulfate ABC enzyme and incubate at 39°C for 7 h. After enzymatic hydrolysis, boil in a 100°C water bath to inactivate the chondroitin sulfate ABC enzyme. Centrifuge at 16,000 rpm for 12 min, collect the supernatant, and lyophilize to obtain low-molecular-weight chondroitin sulfate.

[0076] The above-mentioned partial enzymatic hydrolysis buffer was prepared by mixing 0.1 mol / L sodium acetate (adjusted to pH 7.0 with acetic acid) and 0.01 mol / L calcium acetate in a volume ratio of 22:4.

[0077] 2. Chromatographic separation to obtain chondroitin sulfate octasaccharide

[0078] Same as Example 1.

[0079] 3. Preparation of CS-dp8-de6S by directional desulfation of the sulfate group

[0080] Weigh 6 g of the above chondroitin sulfate octasaccharide and dissolve it in 20 mL of deionized water. + Ion exchange column. Add pyridine to the eluate collected after column flow until the solution reaches pH 7. Stir at room temperature for 1 hour, concentrate under vacuum, and lyophilize to obtain chondroitin sulfate octasaccharide pyridinium salt. Weigh 1 g of chondroitin sulfate octasaccharide pyridinium salt and dissolve it in 6 mL of anhydrous pyridine. Add 1.2 mL of MSTFA. Stir at 110°C for 11 hours, then dilute the solution with 5 mL of purified water. Dialyze the solution against water for 24 hours and lyophilize to obtain the desulfurized product of chondroitin sulfate octasaccharide, namely CS-dp8-de6S.

[0081] Example 4

[0082] In this embodiment, a composition comprising the above-mentioned CS-dp8-de6S is provided. The composition comprises the CS-dp8-de6S prepared in any one of Examples 1-3 and an absorption enhancer SNAC, wherein the mass ratio of the CS-dp8-de6S and the absorption enhancer SNAC is 1:3. The composition is prepared as follows:

[0083] 300 mg of CS-dp8-de6S and 100 mg of SNAC were weighed and ultrasonicated for 40 min until the mixture was uniformly mixed to obtain the above-mentioned composition.

[0084] Example 5

[0085] In this embodiment, an oral tablet is provided, which is prepared as follows:

[0086] The composition in Example 4 was crushed to a particle size of 20 mesh and fully dried. The dried powder was placed in a tablet press to prepare tablets.

[0087] Biological validation

[0088] 1. Bioavailability Verification

[0089] 1. Experimental materials: C57BL / 6J mice, 8 weeks old.

[0090] 2. Experimental Methods

[0091] Eighteen mice each were subcutaneously injected with the CS-dp8-de6S described in Example 1 and orally gavaged with the composition described in Example 4. Doses were administered at a subcutaneous injection of 10 mg / kg and an oral gavage of 100 mg / kg. Blood was collected from the medial canthus at 5, 10, 20, 30, 60, and 120 minutes after administration. The whole blood was centrifuged at 3000 rpm for 15 minutes at 4°C. The supernatant was plasma. The plasma was mixed with methanol in a 1:9 volume ratio and allowed to precipitate at room temperature for 10 minutes. The solution was centrifuged at 1000 g for 10 minutes, and the supernatant was discarded. The precipitate was enzymatically digested with pronase E at 55°C for 24 hours. After enzymatic digestion, the solution was boiled at 100°C for 10 minutes to inactivate the protease. The supernatant was obtained by centrifugation at 14,000 rpm for 10 minutes. The supernatant was then purified by anion exchange column to further remove impurities, yielding relatively pure CS-dp8-de6S. The eluted sugars were desalted using a 2 kDa ultrafiltration tube and freeze-dried. The desalted CS-dp8-de6S was completely hydrolyzed into disaccharides using CS ABC enzyme. The disaccharides were labeled with AMAC and quantitatively analyzed by LC-MS / MS-MRM using a Nexera LC-20A UPLC system and a SCIEX Triple Quad™ 5500+ mass spectrometer.

[0092] 3. Experimental Results

[0093] Figure 4 Table 1 shows the blood concentration curves of the subcutaneous injection and oral compositions, and Table 2 shows the relevant parameters and bioavailability of the subcutaneous injection and oral compositions.

[0094] Table 1 Related parameters and bioavailability of subcutaneous injection and oral compositions

[0095]

[0096] Quantitative results indicate that the bioavailability of the subcutaneous injection group (control group) was 90.56%, while that of the oral composition group reached 8.19%. Generally speaking, the oral absorption rate of chondroitin sulfate drugs does not exceed 1%. Furthermore, the concentration of drug actually absorbed into the blood (i.e., the AUC value) is the effective concentration for drug efficacy. Therefore, increasing the oral dose can improve the AUC value. In this study, an oral dose 10 times the injection dose was used. At this dose, the drug concentration in the composition group was 204.7, approximately 90% of the concentration in the subcutaneous injection group. This demonstrates that the prepared composition exhibits good oral absorption after administration, effectively improving the oral absorption rate of CS and possessing great therapeutic potential.

[0097] 2. Verification of Parkinson's disease treatment effects

[0098] 1. Experimental materials: C57BL / 6J mice, 8 weeks old.

[0099] 2. Experimental Methods

[0100] 2.1 Experimental Grouping

[0101] The mice were randomly divided into three groups: (A) normal group, (B) model group, and (C) drug-treated group. The drug was the composition in Example 4.

[0102] 2.2 Modeling method

[0103] Groups B and C were subcutaneously injected with MPTP 20 mg / kg / day for 5 consecutive days to establish Parkinson's disease models; group A was intraperitoneally injected with an equal amount of normal saline as a control.

[0104] 2.3 Administration

[0105] After successful modeling, drug treatment began. Group C was gavaged with the composition of Example 4 for 7 consecutive days. Groups A and B were gavaged with an equal volume of saline as a control. Group C was administered CS-dp8-de6S at a dose of 300 mg / kg / day and SNAC at a dose of 100 mg / kg / day.

[0106] 3. Experimental Results

[0107] After the model was established, tyrosine hydroxylase (TH) immunohistochemical staining was performed on the substantia nigra sections of the brain tissue, and the results were quantified using the average optical density (AOD). Figure 5 、 Figure 6 From the results of statistical analysis, after treatment with the neurotoxin MTPT, the number of TH-positive cells in the model group was significantly lower than that in the normal group, and the average optical density value was statistically significantly different ( P <0.05), indicating that the Parkinson's model was successfully established. After the treatment with the composition in Example 4, the number of TH positive cells increased significantly compared with the model group ( P <0.001), indicating that CS-dp8-de6S has good neuroprotective activity and can be used to treat Parkinson's disease.

[0108] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A use of chondroitin sulfate oligosaccharide in preparing a drug for treating Parkinson's disease, characterized in that: The preparation method of the chondroitin sulfate oligosaccharide is as follows: (1) Enzymatic hydrolysis of chondroitin sulfate into low molecular weight chondroitin sulfate; (2) The low molecular weight chondroitin sulfate prepared in step (1) is separated by chromatography to obtain chondroitin sulfate octasaccharide; (3) The chondroitin sulfate octasaccharide prepared in step (2) was subjected to H + Ion exchange column, adding pyridine to the product of the column to obtain chondroitin sulfate octasaccharide pyridinium salt, and mixing it with N -methyl- N -Trimethylsilyltrifluoroacetamide is stirred at 90-110°C for 10-14h to obtain the product.

2. Use of the pharmaceutical composition in preparing a drug for treating Parkinson's disease, characterized in that: The pharmaceutical composition is composed of the chondroitin sulfate oligosaccharide according to claim 1 and an absorption enhancer, wherein the absorption enhancer is N -Sodium (8-[2-hydroxybenzoyl]-amino) caprylate.

3. Use of the pharmaceutical composition according to claim 2 in preparing a drug for treating Parkinson's disease, characterized in that: The chondroitin sulfate oligosaccharide and N The mass ratio of sodium 2-(8-[2-hydroxybenzoyl]-amino)caprylate is 2~4:

1.

4. Use of the pharmaceutical composition according to claim 3 in preparing a drug for treating Parkinson's disease, characterized in that: The chondroitin sulfate oligosaccharide and N The mass ratio of sodium -(8-[2-hydroxybenzoyl]-amino)caprylate is 3:

1.

5. The use according to claim 1 or 2, characterized in that The dosage form of the Parkinson's disease treatment drug is selected from intramuscular injection, subcutaneous injection, intravenous injection, oral preparation or interventional preparation taken by surgical means.

6. The use according to claim 5, characterized in that The Parkinson's disease treatment drug is an oral preparation.

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