阅读说明：本技术 罗丹宁衍生物在治疗代谢性疾病中的应用 (Use of rhodanine derivatives for the treatment of metabolic disorders ) 是由 丁秋蓉 冯庄慧 于 2019-01-14 设计创作，主要内容包括：本发明首次揭示罗丹宁衍生物可以激活STAT3、CREB和PPARs信号通路,显著性提高UCP1的表达,增强线粒体氧化磷酸化基因表达,有效提高产热,可用于治疗代谢性疾病。因此,本发明涉及罗丹宁衍生物在治疗代谢性疾病中的应用。本发明还涉及含有罗丹宁衍生物作为活性成分的药物组合物,用于治疗代谢性疾病。(The invention discloses that the rhodanine derivative can activate STAT3, CREB and PPARs signal channels for the first time, remarkably improve the expression of UCP1, enhance the expression of mitochondrial oxidative phosphorylation genes, effectively improve heat production, and can be used for treating metabolic diseases. Accordingly, the present invention relates to the use of rhodanine derivatives in the treatment of metabolic disorders. The present invention also relates to pharmaceutical compositions containing the rhodanine derivatives as active ingredients for the treatment of metabolic disorders.)

1. Use of a rhodanine derivative represented by the following formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a metabolic disease:

in the formula (I), the compound is shown in the specification,

R¹selected from phenyl or phenyl-C1-C4 alkyl, wherein said phenyl is optionally substituted by 1 or 2 groups selected from-COOH, C1-C4 alkyl, C1-C4 alkylcarboxy, hydroxy, -SO₃H and-C1-C4 alkyl-H₂PO₃Substituted with the substituent(s);

R²selected from C1-C4 alkyl, C3-C8 cycloalkyl and optionally substituted by 1 or 2 groups selected from hydroxy, C1-C4 alkyl, C1-C4 alkoxyNitro and halogen substituted phenyl.

2. The use according to claim 1, wherein R is¹Selected from:

3. use according to claim 1 or 2, wherein R is²Selected from:

4. the use according to claim 1, wherein R is¹Selected from:

R²selected from:

5. the use according to claim 1, wherein R is¹Selected from:

R²is phenyl.

6. Use of a rhodanine derivative having a structure represented by formula II below or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of metabolic disorders such as obesity and obesity-related disorders:

wherein Ra is carboxyl, Rb is H, nitro, halogen or C1-C4 alkyl; preferably, Ra is in the para or meta position; preferably, Rb is in the para or meta position.

7. Use of a compound having the formula:

8. a pharmaceutical composition comprising a first active ingredient, a second active ingredient and a pharmaceutically acceptable carrier, wherein the first active ingredient is a compound of formula I as defined in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or a compound of formula II as defined in claim 6; the second active ingredient is a different drug from the first active ingredient for the treatment of obesity and obesity-related diseases.

9. A pharmaceutical composition comprising a first active ingredient, a second active ingredient, and a pharmaceutically acceptable carrier, wherein the first active ingredient is a compound of the formula:

the second active ingredient is a different drug from the first active ingredient for the treatment of obesity and obesity-related diseases.

10. Use of a compound of formula I as defined in any one of claims 1 to 5 or a compound of formula II as defined in claim 6 or a compound as defined in claim 7 as lead compound for the manufacture of a medicament for the treatment or prevention of metabolic disorders such as obesity and obesity related disorders.

Technical Field

The present invention relates to the use of rhodanine derivatives in the treatment of metabolic disorders.

Background

Obesity is a chronic metabolic disease caused by various factors and is characterized by abnormal increase in the percentage of body fat to body weight and excessive deposition of fat in some parts due to increase in the volume and number of fat cells in the body. Current anti-obesity drugs limit energy intake mainly by reducing fat absorption or suppressing appetite, however clinical effects are not obvious and have side effects. There is no medicine for treating obesity by increasing energy utilization in clinic, and the energy of body is consumed in the form of heat energy by increasing thermogenesis function of brown adipose tissue BAT, so that it can be used as target for treating obesity and related metabolic diseases.

Disclosure of Invention

The invention provides application of a rhodanine derivative or a pharmaceutically acceptable salt thereof in preparing medicines for treating or preventing metabolic diseases.

The rhodanine derivatives of the present invention may have the structure shown in formula I below:

in the formula (I), the compound is shown in the specification,

R¹selected from phenyl or phenyl-C1-C4 alkyl, wherein said phenyl is optionally substituted by 1 or 2 groups selected from-COOH, C1-C4 alkyl, C1-C4 alkylcarboxy, hydroxy, -SO₃H and-C1-C4 alkyl-H₂PO₃Substituted with the substituent(s);

R²selected from the group consisting of C1-C4 alkyl, C3-C8 cycloalkyl and phenyl optionally substituted with 1 or 2 groups selected from the group consisting of hydroxy, C1-C4 alkyl, C1-C4 alkoxy, nitro and halogen.

In certain embodiments, in formula I, R¹Selected from:

in certain embodiments, in formula I, R²Selected from:

in certain embodiments, in formula I, R¹Selected from:

R²selected from:

in certain embodiments, in formula I, R¹Selected from:

R²is phenyl.

In certain embodiments, the rhodanine derivative is BM L-260:

in certain embodiments, the present invention provides the use of a rhodanine derivative of formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of obesity and obesity-related diseases.

In certain embodiments, the present invention provides the use of a rhodanine derivative of formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a product for inhibiting weight gain, reducing body fat content, and/or improving metabolism in a subject.

The invention also provides a pharmaceutical composition, which comprises a first active ingredient, a second active ingredient and a pharmaceutically acceptable carrier, wherein the first active ingredient is the compound of the formula I or the pharmaceutically acceptable salt thereof; the second active ingredient is a different drug from the first active ingredient for the treatment of obesity and obesity-related diseases.

The invention also provides a pharmaceutical composition, which comprises a first active ingredient, a second active ingredient and a pharmaceutically acceptable carrier, wherein the first active ingredient is a compound shown as the following formula or a pharmaceutically acceptable salt thereof:

the second active ingredient is a different drug from the first active ingredient for the treatment of obesity and obesity-related diseases.

The invention also provides the use of a compound of formula I or a compound of formula II as described herein or BM L-260 as a lead compound in the manufacture of a medicament for the treatment of metabolic disorders such as obesity and obesity related disorders.

Drawings

FIG. 1 is a representative picture of GFP fluorescence intensity of mature brown adipocytes treated by BM L-260 for 1 day, 2 days, and 3 days (left panel) and Western blot assay for UCP1 (right panel). Scale bar: 200 μm.

FIG. 2 shows that the mRNA expression level of UCP1 of mature brown adipocytes treated by BM L-260 for 1 day, 2 days and 3 days is significantly up-regulated compared with that of a negative control group and is increased along with the increase of the treatment time, and the mRNA expression level of UCP1 of an experimental group treated by BM L-260 for 3 days is similar to that of a positive control ISO-treated group.

FIG. 3 is a graph representing the GFP fluorescence intensity of experimental groups sampled from day 1 of differentiation plus BM L-260 until day 10, and a control group (upper panel), and real-time fluorescence quantitative PCR results of Ucp1 and its closely related transcription factors (lower panel).

FIG. 4 is a graph showing representative GFP fluorescence intensities of each group when BM L-260 was used for the induction of differentiation stage alone, in a scale of 200. mu.m.

FIG. 5 GFP obtained 3 days after DMSO or BM L-260 treatment after flow sorting based on GFP fluorescence intensity⁺Analysis of cellular mitochondrial Activity (APC intensity) (left panel), Electron microscopy of mature brown adipocytes treated with DMSO and BM L-260 for 3 days (right panel, L D for lipid droplets, M for mitochondria). Scale bars, top 2 μ M, bottom 0.5 μ M.

FIG. 6 MitoDNA/NuDNA results of 3-day treatment of brown adipocytes with DMSO and BM L-260 (left panel), while ATP production was significantly reduced (right panel).

FIG. 7 real-time fluorescent quantitative PCR results (left panel) and Western immunoblot results (right panel) of mature white adipocytes treated with DMSO or BM L-260 for 5 days.

FIG. 8: JSP-1 protein expression analysis (left panel) and UCP1 protein expression analysis of differently treated white adipocytes (right panel).

FIG. 9: and (5) RNA sequencing analysis results.

FIG. 10: immunoblot detection of brown adipocytes.

FIG. 11: graphical illustration and results of connected Map analysis.

FIG. 12: representative pictures of GFP fluorescence intensity (left panel) and detection of UCP1 protein expression levels (right panel) for different treatment groups. Scale bar: 250 μm.

FIG. 13: mouse subcutaneous white adipose tissue UCP1 protein expression after in situ injection.

FIG. 14, BM L-260 injected mice showed higher rectal temperature in cold stimulation (left panel), a tendency for weight loss (middle panel), and a significant decrease in subcutaneous white adipose tissue (iWAT) (right panel).

FIG. 15, it can be seen from the hematoxylin and eosin staining (upper row) that iWAT in the BM L-260 injected group had a distinct browning phenotype with smaller lipid droplets and significantly enhanced UCP1 expression (lower row).

FIG. 16 shows that p-STAT3 and p-CREB protein signals of BM L-260 injection group are obviously improved compared with those of a control group, which shows that BM L-260 can directly or indirectly activate STAT3 and CREB signal pathways and is part of the reason of downstream activation of UCP 1.

FIG. 17 RNA sequencing analysis results in GO enrichment analysis (left panel), genes significantly upregulated in subcutaneous white adipose tissue of BM L-260 injection group were enriched in physiological processes such as myocyte differentiation, myostructural development, and myosystem process control, which is similar to UCP1 in white adipose tissue⁺The cells have consistent smooth muscle-like characteristics and the heat map shows (right panel) that the BM L-260 injected group also has significant upregulation of many thermogenic genes.

Detailed Description

It is to be understood that within the scope of the present invention, the above-described technical features of the present invention and the technical features described in detail below (e.g., the embodiments) may be combined with each other to constitute a preferred embodiment.

The invention discloses a medicine for treating or preventing metabolic diseases, in particular metabolic diseases benefiting from non-shivering heat production of an organism, which is prepared by modifying or modifying the chemical structure of rhodanine based on the discovery of the invention, and the compounds of formula I and formula II, including BM L-260, can be used as lead compounds (lead compounds) to obtain the medicine for treating or preventing metabolic diseases through modification and modification.

The rhodanine derivatives of the present invention may generally have the structure shown in formula I below:

in the formula, R¹Selected from phenyl or phenyl-C1-C4 alkyl, wherein said phenyl is optionally substituted by 1 or 2 groups selected from-COOH, C1-C4 alkyl, C1-C4 alkylcarboxy, hydroxy, -SO₃H and-C1-C4 alkyl-H₂PO₃Substituted with the substituent(s); r²Selected from the group consisting of C1-C4 alkyl, C3-C8 cycloalkyl and phenyl optionally substituted with 1 or 2 groups selected from the group consisting of hydroxy, C1-C4 alkyl, C1-C4 alkoxy, nitro and halogen.

Preferred R¹Is phenyl substituted with carboxy and optionally hydroxy. Preferably, R¹Is phenyl substituted by carboxyl at meta-or para-position.

Preferred R²Is C3-C8 cycloalkyl and phenyl optionally substituted with nitro, halogen or C1-C4 alkyl; preferably, the substitution occurs in the para position.

In certain embodiments, the rhodanine derivatives suitable for use in the present invention have the structure shown in formula II below:

wherein Ra is carboxyl, and Rb is H, nitro, halogen or C1-C4 alkyl. Preferably, Ra is in the para or meta position; preferably, Rb is in the para or meta position.

More preferably, the rhodanine derivative is BM L-260 having the formula:

the rhodanine derivatives can be synthesized by conventional methods, for example, reference can be made to the methods disclosed in Neil S.Cutshall et al (Bioorganic & Medicinal Chemistry L etters 15 (2005)) 3374-3379.

The invention provides application of a rhodanine derivative or a pharmaceutically acceptable salt thereof in preparing a medicament for treating or preventing metabolic diseases, particularly obesity and obesity-related diseases. In certain embodiments, the metabolic disorders of the present invention are those that benefit from non-tremor thermogenesis in the body, including obesity and obesity-related disorders.

Herein, obesity is a chronic metabolic disease characterized by an increase in the volume and cell number of fat cells in the body, resulting in an abnormally high percentage of body fat to body weight and excessive deposition of fat in some regions. Generally, whether a subject is obese can be assessed by Body Mass Index (BMI): BMI is the square of body weight (kg)/height (m). If the BMI is greater than 30, the subject is considered obese. Alternatively, Waist Circumference (WC) or waist-to-hip ratio (WHR) may be used to determine whether a subject is obese. Obesity in addition to a large amount of fat stored in the body, there is also obesity in which body fat is distributed in the internal organs and abdominal walls, manifesting as a large abdominal stool, which is called central obesity or abdominal obesity. The waist circumference of a male exceeds 94cm, and the waist circumference of a female exceeds 80cm, and can be used as the standard of obesity; waist-hip ratio of more than 0.9 for men and more than 0.8 for women may be considered central obesity.

Herein, obesity-related diseases include, but are not limited to, type 2 diabetes, hypertriglyceridemia, hypercholesterolemia, hypo-high density lipoprotein cholesterolemia, hyperuricemia (ventilation), coronary heart disease, hypertension, cholelithiasis, fatty liver, obesity hypoventilation syndrome, pulmonary heart syndrome, osteoarthritis, osteoporosis, endocrine disorders, and the like. In certain embodiments, the obesity-related disease is a metabolic disease resulting from obesity, such as type 2 diabetes, hypertriglyceridemia, hypercholesterolemia, hypo-high density lipoprotein cholesterolemia, hyperuricemia, fatty liver, and the like.

Herein, halogen includes F, Cl, Br and I, and alkyl includes straight chain and branched chain alkyl groups.

As used herein, pharmaceutically acceptable salts refer to those salts which retain the biological potency and properties of the parent compound, including the acid addition salts obtained by reacting the free base of the parent compound with an inorganic or organic acid, examples of which include hydrochloric, hydrobromic, nitric, phosphoric, sulfuric, perchloric, and the like, examples of which include acetic, oxalic, (D) or (L) malic, maleic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, salicylic, tartaric, citric, succinic, or malonic acid and the like, preferably hydrochloric or (L) -malic acid, or salts formed when the acid proton present in the parent compound is replaced with a metal ion, such as an alkali metal ion, alkaline earth ion, or aluminum ion, or a coordination compound formed with an organic base, such as ethanolamine, diethanolamine, triethanolamine, tris, N-methylglucamine, and the like.

Herein, a "pharmaceutical composition" is a composition comprising a rhodanine derivative or a pharmaceutically acceptable salt thereof as described herein, together with physiologically/pharmaceutically acceptable carriers and excipients. "physiologically/pharmaceutically acceptable carriers" means those carriers and diluents which do not significantly irritate the organism and which do not impair the biological activity and properties of the administered rhodanine derivative or a pharmaceutically acceptable salt thereof. "physiologically/pharmaceutically acceptable excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a rhodanine derivative or a pharmaceutically acceptable salt thereof. Non-limiting examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

The rhodanine derivative or a pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a therapeutically effective amount or a prophylactically effective amount. An effective amount is an amount administered sufficient to ameliorate or in some way reduce the symptoms associated with the disease. The amount administered is an amount effective to ameliorate or eliminate one or more symptoms and can be determined by one of ordinary skill in the art based on the age, sex, physical condition, etc. of the subject. The amount administered may be sufficient to cure the disease, but is generally administered to ameliorate the symptoms of the disease. Repeated administration is generally required to achieve the desired improvement in symptoms. Generally, a unit oral dosage may include about 0.01 to 50 mg, preferably about 0.1 to 10 mg, of a rhodanine derivative or a pharmaceutically acceptable salt thereof. The unit dose may be administered one or more times, for example, in one or more tablets per day, each tablet containing about 0.01 to 50 mg of the rhodanine derivative or pharmaceutically acceptable salt thereof.

The pharmaceutical compositions of the present invention may be formulated in any suitable dosage form for oral, intravenous, topical or topical administration.

In certain embodiments, the active ingredient for treating or preventing metabolic diseases such as obesity or obesity-related diseases in the pharmaceutical composition of the present invention may further comprise other drugs for treating obesity or obesity-related diseases, such as other currently known anti-obesity drugs or drugs for treating type 2 diabetes, hypertriglyceridemia, hypercholesterolemia, hypo-high density lipoprotein cholesterolemia, hyperuricemia (ventilation), coronary heart disease, hypertension, cholelithiasis, fatty liver, obesity hypoventilation syndrome, pulmonary heart syndrome, osteoarthritis, osteoporosis and/or endocrine disorders (referred to as active ingredients), in addition to the rhodanine derivative or pharmaceutically acceptable salt thereof according to the present invention.

Herein, a product refers to a product other than a medicament, such as a food or health care product. For example, in the case of a high-fat diet subject, a suitable amount of a rhodanine derivative or a pharmaceutically acceptable salt thereof may be added to the diet thereof, thereby playing a role in preventing obesity or obesity-related diseases. Of course, the amount of the rhodanine derivative or its pharmaceutically acceptable salt in the food or health care product should be controlled, which can be determined by a person skilled in the art by taking conventional means.

In certain embodiments, the present invention also provides a method of treating or preventing obesity or an obesity-related disease, the method comprising the step of administering to a subject in need thereof a therapeutically or prophylactically effective amount of a rhodanine derivative, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

Herein, the subject is a mammal, especially a human.

Also included are rhodanine derivatives or pharmaceutically acceptable salts thereof, especially the compounds of formula I or pharmaceutically acceptable salts thereof, or the compounds of formula II or pharmaceutically acceptable salts thereof, or BM L-260 or pharmaceutically acceptable salts thereof, as described herein, for use in the treatment or prevention of metabolic disorders, especially obesity or obesity related disorders.

The invention also provides the use of a compound of formula I or a compound of formula II as described herein or BM L-260 as a lead compound in the manufacture of a medicament for use in the treatment of metabolic disorders such as obesity and obesity related disorders.

The present invention will be illustrated below by way of specific examples. It should be understood that these examples are illustrative only and are not intended to limit the scope of the present invention. The methods and reagents used in the examples are, unless otherwise indicated, conventional in the art.