阅读说明：本技术 化合物作为或制备***素受体激动剂或拮抗剂中的应用 (Use of compounds as or for the preparation of cannabinoid receptor agonists or antagonists ) 是由 梁鑫淼 周晗 彭惺蓉 侯滔 张秀莉 邱明华 于 2019-04-09 设计创作，主要内容包括：本发明涉及一种大麻素受体新激动剂和三种新拮抗剂的发现及应用,激动剂为Kfb77,拮抗剂为Kfb28,Kfb68和Kga1。体外细胞实验表明,化合物Kfb77对大麻素I型受体(CB1)有激动作用,Kga1对CB1有拮抗作用,Kfb28对大麻素II型受体(CB2)有拮抗作用,及Kfb68对CB1和CB2有拮抗作用。据此,本发明提到的CB1和CB2的新天然激动剂和拮抗剂为治疗精神性疾病、肥胖/代谢失常相关疾病、免疫系统调节、及多发性硬化等神经退行性疾病的药物研发提供了候选化合物。(The invention relates to discovery and application of a cannabinoid receptor new agonist and three new antagonists, wherein the agonist is Kfb77, and the antagonists are Kfb28, Kfb68 and Kga 1. In vitro cell experiments showed that compound Kfb77 has agonistic effects on cannabinoid I-type receptor (CB1), Kga1 has antagonistic effects on CB1, Kfb28 has antagonistic effects on cannabinoid II-type receptor (CB2), and Kfb68 has antagonistic effects on CB1 and CB 2. Therefore, the novel natural agonists and antagonists of CB1 and CB2 provided by the invention provide candidate compounds for the development of medicaments for treating neurodegenerative diseases such as mental diseases, obesity/metabolic disorder related diseases, immune system regulation, multiple sclerosis and the like.)

1. Use of a compound as or in the manufacture of a cannabinoid receptor agonist or a cannabinoid antagonist, characterized in that: the cannabinoid receptor agonist contains Kfb77 as an active ingredient; the cannabinoid antagonist contains one or more than two of the following substances as active ingredients: kfb68, Kfb28 or Kga 1; the structural formula of the compound is shown as follows;

2. use according to claim 1, characterized in that: the agonist further comprises a pharmaceutically acceptable carrier and/or excipient.

3. Use according to claim 1, characterized in that: the antagonist also comprises a pharmaceutically acceptable carrier and/or excipient.

4. Use according to claim 1, characterized in that: the compound Kfb77 has agonistic activity on CB1 and has CB1 subtype selectivity; the compound with subtype selectivity can distinguish different subtypes of the same series of receptors, namely cannabinoid I type receptors and cannabinoid II type receptors in the cannabinoid receptors, and has better specificity; CB1 selective agonists are useful for the treatment of one or more of pain, neurodegenerative diseases, anorexia, and the like.

5. Use according to claim 1, characterized in that: the antagonism of the antagonist on CB1 is Kga1> Kfb68, and the antagonism on CB2 is Kfb28 which is approximately equal to Kfb 68.

6. Use according to claim 1, characterized in that: EC of agonist Kfb77 on CB1 at cellular level₅₀The value was 21.39. + -. 0.69. mu.M.

7. Use according to claim 1, characterized in that: IC of antagonistic activity of the antagonist Kfb68 on CB1 and CB2 at cellular level₅₀The values are 33.79 + -1.72 μ M and 24.30 + -1.56 μ M, respectively; kga1 antagonistic Activity on CB1 at cellular level IC₅₀The value was 22.90. + -. 0.54. mu.M; IC at cellular level of antagonistic Activity of Kfb28 on CB2₅₀The value was 24.15. + -. 6.16. mu.M.

8. Use according to claim 1, characterized in that: the compounds Kfb77, Kfb68, Kfb28 and Kga1 are all from natural fungi ganoderma lucidum and are prepared from ganoderma lucidum.

9. Use of a cannabinoid receptor agonist or antagonist in the manufacture of a medicament for the treatment of: comprises one or more of neurasthenia, insomnia, amnesia, intractable pain, motor neuropathy, cardiovascular and cerebrovascular diseases (such as one or more of hypertension and hyperlipidemia), inflammation, oxidative aging, obesity/metabolism disorder, etc.;

the cannabinoid receptor agonist contains Kfb77 as an active ingredient;

the cannabinoid receptor antagonist contains one or more than two of the following substances as active ingredients: kfb68, Kfb28 or Kga 1;

the compound has the following structure:

Technical Field

The invention relates to discovery and application of targeted active molecules in fungi and plants, in particular to discovery and application of a new cannabinoid receptor agonist and three new antagonists in ganoderma tsugae and ganoderma bambusicola, wherein the new natural cannabinoid receptor agonist is Kfb77, and the new natural antagonists are Kfb28, Kfb68 and Kga 1. The application comprises treating one or more of neurasthenia, insomnia, amnesia, cardiovascular and cerebrovascular diseases (such as hypertension and hyperlipidemia), inflammation, oxidative aging, obesity/metabolism disorder, etc.

Background

The cannabinoid type I and type II receptors (cannabinoid drecepters, CB1 and CB2), two important receptors in the endocannabinoid system, belong to the class a rhodopsin-like family of G protein-coupled receptors, are widely distributed in various tissues, and mediate various physiological functions. CB1 is distributed mainly in the central nervous system (e.g., brain) and regulates neurotransmitter release; CB2 is mainly distributed in immune tissues and cells and plays a role in immune regulation. In terms of neuromodulation, the endocannabinoid system underlies emotional homeostasis and cognitive function¹The research of a mouse model proves that the deletion of the CB1 receptor is related to the symptoms of nervous disorder such as depression, anxiety and the like^2-4. In the aspect of treating obesity/metabolic disorder, a plurality of CB1 target point inhibitors have been approved as clinical drugs for treating obesity, but are stopped to be used due to side effects caused by nerves⁵However, agonism of the CB2 receptor has been shownBecome the focus of treatment⁶. In terms of immunomodulation, a large body of data demonstrates that the CB2 receptor is involved in maintaining immune balance in vivo⁷The function of regulating immune cells in inflammatory diseases and animal models; in a CB2 knockout mouse model, mice show dysplasia of T cell and B cell subsets⁸In addition, the number of osteoblasts in the mice decreases, and the bone mass decreases with age⁹. In the aspect of treating cardiovascular diseases, the CB1 receptor antagonist can remarkably reduce blood pressure on a hypertensive rat model dependent on a renin-angiotensin system¹⁰(ii) a CB2 receptors are involved in the protection of the cardiovascular system¹¹The selective activation of the CB2 receptor has anti-inflammatory and protective effects in atherosclerotic blood vessels¹². As important targets for a variety of physiological and pathological processes, the CB1 receptor and the CB2 receptor are closely related to human health and disease.

Natural products have been the most major and effective source in drug development. The compounds in plants and fungi in the nature are various in types and rich in structure, and provide a basis with creativity for finding lead compounds and researching and developing medicaments. In the case of the endocannabinoid system, the system and the endocannabinoids and receptors it contains are named after the plant Cannabis sativa (Cannabis sativa). Cannabis sativa is rich in various cannabinoids, of which Tetrahydrocannabinol (Tetrahydrocannabinol) is the main psychoactive substance in Cannabis sativa¹³The long-term taking of the health-care product has great harm to health; cannabidiol (Cannabidiol) has no mental activity, and can protect nerve, relieve inflammation, and relieve side effects (such as depression and anxiety) caused by mental active substance¹⁴Has medical value¹⁵. In addition to the plant cannabis, natural cannabinoids are also present in non-cannabinoids species¹⁶Such as resinol, which is widely available in nature, has very high activity on CB1 receptor (K)_i＝0.133nM)¹⁷. The discovery of more and more natural cannabinoids helps us to understand cannabinoid receptors, explore the relationship between receptors and diseases, and provide a structural template for the development of new highly active and potent drugs¹⁸。

Plants and fungi have long been used flexibly in traditionsHas remarkable treatment effect on maintaining mental health, improving immunity and enhancing physical performance. Such as Ganoderma, and can be used for resisting aging and protecting cardiovascular diseases¹⁹(ii) a Plant Lepidium meyenii Walp used for enhancing physical performance, improving female hormone balance, relieving anxiety, etc²⁰. Whether cannabinoid receptors are involved in the above neurological and immunological aspects is not known. The invention searches for new natural cannabinoids from 82 natural compounds in ten species (five plants and five fungi), and lays a foundation for the safe and efficient drug development of nervous system diseases, immune system diseases, metabolic diseases and cardiovascular diseases. The five plants are safflower seed oil cake (Carthamus tinctorius L), Cynanchum otophyllum Schneid, Coffee (Coffee Arabica), Prinsepia utilis Royle, and Lepidium meyenii Walp (maca); the five fungi are Ganoderma bambusae (Ganoderma hayenense), Ganoderma capense (Ganoderma capense), Ganoderma sinense (Ganoderma cochleariae), Ganoderma sessiliflorum (Ganoderma resinatum) and Ganoderma applanatum (Ganoderma applanatum).

At present, the activity and function of four natural compounds in the ganoderma lucidum discovered by the invention at cannabinoid type I and cannabinoid type II receptors have not been reported.

Reference to the literature

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Disclosure of Invention

The present invention provides 4 cannabinoids related to cannabinoid receptors in natural products, having the structure:

the use of a compound as or for the preparation of a cannabinoid receptor agonist or a cannabinoid antagonist, said cannabinoid receptor agonist having Kfb77 as active ingredient; the cannabinoid antagonist contains one or more than two of the following substances as active ingredients: kfb68, Kfb28 or Kga 1; the structural formula of the compound is shown as above.

The agonist further comprises a pharmaceutically acceptable carrier and/or excipient.

The antagonist also comprises a pharmaceutically acceptable carrier and/or excipient.

The compound Kfb77 has agonistic activity on CB1 and has CB1 subtype selectivity; the compound with subtype selectivity can distinguish different subtypes of the same series of receptors, namely cannabinoid I type receptors and cannabinoid II type receptors in the cannabinoid receptors, and has better specificity; CB1 selective agonists are useful for the treatment of one or more of pain, neurodegenerative diseases, anorexia, and the like.

The antagonism of the antagonist on CB1 is Kga1> Kfb68, and the antagonism on CB2 is Kfb28 which is approximately equal to Kfb 68.

EC of agonist Kfb77 on CB1 at cellular level₅₀The value was 21.39. + -. 0.69. mu.M.

IC of antagonistic activity of the antagonist Kfb68 on CB1 and CB2 at cellular level₅₀The values are 33.79 + -1.72 μ M and 24.30 + -1.56 μ M, respectively; kga1 antagonistic Activity on CB1 at cellular level IC₅₀The value was 22.90. + -. 0.54. mu.M; IC at cellular level of antagonistic Activity of Kfb28 on CB2₅₀The value was 24.15. + -. 6.16. mu.M.

The compounds Kfb77, Kfb68, Kfb28 and Kga1 are all from natural fungi ganoderma lucidum and are prepared from ganoderma lucidum.

Use of a cannabinoid receptor agonist or antagonist in the manufacture of a medicament for the treatment of: comprises one or more of neurasthenia, insomnia, amnesia, intractable pain, motor neuropathy, cardiovascular and cerebrovascular diseases (such as one or more of hypertension and hyperlipidemia), inflammation, oxidative aging, obesity/metabolism disorder, etc.;

the cannabinoid receptor agonist contains Kfb77 as an active ingredient;

the cannabinoid receptor antagonist contains one or more than two of the following substances as active ingredients: kfb68, Kfb28 or Kga 1;

the compound has the following structure:

the invention also provides the activity and function of these several natural compounds at the cannabinoid type I and type II receptors. The lucid ganoderma has the effects of regulating the nervous system, relieving insomnia and amnesia and regulating neurasthenia; has effects in regulating immune system, improving immunity, treating cardiovascular and cerebrovascular diseases (such as hypertension and hyperlipidemia), resisting oxidation, and relieving inflammation. The current research shows that CB1 is highly expressed in the central nervous system and has modulation effects on motor function, learning and memory, neuroendocrine and the like; CB2 is mainly expressed in peripheral immune system, and has anti-inflammatory, immunosuppressive, immunoregulatory effects; the cannabinoid system is involved in the treatment of psychiatric disorders, obesity/metabolic disorder related disorders, immune system modulation, and cardiovascular disease. Accordingly, the novel natural agonists and antagonists of CB1 and CB2 mentioned in the present invention provide candidate compounds for drug development of these diseases.

Drawings

FIG. 1A target model of CHO-CB1 cells characterizing the DMR response signal of A-agonist CP55,940; DMR response signals for B-agonist WIN55, 212-2; dose response curves for C-CP55,940 (filled squares), and 100nM CP55,940 (open squares) one hour after pretreatment; dose response curves for D-WIN55,212-2 (filled squares), and 1 μ M WIN55,212-2 (open squares) after one hour of pretreatment; DMR response signals for E-antagonist SR 141716A; one hour after pretreatment with F-SR141716A, dose response curves for 100nM CP55,940 (open lower triangle) and 1 μ M WIN55,212-2 (open upper triangle)

FIG. 2A-agonist CP55,940 DMR response signals characterized by the target model of CHO-CB2 cells; DMR response signals for B-agonist WIN55, 212-2; dose response curves for C-CP55,940 (filled circles), and 100nM CP55,940 (open circles) one hour after pretreatment; dose response curves for D-WIN55,212-2 (filled circles), and for WIN55,212-2 at 200nM (open circles) after one hour of pretreatment; DMR response signal for E-antagonist SR 144528; one hour after F-SR144528 pretreatment, dose response curves for 100nM CP55,940 (open lower triangle) and 200nM WIN55,212-2 (open upper triangle)

FIG. 3 Structure of Natural Compounds (first portion 36)

FIG. 4 Structure of Natural Compounds (second portion 20)

FIG. 5 Structure of Natural Compounds (third part 26)

FIG. 6 preliminary screening of Natural Compounds on CHO-CB1 and CHO-CB2 cells for DMR response signals of A-compounds on CHO-CB1 cells; DMR response signals at CHO-CB1 cells at 50nM of CP55,940 after one hour of B-compound pretreatment; DMR response signals of C-compounds in CHO-CB2 cells;

DMR response signals of 50nM CP55,940 in CHO-CB2 cells after one hour of D-compound pretreatment fig. 7 activity of active compounds on CHO-CB1 cells and CHO-CB2 cells vs dose response curves of functional a-Kfb77 in CHO-CB1 cells (filled circles), and Kga1 (open squares), Kfb68 (open triangles) and Kfb77 (open circles) after one hour of pretreatment, 50nM CP55,940 in CHO-CB1 cells; dose response curve of 100 μ M Kfb77 in CHO-CB1 cells one hour after pretreatment with the B-antagonist SR 141716A; dose response curves for C-A-Kfb77 in CHO-CB2 cells (filled circles), and for 200nM WIN55,212-2 in CHO-CB2 cells one hour after pretreatment with Kfb28 (open diamonds), Kfb68 (open triangles) and Kfb77 (open circles); dose response curves for WIN55,212-2 (open upper triangle) and CP55,940 (open lower triangle) in CHO-CB2 cells one hour after pretreatment with D-Kfb 68.

Detailed Description

The present invention will now be further described with reference to examples. The examples illustrate the invention in more detail, but do not limit the invention in any way.