Application of fructus cnidii or fried fructus cnidii aqueous extract in preparation of drug for treating Duchenue type muscular dystrophy
By using snake bed or fried snake bed water extract, the deficiency of the middle and late stage treatment of Duchenne muscular dystrophy was solved, and the effect of reducing serum creatine kinase content and restoring muscle fibers was achieved, which significantly improved the middle and late stage symptoms of the disease.
Patent Information
- Application Number
- CN202510582497.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-07
- Publication Date
- 2025-06-17
AI Technical Summary
There is no effective drug for treating the middle and late stages of Duchenne muscular dystrophy in the prior art, so new treatment plans are urgently needed.
In the preparation of Duchenue-type muscular dystrophy drugs, the preparation method of stir-frying and water extracts is obtained and used as a pharmaceutical ingredient.
The water extract of Snake or Fried Snake significantly reduces the serum creatine kinase (CK). The broken and broken muscle fibers tend to recover significantly, and the infiltration of inflammatory cells is significantly reduced, which can effectively treat Duchenne muscular dystrophy in the middle and late stages.
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Figure CN120154650A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the application of the aqueous extract of Fructus Cnidii or stir-fried Fructus Cnidii in the preparation of drugs for treating Duchenue muscular dystrophy, belonging to the technical field of natural drugs. Background Art
[0002] Duchenne muscular dystrophy (DMD) is a severe, progressive, and disabling X-linked disease characterized by muscle atrophy. This disease causes difficulty in movement, ultimately requiring assisted ventilation and often leading to premature death. The main cause of DMD is the mutation of the dystrophin gene, which is responsible for encoding dystrophin. These mutations result in a decrease or absence of dystrophin production in muscle tissue, triggering muscle atrophy and a series of complications. The absence of dystrophin leads to the breakdown of the dystrophin-associated protein complex (DAPC) within the muscle membrane, and this breakdown disrupts the interaction between actin and the extracellular matrix, making muscles lacking dystrophin more vulnerable to injury. This susceptibility leads to the gradual loss of muscle tissue and function, as well as the development of cardiomyopathy.
[0003] Duchenne muscular dystrophy (DMD) has the following case characteristics, and bioinformatics analysis and bibliometric analysis are carried out from the following five aspects:
[0004] Genetic basis: An X-linked recessive genetic disease, resulting in the absence of dystrophin encoding due to gene defects.
[0005] Muscle structure damage: Muscle fibers are gradually replaced by fat and connective tissue, triggering progressive muscle atrophy and fibrosis.
[0006] Dysfunction: Patients present with muscle weakness and gradually lose motor function (such as walking ability), and in the later stage, the respiratory muscles and myocardium are involved, leading to respiratory failure and cardiomyopathy.
[0007] Disease progression: Early glucocorticoid treatment can delay the disease process, but the intervention effect is limited after the disease enters the middle and late stages.
[0008] Associated pathology: Muscle stem cells may actively eliminate abnormal cells through apoptosis or programmed necrosis during the degeneration process, and this compensatory mechanism plays a role in slowing down the pathological progression in the early stage of the disease.
[0009] Duchenne muscular dystrophy can be treated or alleviated with glucocorticoids in the early stage. Currently, there are no drugs for treating the middle and late stages of Duchenne muscular dystrophy, so there is an urgent need for the application of the aqueous extract of Fructus Cnidii and stir-fried Fructus Cnidii in the preparation of drugs for treating Duchenue muscular dystrophy. Summary of the Invention
[0010] In view of the above problems, the object of the present invention is to provide the use of the water extract of Cnidium monnieri (L.) Cuss. or stir-fried Cnidium monnieri (L.) Cuss. in the preparation of a drug for treating Duchenne muscular dystrophy.
[0011] To solve the above technical problems, the technical solution adopted by the present invention is as follows:
[0012] The use of the water extract of Cnidium monnieri (L.) Cuss. or stir-fried Cnidium monnieri (L.) Cuss. in the preparation of a drug for treating Duchenne muscular dystrophy.
[0013] The preparation method of stir-fried Cnidium monnieri (L.) Cuss. is: stir-fry at 103°C for 25 minutes, take out and cool, then it is obtained.
[0014] The preparation method of the water extract of stir-fried Cnidium monnieri (L.) Cuss. is: take 100 g of stir-fried Cnidium monnieri (L.) Cuss., place it in a traditional Chinese medicine decocting earthenware pot, add water and decoct twice. For the first decoction, add 800 mL of water, soak for 30 minutes, then bring to a boil over high heat, and then simmer for 30 minutes. For the second decoction, add 600 mL of water, bring to a boil over high heat, and then simmer gently for 20 minutes; combine the decoction liquids, filter while it is hot, and then concentrate under reduced pressure to a thick extract, place it in a glass evaporating dish and freeze-dry to obtain the freeze-dried powder of the water extract of stir-fried Cnidium monnieri (L.) Cuss., and set aside. Preferably, the filtration is through a 300-mesh sieve; the temperature of concentration under reduced pressure is 60°C, and the pressure is -0.08 MPa.
[0015] The preparation method of the water extract of Cnidium monnieri (L.) Cuss. is: take 100 g of raw Cnidium monnieri (L.) Cuss., place it in a traditional Chinese medicine decocting earthenware pot, add water and decoct twice. For the first decoction, add 800 mL of water, soak for 30 minutes, then bring to a boil over high heat, and then simmer for 30 minutes. For the second decoction, add 600 mL of water, bring to a boil over high heat, and then simmer gently for 20 minutes; combine the decoction liquids, filter while it is hot, and then concentrate under reduced pressure to a thick extract, place it in a glass evaporating dish and freeze-dry to obtain the freeze-dried powder of the water extract of Cnidium monnieri (L.) Cuss., and set aside. Preferably, the filtration is through a 300-mesh sieve; the temperature of concentration under reduced pressure is 60°C, and the pressure is -0.08 MPa.
[0016] A drug for treating Duchenne muscular dystrophy, comprising the water extract of Cnidium monnieri (L.) Cuss. or stir-fried Cnidium monnieri (L.) Cuss.
[0017] A drug for treating Duchenne muscular dystrophy, further comprising pharmaceutical excipients.
[0018] Preferably, the pharmaceutical excipients include any one or more of solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, antiadhesives, chelating agents, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, clathrates, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retardants.
[0019] A preparation of a drug for treating Duchenue muscular dystrophy, the preparation including powder, tablets, granules, capsules, injections or oral liquids.
[0020] Compared with the prior art, the present invention has the following advantages:
[0021] In the early stage of Duchenne muscular dystrophy, glucocorticoids can be used for treatment or alleviation. However, at present, there are no drugs for treating the middle and late stages of Duchenne muscular dystrophy. The aqueous extract of Cnidium monnieri or stir-fried Cnidium monnieri of the present invention, especially the aqueous extract of stir-fried Cnidium monnieri, has a significant therapeutic effect on Duchenne muscular dystrophy and has the effect of reducing the content of serum creatine kinase (CK); after the muscle fibers of the Duchenne muscular dystrophy group (DMD) after being broken and fragmented are intervened with the aqueous extract of Cnidium monnieri or stir-fried Cnidium monnieri, the muscle fibers have an obvious tendency to recover, and the degree of infiltration of inflammatory cells is significantly reduced, indicating that the aqueous extract of Cnidium monnieri or stir-fried Cnidium monnieri of the present invention, especially the aqueous extract of stir-fried Cnidium monnieri, can not only treat the early stage of Duchenne muscular dystrophy, but also treat the middle and late stages of Duchenne muscular dystrophy. Description of the Drawings
[0022] Figure 1 For the content of creatine kinase in the serum of mice of the present invention, ** P < 0.01;
[0023] Figure 2 For the histological analysis of muscle pathology of the present invention (100×). Detailed Embodiments
[0024] The present invention will be further described in detail below with reference to the drawings and specific embodiments. The following embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.
[0025] Application of the aqueous extract of Cnidium monnieri or stir-fried Cnidium monnieri in the preparation of a drug for treating Duchenue muscular dystrophy.
[0026] 1. Preparation of Test Samples
[0027] 1.1 Preparation of stir-fried Cnidium monnieri samples: In a small-scale medicine frying machine with a capacity of 3L, 30g of Cnidium monnieri each time, stir-fry at 103 °C for 25 min, take out and let cool, and you will get it.
[0028] 1.2 Preparation of standard decoction: Weigh 100 g of Fructus Cnidii and stir-fried Fructus Cnidii respectively, place them in a decocting earthenware pot, add water and decoct twice. For the first decoction, add 800 mL of water, soak for 30 minutes, then bring to a boil over high heat and then simmer for 30 minutes. For the second decoction, add 600 mL of water, bring to a boil over high heat, and then simmer gently for 20 minutes. Combine the decoction liquids, filter while it is hot (through a 300-mesh sieve), and then concentrate under reduced pressure (at 60 °C, -0.08 MPa) to a thick extract with an appropriate relative density. Place it in a glass evaporating dish and freeze-dry to obtain the freeze-dried powder of the standard decoction of Fructus Cnidii and stir-fried Fructus Cnidii for standby.
[0029] A drug for treating Duchenue muscular dystrophy, comprising the water extract of the above-mentioned Fructus Cnidii or stir-fried Fructus Cnidii.
[0030] A drug for treating Duchenue muscular dystrophy further comprises pharmaceutical excipients.
[0031] Pharmaceutical excipients include any one or more of solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, antiadhesives, chelating agents, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, clathrates, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, and release retardants.
[0032] The preparation of a drug for treating Duchenue muscular dystrophy according to this example, the preparation includes powder, tablets, granules, capsules, injections or oral liquids.
[0033] 2. Experimental process:
[0034] The modeling method of DMD model mice (C57 DMD(- / -) ) is as follows:
[0035] Introduce a C3197T point mutation into exon 23 of the DMD gene of wild-type C57 mice to obtain the DMD-C57 DMD (- / -) mouse model.
[0036] 15-week-old DMD model mice (C57 DMD(- / -) ) were respectively continuously intragastrically administered the freeze-dried powder of the standard decoction of Fructus Cnidii and stir-fried Fructus Cnidii (0.2 g / 20 g (body weight)) for 7 days, twice a day; at the same time, 15-week-old wild-type C57 and DMD model mice (C57 DMD(- / -) ) were continuously intragastrically administered an equal dose of normal saline.
[0037] 2.1 Detection of serum creatine kinase (CK)
[0038] The CK experiment for Duchenne muscular dystrophy (DMD), namely the creatine kinase experiment, plays an important role in the diagnosis and disease monitoring of DMD. When muscle cells are damaged, CK is released into the blood, leading to an increase in the CK level in the blood. Due to the defect in the muscle cell membrane of DMD patients, the contents of muscle cells leak out, including CK. Therefore, by detecting the content of CK in the blood, it can assist in the diagnosis of DMD. Usually, the enzyme-linked immunosorbent assay is used to detect the CK level in serum. Figure 1 The OD value therein represents the content value of CK detected by the enzyme-linked immunosorbent assay.
[0039] After the mice were euthanized, fresh blood was collected by eye bleeding immediately. The blood samples were centrifuged at 1500 r at 4 °C for 10 min and stored in a -80 °C refrigerator. The blood samples were taken out from the refrigerator and placed on ice to wait for thawing. According to the operation instructions of the creatine kinase assay kit (N-acetylcysteine method), the CK content in the blood samples was measured and the OD value was calculated. The results are as Figure 1 shown. The OD value of the Duchenne muscular dystrophy group (DMD) was significantly higher than that of the normal group (WT), the cnidium fruit group (CF), and the stir-fried cnidium fruit group (FCF), ( ** P < 0.01). There was no significant difference in OD between the normal group (WT) and the stir-fried cnidium fruit group (FCF). It indicated that both cnidium fruit (CF) and stir-fried cnidium fruit (FCF) could reduce the content of serum creatine kinase (CK), and the effect of FCF was better.
[0040] 2.2 Pathological injury of mouse skeletal muscle
[0041] The gastrocnemius muscles of the mice were fixed with 4% paraformaldehyde for HE staining experiments, and pathological histological examinations were performed to observe the muscle lesion conditions. The results are as Figure 2 : In the Duchenne muscular dystrophy group (DMD), the morphology of muscle fibers was no longer regular. The polygonal muscle fibers became distorted and deformed, with uneven edges. Some muscle fibers also showed rupture and fragmentation. A large number of irregular and diffuse inflammatory cell infiltrations were visible between muscle fibers; in the normal group (WT), that is, the muscle fibers of wild-type C57 mice were regular in morphology, without rupture and fragmentation, and the immune cells were evenly distributed; in the DMD model mice intervened with the freeze-dried powder of the standard decoction of cnidium fruit (CF) and stir-fried cnidium fruit (FCF) respectively, that is, in the Duchenne muscular dystrophy + cnidium fruit group (CF) and the Duchenne muscular dystrophy + stir-fried cnidium fruit group (FCF), the morphology of muscle fibers was as regular as that of the normal group, showing a tendency to recover compared with C57 DMD(- / -) and the degree of inflammatory cell infiltration decreased.
[0042] It should be understood that, in order to streamline the present disclosure and assist in understanding one or more of the various inventive aspects, in the foregoing description of the exemplary embodiments of the present invention, the various features of the present invention are sometimes grouped together into a single embodiment or the description thereof. However, the disclosed method should not be construed as reflecting an intention that the claimed invention requires more features than are expressly recited in each claim. Rather, as reflected by the claims, the inventive aspects lie in less than all the features of the previously disclosed embodiments. Thus, the claims following the detailed description hereby expressly incorporate the detailed description, where each claim itself serves as a separate embodiment of the present invention.
[0043] Although the present invention has been described in terms of a limited number of embodiments, those skilled in the art, having the benefit of the foregoing description, will appreciate that other embodiments can be contemplated within the scope of the invention as thus described. In addition, it should be noted that the language used in this specification has been principally selected for readability and instructional purposes and not for the purpose of explaining or limiting the subject matter of the invention. Accordingly, many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the appended claims. For the scope of the present invention, the disclosure of the present invention is illustrative, not restrictive, and the scope of the present invention is defined by the appended claims.
[0044] The foregoing are only the preferred embodiments of the present invention, and it should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.
Claims
1. Application of Cnidium monnieri or fried Cnidium monnieri water extract in the preparation of drugs for treating Duchenue muscular dystrophy.
2. The use according to claim 1, characterized in that: The preparation method of fried Cnidium monnieri is as follows: fry at 103°C for 25 minutes, take out and let cool.
3. The use according to claim 2, characterized in that: The preparation method of the fried Cnidium monnieri water extract is as follows: 100 g of the fried Cnidium monnieri is taken, placed in a decoction casserole, and water is added and decocted twice, 800 ml of water is added for the first decoction, and the mixture is boiled over high heat after soaking for 30 minutes, and then decocted over low heat for 30 minutes, and 600 ml of water is added for the second decoction, and the mixture is boiled over high heat and then simmered over low heat for 20 minutes; the decoctions are combined, filtered while hot, and then concentrated under reduced pressure to a thick extract, and the mixture is freeze-dried in a glass evaporating dish to obtain freeze-dried powder of the fried Cnidium monnieri water extract for standby use.
4. The use according to claim 3, characterized in that: The filtration was through a 300-mesh screen; the temperature for reduced pressure concentration was 60°C and the pressure was -0.08 MPa.
5. The use according to claim 1, characterized in that: The preparation method of the Cnidium monnieri water extract is as follows: 100 g of raw Cnidium monnieri is taken, placed in a decoction casserole, and water is added and decocted twice, 800 mL of water is added for the first decoction, and the mixture is boiled over high heat after soaking for 30 minutes, and then decocted over low heat for 30 minutes, and 600 mL of water is added for the second decoction, and the mixture is boiled over high heat and then simmered over low heat for 20 minutes; the decoctions are combined, filtered while hot, and then concentrated under reduced pressure to a thick extract, and freeze-dried in a glass evaporating dish to obtain freeze-dried powder of the Cnidium monnieri water extract for later use.
6. The use according to claim 5, characterized in that: The filtration was through a 300-mesh screen; the temperature for reduced pressure concentration was 60°C and the pressure was -0.08 MPa.
7. A drug for treating Duchenue muscular dystrophy, characterized in that: Contains water extract of Cnidium monnieri or stir-fried Cnidium monnieri.
8. A drug for treating Duchenue muscular dystrophy according to claim 7, characterized in that: Also contains pharmaceutical excipients.
9. A drug for treating Duchenue muscular dystrophy according to claim 8, characterized in that: Pharmaceutical excipients include any one or more of solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, and release retardants.
10. A preparation comprising the drug for treating Duchenue muscular dystrophy according to claim 7, characterized in that: Preparations include powders, tablets, granules, capsules, injections or oral solutions.