阅读说明：本技术 甲基黄连碱用作jnk2激酶抑制剂和治疗银屑病的用途 (Use of methylprednisolone coptisine as JNK2 kinase inhibitor and for treating psoriasis ) 是由 朱峰 段秋红 路慧 马腾飞 倪小芳 于 2019-12-09 设计创作，主要内容包括：本发明公开了甲基黄连碱用作JNK2激酶抑制剂的用途,甲基黄连碱能特异性结合JNK2激酶且二者具有较强的结合能力,同时呈剂量依赖性抑制JNK2的活性。甲基黄连碱对银屑病具有很好的治疗活性,该活性与甲基黄连碱抑制JNK、STAT3信号通路的活化以及银屑病标志分子S1007的表达密切相关,因此可作为活性成分用于制备治疗银屑病的药物。(The invention discloses application of methylprednisolone coptisine as a JNK2 kinase inhibitor, wherein the methylprednisolone coptisine can be specifically combined with JNK2 kinase, has stronger combining capacity and simultaneously inhibits the activity of JNK2 in a dose-dependent manner. The methylprednisolone coptisine has good therapeutic activity on psoriasis, and the activity is closely related to the inhibition of the activation of JNK and STAT3 signal pathways and the expression of psoriasis marker molecule S1007 by the methylprednisolone, so that the methylprednisolone coptisine can be used as an active ingredient for preparing a medicament for treating psoriasis.)

1. Use of methylprednisolone as JNK2 kinase inhibitor.

2. A JNK2 kinase inhibitor contains methylprednisolone.

3. Application of Meadowrine in preparing medicine for treating psoriasis is provided.

4. A medicine for treating psoriasis contains methyl coptisine as active component.

5. The medicament for treating psoriasis according to claim 4 wherein: the medicine is an external medicine.

Technical Field

The invention relates to a new application of methylprednisolone as JNK2 kinase inhibitor, and also relates to an application of methylprednisolone in preparing medicine for treating psoriasis and a medicine for treating psoriasis.

Background

c-jun amino-terminal kinases [ c-jun N-terminal kinases (JNK) ] were first defined as stress-activated protein kinases [ stress-activated protein kinases (SAPK) ], a serine kinase, a member of the mitogen-activated protein kinase (MAPK) ] family. Mammalian JNKs are encoded by three different genes, JNK1, JNK2 and JNK3, and have more than ten protein products through alternative splicing, and the molecular weight is from 46kDa to 55kDa [1 ]. JNK1 and JNK2 were expressed in most tissues, and JNK3 expression was mainly restricted to brain, heart and testis. The different distribution of JNK subtypes in organ tissues indicates that the JNK function is different in different cells.

JNK2 has 84% sequence similarity with JNK3 and 80% sequence similarity with JNK1 the crystal structure of JNK2 protein shows that it has a typical protein kinase double-leaf structure, i.e. N-terminal domain (containing lots of β -fold) and C-terminal domain (containing lots of α -helix), and hinge region (109 and 113 amino acids) between two leaves, and provides binding site for ATP and competitive inhibitor of ATP.

JNKs are members of MAPK, and like the activation of MAPK cascade, the activation of JNK can activate JNK signal pathways by extracellular stimulation (such as growth factors and cytokines) and cell pressure (such as heat shock, high osmotic pressure, UV-radiation and ischemia/reperfusion), participate in the regulation of cell proliferation, differentiation, mitosis, cell survival, apoptosis, autophagy and other vital activities, and play an important role in the pathological processes of tumor growth and drug resistance, inflammatory response, obesity and insulin resistance, neurosynaptic plasticity, memory injury and the like [3-19 ].

Therefore, the inhibitor designed or discovered by taking the JNKs as the target is expected to treat related diseases caused by the abnormal JNKs. Currently, inhibitors of JNKs are divided into two classes, ATP competitive inhibitors and ATP non-competitive inhibitors, depending on the mode of action. Studies on ATP competitive inhibitors targeting JNKs have been extensive, mainly including some indazoles, aminopyrazoles, aminopyridines, picolinamides, quinoline derivatives, and aminopyrimidines. However, these inhibitors have not been widely used as clinical drugs, and have been the only subjects of study in animal or cell disease models. For example, SP600125, which is the earliest and most extensively studied ATP competitive JNK inhibitor, inhibits three JNK kinases simultaneously at low concentrations and ERKs and p38 MAPKs at high concentrations [20, 21], with good results in a mouse Parkinson disease model [22 ]. AS601245 is another inhibitor of JNKs, can inhibit the transcription factor c-jun even at high concentration, and shows good neuroprotective effect in a mouse ischemia-reperfusion model [23, 24 ]. JNK-IN-7 and JNK-IN-8 are potent specific inhibitors of JNKs, covalently bound to conserved cysteine groups IN the ATP binding site, which produce irreversible inhibition of JNKs [25 ]. Another strategy for inhibiting JNK activity is to inhibit JNK enzyme activity by ATP-noncompetitive peptides and small compounds, such as pepJIP1, which is an 11-amino acid peptide cut from the binding region of JIP1, which is a JNK binding protein, and in vitro studies have shown that pepJIP1 specifically and effectively inhibits JNK activity [26, 27 ]. In addition, the inhibitory peptide PYC71N also prevented JNK interaction with c-jun and thus inhibited the action of JNKs [28, 29 ].

Although the development and research of inhibitors of JNKs are continuously developing and mature, and the action mechanism of the corresponding inhibitors is becoming clearer in cells and animal models, the cases of JNKs inhibitors as clinical drugs do not appear, and one important problem is that the selectivity of JNKs inhibitors against different subtypes of JNK is poor, and JNK1 and JNK2 often play mutually opposite roles in the cell growth process. Therefore, in the research of inhibitors of JNKs, the development of specific inhibitors of JNKs specifically targeting different subtypes for different disease mechanisms is required to achieve better therapeutic effects.

The coptis chinensis is used as a traditional Chinese herbal medicine in China and is applied to treating inflammation very early. Various alkaloids have been isolated from coptis chinensis, including african tetrandrine, berberine, epiberberine, jateorhizine, coptisine, phellodendrine, methyberberine, berrubine, chelerythrine, nitidine and the like [30 ]. Researches show that various components of the coptis chinensis have biological activity and have various functions of oxidation resistance, inflammation resistance, hypertension resistance, cancer resistance and the like [31, 32 ]. Although the pharmacological action of coptis is clear, the coptis has a plurality of effective components, unclear molecular targets and unclear action mechanism, so that the wide application of the coptis is limited. Methylpcoptisine (workine) is one of the effective components of coptis, belongs to isoquinoline alkaloid [33], and the activity and function of the methylpcoptisine are not studied in detail. By adopting a computer-aided drug screening technology, a candidate action target of the methylprednisolone is screened, and the methylprednisolone is found to be capable of being used as an inhibitor of JNK2 and used for treating psoriasis in cell and animal experiments.

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disclosure of Invention

The invention aims to provide a new application of a traditional Chinese medicine monomer drug, namely the methylprednisolone alkaloid in inhibiting JNK2 kinase and treating psoriasis.

The above object is achieved on the basis of the following studies:

1. the Meadoline is proved to be capable of binding JNK2 kinase and inhibiting the enzymatic activity of the JNK2 kinase.

MST and in vitro kinase experiments are carried out on JNK2 kinase, and the result shows that the methylprednisolone goldthread base can be specifically combined with JNK2 kinase and has strong combining capacity. Meanwhile, the Medohedrine is in dose-dependent inhibition on the activity of JNK2, so that phosphorylation modification of JNK2 on a substrate H2AX is inhibited.

Therefore, the methylpacharidine can be used as a JNK2 kinase inhibitor, has wide application prospects in the aspects of treatment and prevention of diseases caused by abnormal JNK2 kinase activity in human bodies or animals, and has the potential of corresponding detection or diagnosis and other applications by inhibiting enzyme activity in vitro.

2. The action mechanism of the methylprednisolone hance alkaloid is researched in a psoriasis cell model, and the effect mechanism of the methylprednisolone hance alkaloid is found to inhibit a JNK2 signal path so as to inhibit psoriasis-related inflammatory reaction.

The high activation of the S1007, STAT3, JNK and p38 signaling pathways are the most prominent molecular alterations characteristic of psoriasis. M5(10ng/ml TNF-a, IL-17A, IL-22, IL-1a and Oncostatin-M) (references: Teng X, Hu Z, Wei X, Wang Z, Guan T, Liu N, Liu X, Ye N, Deng G, Luo C, Huang N, Sun C, Xu M, Zhou X, Deng H, Edwards CK 3rd, Chen X, Wang X, Cui K, Wei Y, Li J.IL-37 ameliorantes and processes in psoriasis by treating psoriasis cytokinase. J. 2014feb 15; 192(4) 1815-23. epub.22. Jab 22) Jab cells are a model of psoriasis stimulation, and we further use a model of the effects of methyl-p-S38 and STAT K on the mechanisms of psoriasis. The results show that the methylprednisolone alkaloid obviously inhibits the activation of JNK and STAT3 signal pathways induced by M5 and the expression of psoriasis marker molecule S1007.

3. The effect of the methyl coptisine is observed in an animal model of the psoriasis, and the methyl coptisine is found to effectively inhibit the clinical symptoms and pathological changes of the psoriasis.

A classical animal model of psoriasis is 5% imidazolemote (Imiquimod, IMQ) applied to the skin of the back and ears of adult Balb/c mice (nine weeks old) for 5 consecutive days, inducing skin lesions. We adopt the model to observe the therapeutic effect of the Medohedrine on psoriasis. The results show that the methylprednisolone alkaloid effectively inhibits 5% of the skin symptoms and pathological changes induced by the imidazolate.

The researches are combined to discover that the methylparaben has good therapeutic activity on psoriasis, the activity is closely related to the inhibition of the activation of JNK and STAT3 signal pathways by the methylparaben and the expression of psoriasis marker molecule S1007, so that the methylparaben can be used as an active ingredient for preparing a medicament for treating psoriasis, the methylparaben and other active ingredients can be compounded to prepare the medicament for treating psoriasis in order to further improve the curative effect, and pharmaceutically acceptable carriers can be added during the preparation of the medicaments.

According to one embodiment of the invention, the medicament is for topical administration.

Drawings

FIG. 1 shows the results of in vitro kinase assays for inhibition of JNK2 activity by methylprednisolone.

FIG. 2 establishment of psoriasis cell model.

FIG. 3 uses Western blot to detect signal molecules to evaluate inhibitory effect of methylprednisolone on JNK2 signal pathway in psoriasis cell model.

Figure 4 effect of methylccoptisine on imiquimod-induced skin pathological changes in animal models of psoriasis.

Detailed Description

The present invention will be described in detail below with reference to specific examples.