CN114831990A - Application of sinomenine in preparing pharmaceutical composition for treating Parkinson's disease - Google Patents
Application of sinomenine in preparing pharmaceutical composition for treating Parkinson's disease Download PDFInfo
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- CN114831990A CN114831990A CN202210284708.9A CN202210284708A CN114831990A CN 114831990 A CN114831990 A CN 114831990A CN 202210284708 A CN202210284708 A CN 202210284708A CN 114831990 A CN114831990 A CN 114831990A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/485—Morphinan derivatives, e.g. morphine, codeine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
- A61P25/16—Anti-Parkinson drugs
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Abstract
The invention belongs to the technical field of medicines, and particularly discloses application of sinomenine in preparing a pharmaceutical composition for treating Parkinson's disease, in particular to application of sinomenine in activating an Nrf2/ARE channel to promote expression of antioxidant enzyme to play a role in resisting mitochondrial damage and improving Parkinson's motor function. Experimental research proves that sinomenine can be used for treating Parkinson's disease, a certain concentration of sinomenine can effectively activate an Nrf2/ARE channel, expression of antioxidant enzyme is promoted to play a role in resisting mitochondrial damage, and the movement function of Parkinson's disease is improved.
Description
[ technical field ] A method for producing a semiconductor device
The invention belongs to the technical field of medicines, and particularly relates to application of sinomenine in preparation of a pharmaceutical composition for treating Parkinson's disease.
[ background of the invention ]
Parkinson's Disease (PD) is a common nervous system degenerative disease, and is common in the elderly, with the average age of about 60 years, and the onset of juvenile Parkinson's disease below 40 years being rare. The prevalence rate of PD in people over 65 years old in China is about 1.7%. Most parkinson's disease patients are sporadic cases, with less than 10% of patients having a family history. The most prominent pathological change of parkinson's disease is the degenerative death of mesocerebral Dopaminergic (DA) neurons, which causes a marked reduction in striatal DA content and causes disease. The exact etiology of this pathological change remains unclear, and genetic factors, environmental factors, age-related aging, oxidative stress, etc. may all be involved in the degenerative death process of PD dopaminergic neurons.
Parkinson is the second largest neurodegenerative disease next to alzheimer's disease, which is dominated by motor dysfunction. At present, dopamine preparations are mostly used for treatment, but long-time dopamine treatment can cause motor complications such as symptom fluctuation, dyskinesia and the like, and a new therapeutic medicament is urgently needed.
Mitochondrial damage is considered to be one of the major pathogenesis of parkinson. The Nrf2-ARE signal path is the most important antioxidant path, and the antioxidant enzyme such as HO-1 generated by the path can eliminate excessive active oxygen generated by mitochondria, protect the mitochondria and maintain the functions of the mitochondria.
In conclusion, the research on the medicine for treating the Parkinson's disease with good curative effect and small side effect is a hot point of research. No report related to the application of the sinomenine in the preparation of the Parkinson disease medicine or how to treat the Parkinson disease exists in the prior art.
[ summary of the invention ]
The invention aims to: aiming at the existing problems, the invention provides the application of sinomenine in preparing the pharmaceutical composition for treating the Parkinson disease, in particular to the application of the sinomenine in activating an Nrf2/ARE channel, promoting the expression of antioxidant enzymes to play a role in resisting mitochondrial damage and improving the parkinsonism motor function. Experimental research proves that sinomenine can be used for treating Parkinson's disease, a certain concentration of sinomenine can effectively activate an Nrf2/ARE channel, expression of antioxidant enzyme is promoted to play a role in resisting mitochondrial damage, and the movement function of Parkinson's disease is improved.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the invention relates to an application of sinomenine in preparing a pharmaceutical composition for treating Parkinson's disease, in particular to an application of sinomenine in activating an Nrf2/ARE passage to promote expression of antioxidant enzymes to play a role in resisting mitochondrial damage and improving Parkinson's motor function.
In the invention, the pharmaceutical composition is further illustrated to be prepared into a clinically acceptable pharmaceutical preparation by taking sinomenine as a main component and adding pharmaceutically acceptable auxiliary materials or auxiliary components.
In the present invention, further, the pharmaceutical preparation includes a tablet, a capsule, an injection or a powder.
In the invention, the content of the sinomenine in the pharmaceutical composition is further illustrated to be 2.5 mg/kg-40 mg/kg.
In the present invention, it is further explained that, when the pharmaceutical composition is applied to human, the content of the sinomenine in the pharmaceutical composition is 0.6mg/kg to 10mg/kg, depending on the ratio of the body surface area of the animal to the human.
In the invention, the content of the sinomenine in the pharmaceutical composition is further explained to be 5 mg/kg-20 mg/kg.
In the invention, it is further explained that when the pharmaceutical composition is applied to human, the content of the sinomenine in the pharmaceutical composition is 0.8 mg/kg-5 mg/kg according to the ratio of the body surface area of the animals to the body surface area of the human.
Generally, the drugs are clinically applied after being prepared into preparations. The pharmaceutical composition of the present invention can be prepared according to a method known in the art as a pharmaceutical composition. The pharmaceutical compositions of the present invention may be formulated into any dosage form suitable for human or animal use by combining them with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants.
The pharmaceutical composition of the invention or the pharmaceutical composition containing the same can be administered in unit dosage form, and the administration route can be intestinal or parenteral, such as oral administration, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum and the like.
The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.
The pharmaceutical composition can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle delivery systems. For tableting the pharmaceutical composition of the present invention, a wide variety of excipients known in the art may be used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent can be water, ethanol, isopropanol, etc.; the adhesive can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, and the like.
The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.
In order to encapsulate the administration unit, the pharmaceutical composition of the present invention as an active ingredient may be mixed with a diluent and a glidant, and the mixture may be directly placed in a hard capsule or a soft capsule. Or the active ingredients of the pharmaceutical composition of the invention can be prepared into granules or pellets with diluent, adhesive and disintegrating agent, and then placed into hard capsules or soft capsules. The various diluents, binders, wetting agents, disintegrants, glidants used for preparing the pharmaceutical composition tablets of the invention can also be used for preparing capsules of the pharmaceutical composition of the invention.
In order to prepare the pharmaceutical composition of the invention into injection, water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent, and a proper amount of solubilizer, cosolvent, pH regulator and osmotic pressure regulator which are commonly used in the field can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.
In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
experiments show that sinomenine can obviously reduce the circling behavior caused by apomorphine, enhance the activity of SY5Y cells after 6-OHDA treatment and enhance the transcription levels of Nrf2 and HO-1 in striatum; it is obvious through research that sinomenine promotes the expression of antioxidase such as HO-1 and the like to play a role of resisting mitochondrial damage by activating an Nrf2/ARE channel, and finally improves the parkinsonism motor function, so that the sinomenine has a good clinical application prospect.
[ description of the drawings ]
FIG. 1 is a statistical chart of the effect of apomorphine-induced circling behavior in rats according to the present invention. N ═ 6, compared to model group, P < 0.05; #, P <0.05 compared to sham.
FIG. 2 is a statistical graph of the drop latency of the ethological suspension test of rats according to the invention. N ═ 5, compared to model group, P < 0.05; # P <0.05 compared to sham.
FIG. 3 is a graph showing the effect of various concentrations of sinomenine on the activity of SY5Y cells in the present invention. P <0.05 compared to model group; p <0.01, N ═ 2, compared to model groups.
FIG. 4 is a PCR quantitative graph of the Nrf2, HO-1mRNA of the present invention. # P <0.05 compared to model group; p <0.05, N ═ 3, compared to sham group.
FIG. 5 is a quantitative graph of Nrf2 and HO-1Western Blot protein. A is a representative graph, and B is a quantitative result. # P <0.05 compared to model group; p <0.05, N ═ 3, compared to sham group.
[ detailed description ] embodiments
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but rather should be construed as broadly as the present invention is capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Example (b):
the application of sinomenine in preparing the pharmaceutical composition for treating the Parkinson disease comprises the following steps:
1. preparation of Parkinson rat model
Taking out the experimental rat, weighing, injecting phenobarbital into abdominal cavity for anesthesia, cutting off the hair of the head of the rat, fixing the head on a stereotaxic apparatus, carrying out skin disinfection, cutting off the scalp, and removing the tissue membrane. Positioning to right striatum with coordinates of-1.0 mm and DL of 3mm relative to bregma AP, drilling skull with electric skull drill, positioning injector to 4mm under dura mater, injecting 6-OHDA 4 μ g/μ L2 μ L at administration speed of 1 μ L/min, after injection, slowly lowering needle to 6mm and injecting 2 μ L, after needle retention for 5min, slowly pulling out needle, suturing, sterilizing with iodine tincture, injecting penicillin to prevent wound infection, placing rat back in cage, and feeding after waking.
2. Grouping of laboratory animals
A group of false operations comprises a plurality of operations,
model group: the normal saline is injected into the abdominal cavity,
low dose group: injecting 10mg/kg of sinomenine into the abdominal cavity,
the medium dose group: injecting sinomenine 20mg/kg into abdominal cavity,
high dose group: injecting sinomenine 40mg/kg into abdominal cavity,
the medicine is taken one week earlier, and the subsequent experiments are carried out 1 week after the model is made,
a positive drug group: medoba, administered 1 week prior to behavioural testing.
3. Behavioural testing
The rotation was performed by inducing rotation of rats by subcutaneous injection of Apomorphine (APO) (0.05mg/kg) and the number of revolutions and direction of rotation were recorded for 30 minutes.
Suspension behavior detection, the rat is suspended on the iron wire in proper order, and the time that the rat hung to drop from beginning is recorded, and is repeated 3 times, gets the average.
4. Cell experiments
Placing SH-SY5Y cells on culture plate, replacing culture solution once after 24h, adding sinomenine with different concentrations, adding 5% CO 2 The culture solution is placed in an incubator at 37 ℃ for standing culture for 8 hours under a moist and sterile environment. 6-OHDA 100. mu.M was added to stimulate SH-SY5Y cells for 24 h. Cell viability was measured using the CCK8 kit.
RT-PCR method for mRNA detection
Taking animal striatum tissue, extracting total RNA by TRIZOL, performing RT-PCR design after reverse transcription into cDNA to synthesize Nrf2, HO-1 primer, performing Realtime-PCR by using beta-actin or GADPH as reference gene and ABI-7300 instrument. The amplification efficiency between targets and the comparison between reference genes was calculated using Δ Δ CT, and the average content of target genes was calculated as 2 -ΔΔCT And (4) showing.
Western Blot protein content detection
Taking striatum tissue of experimental rat, adding protein lysate, fully homogenizing, then cracking on ice for 30min, transferring the cracking mixture into a 1.5ml EP tube, centrifuging at 4 ℃, 12500rpm for 15min, taking supernatant and storing on ice. And (3) quantifying protein by using a BCA kit, taking the same mass of protein, loading, separating SDS-PAGE protein, transferring a PVDF membrane, sealing 5% skimmed milk at room temperature for 1h, adding a primary antibody (anti-Nrf 2, HO-1 antibody), standing overnight at 4 ℃, adding a corresponding secondary antibody, incubating at room temperature for 1h, developing by using an ECL reagent, and taking a picture.
7. Test results
7.1 Effect of Sinomenine on rat behaviourology
In the circling behaviors generated by the sham operation group, the model group, the medosha group, the sinomenine low-dose group, the sinomenine medium-dose group and the sinomenine high-dose group, the number of revolutions is 0, 407.8, 272.5, 219.5, 267.2 and 297.0 circles/30 minutes respectively, and obviously compared with the model group, the sinomenine low-dose group, the sinomenine medium-dose group and the sinomenine high-dose group can obviously reduce the circling behaviors caused by apomorphine. The sinomenine administration can obviously reduce the circling behavior caused by apomorphine and relieve the nerve function damage caused by 6-OHDA.
From the drop latency of the suspension test in fig. 2, the suspension latencies of rats in the sham operation group, the model group, the medoba group, the sinomenine low dose group, the sinomenine medium dose group and the sinomenine high dose group are respectively 10.2, 6.6, 14.2, 16.8, 12.8 and 13.0 minutes, and it is obvious that the suspension latency is significantly improved and has statistical significance (P <0.05) compared with the model group in the sinomenine low dose group, the sinomenine medium dose group and the sinomenine high dose group.
7.2 cell viability assay
As shown in FIG. 3, when the concentration of sinomenine is 1 μm, 2.5 μm and 5 μm, the activity of SH-SY5Y cells can be significantly improved compared with the model group, and the statistical significance is achieved (P is less than 0.05); when the concentration of the sinomenine is 10 mu m, 15 mu m and 20 mu m, the SH-SY5Y cell activity can be obviously improved compared with a model group, and the statistical significance is achieved (P is less than 0.01).
7.3 Western Blot protein content detection
As shown in FIG. 4, the transcriptional level of Nrf2, HO-1 in striatum was statistically enhanced after administering 20mg/kg sinomenine compared to the model group (P < 0.05).
7.4 Western Blot protein content detection.
As shown in A, B in FIG. 5, it is statistically significant (P <0.05) that 20mg/kg sinomenine administered enhances the expression of Nrf2 and HO-1 proteins in striatum, compared to the model group.
In conclusion, sinomenine can obviously reduce the circling behavior caused by apomorphine, prolong the drop latency period of a suspension test, enhance the activity of SY5Y cells treated by 6-OHDA and enhance the transcription and protein level of Nrf2 and HO-1 in striatum. According to researches, the sinomenine promotes the expression of antioxidase such as HO-1 and the like by activating an Nrf2/ARE channel, so that the effect of resisting mitochondrial damage is exerted, the Parkinson's motor function is finally improved, and the sinomenine has a good clinical application prospect.
The above examples merely represent some embodiments of the present invention and are described in more detail and detail but are not to be construed as limiting the scope of the invention. It should be noted that it is within the scope of the present invention for a person skilled in the art to make several variations and modifications without departing from the spirit of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.
Claims (7)
1. The application of the sinomenine in preparing the pharmaceutical composition for treating the Parkinson disease is characterized in that: the sinomenine promotes the expression of antioxidant enzyme to play a role in resisting mitochondrial damage and improving the application of the pharmaceutical composition with the Parkinson's motor function by activating an Nrf2/ARE channel.
2. The use of sinomenine according to claim 1 in the preparation of a pharmaceutical composition for the treatment of parkinson's disease, wherein: the pharmaceutical composition is a clinically acceptable pharmaceutical preparation prepared by taking sinomenine as a main component and adding pharmaceutically acceptable auxiliary materials or auxiliary components.
3. The use of sinomenine in the preparation of a pharmaceutical composition for the treatment of parkinson's disease according to claim 1, wherein: the pharmaceutical preparation comprises tablets, capsules, injections or powders.
4. The use of sinomenine according to claim 1 in the preparation of a pharmaceutical composition for the treatment of parkinson's disease, wherein: the content of the sinomenine in the pharmaceutical composition is 2.5 mg/kg-40 mg/kg.
5. The use of sinomenine in the preparation of a pharmaceutical composition for the treatment of parkinson's disease according to claim 1, wherein: when the pharmaceutical composition is applied to human beings, the content of the sinomenine in the pharmaceutical composition is 0.6-10 mg/kg according to the ratio of the body surface area of the animals to the body surface area of the human beings.
6. The use of sinomenine in the preparation of a pharmaceutical composition for the treatment of parkinson's disease according to claim 1, wherein: the content of the sinomenine in the pharmaceutical composition is 5 mg/kg-20 mg/kg.
7. The use of sinomenine in the preparation of a pharmaceutical composition for the treatment of parkinson's disease according to claim 1, wherein: when the pharmaceutical composition is applied to human beings, the content of the sinomenine in the pharmaceutical composition is 0.8-5 mg/kg according to the ratio of the body surface area of the animals to the body surface area of the human beings.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210284708.9A CN114831990A (en) | 2022-03-22 | 2022-03-22 | Application of sinomenine in preparing pharmaceutical composition for treating Parkinson's disease |
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| CN202210284708.9A CN114831990A (en) | 2022-03-22 | 2022-03-22 | Application of sinomenine in preparing pharmaceutical composition for treating Parkinson's disease |
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| CN202210284708.9A Pending CN114831990A (en) | 2022-03-22 | 2022-03-22 | Application of sinomenine in preparing pharmaceutical composition for treating Parkinson's disease |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004048340A1 (en) * | 2002-11-28 | 2004-06-10 | Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences | Sinomenine and sinomenine compounds, synthesis and use |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004048340A1 (en) * | 2002-11-28 | 2004-06-10 | Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences | Sinomenine and sinomenine compounds, synthesis and use |
Non-Patent Citations (2)
| Title |
|---|
| LI QIAN ET AL.: ""Sinomenine, a natural dextrorotatory morphinan or analog, is anti-inflammatory and neuroprotective through inhibition of microglial NADPH oxidase"", vol. 4, no. 23, pages 1 - 14 * |
| 孙凤艳主编: "《医学神经生物学》", 上海科学技术出版社, pages: 129 - 130 * |
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