Application of vasicine in preparation of medicine for intervening psoriasis
By using drugs prepared by sausagenine, the skin damage and adverse reactions of existing psoriasis treatment methods have been solved, and effective treatment of psoriasis is achieved, and high safety is achieved.
Patent Information
- Application Number
- CN202411653532.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-19
- Publication Date
- 2025-02-21
AI Technical Summary
The existing treatment methods for psoriasis have damage to the skin and adverse reactions, and no drugs or methods can be found that can cure psoriasis.
Using pyrosine as the main component, high-purity pyrosine products are obtained by preparing drugs for psoriasis, including extraction with organic solvents, macroporous resin enrichment and silica gel column chromatography purification.
Duckling-billed solanine can significantly improve the pathological symptoms of psoriasis and inhibit the overexpression of inflammatory cytokines, thus having a therapeutic effect on psoriasis. Compared with traditional drugs, its therapeutic effect and safety are close to that of the patient.
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Figure CN119488526A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of drug application, and particularly relates to the application of dapoxetine in the preparation of drugs for intervening in psoriasis. Background Art
[0002] Psoriasis, commonly known as psoriasis, is a common chronic inflammatory, erythematous and scaly skin disease in dermatology. It is prone to recurrence and difficult to cure. The onset time is long. Some patients are almost never cured. Anyone who stops taking the medicine may relapse. It is often aggravated in winter and significantly relieved in summer, that is, it is severe in winter and mild in summer. The cumulative population is relatively wide. Because of its easy recurrence and erythematous and scaly skin lesions, it seriously affects the quality of life and beauty of patients, and has an impact on the physical health and mental state of patients. The pathogenesis of psoriasis is very complex, including genetic factors, immune mechanisms, environmental factors, and traditional Chinese medicine etiology. Human understanding of psoriasis has gone through a relatively long and tortuous process. So far, no drugs and methods that can cure psoriasis have been found. The most common treatment measures are biological agents, phototherapy, immune regulation, anti-inflammatory, etc. The existing schemes include sequential therapy, alternation and combination. Although these methods have been affirmed in clinical treatment, they all have damage to the skin and corresponding adverse reactions to a certain extent. Therefore, the development of safe and effective new drugs is still the focus of medical attention.
[0003] The pathological manifestations of psoriasis are mainly based on the clear boundary of infiltrative erythema, with hyperkeratosis of the epidermis accompanied by incomplete angles, covered with silvery white scales, thick scales, and generally no obvious itching. There are many factors that induce and aggravate it, which have not yet been fully clarified. In the theory of traditional Chinese medicine, psoriasis is considered to be in the category of "pine skin ringworm", "white scaly skin", "scaly skin wind", etc. Its causes include blood stasis, dampness, blood heat, external wind, etc. Among them, the most common pathogenic factor of psoriasis is blood heat and wind, which runs through the entire pathological process of psoriasis. The evil of blood heat gathers in the skin. People with psoriasis will feel heat accumulation in the spleen and stomach and excessive internal heat, or exogenous wind heat, heat enters the blood, local qi and blood flow is blocked, blood heat causes skin erythema, and heat and blood dryness cause symptoms such as scales and desquamation. Therefore, traditional Chinese medicine can also treat psoriasis from the perspective of clearing heat and cooling blood. From the perspective of modern medicine, it is a skin inflammatory reaction caused by excessive proliferation of epidermal cells caused by genetic inheritance, immune dysfunction and T cell abnormalities.
[0004] Vasicine is a pyrrolo[2,1-b]quinazoline alkaloid with a chiral center isolated from Adhatodavasica Nees. of the Acanthaceae family and Peganumharmala Linn. of the Tribulaceae family. It is mainly distributed in Yunnan and Sichuan, my country. The classic Tibetan medicine book "Jingzhu Bencao" records that vasicine is bitter and cool in nature, and has the effect of treating blood heat disease, liver heat, and tribal heat. It is a commonly used drug for rheumatoid arthritis. Among them, a series of compounds with strong anti-inflammatory activity are extracted from the dichloromethane part of vasicine, including vasicone, vasicine, 7-methoxyvasicine, syringaresinol, 3-indolecarboxaldehyde and other ingredients. Subsequent experiments confirmed that these ingredients are indeed effective in improving the pathological condition of skin lesions and can reduce the number of mast cells and inflammatory cytokines in peripheral blood. This led to the idea that the ingredient daffodil in daffodil may have a therapeutic effect on skin inflammatory psoriasis. Summary of the invention
[0005] In order to solve the above technical problems, the present invention proposes the use of gentianine in the preparation of drugs for intervening in psoriasis.
[0006] In order to achieve the above-mentioned object, the present invention provides, on one hand, the use of dapoxetine in the preparation of a drug for intervening in psoriasis.
[0007] The drug for intervening psoriasis is in any dosage form acceptable in the pharmaceutical field, and the dosage form is powder, granule, injection, capsule, tablet or oral solution.
[0008] Another aspect of the present invention provides a method for preparing daffodil, comprising the following steps:
[0009] S1, drying and crushing the plant material containing chloranthine to obtain a raw material powder;
[0010] S2, using an organic solvent to perform multiple dynamic extractions on the raw material powder, combining the extracts, filtering to remove impurities, and performing reduced pressure concentration to obtain a preliminary extract;
[0011] S3, using a macroporous resin to enrich the preliminary extract, and using an eluent to separate dapoxetine from other impurities; the enriched solution is purified by silica gel column chromatography, the purified dapoxetine eluate is collected, and evaporated to dryness under reduced pressure or freeze-dried to obtain a high-purity dapoxetine product;
[0012] S4. Use high performance liquid chromatography to test the purity of chloranthine products to ensure that the products meet the quality requirements.
[0013] Preferably, the organic solvent in S2 is a mixture of ethanol and water, and the volume ratio of ethanol to water is 1 to 3:1.
[0014] Preferably, in S2, the raw material powder is dynamically extracted 2-4 times using an organic solvent, the extraction temperature is 40-60° C., and the extraction time for each time is 30-60 minutes.
[0015] Preferably, the macroporous resin in S3 is of type D101, and the concentration and pH value of the eluent during the enrichment process are optimized to effectively separate chloranthelline from other impurities.
[0016] The beneficial effects of the present invention are:
[0017] The bioactive ingredients in coleus can significantly improve the pathological symptoms of psoriasis. Coleus alkaloids can inhibit the overexpression of inflammatory cytokines. Drugs prepared from coleus alkaloids can have a therapeutic effect on psoriasis. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for describing the embodiments are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.
[0019] Figure 1 Effects of chloranthine on the appearance of skin tissue in psoriasis mice.
[0020] Figure 2 PASI images of skin tissues of mice in each group.
[0021] Figure 3 A diagram showing the effect of chloranthine on the pathological changes of skin lesions on the back of psoriasis mice.
[0022] Figure 4 Changes of spleen index in each group of mice.
[0023] Figure 5 The effect of chloranthine on the IL-17A content in the peripheral blood of psoriasis model mice.
[0024] Figure 6 The effect of chloranthine on the IL-23 content in the peripheral blood of psoriasis model mice.
[0025] Figure 7 The effect of chloranthine on the IL-25 content in the peripheral blood of psoriasis model mice.
[0026] Figure 8 Effect of chloranthine on IL-17A content in skin tissue homogenate of psoriasis model mice.
[0027] Fig. 9Effect of chloranthine on IL-23 content in skin tissue homogenate of psoriasis model mice.
[0028] Fig.10 Effect of chloranthine on IL-25 content in skin tissue homogenate of psoriasis model mice. DETAILED DESCRIPTION
[0029] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.
[0030] Example 1
[0031] The preparation method of daffodil alkaloids comprises the following steps:
[0032] S1, drying and crushing the plant material containing chloranthine to obtain a raw material powder;
[0033] S2. The raw material powder is subjected to multiple dynamic extractions using an organic solvent, wherein the organic solvent is a mixture of ethanol and water, and the volume ratio of ethanol to water is 1:1. The raw material powder is subjected to 4 dynamic extractions using the organic solvent, the extraction temperature is 60° C., and the extraction time for each extraction is 60 minutes. The extracts are combined, filtered to remove impurities, and concentrated under reduced pressure to obtain a preliminary extract;
[0034] S3, using a macroporous resin to enrich the preliminary extract, and using an eluent to separate dapoxetine from other impurities; the enriched solution is purified by silica gel column chromatography, the purified dapoxetine eluate is collected, and evaporated to dryness under reduced pressure or freeze-dried to obtain a high-purity dapoxetine product;
[0035] S4. Use high performance liquid chromatography to test the purity of chloranthine products to ensure that the products meet the quality requirements.
[0036] Example 2
[0037] Experimental material preparation: Male mice weighing 18-20g and aged 8-12 weeks were provided by Experimental Animal Co., Ltd. They were raised in the SPF-level Animal Experimental Center of Yunnan University of Traditional Chinese Medicine. Drinking water was provided by the experimental center. The breeding temperature was maintained at 18-24℃. They were fed regularly, the bedding was replaced, and the cages were cleaned and disinfected regularly. The experiment began after one week of experimental feeding.
[0038] The experimental drugs include dapoxetine, imiquimod ointment, and positive drugs (methotrexate); the experimental reagents include normal saline, 0.5% CMC-Na, and white vaseline with a specification of 500g / bottle supplied by Yunnan Provincial Hospital of Traditional Chinese Medicine; the experimental instruments include Shengqiang Technology slide scanning image analysis system, centrifuge, and tissue homogenizer.
[0039] The experimental method includes the following steps:
[0040] 1) Animal modeling
[0041] Animal modeling refers to artificially inducing psoriasis symptoms similar to those of humans on the back of animals by some methods. Common artificial methods include controlling the essential fatty acids required by the animal, giving chemical stimulation and drugs, etc. Currently, most of them use imiquimod and propranolol for induction. This experiment used imiquimod to create the model.
[0042] After one week of adaptive feeding, 24 BALB / c male mice were subjected to shaver and depilatory cream to remove hair on the back of all mice to form a 2cm×3cm exposure area. Then, they were randomly divided into groups: Normal group; Model group; MTX-positive control group (3mg / kg); Vas-vastatin group (200mg / kg).
[0043] 2) Animal administration
[0044] The normal group was smeared with 62.5 mg of vaseline on the back; then normal saline was gavaged once a day for 5 consecutive days.
[0045] The model group was smeared with 62.5 mg of imiquimod on the back, and then gavage with normal saline once a day for 5 consecutive days.
[0046] In the pyralidin control group, 62.5 mg of 5% imiquimod ointment was applied to the exposed area; then 200 mg / kg of pyralidin dissolved in 0.5% CMC-Na was administered intragastrically once a day for 5 consecutive days.
[0047] In the positive drug control group, 62.5 mg of 5% imiquimod ointment was applied to the exposed area; then 3 mg / kg of methotrexate dissolved in 0.5% CMC-Na was administered intragastrically once a day for 5 consecutive days.
[0048] 3) Analysis of the tissue condition of the skin lesions on the back of male mice
[0049] PASI evaluation of skin lesions
[0050] During the drug administration process, PASI scoring was performed according to the PASI scoring standard. Since the skin lesions were all on the back, the area percentage was not considered. The erythema, epidermal desquamation and infiltration degree of the mouse skin lesions were recorded as 0-4 points respectively. The total score of the mouse skin lesion index was the sum of erythema, epidermal desquamation and plaque hypertrophy. The specific evaluation criteria are: no erythema, no epidermal desquamation, no tissue hypertrophy or intact skin, recorded as 0 points; light red spots can be seen on the skin epidermis, a few minor epidermal desquamation can be seen with the naked eye, or the degree of skin damage is slightly higher than normal, recorded as 1 point; red patches can be seen on the skin epidermis, the skin lesions have obvious bulges, or the degree of epidermal damage is skin-like epidermal desquamation, recorded as 2 points; obvious lesion tissue bulges can be seen with the naked eye, the skin color is dark red, and the epidermis can be seen with obvious desquamation or obvious infiltration, recorded as 3 points; the skin lesions are extremely obviously thickened and raised, and the color is very dark red patches, the surface is covered with thick and layered desquamation or obvious infiltration is extremely severe, recorded as 4 points. PASI = scale score + erythema score + skin thickening score. The scores of the three groups of mice were averaged, and the trend line was drawn. The trend line was observed to evaluate the psoriasis-like skin lesions of each group.
[0051] Animal sampling and HE staining
[0052] After the last administration and 12 hours of fasting, blood was collected from mice in each group. After blood collection, the mice were centrifuged at 1500r / min for 5 minutes at 4°C, and the upper serum was drawn out and stored at -20°C for cytokine detection. Then the mice were killed by cervical dislocation, and a part of the back skin was removed for pathological sectioning, then fixed in 10% formaldehyde solution, embedded in paraffin, sectioned, and finally stained with hematoxylin-eosin (HE) and toluidine blue. A part of the back skin was removed for tissue homogenization and cytokine detection in the tissue. The pathological damage of the HE-stained skin lesion tissue of the mice was observed by glass slide scanning. The specific evaluation criteria are: hyperkeratosis or incomplete keratinization lesions can be clearly observed under the microscope, the disappearance of particles or significant thinning of the granular layer can be observed, papillary edema and the thinning of the spinous layer on the papilla and the extension of the upward extension, capillary dilation, inflammatory cells exuding from the blood vessels, and a large number of inflammatory cells infiltrated (such as: a large number of mast cells accumulate to form abscesses).
[0053] ELISA was used to detect the expression levels of IL-17A, IL-23 and IL-25 in mouse skin homogenate
[0054] Accurately weigh the mouse skin tissue, add PBS at a ratio of 1:9, and fully homogenize with a tissue grinder. Centrifuge at 4°C, 3000 rpm for 20 min, collect the supernatant, and detect the expression levels of IL-17A, IL-23, and IL-25 inflammatory factors in the skin tissue of each group according to the instructions of the ELISA kit.
[0055] ELISA was used to detect the expression levels of IL-17A, IL-25, and IL-23 in peripheral blood
[0056] The eyeballs of anesthetized mice were removed, blood was collected from the eye sockets, and the blood was allowed to stand for 30 minutes. The blood was centrifuged in a low-temperature high-speed centrifuge for 10 minutes at 4°C and 3000 rpm, and the upper serum was aspirated and stored at -80°C. The expression levels of IL-17A, IL-23, and IL-25 inflammatory factors in the peripheral blood of each group were detected according to the instructions of the ELISA kit.
[0057] General analysis: Observe and record the body weight and spleen weight of the mice, and calculate the spleen index; spleen index = spleen weight / body weight × 100%.
[0058] 4) Process relevant data through statistics and analyze the results
[0059] The obtained data were statistically analyzed, and the data were expressed as x±s. One-way analysis of variance and t-test were used for analysis, and a value of P<0.05 indicated that the difference was statistically significant.
[0060] Effects of chloranthine on skin lesions and PASI scores in mice
[0061] The back skin of the mice in the normal group was smooth without erythema, scaling, thickening and other changes. The back skin lesions of the model group were very obvious, thickened and raised, with obvious erythema, and the epidermis was covered with thick and layered fine scaling. The psoriasis lesion-like symptoms on the back of the mice in the daffodil group and the positive control group were milder than those in the model group. The epidermal thickening, erythema and scaling in the daffodil group were slightly higher than those in the positive control group (see Figure 1 ). PASI score results (such as Figure 2 ) As shown in the figure, the erythema, epidermal desquamation, plaque thickening and infiltration of the skin lesions of the mice in the model group have been on a trend of aggravation since the second day of drug administration. The skin lesion-like symptoms of the positive drug group and the anthracene group were significantly improved compared with the model group, and the PASI scores of the positive group and the anthracene group in the next two days were almost the same, indicating that anthracene does have a therapeutic effect on the symptoms of psoriasis in mice, and the therapeutic effect is almost consistent with the efficacy of the positive drug.
[0062] Effects of chloranthine on pathological degree of skin lesions in mice
[0063] The skin tissue of mice in the normal control group showed no obvious abnormalities, the epidermis was intact and clear, the stratified squamous epithelium was thin, and the cells were arranged more neatly and tightly; the cytoplasm in the dermis was red-stained and eosinophilic, the hair follicles and glands were normal in morphology, and the fat tissue was clearly visible. The stratum corneum of mice in the model group was hyperkeratotic, the granular layer was significantly thickened, the skin processes were significantly elongated and undulating, the spinous layer was thickened, and inflammatory cell infiltration was visible in the dermis. The granular layer of mice in the positive drug (methotrexate) group was significantly thinned, the skin processes were slightly elongated and undulating, a small amount of inflammatory cell infiltration was visible in the dermis, and no other obvious lesions were found. The stratum corneum of mice in the dapoxetine group was hyperkeratotic, and a small amount of inflammatory cell infiltration was visible in the dermis, and no other obvious lesions were found.
[0064] Effect of chloranthine on spleen coefficient of mice
[0065] The spleen index of mice in the normal group was low, below 0.5, while the spleen index of the model group was much higher than that of the normal group, about 1.3. Compared with the model group, the indexes of the pyralidin group and the positive drug group were significantly reduced, indicating that pyralidin can reduce the spleen index of psoriasis model mice.
[0066] Effects of chloranthine on IL-7A, IL-23 and IL-25 cytokines in peripheral blood of mice
[0067] The levels of IL-7A, IL-23, and IL-25 in the peripheral blood of mice in the normal group were low, less than 20pg / ml. Compared with the normal group, the levels of IL-7A, IL-23, and IL-25 in the peripheral blood of mice in the model group increased significantly, reaching 30pg / ml. It can be seen that the mechanism of psoriasis-like symptoms in mice is related to the increased expression of inflammatory cytokines IL-7A, IL-23, and IL-25. Compared with the model group, the levels of IL-7A, IL-23, and IL-25 in the peripheral blood of the daffodil group and the positive group were reduced, indicating that daffodil can inhibit the expression of IL-7A, IL-23, and IL-25 in the peripheral blood of psoriasis model mice. (e.g. Figure 5-7 )
[0068] Effect of calanine on the contents of IL-7A, IL-23 and IL-25 in skin tissue homogenates of psoriasis mice.
[0069] The expression levels of IL-7A, IL-23, and IL-25 in the skin tissue homogenate of mice in the normal group were low. Compared with the normal group, the content of IL-7A, IL-23, and IL-25 in the skin tissue homogenate of mice in the model group increased significantly, indicating that when mice developed psoriasis-like symptoms, the expression levels of inflammatory cytokines IL-7A, IL-23, and IL-25 in the skin tissue homogenate also showed an increasing trend. After the administration of daffodil and positive control drugs, the content of IL-7A, IL-23, and IL-25 in the skin tissue homogenate of mice decreased. The degree of decrease in the expression content in the daffodil group was less than that in the positive control group. It is speculated that daffodil can inhibit the expression of IL-7A, IL-23, and IL-25 in the skin tissue homogenate of psoriasis model mice, but the inhibitory effect is not as good as that of the positive drug. (such as Figure 8-10 )
[0070] Studies have found that psoriasis often occurs together with metabolic syndrome, obesity, diabetes and other diseases. The cause and pathological development of psoriasis are closely related to small molecules such as cytokines. Studies have confirmed that the IL-17A / IL-23 axis is the central inflammatory axis that connects various cytokines in the development of psoriasis. IL-23 mainly comes from monocytes, Langerhans cells, macrophages, and dermal dendritic cells. In the pathological process of psoriasis, it promotes the proliferation of Th17 cells and the secretion of IL-17, IL-25 and other inflammatory factors by binding to related receptors, resulting in overexpression of these inflammatory factors, which in turn causes abnormal proliferation and overactivation of keratinocytes, resulting in a series of specific immune responses, and also aggravates the pathological condition of psoriasis by inducing overexpression of other inflammatory factors such as IL-36 (IL-36α, IL-36β, IL-36γ). IL-17 is particularly important in the pathogenesis of psoriasis. It is a key pathogenic factor of psoriasis and is mainly derived from dermal γδTCRlow T cells. It can activate neutrophils, prevent their apoptosis, promote the formation of various cells such as TNF-α, IL-6, IL-1, etc., leading to keratinocyte activation and abnormal proliferation, prompting neutrophils to gather in the lesions, and causing pathological changes in the epidermal tissue structure. Therefore, to effectively improve the pathological condition of psoriasis, it should be possible to inhibit the expression of inflammatory cytokines IL-23 and IL-17.
[0071] The results of this study showed that the bioactive components in Duckbill Flower can significantly improve the pathological symptoms of psoriasis model mice, and the results of immunohistochemistry showed that Duckbill Flower alkaloids can inhibit the overexpression of inflammatory cytokines and have a therapeutic effect on psoriasis mice.
[0072] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all details in detail, nor do they limit the invention to only the specific embodiments described. Obviously, many modifications and changes can be made according to the content of this specification.
Claims
1. Application of chloranthelline in the preparation of drugs for intervention of psoriasis.
2. The use according to claim 1, characterized in that: The drug for intervening psoriasis is in any dosage form acceptable in the pharmaceutical field, and the dosage form is powder, granule, injection, capsule, tablet or oral solution.
3. The use according to claim 1, characterized in that: The preparation method of the daffodil alkali comprises the following steps: S1, drying and crushing the plant material containing chloranthine to obtain a raw material powder; S2, using an organic solvent to perform multiple dynamic extractions on the raw material powder, combining the extracts, filtering to remove impurities, and performing reduced pressure concentration to obtain a preliminary extract; S3, using a macroporous resin to enrich the preliminary extract, and using an eluent to separate dapoxetine from other impurities; the enriched solution is purified by silica gel column chromatography, the purified dapoxetine eluate is collected, and evaporated to dryness under reduced pressure or freeze-dried to obtain a high-purity dapoxetine product; S4. Use high performance liquid chromatography to test the purity of chloranthine products to ensure that the products meet the quality requirements.
4. The use according to claim 3, characterized in that: The organic solvent in S2 is a mixture of ethanol and water, and the volume ratio of ethanol to water is 1 to 3:
1.
5. The use according to claim 3, characterized in that: In S2, the raw material powder is dynamically extracted 2-4 times using an organic solvent, the extraction temperature is 40-60° C., and the extraction time for each time is 30-60 minutes.
6. The use according to claim 3, characterized in that: The macroporous resin in S3 is of D101 type, and the concentration and pH value of the eluent in the enrichment process are optimized to effectively separate the cyperine from other impurities.